l 111 mu; (will!!! Lu: ll 1m mu Hill :1 III”! ' us u R y 63 2417 Michigan State University $18.60 This is to certify that the thesis entitled THE EFFECTS OF SEVERAL EXERCISE REGIMENS ON RAT MYOCARDIUM presented by Bonnie Lee Smoak has been accepted towards fulfillment of the requirements for M.A. Physical Education degree in QM /é/Z, Major professor Date my 03639 OVERDUE FINES ARE 25¢ PER DAY PER ITEM Return to book drop to remove this checkout from your record. THE EFFECTS OF SEVERAL EXERCISE REGIMENS 0N RAT MYOCARDIUM By Bonnie Lee Smoak A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF ARTS Department of Health, Physical Education and Recreation 1979 ABSTRACT THE EFFECTS OF SEVERAL EXERCISE REGIMENS 0N RAT MYOCARDIUM By Bonnie Lee Smoak This investigation was undertaken to investigate the effects of long-term exhaustive workloads on the myocardium of normal male albino rats. Animals for this study were obtained from two experiments. In the first experiment, the animals were placed in three activity groups, with each group divided into two dietary supplement groups. The second experiment was organized as a two-way design with five activity groups and two dietary supplement groups. Analysis of variance of the dietary supplements in both experi- ments indicated no significant differences in body weight gained, absolute heart weight, and relative heart weight. In experiment one, the absolute heart weight of the control group was significantly larger than the sprint group. No significant differ- ences were observed in relative heart weights. In experiment two, the absolute heart weights of the control group were significantly larger than the high endurance, regular sprint, and regular endurance groups. Again there were no statistically significant differences observed in relative heart weights. A chi-square test indicated there was no significant difference in heart damage between trained and control animals. ACKNOWLEDGEMENTS Deep appreciation is given to Dr. Wayne Van Huss for his continual guidance during my graduate career and during the writing of this thesis. Gratitude is given to Dr. Herbert Olsen and Dr. Kwok-kai Ho for their participation in the preparation of this manuscript. Finally, I would like to thank all of my friends for their support. ii TABLE OF CONTENTS CHAPTER Page LIST OF TABLES ........................................... v LIST OF FIGURES .......................................... V“ I. THE PROBLEM .............................................. l Need for the Study .................................... 3 Statement of Problem .................................. 3 Research Hypothesis ................................... 4 Research Design ....................................... 4 Rationale ............................................. 5 Limitations ........................................... 5 Significance of the Study ............................. 6 II. REVIEW OF RELATED LITERATURE ............................. 7 Catecholamines and Heart Damage ....................... lO Catecholamine Secretion During Exercise ............... 11 III. METHODS AND MATERIALS .................................... 13 Animals In Experiment One ............................. 13 Research Design In Experiment One ..................... 14 Control Group ...................................... 14 Sprint Group ....................................... l4 Endurance Group .................................... 15 Vitamin C Group .................................... 15 Placebo Group ...................................... 16 Animals In Experiment Two ............................. 16 Research Design In Experiment Two ..................... 16 Control Group ...................................... 16 Regular Sprint Group ............................... 16 High Sprint Group .................................. 17 Regular Endurance Group ............................ 17 High Endurance Group ............................... 18 Sugar Group ........................................ 18 Placebo Group ...................................... 18 CHAPTER Training Procedures ................................... Animal Care ........................................... Sacrifice Procedures .................................. Experiment One ..................................... Experiment Two ..................................... Histological Methods ............................... Pathological Evaluations ........................... Statistical Procedures ................................ IV. RESULTS AND DISCUSSION ................................... Training Results of Experiment One .................... Training Results of Experiment Two .................... Body and Heart Weight Results at Sacrifice for Experiment One ..................................... Body and Heart Weight Results at Sacrifice for Experiment Two ..................................... Histopathological Results ............................. Discussion ............................................ V. SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS ................ Summary ............................................... Conclusions ........................................... Recommendations ....................................... REFERENCES ...................................................... APPENDICES A. TRAINING PROGRAMS ........................................ B. BASIC STATISTICS FOR TRAINING DATA ....................... iv 52 58 TABLE LIST OF TABLES . Analysis of Variance for overall training effects and Newman-Keul's tests of paired comparisons for body weight at sacrifice and absolute and relative heart weights, for experiment one ............................................ . Analysis of variance for overall vitamin C effects for body weight at sacrifice and absolute and relative heart weights for experiment one ................................ . Analysis of variance for overall sugar effects for experiment one ............................................ . Analysis of variance for overall training effects and student Newman-Keul's tests of paired comparisons for experiment two ............................................ . Frequency distribution of ratings of myocardial damage in experiment one ............................................ . Frequency distribution of ratings of myocardial damage in experiment two ............................................ . Chi-square test for treatment effects on the degree of myocardial damage ......................................... . Modified eight week sprint training program for postpu- bertal and adult male rats in controlled-running wheels for experiment one ........................................ . Modified eight week sprint training program for postpu- bertal and adult male rats in controlled-running wheels for experiment one ........................................ . Modified eight week regular sprint training program for postpubertal and adult male rats in controlled-running wheels .................................................... . Modified eight week regular endurance training program for postpubertal and adult male rats in controlled- running wheels ............................................ Page 36 38 39 41 42 42 42 52 53 54 55 TABLE A-5. A-6. 8-2. Modified eight week high sprint training program for post- pubertal and adult male rats in controlled-running wheels .................................................... Modified eight week high endurance training program for postpubertal and adult male rats in controlled-running wheels .................................................... . Basic statistics for percentage of body weight loss, environmental factors and performance criteria for experiment one ............................................ Basic statistics for percentage of body weight loss, environmental factors, and performance criteria for experiment two ............................................ vi Page 56 57 58 59 LIST OF FIGURES FIGURE Page 1. Mean Daily Percent Expected Meters Run for SPRINT Animals ................................................. 27 2. Mean Daily Percent Expected Meters Run for ENDURANCE Animals ................................................. 28 3. Mean Daily Percent Expected Meters Run for REGULAR SPRINT Animals .......................................... 3O 4. Mean Daily Percent Expected Meters Run for REGULAR ENDURANCE Animals ....................................... 32 5. Mean Daily Percent Expected Meters Run for HIGH SPRINT Animals ................................................. 33 6. Mean Daily Percent Expected Meters Run for HIGH ENDURANCE Animals ....................................... 34 vii CHAPTER I THE PROBLEM Competition between athletes in any single event has become so intense, that the winner may be determined by one-hundredth of a second. The need for scientific training regimens for athletes has become para- mount. It is often not the natural athlete who wins today, but the athlete who has been willing to endure agonizing, painful workouts. It is not enough in this day and age to train hard only during the com- petitive season. The attainment of high competitive performance levels requires several years of preparation. Uith intensified workloads and prolonged training periods, con- sidering the present state of knowledge, it is probable that mistakes in methods and training regimens occur. In addition, with a social milieu demanding improved performance, it is likely that an athlete may be motivated to train at an intense level when ill or under adverse environ- mental conditions. The athlete also may train so intensively on his regular training regimen that he may surpass his body's ability to adapt to the training load. Physiological changes in metabolism and in neuro- genic responses may be impaired, and damage to various tissues could occur. The phenomenon that this study is designed to explore is of a chronic nature. It is already known that an acute exposure of an indi- vidual to extremely heavy workloads, or even to moderate workloads in extreme environmental conditions, can produce damage to the organism and even death (24,34). The problem that needs to be studied is the effects of repetitive workloads on an individual that are so demanding that the body can no longer respond to this chronic stress with bene- ficial adaptations. A question exists as to whether detrimental effects may result from such excessive training. Relatively little research appears to have been published in this area. Two studies were completed which have reported damage to the myocardium and myocardial function as a result of intense, prolonged exercise (23,27). The lack of research in this area should not be con- strued as a lack of a problem. The marked increase in the intensity and duration of training regimens is only of recent origin. Absence of evidence is not evidence of absence. Several explanations can be given for this dearth of information. Foremost is the difficulty in quantitatively defining the problem. A need exists to identify the sensitive parameters which reflect over- training. In addition, few researchers would purposely try to damage the tissues of a human subject, even though the result of their experi- ments may help prevent similar traumas from occurring in other athletes. For such investigations the use of animal models is indicated. In this study, therefore, male albino rats were used as subjects. They were considered overtrained if they were unable to maintain seventy percent (70%) or less of the expected work performance for several weeks. Previous studies have shown that animals completing 75% of the expected workload had successfully adapted to the training regimen (16,18). An explanation for the scarcity of animal research in this area is the problem associated with motivation of the animals to perform at such high workloads. The development of the controlled-running wheel for the training of animals provided a system of exercise regimens which could be utilized for "overtraining" experiments (44). Need for the Study With increasing numbers of individuals participating in sports and increased pressure to perform at high levels, the effects of pro- longed, intense workouts should be examined. This study may help prevent unnecessary physical damage from occurring as a result of an intense training program. Statement of Problem The purpose of this study was to determine the effects of six different exercise regimens on the morphologic characteristics of the myocardium in the adult male albino rat. Since there was little prior information regarding the type and the intensity of training that might cause deleterious effects, two activity regimens were selected that required mainly aerobic metabolism (endurance running) and two activity regimens were selected that are thought to require mainly an anaerobic response (sprint running). The final two activity regimens were known to produce beneficial adaptations to aerobic and anaerobic workloads (16,18). Research Hypothesis Animals involved in an intense, exhaustive training program will exhibit morphological damage in the myocardium. Major characteristics of the damaged areas will be: 1) loss of striation and histochemical staining qualities, 2) focal myolysis with or without an inflammatory response, and 3) fibrosis. Research Design The animals for this study were obtained from two experiments. Both experiments' primary purpose was to train animals at exhaustive workloads. However, not all animals in the first experiment were avail- able for this study. More data were needed to draw statistical conclu- sions. This was accomplished by using animals from a later experiment. In the first experiment, eighty-four male albino rats were used as subjects. The study was organized as a two-way design with three activity groups and two dietary supplement groups. In the second experi- ment, thirty-eight male albino rats were placed into five activity groups, with each activity group divided into two dietary supplement groups. Experimental protocol and sacrifice procedures were nearly identi- cal in both experiments. The heart was removed from each animal and the tissues prepared for histological and pathological evaluations. Rationale The heart is an extremely important muscle, not only during daily existence but during exercise. It supplies the rest of the body with nutrients and with the removal of waste materials via the blood. Any damage accrued to this muscle will affect the performance of the entire organism. Therefore, the myocardium was examined in this study to observe any microscopic alterations that might result from a prolonged, exhaustive training program. Previous experiments in this laboratory have shown that training for a period of eight weeks is sufficient to produce physiological changes in male albino rats. For this reason an experimental period of eight weeks was chosen for this study (16,18,36). Male albino rats were chosen as the experimental model because of their capability for successfully completing a running exercise program. Limitations l. The results of this study can only be applied to adult male albino rats. 2. The morphological analyses were limited to two cross sections of the heart, one ventricular and one apical. Small areas of damages nay have existed in portions of the heart that were not examined. 3. The shock stimulus used for motivation in the training program may be involved in the production of myocardial changes. 4. There was no quantitative control over diet. 5. while the exercise regimen were designed deliberately to pro- duce an exhaustive overload on the animals, it may not have been diffi- cult enough to produce the phenomenon of overtraining. Significance of the Study Determinations of the possible deleterious effects of prolonged, exhaustive exercise in rat myocardium may provide insight into an organism's ability to adapt to such overloads. Injuries to future athletes may be avoided if it is demonstrated that a system of dimin- ished returns operates in heavy training. CHAPTER II REVIEW OF RELATED LITERATURE A small number of studies have reported detrimental effects to the heart as a result of continuous exhaustive exercise. Stevenson et al., in experiments with male albino rats that were swum or run on a treadmill for four weeks, reported that: ... the heavier and more frequent the exercise, the Poorer was the gain in body weight and the increase in coronary tree size. Stevenson hypothesized that the tissue catabolism that follows strenuous exercise was detrimental to the vascularization and work hypertrophy of the heart (41). The supporting data in Stevenson's work are difficult to inter- pret. The work intensity of the animals running on the treadmill was not given. Only the total distance run is mentioned (1.3 km). Velocity, work time, and rest time were not stated. It is possible that animals in this experiment were still in the process of adapting to a training program. Since the duration of the experimental period was relatively short, the observations may represent the process of adaptation to run- ning rather than effects produced by overtraining. Interpretation of the swimming data is also difficult. It is cur- rently recognized that while rats can swim with relative ease, it is extremely difficult to quantify the amount of work done. Buoyancy is a problem in most swimming programs. It was not stated if the animals were shaved, if weights were attached to their bodies, or if a detergent was added to the water to counteract the animals' natural buoyancy. It was also not reported if the animals swam singly or as one group. When rats were swum as a group, the animals try to climb on each others' back, causing frequent submersion of the weaker animals. Unless the training procedures prevented this from occurring, the effects observed on the rat myocardium may have been due to stresses other than exercise. Further studies are necessary to replicate Stevenson's observa- tions. Morphological signs of myocardial damage attributed to exhaustive exercise have been reported by Kitamura (23) and Letunov (27). In experiments with young mice trained to swim or to run on a motor-driven treadmill, Kitamura found that after ten weeks of training the myocar- dial tissue showed small hemorrhages, infiltration of leukocytes, inter- stitial fibrosis, and myocardial hypertrophy. Six weeks after the completion of fourteen weeks of training, the heart size had decreased and extensive irreversible fibrosis of the myocardium had occurred. Kitamura also studied the effects of eight weeks of intensive exercise on the ultrastructure of cardiac muscle fibers. When compared to the control animals the structure of the muscle fibers appeared dis- arranged. In some cases, vacuolization of the muscle fiber and osmio- philic degeneration within the myocardium nuclei were seen. The ratio between the number of myocardial fibers and capillaries within 100 square microns was also studied. The exercise group showed a marked decrease in fiber to capillary ratios for the first forty days of the training program when compared to the ratio of the control animals. After forty days, the ratios of the exercise group jumped markedly sur- passing the ratios of the control groups. These data may support the view that the detrimental effects of exhaustive exercise as mentioned in the Stevenson's study are part of an adaptive process. Unfortunately, as in the Stevenson's paper, the training regimen of Kitamura was not described. Letunov has summarized the works of several authors on the effects of exhaustive training in a paper presented at the International Congress of Sport Sciences, 1964. Unfortunately, due to the format of publica- tion, no bibliography was given and the original articles could not be located. Therefore, the results of these studies will be presented as described by Letunov. Morphological data on the myocardium from overtraining studies performed by Gudz were discussed by Letunov. Perivascular bleeding in the right and left ventricle as well as microinfractions and micro- necroses in the anterior and posterior wall of the left ventricle were noted. Fatty degeneration and myolysis of the cardiac muscle fibers also were reported. Although the experimental model and training regime were not reported, it is interesting to note the similarities in morpho- logical findings between Kitamura and Gudz. Clinical symptoms of chronic overtraining described by Letunov in humans are hypertension, heart pain, and abnormal EKG patterns. Two per- cent of athletes tested had abnormal ERG patterns of the left ventricles attributed solely to overtraining (27). 10 Cases of sudden death in supposedly fit athletes occurring during or after extremely heavy physical activity have been reported. Many of these deaths were attributed to organic defects of the heart structure (21,22). Other deaths were associated with extreme environmental condi- tions concommitant with exercise (33,38). In addition, a few case studies have reported that small cardiac lesions of the conduction system will cause death (19). This mechanism could explain those cases of sudden death not involving the myocardium, valves, or coronary vessels. Small hemorrhages, like those observed by Gudz and Kitamura, would cause death if they affected the conduction system of the heart. In the data reviewed, exercise was not determined to be the sole causal factor in producing sudden death in any athlete. This is not surprising. Currently there is no syndrome to delineate exercise as the cause of death during an autopsy. The mechanism that causes an apparently healthy heart to develop pathological symptoms from overtraining have not been elucidated. One possible mechanism, suggested by Raab (34), is the intensive absorption of catecholamines by the myocardium during exercise. In the following sections, the effects of catecholamines on the myocardium and the effects of exercise on catecholamine secretion are briefly reviewed. Catecholamines and Heart Damage Various studies have demonstrated that exogenously administered catecholamines can elicit heart damage. Morphological alterations in the myocardium following the injection of epinephrine are essentially of 11 two types: 1) hypertrophy and dilation, and 2) myocardial degenerative changes varying from slight edema, hyaline degeneration, leukocytes infiltration to necroses and scar formation (34). Alterations in the ultrastructure of myocardial cells occur after the injection of minute doses of epinephrine (39). These changes are reversible. Larger doses cause an abnormal distribution of potassium, a depletion of glycogen reserves, and a decline in phosphorylase activ- ity in the subendocardium and apex (l). The spotty distribution of potassium is similar to that produced by experimental myocardial hypoxia. This led Bajusz and Raab (l) to hypothesize that the necrotizing proper- ties of catecholamines is related to their influence on myocardial electrolyte metabolism. Raab had suggested earlier the epinephrine mimics the effects of coronary insufficiency by increasing local oxygen consumption (34). Damage occurs because the increased oxygen consump- tion is only partially compensated for by an increased coronary flow. Catecholamine Secretion During Exercise The release of epinephrine during exercise was observed as early as 1922 by Hartman (15). These early studies measured the secretion of the adrenomedullary hormones in venous plasma from an exteriorized gland. Current methods for the estimation of catecholamines allow measurement of the actual level of catecholamines in the circulating blood. Epinephrine values in venous plasma of cats (5,15), dogs (4), and humans (2,11,13,14,20,25,43) have been shown to increase progressively with heavy exercise. The rise in epinephrine level in venous plasma 12 during heavy exercise is also manifested by an increased epinephrine value in arterial plasma (6,11). There is general agreement that norepinephrine increases during heavy exercise both in venous and arterial plasma. However, the increase is not proportional to workload. There are small increases in norepinephrine levels up to 60-70% of an individual's max V02, where- after it rises relatively quickly. Haggendal has proposed that the norepinephrine level rises exponentially to the workload (12). The levels of catecholamines in venous plasma for untrained sub- jects during a given workload is higher than for trained subjects, even though both groups have similar values at rest (13,14). Training of an individual decreases the catecholamine levels in venous plasma for a given workload (ll,13,l4,20). Decreased, increased, and unchanged myocardial norepinephrine levels in heart tissues of trained animals have been reported. However, these levels have not been closely correlated with resting heart rate (8,26,31,32). Epinephrine and norepinephrine uptake and turnover were found to be decreased in the hearts of trained animals. The results indicated that physical conditioning causes an adaptation of the sympatho-adrenal system to exercise stress. The functional changes that accompany train- ing lead to a better transmitter economy during exercise. It appears that training provides protection to the heart from the necrotizing effects of catecholamines. Both epinephrine and nor- epinephrine secretion and myocardial uptake and turnover are lower for the trained animal during exercise. It is not likely that myocardial damage from exercise can be attributed solely to catecholamines. CHAPTER III METHODS AND MATERIALS Exercise can be viewed as a continuum of various activities. Each level within the continuum requires specific adaptation of the body in order to meet the metabolic requirements of that activity. For example, distance running is dependent on oxidative metabolism and a high oxygen uptake, while sprint running is dependent on anaerobic metabolism and a high oxygen debt. Each of these events requires dif- ferent cellular and cardiovascular adjustments of the body. It would seem likely then, that repetitive exhaustive workloads of differing exercise regimens would lead to different pathological states. This study was designed to investigate different exhaustive training regimens and their effects on the morphological condition of the heart. In order to sample from a variety of training programs, animals for this study were selected from two experiments. Animals In Experiment One Eighty-four normal male albino rats of the Sprague-Dawley strain were obtained from Hormone Assay Inc., Chicago, Illinois.1 They were 1Hormone Assay Laboratory, 8159 S. Spalding, Chicago, Illinois. 13 14 received at weekly intervals in three shipments of 30, 24, and 30 animals respectively. Each shipment was designated as a separate activ- ity group. A standard period of 12 days was allowed for adjustment to the laboratory conditions. Activity was initiated when the animals were 84 days old. Research Design In Experiment One This study was organized as a two-way design with three activity groups and two dietary supplement groups. The duration of the experi- ment was eight weeks. The treatment conditions were as follows. Control Group The 24 animals in the second shipment constituted the sedentary control (CON) group. These animals received no exercise and were forced to remain relatively inactive throughout the experiment. The animals were housed in individual sedentary cages (24 x 18 x 18 cm) during both the adjustment and treatment periods. Sprint Group The sprint running (SPT) group was comprised of the 30 animals in the first shipment. Each of these animals were housed in individual voluntary activity cages (sedentary cages with access to a freely revolv- ing wheel) during the adjustment period and in individual sedentary cages during the experimental period. The SPT animals were subjected to an interval training program of very high-intensity sprint running. The workload of the SPT group was gradually increased until on the 27th 15 day of training and thereafter, the animals were expected to complete six bouts of exercise with 2.5 minutes of inactivity between bouts. Each bout included five lS-second work periods with four 30-second rest periods. During the work periods, the animals were expected to run at a velocity of 108 m/min. Endurance Group The endurance running (END) group was composed of the 30 animals of the third shipment. These animals were housed the same as the SPT animals. The END animals were subjected to a demanding program of dis- tance running. The workload was gradually increased so that on the 30th day of training and thereafter, the animals were expected to complete 60 minutes of continuous running at 36 m/min. Vitamin C Group One-half of the animals in each activity group received Vitamin C supplementation (C group). Vitamin C supplementation was administered orally by syringe with a dosage of 2.4 mg vitamin (Merck)2 in a .1 ml 5% sugar solution per 100 g body weight between 1900 and 2100 hrs daily. The administration of the supplement was begun the day prior to the initiation of activity and terminated the day prior to sacrifice. The dosage used was equivalent to 1680 mg daily for a 70 kg man. 2Merck and Co., Inc., Rathway, N. J. 16 Placebo Group The remaining animals in each of the activity groups (no C group) received an identical quantity of the sugar solution per unit of body weight. Animals In Experiment Two Thirty-eight normal male albino rats of the Sprague-Dawley strain were obtained from Hormone Assay, Inc., Chicago, Illinois. The animals were randomly assigned to treatment groups. A standard period of twelve days was allowed for adjustment to laboratory conditions. Training was initiated when the animals were eighty-four days old. Research Design In Experiment Two This study was organized as a two-way design with five activity groups and two dietary supplement groups. The duration of the treatment period was eight weeks. The treatment conditions were as follows. Control Group Six animals were randomly assigned to the control group. These animals were placed in individual sedentary cages (24 x 18 x 18) through- out the adjustment and treatment period. The animals remained sedentary the entire period. Regular Sprint Grogp Eight animals were subjected to a high-intensity, short duration exercise program. Gradual increases in the intensity of the program 17 were made so that by the 37th day of training the animals were required to run eight bouts at a velocity of 99 m/min. Each bout consisted of six lO-second work intervals alternated with 40-second rest intervals. The animals were allowed to rest 2.5 min between bouts. A description of the complete training program is given in Appendix A. The regular sprint (REG SPT) group animals were housed in individ- ual voluntary activity cages during the adjustment period and in individual sedentary cages during the experimental period. High Sprint Group Eight animals were subjected to a very high-intensity, short dura- tion running program. The training progression was identical to the regimen used by the sprint group. A description of the complete train- ing program is given in Appendix A. Animals in the high sprint (HI SPT) group were housed in a similar manner as the REG SPT. Regular Endurance Group The regular endurance (REG END) group was composed of eight animals which were subjected to a low-intensity, long duration running program. By the 37th day and thereafter, the animals were expected to complete four bouts of exercise with 2.5 minutes of inactivity between bouts. Each bout consisted of one 12.5 minute continuous run at a speed of 36 m/min (see Appendix A). These animals were housed similarly to the REG SPT. 18 High Endurance Group Eight animals were subjected to a moderate intensity, long dura- tion running program. The workload was rapidly increased so that by the 32nd day, the animals were expected to run 60 minutes at a velocity of 45 m/min (see Appendix A). The high endurance (HI END) group were housed like the REG SPT group. Sugar Group One-half of the animals in each activity group received daily sugar supplements. The sugar was administered orally by syringe after completion of the exercise regimen, or for the controls, between 1100 and 1200 daily. Each dose consisted of .1 ml 5% solution per 100 g body weight. The administration of the sugar supplement was initiated on the first day of the treatment period and terminated the day prior to sacrifice. Placebo Group The remaining animals in each activity group received an identical quantity of water per unit of body weight under identical circumstances. Trainipg Procedures All exercise groups in both experiments were trained in a battery of individual controlled running wheels (CRM). The apparatus has been described as: 19 ... a unique animal-powered wheel which is capable of induc— ing small laboratory animals to participate in highly specific programs of controlled, reproducible exercise. (44) Animals learn to run in the CRW by avoidance-response operant conditioning. A low intensity controlled shock current (1.2 ma), applied through alternating grids comprising the running surface, pro- vides the motivation for the animals to run. A light above the wheel signals the start of each work period. The animal is given a pre- determined amount of time (acceleration time) to attain a prescribed running speed. If the animal does not reach the prescribed speed by the end of the acceleration period, the light remains on and shock is admin- istered. As soon as the animal reaches the desired speed, the light is immediately extinguished and shock is avoided. If the animal fails to maintain the prescribed speed throughout the work period, the light- shock sequence is repeated. Most animals learn to react to the light stimulus after only a few days of training. A typical training session consists of alternated work and rest periods. The wheel is braked automatically during all rest periods to prevent spontaneous activity. The brake is released and the wheel is free to turn during the work periods. Performance data are displayed for each animal in terms of the total meters run (TMR) and the cumulative duration of shock (C05). The TMR and the total expected meters (TEM) are used to calculate the per- centage of expected meters (PEM): PEM = 100(TMR/TEM) PEM values are the chief criteria used to evaluate and compare training 20 performance. A secondary criteria is provided by the percentage of shock-free time (PSF) which is calculated from the C05 and the total work time (TWT): PSF = 100 - 100(CDS/TWT) In experiment one, all exercise treatments were administered once a day, Monday through Friday, between 1230 and 1730. In experiment two, all exercise treatments were administered once a day, Monday through Friday, between 0800 and 1230. Animal Care Animal care procedures were identical in both experiments. All housing cages were steam-cleaned every two weeks. Standard procedures for CRW cleaning and maintenance were observed. The animals received food (Wayne Lab Blox)3 and water ag_libitum. A relatively constant environment was maintained for the animal by daily handling as well as by temperature control. The animals were exposed to an automatically regulated daily sequence of 12 hours of light followed by 12 hours without light. Since the rat is normally a nocturnal animal, the light sequence was estab- lished so that the lights were off between 1300 and 0100 and on between 0100 and 1300. This lighting pattern altered the normal day-night schedule for the animals so that they were trained during the active phase of their diurnal cycle. 3Allied Mills, Chicago, Illinois. 21 Body weights of all exercised animals were recorded before and after each training session. The control animals in each experiment were weighed weekly. Sacrifice Procedures Experiment One Anticipated limitations of time and personnel restricted the number of animals that could be handled at sacrifice to 12 in each treatment group. Since one of the inherent purposes of the study was to compare various parameters in two groups of highly trained animals and a group of untrained animals, three extra rats originally were included in the SPT and END groups. Twelve animals were selected for sacrifice from each of these two groups on the basis of their health and their training performance throughout the treatment period. Only animals subjectively determined to be in good health were chosen. Because the training requirements were extremely vigorous, no absolute minimal performance criteria were established. However, individual daily records of PEM and PSF values were examined, and those animals making the best adaptations to the training regimens were selected for sacrifice. All 12 CON animals were judged to be healthy and were sacrificed. Three sacrifice periods of two-days duration (Monday and Tuesday) were established. All animals within a treatment group were killed dur- ing a single sacrifice period (i.e., six animals each day). The trained animals were killed either 72 or 96 hrs after their last exercise bouts 22 were completed. This procedure was followed to eliminate any transient effects of acute exercise. The animals were either 140 or 141 days old at sacrifice. Final body weights were recorded inmediately prior to sacrifice. Each animal was anesthetized by an interperitoneal injection (4 mg/lOO gm body weight) of a 6.48% sodium pentobarbital (Halatal) solution.4 After selected leg muscles were removed for analysis by other investiga- tors, the heart was removed and washed free of blood with Ringer's solu- tion. The great vessels of the heart were trimmed. A dissection along the atrioventricular groove was performed to separate the atria from the ventricles which then were tranversely sectioned approximately one- third of the ventricular length from the apex. The apical portion, mounted apex up, and the ventricles were mounted on cork strips using 5% gum tragacanth. The preparation was held with forceps and lowered, for approximately 20 seconds, into 2-methylbutane (isopentane) pre- cooled to a viscous fluid (-l40° to -185° C) by liquid nitrogen. The apical portion of the heart was sectioned approximately 300p from the apex. Sections, lOu thick, for both portions were cut on a microtome in a cryostat at a temperature of -20°C. Experiment TWo One sacrifice period of two-day duration (Saturday and Sunday) was established. Animals were randomly assigned to sacrifice order. The trained animals were killed 72 hrs after their last exercise bout was completed. The animals were 140 or 141 days old at sacrifice. 4Haver-Lockhart Laboratories, Shawnee, Kansas. 23 Eighteen animals were killed on Saturday and seventeen animals were killed on Sunday. Final body weights were recorded immediately prior to sacrifice. Each animal was decapitated and quartered. The heart was removed and the great vessels trimmed. The heart was sectioned approximately one- third of the ventricular length from the apex. At this time, sacrifice procedures parallel those described in experiment one. Histological Methods The fresh frozen sections from the apical and ventricular portions of the heart were stained with a hematoxylin-eosin (H&E) technique for the evaluation of general cellular structure (28). A modified Gomori‘s trichrome and Van Kossa's stain were used to identify collagen deposi- tion and calcification, respectively (28). Absence of succinic dehydro- genase (SDH) activity was used as an indication of early necrosis (29). Pathological Evaluations The heart sections stained with H&E, SDH, modified Gomori's tri- chrome, and Van Kossa's stain were evaluated subjectively under a light microscope. They were rated on a 0 to 4 scale (29,35) according to the lesions exhibited. Both the severity and the number of lesions were considered. A low rating (0 to l) was assigned to those sections with no damage or only a few small foci of damage. A higher rating was used to indicate more-extensive pathological involvement. 24 Statistical Procedures The body weights and the absolute and relative heart weights were analyzed using a two-way (3 x 2 design in experiment one; a 5 x 2 de- sign in experiment two) fixed effects analysis of variance. Student- Newman-Kuels tests were used to evaluate differences between pairs of means whenever a significant F-ratio is obtained. Chi-square con- tingency tests were used for the pathological data. Levels were com- bined to meet the assumptions of chi-square tests. CHAPTER IV RESULTS AND DISCUSSION The material in this chapter is organized into six main sections. The first part deals with training results from the Controlled-Running Wheel (CRW) programs of experiment one. The second section covers the training results of experiment two. Body and heart weight results at sacrifice are discussed in the third and fourth sections for experiment one and two respectively. Histopathological results are presented next. Finally, a discussion of the more important findings is given at the end of this chapter. Training_Results of Experiment One1 The sprint (SPT) and endurance (END) Controlled-Running Wheel (CRW) training programs are presented in Appendix A. These programs are modi- fied versions of standard regimens routinely used in the Human Energy Research Laboratory, Michigan State University, East Lansing, Michigan. The modifications were incorporated in an attempt to design strenuous exercise programs which would primarily stimulate anaerobic or aerobic 1Part of the material in this section has been adapted from the unpub- lished Ph.D. dissertation of Roland R. Roy (36). 25 26 metabolic processes in the animals. The performances of the animals were evaluated using the percentage of expected meters (PEM) and the percentage of shock-free time (PSF) as criterion measures (44). The performance data for the SPT-C and the SPT-No C groups are presented in Figure l. A runs test indicated that there were no sig- nificant differences in performance between the SPT-C and SPT-No C animals. Progressive increases in the required running velocity were made rapidly. From the beginning of the fourth week of training to the end of the program, the animals were expected to run at velocities rang- ing from 90 to 108 m/min (see Figure l and Appendix A). No comparable exercise programs for small animals have been found in the literature. The results indicate that the animals could not maintain the program requirements. PEM values fell to approximately 45% during the last three weeks of training as contrasted with the usual criteria of 75% for satisfactory completion of an exercise regimen. The training data for the END-C and END-No C groups are shown in Figure 2. PEM values were 70% or higher each day of training in both the C and the No C animals. These results indicate that the animals were able to maintain the daily requirements of the END program rela- tively well. The END animals ran at the relatively slow speed of 36 m/min. Periods of continuous running were progressively increased to 60 min at the end of five weeks of training and were maintained at this level for the remainder of the eight week program (see Figure 2 and Appendix A). The single bout of exercise was determined subjectively to result in daily physical exhaustion of the animals. On the average, the rats lost 27 .m_as_=< czcmam toe cam agape: umuuaaxm “smegma spew: cam: ._ 0v F on pth.__ 9.90 0 02 I 990 o 0110 b h 0 . fi....n..fi.$i...t ___—bP_bPPbP_—h_b—F_—h_~P-FFPp wgsmwd .56)... .._m> :3 .225 .. o J 1 8 o 0 w 1 m .... m .. 0.. %w 1 .w 3 x n.0w Am 3 m I. p w .... : 8 n s I .. oo. 1 8. L 93 28 .m_asc=< muz_cao cam: .N wgamwg _— 1 on LTi 6.5.5.5 ..m> 9v mm on mm ON 9 O. Fb_b__P_.E_P_b__r_.PcpptLEFb.p_c___m_»_ 9.80 0 oz I 9.20 o olo h >40 2.4m: . a 0 ON 1 1 O? Harem poisedxg ;o «y. pemdmoo .. 00. .. 0N. 29 2.55% of their body weight during each training session (see Appendix 8). Body weight data were used to award an unplanned recovery day on Wednesday of each of the last three weeks of training. The animals were run on the 39th and 40th day of the program, but the results were not recorded due to a technician error. Trainipg Results of Experiment Two The regular sprint (REG SPT), the regular endurance (REG END), the high sprint (HI SPT), and the high endurance (HI END) Controlled- Running Wheel training programs are presented in Appendix A. The REG SPT and REG END programs are the standard regimens routinely used at the Human Energy Research Laboratory at Michigan State University to stimu- late anaerobic and aerobic metabolic processes in small animals. The HI SPT and HI END programs are modified versions of the SPT and END programs used in experiment one. The modifications were incorporated to further refine strenuous exercise programs. The performances of the animals were evaluated using the percentage of expected meters (PEM) and the percentage of shock-free time (PSF) as criterion measures. The performance data for the REG SPT-Sugar and the REG SPT-Placebo are presented in Figure 3. Animals in both groups demonstrated similar training patterns throughout the experimental period. A runs test was significant (p< .05). An unknown factor may have influenced the perform- ance of the SPT group. The sugar treatment is most likely the unknown factor. PEM values for the REG SPT groups were 56% or higher each day of training. These values are lower than values from animals previously 30 .m_asc=< cszam m 0.. mm on mm 8 m. o. m I1 ON 3 O W l d .I .1... .l O? 3 0 . . I 9/9 o a o o o o O o o o g o o o '1 00 J.— o o o 3 . . X o 0 l d . 3 . . O . . . 1 8 a O . l w . 3 . m . l 00. a S . I . .1 ON. .096 02 To L ..OODW I . L 0?. 31 trained using the identical protocol (16,18). A PEM value of 75% is the usual criteria for satisfactory completion of an exercise regimen. The small sample size of this experiment required that every animal trained be considered in the data analysis. The correlation between PEM and PSF (.45) for the REG SPT animals was also lower than earlier studies (see Appendix B). This may indi- cate that the current animals did not respond satisfactorily to the training regimen. A small sample size (N=7) may have depressed the correlation. The training data for the REG END-Sugar and REG END-Placebo groups are shown in Figure 4. Animals in these groups were subjected to a low intensity, long duration exercise program designed to stimulate aerobic metabolic processes. Both the Sugar and Placebo groups demon- strated similar training patterns. The data indicates that these animals maintained a PEM of approximately 70% during the later stages of training. While this value is lower than previous studies, it is suffi- ciently high to assume that the majority of animals had successfully completed the requirements of the training program. The training data for the HI SPT-Sugar and HI SPT-Placebo are shown in Figure 5. Animals in both groups had similar training patterns. The HI SPT program in experiment two is very similar to the SPT program in experiment two. The main differences exist in the first seven days of training. In experiment two, the intensity and total work time was increased in the early phase of training to stimulate anaerobic processes (see Appendix A). Progressive increases in the required 32 .mpaec=< muz<¢=ozm m 9. on 0» mm 8 n. o. a >3 2.5: _pc_____.__._~_Lhc___.____________P__~__.lo .v.> . . . .... 88313011 03103:!)(3 :10 'l. 03131651100 beam 02 I .. l Boom I LO! 33 .m_mswc< Hzamam :uH: cot cam mcmuwz umuomaxm “emote; apmmo new: .m mgzmwd T 8. ‘TI mm 1*1 om lle. .m lle we. Ile on linen.— A.=_e>5 ...u> 0% mm om m. o. m on On >8.2. m O? mm Om mN ON 0. O. >40 .223: __b_______7_________P~_r_.__b__________rO l._. -1 ION I 3 O w 3 l l 3 O 18% 0 I ... 3 x I00% 0 II. I 3 0 109m 1 3 .4 w . ION. .103 cooamozollo I8. .0951 . L8. 35 running velocity were made rapidly. From the beginning of the fourth week of training to the end of the program, the animals were expected to run at velocities ranging from 90 to 108 m/min. As in experiment one, PEM values fell to approximately 45% during the last three weeks of training. The response of the HI END-Sugar and HI END-Placebo animals to the training regimen is presented in Figure 6. The HI END program was a modification of the END program from experiment one. Changes in the regimen were made to produce a more exhaustive, long duration exercise program. At the completion of six weeks of training and thereafter, the HI END animals maintained a running velocity of 45 m/min for one hour. During the same period of training the animals in experiment one only maintained a running velocity of 36 m/min for one hour. PEM values indicate that the HI END animals could not maintain the program's requirements. PEM values in experiment two fell to approximately 39% during the last three weeks of training. In experiment one, PEM values were approximately 70% during this time. Body and Heart Weight Results at Sacrifice for Experiment one At the end of eight weeks of exercise, the trained animals were significantly lighter than the sedendary control animals (Table l). The differences in body weight between the SPT and END groups of animals were not statistically significant (Table 1). Both trained groups were approximately 20% lighter than the CON group. These results are in agreement with those of previous studies using the CRW (16,18,42) and 36 .Fm>mp mo.o esp um “emcee newspmmcu Ppmcm>o ucmowmwcmwm * Am-o_v Scars: NNN. mvm.p N_.e o~.m mm.m acme: m>wumpmm Amy peace: Hzm A zoo mmo. nnm.m mmo.p mw¢.— mew.P acme: muzpomn< zoo v ozm Amv muccccuam zoo v ham amooo.v Pmm.om m.wov m.~mm F.mpm am weave: xuom xzm mapm> mzpm> ozm ham zoo mpnmwce> a a meme: pcmsummcp ucmucmamo .mco pewsmcmaxm toe .mucmwwz “Low; m>wpm~mc use muapounm use mowevcumm an Scmwmz xuoa Loy mcompgmaeoo umcwma co mummy m_Fzmx-cmEzwz use muommcm mcwcwmcu ppmcw>o LOO mocwwcm> co mpmxpmc< .P mFQm» 37 and support the general observation that strenuous exercise slows the usual gain in body weight seen in the male rat over time (7). The slower rate of weight gain usually is attributed to an increase in caloric expenditure associated with exercise. In some instances the growth impairment has been ascribed to a significant reduction in food intake (7,30). However, these parameters were not monitored in the present study. Animals in the control group had a statistically significant larger absolute heart weight than the animals in the sprint group. The absolute heart weights of the control animals were not different from the endurance animals. The exercised animals had larger relative heart weights than the control animals and the END animals had larger relative weights than the SPT animals. None of the differences in relative heart weight were statistically significant. Vitamin C supplementation did not appear to affect body weight, absolute heart weight, or relative heart weight. Mean values and the two-way ANOVA results are shown in Table 2. Body and Heart Weight Results at Sacrifice for Experiment Two Training and diet had an interaction effect on body weight at sacrifice (see Table 3). This interaction may have been caused by a significant diet effect on body weight at the start of the exercise program. An analysis of the weight gained data indicated that there was no true diet effect. Animals in the control group gained signifi- cantly more weight during the experimental period than did animals in 38 Table 2. Analysis of variance for overall vitamin C effects for body weight at sacrifice and absolute and relative heart weights for experiment one. Dependent Treatment Means F P Variable C No C Values Value Body Weight 433.367 447.033 3.202 .079 at sacrifice (9) Absolute Heart 1.641 1.685 .157 .694 Weight (9) Relative Heart 3.83 3.78 .036 .851 Weight (10'3) 39 Table 3. Analysis of variance for overall sugar effects for experiment one. Dependent Treatment Means F P Variable Sugar Placebo Value Value Body Weight* 423.824 440.778 4.870 .035 at sacrifice (9) Beginning Body 321.471 339.333 17.530 .001 Weight (9) Weight Gained (9) 101.444 102.353 .070 .999 Absolute Heart 1.434 1.411 .029 .999 Weight (9) Relative Heart 3.261 3.329 .921 .999 Weight (10-3) *A two-way interaction effect between Diet and Training was observed with body weight at sacrifice (F value = 3.14, P value = .032). There were no other significant two-way interactions among the dependent variables. 40 the running programs (see Table 4). The trained animals were 35% lighter than the control animals. There was no difference in the amount of weight gained between any of the exercised animals. Animals in the control group also had significantly larger abso- lute heart weights than animals in the HI END, REG SHT, and REG END groups. These differences were not significant when relative heart weights were considered. However, the means of relative heart weights for all exercise animals were higher than the control animals. I Histppathological Results Microscopic evaluations of the hearts as previously described revealed a small number of animals who developed heart damage. In order to meet the criteria of Chi-square test, it was necessary to pool the heart damage ratings and the training treatment. Animals with the rat- ing of 0 formed the no myocardial damage group. Animals with a rating of l or higher formed the myocardial damage group. As these groups indicate at least some degree of myocardial damage, the high number of animals classified as having myocardial damage may be misleading. The majority of animals in this group showed very slight damage. The heart damage in most animals consisted of small focal areas of inflammatory reaction. Moderate heart damage was observed in four of the thirty-six trained animals, while only one of fifteen control animals showed such damage (see Tables 5 and 6, on the following page). A Chi-square contingency test showed that there was no statistical- ly significant difference in heart damage between the exercised and control animals (see Table 7). -- mma. mac. aoe.m~m oma.omm coo.mmm ooo.e~m mmm.emm Amy agave: xuom mcpccpmmm Am-o.v peace: -- mam. wee. Nmm.m mmm.m Noe.m amm.m m_N.m cam: m>cpa_mm 1.ozm wmmxomm 4.c:m wmmxomm Amy peace: ozm Hzxamm m_o. _mo.e wmm._ Nam.” mma._ mam.P mmm._ Scam: ap=FOma< cam amzxomm ozu wmzxomm cam Hzxamm _oo. ova.“ o.mo mam.aw mam.em m~¢.om Nep.ae_ “my caccam ugm_a3 azm Hzxomm Amy mowcweomm um _oo. Nee.w ao©.oma mNe.mPe mam.mm¢ www.mma ooo.m~¢ peace: seam “mac a=Pa> a=Pa> mzm HI oz“ mum ..ezm H: .czm gum .zup mpaacca> xzm a a meme: acmEummE» ucmucmamo .ozg pcmEPLmaxm com mcomwcmaeou umcwma we mummp m.P=mx-:mezmz ucmczum use muomccm mcwcwmcp Ppmcm>o so» mocepcm> we mwmxpmc< .e open» 42 Table 5. Frequency distribution of ratings of myocardial damage in experiment one. Trainin Group_g st. Rating CON END boom—Jo ooomoo DONNC‘ O--'-l—|\l Table 6. Frequency distribution of ratings of myocardial damage in experiment two. Trainin Grou Rating CON REG SPT_' HI SPT REG END HI END boom—'0 COO-‘01 OOONU‘I OOO—‘V OOOOJU'I OO—‘OU‘I Table 7. Chi-square test for treatment effects on the degree of myo- cardial damage. CON TRAINED N No Damage 13 35 48 Damage _;3 14_ ll. 16 49 65 2 .60 p> .05 X 11 43 Discussion Several possible explanations could account for the relatively poor performance data of the SPT, HI SPT, and HI END groups. The required running velocities may have been too fast. Observations during the training sessions, however, showed that the animals were capable of running at the desired speeds. Low PEM and PSF values might suggest that the animals responded to the unconditioned shock stimulus rather than to the conditioned light stimulus. Improper initial training and defects in the CRW equipment could lead to such a learning problem, but the END, REG SPT, and REG END groups learned to run under the same conditions and had no such difficulties. A lack of control of environ- mental factors affecting training performance might have accounted for the poor results of the SPT group. This is particularly true for air temperature and percent humidity, but again the END data make this explanation improbable (see Appendix B). The environmental factors during experiment two did not vary as drastically as in experiment one. The most likely cause of the low PEM and PSF values is that the SPT, HI SPT, and HI END regimens may have produced a state of overtraining. The data in Figures 1, 5 and 6 support this hypothesis. Repeated in- creases in the required velocity may have been expected from the animals before they were fully adapted to the previous velocity. The constant additional stress could have resulted in overtraining. Bone data from the animals in experiment one strongly reinforce the hypothesis that overtraining was produced by these exercise regimens (40). A retardation of long bone growth occurred with the trained 44 animals. In addition the SPT group had significantly lighter tibia weights when compared to the bones of the CON group. These results were not anticipated in light of Wolff's Law. The body weight data from experiment one and experiment two are consistent among themselves and with previous studies (16,17). The control animals gained significantly more weight than the trained animals. Larger animals may have larger hearts. From the body weight data, it was not surprising that the CON animals had larger absolute heart weight than did the exercised animals. Although not statistically significant, when body weight was controlled for mathematically by a heart weight/ body weight ratio, the trained animals had relatively larger hearts to support relatively less body mass. There was not a statistically significant degree of myocardial damage associated with strenuous training. Indeed, four of the CON animals showed some degree of myocardial damage. The heart damage seen in control and trained animals may have been caused by unknown stress factors or by an advanced degree of atherosclerosis in the coronary arteries. Since the degree of atherosclerosis was not measured in this experiment, there is no way of determining if either of the above factors were instrumental in the production of myocardial damage. From the training data, it appeared that the trained animals were subjected to exhaustive workloads. It may be that these exercise regi- mens were not sufficiently intense or of a long enough duration to produce myocardial damage. A more likely explanation is that exercise. as a sole factor in a healthy animal, will not produce myocardial damage. CHAPTER V SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS Summar This study was undertaken to investigate the effects of long-term, exhaustive workloads on the myocardium of normal male adult rats (Sprague-Dawley strain). Six different exercise regimens were selected. The programs included two regimens that required mainly aerobic metabol- ism (endurance running), two regimens that are thought to require mainly anaerobic metabolism (sprint running), and two activity regimens that were known to produce beneficial adaptations to aerobic and anaerobic workloads (16,18). Animals for this study were obtained from two experi- ments. In the first experiment, eighty-four male albino rats were placed into three activity groups, with each activity group divided into two dietary supplement groups. The second experiment was organized as a two-way design with five activity groups and two dietary supplement groups. Experimental protocol and sacrifice procedures were nearly identical in both experiments. The heart was removed from each animal and the tissues prepared for histological and pathological evaluations. Analysis of variance of the dietary supplements in both experi- ments indicated no significant differences in body weight gained, absolute heart weight, and relative heart weight. 45 46 In experiment one, the absolute heart weight of the CON group was significantly larger than the SPT group. No significant differences were observed in relative heart weights. In experiment two, the absolute heart weights of the CON group were significantly larger than the HI END, REG SPT, and REG END groups. Again there were no statistically significant differences observed in relative heart weights. A chi-square test indicated there were no significant differences in heart damage between trained and control animals. Conclusions The results of this study have led to the following conclusions: 1) Sedentary animals had significantly larger absolute heart weights than the SPT, REG SPT, REG END, and HI END animals. 2) The inclusion of vitamin C in experiment one and of sugar in experiment two did not affect body weight or heart weight of the animals. 3) The exercise imposed in this study did not affect the amount of heart damage observed. Recommendations 1) Studies are needed to further refine exercise regimens involv- ing repetitive, exhaustive workloads. 2) Exercise regimens that involve power events such as weight- lifting and high jumping should be developed. The effects of these 47 training programs on the myocardium should be investigated. 3) Further studies are needed to delineate the phenomenon of overtraining. These studies should consider serum catecholamine levels, heart catecholamine levels, and adrenal and thyroid responses to chronic exhaustive exercise. 4) Histochemical analysis of the extent of atherosclerosis in coronary arteries of animals with heart damage should be investigated. REFERENCES 10. 11. 12. REFERENCES . Bajusz, E. and W. Raab. Early metabolic aberrations through which epinephrine elicits myocardial necrosis. In Prevention of Ischemic Heggt Disease, W. Raab, Ed., Springfield, 111.: ChaFles C. Thomas, 19 . . Banister, E. W. and J. Griffiths. Blood levels of adrenergic amines during exercise. J. Appl. Phy. 33: 674, 1972. . Barka, T. and P. J. Anderson. Histochemistry Theory, Practices and Bibliography. New York: Harper and raw, 1963, p. 313. . Brzezinska, Z. and K. Nazar. 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J. Physiol. 8: 203, 1961. 36. 37. 38. 39. 40. 41. 42. 43. 44. 51 Roy, R. R. Specific changes in a histochemical profile of rat hind- limb muscle induced by two exercise regimens. Unpublished Ph.D. Thesis, Department of Health, Physical Education, and Recreation, Michigan State University, East Lansing, Michigan, 1976. Salzman, S. H., E. Z. Hirsch, H. K. Hellerstein, and J. H. Bruell. Adaptation to muscular exercise: myocardial epinephrine-3H uptake. J. App1. Physiol. 29: 92, 1970. Schrier, R. W., H. S. Henderson, C. C. Fisher and R. L. Tanner. Nephropathy associated with heat stress and exercise. In Exercise and Cardiac Death, E. Jokl and J. T. McClellan, Eds., Baltimore, Maryland?* University Park Press, 1971. Shimamoto, T. and Y. Hiramoto. Electron microscopic observations on edematous myocardial response to epinephrine. In Prevention of Ischemic Heart Disease, W. Raab, Ed., Springfield, 1111: Charles C. Thomas, 1966. Sive, L. A. The effects of two exercise regimens and supplemental vitamin C intake upon bone growth in albino rats. Unpublished M. A. Thesis, Department of Health, Physical Education and Recrea- tion, Michigan State University, 1978. Stevenson, J. A. F., V. Feleki, P. Rechnitzer and J. R. Beaton. Effect of exercise on coronary tree size in the rat. Circ. Res. 15: 265, 1964. Taylor, J. Histochemical profiles of rat triceps surae and plantaris after seven exercise regimens. Unpublished Ph.D. Thesis, Department of Anatomy, Michigan State University, East Lansing, Michigan, 1971. Vendsalu, A. Studies on adrenaline and noradrenaline in human plasma. Acta. Physiol. Scand. 49: Suppl. 173, 1960. Wells, R. L. and W. W. Heusner. A controlled-running wheel for small animals. Lab. Anim. Sci. 21: 904, 1971. APPENDICES 52 APPENDIX A TRAINING PROGRAMS Table A-1. Modified Eight Week Sprint Training Program for Postpubertal and Adult Male Rats in Controlled-Running Wheels for Experiment One Total Ac- Time Time Total celer- Work Repeti- Be- of Total Work Day Day ation Time Rest tions No. tween Run Prog. Exp. Tine of of Time (min: Time per of Bouts Shock Speed (min: Meters (sec) Wk. Wk. Tr. (sec) sec) (sec) Bout Bouts (min) (ma) (m/min) sec) TEM TWT O 4‘T -2 3.0 40:00 10 l l 5.0 0.0 27 40:00 --- --- 5=F -1 3.0 40:00 10 l 1 5.0 0.0 27 40:00 --- --- l 1=M l 2.0 00:10 10 10 8 2.5 1.2 36 42:50 480 800 2=T 2 2.0 00:10 10 10 8 2.5 1.2 36 42:50 480 800 3=W 3 1.5 00:10 15 10 8 2.5 1.2 54 49:50 720 800 4=T 4 1.5 00:10 15 10 8 2.5 1.2 54 49:50 720 800 5=F 5 1.5 00:10 15 10 8 2.5 1.2 54 49:50 720 800 2 1=M 6 1.5 00 10 15 10 8 2.5 1.2 54 49:50 720 800 2=T 7 1.5 00:10 15 10 8 2.5 1.2 54 49:50 720 800 3=W 8 1.5 00:15 30 6 7 2.5 1.2 72 43:00 756 630 4=T 9 1.5 00:15 30 6 7 2.5 1.2 72 43300 756 630 5=F 10 1.5 00:15 30 6 7 2.5 1.2 72 43:00 756 630 3 1=M 11 1.5 00 15 30 6 7 2.5 1.2 72 43:00 756 630 2=T 12 1.5 00 15 30 6 6 2.5 1.2 81 36:30 729 540 3=W 13 1.5 00 15 30 6 6 2.5 1.2 81 36:30 729 540 4=T 14 1.5 00 15 3O 6 6 2.5 1.2 81 36:30 729 540 5=F 15 1.5 00 15 30 6 6 2.5 1.2 81 36:30 729 540 4 1=M 16 1.5 00 15 3O 6 6 2.5 1.2 81 36:30 729 540 2=T 17 2.0 00 15 30 5 6 2.5 1.2 90 32:00 675 450 3=W 18 2.0 00 15 30 5 6 2.5 1.2 90 32:00 675 450 4=T 19 2.0 00 15 30 5 6 2.5 1.2 90 32.00 675 450 5=F 20 2.0 00 15 30 5 6 2.5 1.2 90 32:00 675 450 5 1=M 21 2.0 00 15 30 5 6 2.5 1.2 90 32:00 675 450 2=T 22 2.0 00 15 30 5 6 2.5 1.2 99 32:00 743 450 3=W 23 2.0 00 15 30 5 6 2.5 1.2 99 32:00 743 450 4=T 24 2.0 00 15 30 5 6 2.5 1.2 99 32:00 743 450 5=F 25 2.0 00 15 30 5 6 2.5 1.2 99 32:00 743 450 6 1=M 26 2.0 00 15 30 5 6 2.5 1.2 99 32:00 743 450 2-T 27 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 3=W 28 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 4=T 29 2.0 00 15 3O 5 6 2.5 1.2 108 32:00 810 450 5=F 30 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 7 18M 31 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 2=T 32 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 3=W 33 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 4=T 34 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 5=F 35 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 8 1=M 36 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 2=T 37 2.0 00 15 3O 5 6 2.5 1.2 108 32:00 810 450 3=W 38 2.0 00 15 3O 5 6 2.5 1.2 108 32:00 810 450 4'T 39 2.0 00 15 3O 5 6 2.5 1.2 108 32:00 810 450 5=F 40 2.0 00 15 30 5 6 2.5 1.2 108 32:00 810 450 This training program is a modified version of a standard program designed using male rats of the Sprague-Dawley strain (16,42). All animals should be exposed to a minimum of one week of voluntary running in a wheel prior to the start of the program. Failure to provide this adjustment period will impose a double learning Situation on the animals and will seriously impair the effectiveness of the training program. 53 APPENDIX A--continued Table A-2. Modified Eight Week Sprint Training Program for Postpubertal and Adult Male Rats in Controlled-Running Wheels for Experiment One Total Ac- No. Par- Time Time Total celer- Work Repeti- of tial Be- of Total Work Day Day ation Time Rest tions Com- Bouts tween Run Prog. Exp. Time of of Time (min: Time per plete (min: Bouts Shock Speed (min: Meters (sec) Wk. Wk. Tr. (sec) sec) (sec) Bout Bouts sec) (min) (ma) (m/min) sec) TEM TWT 0 4=T -2 3.0 40:00 10 l l 5.0 0.0 27 40:00 --- --- 5=F -l 3.0 40:00 10 1 1 5.0 0.0 27 40:00 --- --- l 1=M 1 2.0 02:30 0 l 6 2.5 1.2 27 27:30 405 900 2=T 2 2.0 02:30 0 l 6 2.5 1.2 27 27:30 405 900 3=W 3 1.5 05:00 0 l 3 5.0 1.2 36 25:00 540 900 4=T 4 1.5 05:00 0 1 3 5.0 1.2 36 2 :00 540 900 5=F 5 1.5 05:00 O 1 3 5.0 1.2 36 25:00 540 900 2 1=M 6 1.5 05:00 O l 3 5.0 1.2 36 25:00 540 900 2=T 7 1.0 07:30 O l 2 5.0 1.2 36 20:00 540 900 3=W 8 1.0 07:30 0 l 2 2.5 1.2 36 17:30 540 900 4=T 9 1.0 07 30 O l 2 1.0 1.2 36 16:00 540 900 5=F 10 1.0 15:00 O l 1 0.0 1.2 36 15:00 540 900 3 1=M 11 1.0 15:00 0 1 l 05:00 1.0 1.2 35 21:00 720 1200 2=T 12 1.0 15:00 0 l 1 07:30 1.0 1.0 36 23:30 810 1350 3=W 13 1.0 15:00 0 l 1 10:00 1.0 1.0 36 26:00 900 1500 4=T 14 1.0 15:00 0 l l 12:30 1.0 1.0 36 28:30 990 1650 5=F 15 1.0 15:00 0 l 2 1.0 1.0 36 31:00 1080 1800 4 1=M 16 1.0 15:00 0 1 2 05:00 1.0 1.0 36 37:00 1260 2100 2=T 17 1.0 15:00 0 l 2 07:30 1.0 1.0 36 39:30 1350 2250 3=W 18 1.0 15:00 O l 2 10:00 1.0 1.0 36 42:00 1440 2400 4=T 19 1.0 15:00 0 l 2 12 30 1.0 1.0 36 44:30 1530 2550 5=F 20 1.0 15:00 0 l 3 1.0 1.0 36 47:00 1620 2700 5 1=M 21 1.0 15:00 0 l 3 05:00 1.0 1.0 36 52:00 1800 3000 2=T 22 1.0 15:00 0 l 3 07:30 1.0 1.0 36 54:30 1890 3150 3=W 23 1.0 15:00 0 l 3 10:00 1.0 1.0 36 57:00 1980 3300 4=T 24 1.0 15:00 0 l 3 12:30 1.0 1.0 36 59:30 2070 3450 5=F 25 1.0 15:00 O l 4 1.0 1.0 36 63:00 2160 3600 6 1=H 26 1.0 15:00 0 l 4 1.0 1.0 36 64.00 2160 3600 2=T 27 1.0 30:00 0 1 2 5.0 1.0 36 65:00 2160 3600 3=W 28 1.0 30 00 O 1 2 2.5 1.0 36 62:30 2160 3600 4=T 29 1.0 30:00 0 l 2 1.0 1.0 36 61:00 2160 3600 5=F 30 1.0 60:00 0 l 1 0.0 1.0 36 60:00 2160 3600 7 18M 31 1.0 60:00 0 l l 0.0 1.0 36 60:00 2160 3600 2=T 32 1.0 60:00 0 l l 0.0 1.0 36 60:00 2160 3600 3=W 33 1.0 60:00 O l l 0.0 1.0 36 60:00 2160 3600 4=T 34 1.0 60:00 O l 1 0.0 1.0 36 60:00 2160 3600 5=F 35 1.0 60:00 O l l 0.0 1.0 35 60:00 2160 3600 8 1=H 36 1.0 60:00 0 l l 0.0 1.0 36 60:00 2160 3630 2=T 37 1.0 60:00 0 1 1 0.0 1.0 36 60:00 2160 3600 3=W 38 1.0 60:00 O l 1 0.0 1.0 36 60 00 2160 3600 4=T 39 1.0 60:00 0 1 1 0.0 1.0 36 60:00 2160 3600 5=F 40 1.0 60:00 O l l 0.0 1.0 36 60:00 2160 3600 This training program is a modified version of a standard program designed using male rats of the Sprague-Dawley strain (16,42). ;pcguDIX A--continued 54 Table 4-3. Uodified Eight Heek Regular Sprint Training Program for Postpubertal and Adult Hale Rats in Controlled-Running Wheels Total Acc- Repe- Time Time Total eler- Work ti- Be- Run of Total Work Day Day ation Time Rest tions No. tween Speed Prog. Exp. Time of of Time (min: Time per of Bouts Shock (m/ (min: Meters (sec) Nk. Wk. Tr. (sec) sec) (sec) Bout Bouts (min) (ma) min) sec) TEM TRT 0 4=T -2 3.0 40:00 10 l 1 5.0 0.0 27 40:00 --- --- 5=T -i 3.0 40:00 10 1 5.0 0.0 27 40:00 --- --- 1 1=M l 3.0 00:15 15 25 3 5.0 1.2 27 46:45 506 1125 2=T 2 3.0 00:15 15 25 3 5.0 1.2 27 46:45 506 1125 3=W 3 3.0 00:15 15 25 3 5.0 1.2 27 46:45 506 1125 4=T 4 2.5 00:10 10 40 3 5.0 1.2 36 49:30 720 1200 5=F 5 2.0 00:10 10 40 3 5.0 1.2 36 49:30 720 1200 2 l=M 6 1.5 00:10 10 28 4 5.0 1.2 45 51:40 840 1120 2=T 7 1.5 00:10 15 27 4 5.0 1.2 54 59:00 972 1080 3=W 8 1.5 00:10 15 27 4 5.0 1.2 54 59:00 972 1080 4=T 9 1.5 00:10 15 27 4 5.0 1.2 54 59:00 972 1080 5=F 10 1.5 00:10 15 27 4 5.0 1.2 54 59:00 972 1080 3 1=M 11 1.5 00:10 15 27 4 5.0 1.2 54 59:00 972 1080 2=T 12 1.5 00:10 20 23 4 5.0 '1.2 63 59:40 966 920 3=W 13 1.5 00:10 20 23 4 5.0 1.2 63 59:40 966 920 4=T 14 1.5 00:10 20 23 4 5.0 1.2 63 59:40 966 920 5=F 15 1.5 00:10 20 23 4 5.0 1.2 63 59:40 966 920 4 1=M 16 1.5 00:10 20 23 4 5.0 1.2 63 59:40 966 920 2=T 17 1.5 00:10 25 20 4 5.0 1.0 72 60:00 960 800 3=W 18 1.5 00:10 25 20 4 5.0 1.0 72 60:00 960 800 4=T 19 1.5 00 10 25 20 4 5.0 1.0 72 60:00 960 800 5=F 20 1.5 00:10 25 20 4 5.0 1.0 72 60:00 960 800 5 1=M 21 1.5 00:10 25 20 4 5.0 1.0 72 60:00 960 800 2=T 22 1.5 00:10 30 16 4 5.0 1.0 81 55:40 864 640 3=W 23 1.5 00:10 30 16 4 5.0 1.0 81 55:40 864 640 4=T 24 1.5 00:10 30 16 4 5.0 1.0 81 55:40 864 640 5=F 25 1.5 00:10 30 16 4 5.0 1.0 81 55:40 864 640 6 1=M 26 1.5 00:10 30 16 4 5.0 1.0 81 55:40 864 640 2=T 27 2.0 00:10 35 10 5 5.0 1.0 90 54:35 750 500 3=W 28 2.0 00:10 35 10 5 5.0 1.0 90 54:35 750 500 4=T 29 2.0 00:10 35 10 5 5.0 1.0 90 54:35 750 500 5=F 30 2.0 00:10 35 10 5 5.0 1.0 90 54:35 750 500 7 l=M 31 2.0 00:10 35 10 5 5.0 1.0 90 54:35 750 500 2=T 32 2.0 00:10 35 7 8 2.5 1.0 90 54:50 840 560 3=W 33 2.0 00:10 35 7 8 2.5 1.0 90 54:50 840 560 4=T 34 2.0 00:10 35 7 8 2.5 1.0 90 54:50 840 560 5=F 35 2.0 00:10 35 7 8 2.5 1.0 90 54:50 840 560 8 1=M 36 2.0 00:10 35 7 8 2.5 1.0 90 54:50 840 560 2=T 37 2.0 00:10 40 6 8 2.5 1.0 99 52:10 792 480 3=W 38 2.0 00:10 40 6 8 2.5 1.8 33 23:18 73% zgg 4=T 39 2.0 00:10 40 6 8 2.5 l. - S-F 40 2.0 00:10 40 6 8 2.5 1.0 99 52:10 792 480 APPENDIX A--continued 55 Table A-4. Modified Eight Week Regular Endurance Training Program for Postpubertal and Adult Male Rats in Controlled-Running Wheels acc- Repe- Time Time Total eler- Jork ti- Be- Run of Work Day Day ation Time Rest tions No. tween Speed Prog. Exp. Time of of Time (min: Time per of Bouts Shock (m/ (min: Meters (sec) wk. Hk. Tr. (sec) sec) (sec) Bout Bouts (min) (ma) min) sec) TEM TRT 0 4=T -2 3.0 40:00 10 l l 5.0 0.0 27 40:00 --- --- 5=F -l 3.0 40:00 10 1 1 5.0 0.0 27 40:00 --- --- 1 1=M 1 3.0 00:15 15 25 3 5.0 1.2 27 46:45 506 1125 2=T 2 3.0 00:15 15 25 3 5.0 1.2 27 46:45 506 1125 3=W 3 3.0 00:15 15 25 3 5.0 1.2 27 46:45 506 1125 4=T 4 2.5 00:30 15 20 2 5.0 1.2 27 34:30 540 1200 5=F 5 2.5 00:30 15 20 2 5.0 1.2 27 34:30 540 1200 2 1=M 6 2.0 00:40 20 15 2 5.0 1.2 36 34:20 720 1200 2=T 7 2.0 00:50 25 12 2 5.0 1.2 36 34:10 720 1200 3=W 8 1.5 01:00 30 10 2 5.0 1.2 36 34:00 720 1200 4=T 9 1.5 02:30 60 4 2 5.0 1.2 36 31:00 720 1200 5=F 10 1.0 02:30 60 4 2 5.0 1.2 36 31:00 720 1200 3 1=M 11 1.0 02:30 60 4 2 5.0 1.2 36 31:00 720 1200 2=T 12 1.0 05:00 10 1 5 2.5 1.2 36 35:00 900 1500 3=W 13 1.0 05:00 10 l 5 2.5 1.2 36 35:00 900 1500 4=T 14 1.0 05:00 10 l 5 2.5 1.2 36 35:00 900 1500 5=F 15 1.0 05:00 10 1 5 2.5 1.2 36 35:00 900 1500 4 1=M 16 1.0 05:00 10 l 5 2.5 1.2 36 35:00 900 1500 2=T 17 1.0 07:30 10 l 4 2.5 1.0 36 37:30 1080 1800 3=W 18 1.0 07:30 10 l 4 2.5 1.0 36 37:30 1080 1800 4=T 19 1.0 07:30 10 l 4 2.5 1.0 36 37:30 1080 1800 5=F 20 1.0 07:30 10 l 4 2.5 1.0 36 37:30 1080 1800 5 1=M 21 1.0 07:30 10 1 4 2.5 1.0 36 37:30 1080 1800 2=T 22 1.0 07:30 10 l 5 2.5 1.0 36 47:30 1350 2250 3=W 23 1.0 07:30 10 l 5 2.5 1.0 36 47:30 1350 2250 4=T 24 1.0 07:30 10 l 5 2.5 1.0 36 47:30 1350 2250 5=F 25 1.0 07:30 10 l 5 2.5 1.0 36 47:30 1350 2250 6 1=M 26 1.0 07:30 10 1 5 2.5 1.0 36 47:30 1350 2250 2=T 27 1.0 10:00 10 l 4 2.5 1.0 36 47:30 1440 2400 3=W 28 1.0 10:00 10 l 4 2.5 1.0 36 47:30 1440 2400 4=T 29 1.0 10:00 10 l 4 2.5 1.0 36 47:30 1440 2400 5=F 30 1.0 10:00 10 1 4 2.5 1.0 36 47:30 1440 2400 7 1-M 31 1.0 10:00 10 l 4 2.5 1.0 36 47:30 1440 2400 2=T 32 1.0 10:00 10 1 5 2.5 1.0 36 60:00 1800 3000 3=W 33 1.0 10:00 10 l 5 2.5 1.0 36 60:00 1800 3000 4sT 34 1.0 10:00 10 1 5 2.5 1.0 36 60:00 1800 3000 5=F 35 1.0 10:00 10 l 5 2.5 1.0 36 60:00 1800 3000 8 18M 36 1.0 10:00 10 1 5 2.5 1.0 36 60:00 1800 3000 2=T 37 1.0 12:30 10 1 4 2.5 1.0 36 57:30 1800 3000 3=W 38 1.0 12:30 10 1 4 2.5 1.0 36 57:30 1800 3000 4=T 39 1.0 12:30 10 l 4 2.5 1.0 36 57:30 1800 3000 t-F 40 1.0 12:30 10 l 4 2.5 1.0 36 57:30 1800 3000 56 APPENDIX A--continued Table A-5. Modified Eight Meek High Sprint Training Program for Postpubertal and Adult Male Rats in Controlled-Running wheels Total Acc- Repe- No. Par- Time Time Total eler- work ti- of tial be- Run of Total Hork Ea. Day ation Time Rest tions Com- Bouts tween Speed Prog. Exp. Time (J o O ..., Time (min: Time per plete (min: Bouts Shock (m/ (min: Meters (sec) (sec) sec) (sec) Bout Bouts sec) (min) (ma) min) sec) TEM TNT I . 4 3 3 4=T -2 3.0 40:00 10 1 1 5.0 0.0 27 40:00 --- --- 5=F -l 3.0 40:00 10 1 1 5.0 0.0 27 40:00 --- --- l 1=M 1 2.0 00:15 15 8 7 2.5 1.2 36 41:15 504 840 2=T 2 2.0 00:15 15 8 7 2.5 1.2 36 41:15 504 840 3=H 3 1.5 00:15 25 8 7 2.5 1.2 54 49:25 756 840 4=T 4 1.5 00:15 25 8 7 2.5 1.2 54 .49:25 756 840 5=F 5 1.5 00:15 25 8 7 2.5 1.2 54 49:25 756 840 2 1=M 6 1.5 00:15 25 8 7 2.5 1.2 54 49:25 756 840 2=T 7 1.5 00:15 25 8 7 2.5 1.2 54 49:25 756 840 3=H 8 1.5 00:15 30 6 7 2.5 1.2 72 43:00 756 630 4=T 9 1.5 00:15 30 6 7 2.5 1.2 72 43:00 756 630 5=F 10 1.5 00:15 30 6 7 2.5 1.2 72 43:00 756 630 3 1=M 11 1.5 00:15 30 6 7 2.5 1.2 72 43:00 756 630 2=T 12 1.5 00:15 30 6 6 2.5 1.2 81 36:30 729 540 3=W 13 1.5 00:15 30 6 6 2.5 1.2 81 36:30 729 540 4=T 14 1.5 00:15 30 6 6 2.5 1.2 81 36:30 729 540 5=F 15 1.5 00:15 30 6 6 2.5 1.2 81 36:30 729 540 4 1=M 16 1.5 00:15 30 6 6 2.5 1.2 81 36:30 729 540 2=T 17 2.0 00:15 30 5 6 2.5 1.2 90 32:00 675 450 3=” 18 2.0 00:15 30 5 6 2.5 1.2 90 32:00 675 450 4=T 19 2.0 00:15 30 5 6 2.5 1.2 90 32:00 675 450 5=F 20 2.0 00:15 30 5 6 2.5 1.2 90 32:00 675 450 5 1:? 21 2.0 00:15 30 5 6 2.5 1.2 90 32:00 675 450 2=T 22 2.0 00:15 30 5 6 2.5 1.2 99 32:00 743 450 3:9 23 2.0 00:15 30 5 6 2.5 1.2 99 32:00 743 450 4=T 24 2.0 00:15 30 5 6 2.5 1.2 99 32:00 743 450 5=F 25 2.0 00:15 30 5 6 2.5 1.2 99 32:00 743 450 6 1=M 26 2.0 00:15 30 5 6 2.5 1.2 99 32:00 743 450 2=T 27 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 3=U 28 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 4=T 29 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 5=F 30 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 7 1=M 31 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 2=T 32 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 38H 33 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 4=T 34 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 5=F 35 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 8 1=M 36 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 2=T 37 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 3=W 38 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 4=T 39 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 5=F 40 2.0 00:15 30 5 6 2.5 1.2 108 32:00 810 450 This training program is a modified version of a standard program designed using male rats of the Sprague-Dawley strain (16,42). All animals should be exposed to a minimum of one week of voluntary running in a wheel prior to the start of the program. Failure to provide this adjustment period will impose a double 1earning situation on the animals and will seriously impair the effectiveness of the training program. 57 APPENDIX A--C0ntinued Tabie A-6. Modified Eight Week High Endurance Training Program for Postpubertal and Adu1t Male Rats in Controlled-Running Wheels Total Acc- Repe- No. Par- Time Time Total eler- Work ti- of tial Be- of Total work Cay Jay ation Time Rest tions Com- Bouts tween Run Prog. Exp. Time of of Time (min: Time per plete (min: Bouts Shock Speed (min: Meters (sec) :1. xx. Tr. (sec) sec) (sec) Bout Bouts sec) (min) (ma) (m/min) sec) TEM TNT 0 4=T -2 3.0 40 00 10 l 1 5.0 0.0 27 40:00 --- --- 5=F -1 3.0 40 00 10 1 1 5.0 0.0 27 40:00 --- --- 1 1=M 1 2.0 1:00 0 1 15 2.5 1.2 27 50:00 405 900 2=T 2 2.0 1:00 0 1 15 2.5 1.2 27 50:00 405 900 3=w 3 1.5 1:00 0 l 15 2.5 1.2 36 50:00 540 900 4=T 4 1.5 1:00 0 l 15 2.5 1.2 36 50:00 540 900 5=F 5 1.5 1:00 0 l 15 2.5 1.2 36 50:00 540 900 2 1=M 6 1.5 1:00 0 1 15 2.5 1.2 36 50:00 540 900 2=T 7 1.0 1:00 0 1 15 2.5 1.2 45 50:00 675 900 3=W 8 1.0 1:00 0 1 15 2.5 1.2 45 50:00 675 900 4=T 9 1.0 1:00 ' 0 1 15 2.5 1.2 45 50:00 675 900 5=F 10 1.0 1:00 0 1 15 2.5 1.0 45 50:00 675 900 3 1=M 11 1.0 1:00 0 1 15 2.5 1.0 45 50:00 675 900 2=T 12 1.0 2:30 0 1 9 2.5 1.0 45 43:30 1012 1350 3=A 13 1.0 ' 2:30 0 l 9 1.0 1.0 45 30:30 1012 1350 4=T 14 1.0 5:00 0 1 7 2.5 1.0 45 50:00 1575 2100 5=F 15 1.0 5:00 0 1 7 1.0 1.0 45 41:00 1575 2100 4 1=M 16 1.0 5:00 0 1 7 2.5 1.0 45 50:00 1575 2100 2=T 17 1.0 7:30 0 l 6 2.5 1.0 45 57:30 2025 2700 3=H 18 1.0 7:30 0 1 6 1.0 1.0 45 50:00 2025 2700 4=T 19 1.0 10 00 0 1 5 2.5 1.0 45 60:00 2250 3000 5=F 20 1.0 10:00 0 1 5 1.0 1.0 45 54:00 2250 3000 5 1=M 21 1.0 10:00 0 1 5 2.5 1.0 45 60:00 2250 3000 2=T 22 1.0 12 30 0 l 4 2.5 1.0 45 57:30 2250 3000 3=W 23 1.0 12:30 0 l 4 1.0 1.0 45 53:00 2250 3000 4=T 24 1.0 15:00 0 1 4 2.5 1.0 45 67:30 2700 3600 5=F 25 1.0 15 00 0 1 4 1.0 1.0 45 63:00 2700 3600 6 1=M 26 1.0 15:00 0 1 4 2.5 1.0 45 67:30 2700 3600 2=T 27 1.0 20:00 0 l 3 2.5 1.0 45 65:00 2700 3600 3=9 28 1.0 20:00 0 l 3 1.0 1.0 45 62:00 2700 3600 4=T 29 1.0 30:00 0 l 2 2.5 1.0 45 62:30 2700 3600 5=F 30 1.0 30:00 0 l 2 1.0 1.0 45 61:00 2700 3600 7 1=M 31 1.0 30 00 0 1 2 2.5 1.0 45 62:30 2700 3600 2=T 32 1.0 60:00 0 1 l 0.0 1.0 45 60:00 2700 3600 3=W 33 1.0 60:00 0 1 l 0.0 1.0 45 60:00 2700 3600 4=T 34 1.0 60:00 0 1 1 0.0 1.0 45 60:00 2700 3600 5=F 35 1.0 60 00 0 l l 0.0 1.0 45 60:00 2700 3600 9 1=M 36 1.0 60:00 0 1 l 0.0 1.0 45 60:00 2700 3600 2=T 37 1.0 60 00 0 1 1 0.0 1.0 45 60:00 2700 3600 3=W 38 1.0 60 00 0 1 1 0.0 1.0 45 60:00 2700 3600 4=T 39 1.0 60:00 0 l l 0.0 1.0 45 60:00 2700 3600 5=F 40 1.0 60 00 0 1 l 0.0 1.0 45 60:00 2700 3600 58 APPENDIX 8 BASIC STATISTICS FOR TRAINING DATA Table B-1. Basic Statistics for Percentage of Body Weight Loss, Environmenta1 Factors and Perfornance Criteria for Experiment One Simple Correlations a Standard Air Per Bar Per Body Variable N Mean Deviation Temp Humid Press Ht Loss PEM 21g Air Temp (F) 367 72.9 4.8 Per Humid 367 39.0 12.1 .110 Bar Press (mmHg) 367 740.7 4.3 -.276 -.713 Per Body wt 1055 367 1.7 .5 -.066 -.206 .044 PEM 367 55.4 20.2 -.199 -.359 .121 .258 PSF 367 56.8 19.5 -.398 -_312 .154 .196 .372 EEEQE Air Temp (F) 376 73.1 4.6 Per Humid 376 38.5 12.3 .125 Bar Press (mmHg) 376 740.8 4.2 -.263 -.715 Per Body wt less 376 1.7 .6 .000 -.200 .046 PEM 376 61.6 25.9 -.165 -.383 .210 .115 PSF 376 62.6 23.4 -.260 -.271 .129 .032 .868 9.2; Air temp (F) 340 73.9 4.0 Per Humid 340 47.1 10.7 .134 Bar Dress (rnMg) 340 739.5 3.8 -.288 -.675 Per Bocy wt loss 340 2.5 1.0 .374 .139 -.240 PEM 340 82.8 24.7 -.279 -.173 .232 -.069 PSF 340 70.7 18.6 -.403 -.083 .159 -.118 .685 gimme Air Temp (F) 348 73.9 4.0 Per Humid 348 47.0 10.6 .151 Bar Press (nnmg) 348 739.5 3.8 - 294 - 677 Per Body wt loss 348 2.6 1.0 .439 .114 -.159 PEM 348 82.2 16.7 -.231 -.253 .286 -.003 PSF 348 69.6 19.2 -.254 -.102 .153 .021 .748 Na = total days training. all animals 59 APDENDIX B--Cont1nued Table 8-2. Basic Statistics for Percentage of Body Height Loss. Environ- mental Factors. and Performance Criteria for Experiment Two Standard ’Simple CorreTations a Devia- Air Per Bar Pér Body Variab1e N Mean tions Temp. Humid Press Ht Loss PEM REG SPT - Su ar Air Temp IF} 105 75.32 2.99 Per Humid 105 45.70 6.20 .638 Bar Press (mmHg) 105 742.78 2.56 -.414 -.711 Per Body wt. Loss 105 2.42 .59 .044 .086 -.176 PEM 105 79.57 14.85 .293 .309 -.362 .173 PSF 105 89.31 7.40 .162 .178 -.143 -.219 .464 REG SPT - No Su ar Air Iemp IFI 146 75.36 3.02 Per Humid 146 45.78 6.34 .648 Bar Press (mmHg) 146 742.78 2.55 -.427 -.355 Per Body Ht. Loss 146 2.00 .56 .183 .158 -.186 PEM 146 74.43 23.49 .344 .329 -.337 .106 PSF 146 83.71 8.33 .275 .254 -.219 .034 .447 HT SPT - Su ar Air Temp (PI 137 75.39 2.89 Per Humid 137 45.79 6.24 .397 Bar Press (mmHg) 137 742.73 2.52 .178 .505 Per Body Ht. Loss 137 2.14 .68 .014 .028 .024 PEM 137 63.46 23.99 .133 .099 .112 .127 PSF 137 70.28 24.57 .122 .063 .088 .069 .611 HI SPT - No Sugar Air’Temp (F) 141 75.42 3.02 Per Humid 141 45.87 6.42 .630 Bar Press (unMg) 141 742.85 2.53 -.422 -.710 Per Body Mt. Loss 141 1.98 .70 .119 .169 -.116 PEM 141 59.84 30.24 .364 .315 -.335 .356 PSF 141 68.33 25.54 .350 .252 -.296 .263 .782 REG END - Sugar Air Temp (F) 145 75.44 2.99 Per Humid 145 45.99 6.31 .638 Bar Press (mmHg) 145 742.70 2.55 -.407 -.708 Per Body Mt. Loss 145 2.59 .83 .149 .016 .036 PEM 145 95.86 18.90 .286 .187 -.087 .207 PSF 145 89.11 9.85 .171 .068 -.080 .159 .311 continued 60 APPENDIX B--Table 8-2--continued Standard Simple Correlations a Devia- AT? Per Bar ’Per Body Variable N Mean tions Temp. Humid Press Ht. Loss PEM REG END - No Sugar Air Temp (F1 140 75.39 3.02 Per Humid 140 45.79 6.47 .649 Bar Press (mmHg) 140 742.79 2.56 -.417 -.711 Per Body Ht. Loss 140 2.43 .63 .321 .202 .112 PEM 140 80.80 24.22 .283 .312 -.265 .351 PSF 140 77.99 17.16 .075 .086 .097 .157 .633 HI END - Suggg, 87 75.57 3.10 Per Humid 87 46.34 6.65 .674 Bar Press (mmHg) 87 742.64 2.61 -.469 -.716 Per Body Mt. Loss 87 2.90 .75 -.058 -.096 .166 PEM 87 58.34 34.99 .200 .224 .255 -.l4l PSF 87 50.71 30.48 .122 .147 .245 -.193 .918 HI END - No Sugar Air Temp (FT' 108 75.50 3.00 Per Humid 108 46.02 6.30 .662 Bar Press (mmHg) 108 742.75 2.56 -.450 -.715 Per Body Ht. Loss 108 2.84 .68 .191 .070 .074 PEM 108 55.67 31.11 .212 .180 .151 .043 PSF 108 47.92 29.37 .155 .145 -.190 -.006 .929 "IIIIIII‘lllllfllllllllllg