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ABSTRACT

BLOOD VASCULAR PATTERN TO AND THROUGH THE
GASTROCNEMIUS OF THE SEDENTARY MALE RAT

by Barry 80 Brown

Twenty-four male, post—puberty rats (Sprague-Dawley)
were placed in individual sedentary cageso The rats were
divided into three groups, which were injected under
regulated temperature and pressure at the aorta Just above
the bifurcation with either latex, india ink or vinyl
acetate. Analysis of the various injectional media for
the purposes of gross and intramuscular arterial deter-
minations were performed and the blood vascular pattern
was identified via dissection and microphotography.

A 50% latex ammonium solution was found to be
superior for dissection purposes, The intramuscular
arterial pattern, although variable, can be reduced to

a generalized architecture common to most sedentary ratso

BLOOD VASCULAR PATTERN TO AND THROUGH THE
GASTROCNEMIUS OF THE SEDENTARY MALE RAT

By

Barry S; Brown

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

MASTER OF ARTS

Department of Physical Education
College of Education

1968

Dedicated
to

my wife
and

forthcoming child

11

ACKNOWLEDGMENTS

I wish to acknowledge Dr. Wayne Van Huss, my major
advisor, and Dr. Rexford Carrow, Assistant Professor in
the Anatomy Department, for the assistance and encourage-
ment they have given me during my plight. Additional
expression of gratitude is extended to Mrs. Geraldine
Wilson, Mr. Robert Ruhling, and Mr. Carl Oehling for their
valuable assistance during the injectional phase of the
experiment.

The principles of equality of opportunity and
dedication to one's work fostered by my parents and
brother have contributed in great measure to the final

outcome of this thesis.

iii

FOREWORD

The format of this thesis differs somewhat from the
conventional four or five chapter approach used so fre-
quently. It has been mutually decided by Dr. Van Huss
and myself to present the material in the same manner as
it was submitted for publication. I am of the Opinion
greater benefit will be derived by presenting this thesis
in publication form, being that it represents a more
realistic approach to the presentation of scientific
research.

Within the framework of the thesis will be found
scattered reviews of pertinent literature as they relate

to the topic being discussed.

iv

TABLE OF CONTENTS

Page
ACKNOWLEDGMENTS . . . . . .* . . . . . . . iii
FOREWORD o o o o o o o o o o o o o o ' o 0 1V

LIST OF FIGURES . . . . . . . . . . . . . vi

INTRODUCTION . . . . . . . . . . . . . . 1
METHOD 0 O O O O O O O O O 0 0 I 0 O O 2
OBSERVATIONS . . . . . . . . . . . . . . 7

. . . . . . . . . . . . 7

Gross Supply

Intramuscular Design . . . . . . . . . . 11
DISCUSSION 0 O 0 O 0 0 I O O O O 0 0 O 0 19

REFERENCES 0 O O O O O ‘ O O O O O O O O O 20

LIST OF FIGURES

Figure Page

1. Injection apparatus . . . . . . . . . . 3

2. Gross arterial supply to the-gastrocnemius.
Axillary view. Left leg. Diagrammatic . . 8

3. Major intramuscular arterial pattern to the
gastrocnemius. Arterial diameter range:
.15-.32mm. Axillary view. Left leg.
Diagrammatic. . . . . . . . . . . . 12

A. Minor intramuscular arterial supply.
Arterial diameter range: Less than
.lSmm. Axillary view. Left leg.
Diagrammatic . . . . . . . . . . . 1A

5. Major intramuscular arterial supply of
the medial head of the gastrocnemius. x25 . . 17

vi

INTRODUCTION

Research involving the gastrocnemius of the rat
(Carrow 33 al., 1967; Delasante, 196“) has been undertaken
without regard to the major blood vascular pattern within
the muscle. The gross arterial supply may represent an
essential factor in determining capillary to fiber ratios
(Krogh, 1919; Paff, 1930, Carrow g£_al., 1967) as well as
being of interest in ligation studies.

A survey of literature both past and present indi-
cates the classification of muscle according to its blood
supply into three main classes. Both the medial and
lateral heads of the gastrocnemius have been reported
(Campbell, 1919) to receive their blood supply from one
source with little potential for collateral channels.
With respect to the rat, one author (Jaya, 1958) classi-
fies a "spindle shaped muscle" as having a "dense basket
type" of circulatory pattern, this being most indicative
of the gastrocnemius.

Aside from the abovementioned references, only one
atlas (Greene, 1959) contains the major arterial supply
to the gastrocnemius of the white albino rat with any
degree of accuracy. To my knowledge, our endeavor to map
out the intramuscular pattern to the gastrocnemius of the
sedentary male rat represents an initial exploration in

this area.

METHOD

Twenty-four male albino rats (Sprague-Dawley) were
placed in sedentary individual cages until they had
attained an age of 120 days. Four rats per day, over a
total of six days were administered a dosage of 60 mg/kg
of sodium pentobarbital intraperitoneally. The observed
variability of onset of general anesthesia can be
explained by the-differing rates of drug metabolism by
the rats' liver. No further anesthetic was required
either before or during surgery to maintain a surgical
level of anesthesia. Pentobarbital was chosen in deference
to ether due to its ability to maintain a constant level
of anesthesia without the need for an anesthetist, in
addition to the reports (Olmstead and Page, 1966) of no
significant change in mean arterial pressure that might
tend to bias the arterial pattern.

The equipment utilized in the perfusion and injection
technique is illustrated in Figure 1. Although of a make-
shift variety, the apparatus proved to be experimentally
sound and extremely inexpensive. Allowing for a ten-
minute cardiovascular period of adjustment following
anesthesia, a laparotomy was performed. The aorta was
carefully isolated from the exit of the superior mesenteric

artery to the bifurcation of the common iliac arteries.

Figure l.--Injection apparatus.

MAMMALIAN
RINGERS.

 

 

38°C

 

 

 

INDIA INK

 

 

 

 

 

 

8 PI 10. OIAIOIITII

S?

 

 

j

J

 

I

 

 

L... N
_AORTA

 

A polyethylene tube (P.E. = 20) fastened around a 20 gauge
1% inch disposable needle was inserted into the aorta, one-
half inch caudal to the right renal artery and secured with
non-absorbable silk suture.

Two animals were used for the injection of vinyl
acetate in an attempt to produce a corrosion Specimen
(Ward's Service Bulletin, No. 5). Acetone was used in
place of Mammalian-Ringer's solution to prevent the coagu-
lation of vinyl acetate which would occur with water.

Eight animals were injected with india ink and
fourteen with latex as outlined below.

The hindlimbs were exsanguinated with Mammalian
Ringer's solution at 38°C. ‘When the fluid issuing forth
from the cut Inferior Vena Cava appeared clear, the per-
fusate (india ink, latex or vinyl acetate) was introduced
by the turning of a valve. Heparin (l cc/liter) was added
to prevent clotting. India ink and latex were allowed
to run two minutes, whereas, the vinyl acetate perfused
the lower limb for two hours to assure entrance of the
plastic into the arterioles. All three injection media
were perfused at a pressure of 120 mmHg commensurate with
the normal blood pressure of the rat (Durant, 1927).

Following the injection, the skin and overlying
muscle (Semimembranosus and Biceps Femoris) was removed
and a sketch of the major arterial pattern to the

gastrocnemius was recorded., The right and left limbs

were then removed at the acetabulofemoral joint and
treating according to the perfusage used as follows:

The muscle of the arteries injected with vinyl
acetate were severed at the achillis tendon, tied to a
ten-gram weight and suspended in papain.

The india ink specimens were fixed in 10% formalin
for 2A-A8 hours, dehydrated in three changes of acetone
(30 minutes each), and cleared in methyl benzoate.

The latex injected artieries were fixed in 10%
formalin and remained there until dissected.

MicrOphotographs of the dissected specimens were
taken with a 35mm camera attached to a Leitz Ortholux
microscope magnified 25 times.

Presentation of the arterial pattern from diagrams
instead of micrographs was arrived at by personal consul-
tation with Alfred M. Lucas (Research Zoologist and Project
Leader, USDA Avian Anatomy Project). The sacrifice of
realism for diagrammatism appears to be supportable on the
grounds of presenting a clearer, understandable analysis

of an otherwise confusing, non—informative pattern.

OBSERVATIONS

The diagrammatic illustrations represent generalized
vascular patterns to be found in sedentary male albino
rats.

According to Campbell and Pennefather (1919), the
human gastrocnemius has been classified into the third
main class with respect to its blood supply. The vascular
bed is derived exclusively from one source with almost
total absence of potential collateral channels (Campbell
32 al., 1919). Abramson (1962) adds that the variability
of arterial architecture found within muscles is dependent
upon the availability of alternate arterial sources. It
might then be argued that there is little variability
found in vascular patterns of muscles supplied from a
single source. Campbell 32 El: (1919) would include the
human gastrocnemius in this category, but does not

generalize beyond this species.

Gross Supply

 

This author agrees with the generalization that
the bulk of the vascular supply to the medial and lateral
heads of the gastrocnemius stems from the internal and
external sural aa., respectively (see Figure 2). There

appears to be, however, the presence of one additional

Figure 2.--Gross arterial supply to the
gastrocnemius. Axillary view. Left leg.
Diagrammatic.

 

     
    

 

FEMORAL A.
SAPHENOUS
BRANCH OF
FEMORAL A.

POPLITEAL A.

FEMORO-POPLITEAL A .

 
     
   

 

 

MIDDLE
GENICULAR A.

_.___._LATERAL GENICULAR A.
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POSTERIOR
URAL A , ~«
3 -. , -7 -. TIBIAL A.
w E} SURAL A.
:
GASTROCNEMIUS, I; " ,3 ~ e - GASTROCNEMIUS,

 

SOLEUS

lO

branch to the medial head arising from the middle genicular
artery as supported by Greene (1959), and from three to six
additional vessels to the lateral head arising in the
following manner:

A variable distance cephalad tc> the bifurcation of
the popliteal artery into the external and internal sural
aa., a medial (posterior tibial) and lateral (superficial
sural) branch diverge at acute angles to the parent stem.
Before the posterior tibial artery dips underneath the
medial head, the middle genicular artery emerges at a
right angle and passes to the gastrocnemius close to its
point of attachment to the femur. One of two branches may
spring forth from the middle genicular artery, passing
directly or sending side branches to the lateral head near
its point of attachment to the femur.

The superficial sural artery, having emerged from
the popliteal artery, continues down the lateral head of
the muscle, parallel to its longitudinal axis along the
dorsal border. A variable distance from its exit a
recurrent branch, the femoro-popliteal artery, is given off
at right angles, and travels along the upper border of the
lateral head to supply the biceps femoris muscle. As it
passes the point of attachment two small branches may
emerge and pass to the lateral head near the femur.

Toward the caudal end of the lateral head, a small branch

from the superficial sural has been constantly observed

11

to penetrate the muscle substance and begin its intra-

muscular design.

Intramuscular Design

 

This author did not observe.the numerous arterial
anastomoses of the primary intramuscular vessels as
described by Krogh (1936). As can be demonstrated in
Figure 3, three main intramuscular arteries usually arise
from the external sural, and two major arteries from the
internal sural artery. Each of the arteries immediately
diverge upon entering the muscle substance and travel to
their respective areas outlined in Figure 3. On the
basis of its major* intramuscular supply, the lateral
head can be divided into upper, middle and lower portions
as there is little overlap of the basic pattern. The
medial head may be divided, using the same criteria, into
an upper portion comprising approximately one third of
the total area, and a lower portion making up the
remainder (see Figure A).

A constant vessel arises from the superficial sural
artery approximately three—quarters of the distance from
its exit off the popliteal artery and supplies the lower

lateral quadrant of the lateral head.

 

*The major intramuscular arteries were designated
as those vessels maintaining a diameter between .15 and
.32mm (or one to two units as observed at 25 magnification
on the Leitz microscope).

12

Figure 3.--Major intramuscular arterial
pattern to the gastrocnemius. Arterial
diameter range: .15-.32mm. Axillary view.
Left leg. Diagrammatic.

l3

 

 

 

 

 

 

 

 

14

Figure A.--Minor intramuscular arterial
supply. Arterial diameter range: Less than
.15mm. Axillary view. Left leg. Diagrammatic.

16

The cephalic ends of both heads are supplied by the
branches of the genicular arteries and occasionally by the
recurrent vessels of the femoro-popliteal artery. Having
seen no reference in the literature to these recurrent
vessels, it may well be apprOpriate to designate these
branches as the "recurrent femoro-pOpliteal aa."

There is a lack of certainty concerning the level
to which the dissection was performed. On the basis of
size, vessels were dissected with a diameter of less than
150 u, therby, categorizing them as arterioles. Yet one
author (Wiedeman, 1962, 1965) disclaims the defining of
an arteriole on the basis of its diameter. It is my sub-
jective Opinion that insufficient branching had taken
place to reach arteriole level and that arteriolar size

studies should be undertaken to establish norms.

17

Figure 5.--Major intramuscular arterial
supply of the medial head of the gastrocnemius.
X 25.

l8

 

DISCUSSION

The clinical importance of the gross arterial
pattern has been mentioned (Le Gros Clark and Blomfoeld,
19A5; Abramson, 1962) as being of surgical significance
in the prevention of anaerobic infection as well as
muscle degeneration.

Studies on the presence of arterial anastomoses and
the effects of devascularization of muscle as suggested
by Le Gros Clark 32 31. (19A5) may shed credibility to
the claim that the gastrocnemius is liable to the devel-
opment of necrosis (Pearson, 1965).

It is the author's expectation that greater emphasis
be drawn to the effects of pathology and physical stress
upon the blood vascular pattern to and within skeletal

muscle.

19

REFERENCES

Abramson, David. 1962. Blood vessels and Lymphatics.
Academic Press, New York and London. Chapter XVII,
521-525.

Cambell, J., and c. M. Pennefather.' 1919. The blood
supply of muscles. The Lancet, 1: 29A-296.

Carrow, R. E., R. E. Brown and W. D. Van Huss. 1967.
Fiber sizes and capillary to fiber ratios in
skeletal muscle of exercised rats. Anat. Re.,
159: 33-40.

Dellasanta, Louis R. 1964. Muscle fiber size and fiber
to capillary ratio of the gastrocnemius muscle of
sedentary adult rats. M.S. Thesis. Michigan
State University, East Lansing, Michigan.

Durant, R. R. 1927. Blood pressure in the rat. Am.
J. Physiol., _1: 679-685.

Greene, E. C. 1959. The Anatomy of the Rat. New
York, Hafner Pub. c.; 210-212, 312-322.

Hyman, Chester. 1963. The circulation of blood through
skeletal muscle. Pediatrics, 33: 671-679.

Jaya, Y. (Med. Coll. Hyderabad, India) 1958. The
arterial pattern of skeletal muscles. Jour.
Anat. Soc. India 7(1): 30-3“.

Krogh, A. 1936. Anatomy and Physiology of Capil-
laries. Yale University Press, New Haven; Lecture
II, 22-47, 369-392.

Le Gros Clark, W. E., and L. B. Blomfield. 19A5. The
efficiency of intramuscular anastomoses with
observations on the regeneration of devascularized
muscle. J. Anat. 12: 15-38.

Olmstead, F., and I. H. Page. 1966. Hemodynamic changes

in dogs caused by sodium pentobarbital anesthesia.
Am. J. Physiol., 210(4): 817-820.

20

21

Personal communication with Alfred M. Lucas, USDA Avian
Anatomy Project; research zoologist and project
leader.

Spalteholz, W. 1888. Die Vertheilung der Blutgefasse
in Muskel. Abhandl. Math. Phys. K1. Sachs. Ges.

Ward's Service Bulletin No. 5. 1950. The use of
Latex and Vinyl Acetate in the injection of the
circulatory system. Ward's Natural Science
Establishment, Inc.

West, W. T., and L. W. Gorhaml. 1968. Injection of the
arterial system of the mouse. Stain Technology:
37(2): 9-103.

Wiedeman, M. P. 196A. Dimensions of blood vessels
from distributing artery to collecting vein,
Circ. Res., 13: 375-378.

. 1962. Lengths and diameter of peripheral
arterial vessels in the living animal. Circ.
Res., ;-_O-O

MICHIGAN STATE UNIVERSITY

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