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b+/!F?89:R!.>:F:!8:F?H;F!BAGH6!;>7;!<:?8= '!7:!I:A7H:!87;!(X0X!7I;:8!G?@:8;6!D=<;8B@?;:!;=!;>:!A7B<;:<7BFAFR! !!!!!d`!!!!!!!'!Table 1. Mean Plasma [LH] in Control vs. AraC-treated Rats Time Control (n = 4) AraC (n=3) Hedges' gControl (n = 4) AraC (n=4) Hedges' g1100 1.35 ± 0.31 1.22 ± 0.420.2* 0.97 ± 0.16 0.8 ± 0 0.7**120014.83 ± 5.79 1.58 ± 0.441.2*** 2.89 ± 1.30 1.43 ± 0.390.7**130035.90 ± 7.2012.14 ± 4.101.6*** 21.69 ± 11.15 9.03 ± 1.410.7**1350 46.29 ± 10.1423.01 ± 4.971.1*** 38.57 ± 11.59 14.06 ± 2.271.3***140043.70 ± 9.0929.36 ± 8.250.7**34.08 ± 8.95 16.22 ± 2.541.2***145032.42 ± 6.0527.85 ± 7.410.3*40.02 ± 12.0915.99 ± 2.931.2***150029.44 ± 7.6225.91 ± 4.500.2*37.33 ± 10.0916.02 ± 2.951.2***160025.74 ± 5.7120.19 ± 5.380.4*26.84 ± 7.8312.29 ± 4.171.0***170014.60 ± 4.0714.12 ± 2.390.112.96 ± 1.34 9.54 ± 2.810.7**1800 5.32 ± 1.14 9.33 ± 3.34-0.8*** 3.48 ± 0.58 4.59 ± 1.98-0.3** 1900 2.47 ± 0.31 5.03 ± 1.35-1.3*** 1.45 ± 0.23 1.83 ± 0.53-0.4** AdultMeans are expressed as mean plasma LH concentration ± SEM. * denotes a small effect size (>0.2), ** a medium effect (>0.5), and *** a large effect (>0.8) Pubertal !!da!'+0JCD:'O.8 Q')3F:D0K:A>0K:G0A:'@E':3F:D0K:A>'O['>@'B:>:DK0A:'0E'?:GG='M@DA'BCD0AJ'FCM:D>H' 0/:'BCD0AJ'>9:'>0K:'@E'<'9@DK@A: L0ABC?:B'(I'=CDJ:. ''5:A7H:!87;F!U!18C$!UaRO! r9brH!B:!:R!&R!-BQ!>=?8F!7I;:8!;>:!0!B<^:D;B=BDH:!D=<;8=H!78::!C76F!B0K:G 0A:'@E':3F:D0K:A>'T['>@'B:>:DK0A:'0E'0A90M0>0@A'@E'?:GG'0@A' BCD0AJ'@D':D'FCM:D>H'='>9:'H'>@'J:A:D<>:'<'9@DK@A: L0ABC?:B'(I'=CDJ:. ''5:A7H:!87;F! 8:D:BE:C!=FA=;BD!AB! (874!U``!r 9brH!BBDH:V! PB;>!E:>BDH:! F=H?;B=:!::< !G8BA:C!PB;> !:F;87CB=H!@:W!7:< !G8=9:F;:8=<:!U0Y! OMM!r9!B:! ;>B8C!C76! 7;!NMMM>R! 1H==C!F7AGHB<9!=DD?88:C! =:!F7A:!C76!7F!0!B<^:D;B=!;=! Nf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p!`ORcdc W!#!MRMMMNVR! *87G>!8:G8:F:<;F!A:7'A@A:':3FD:==';% .'U(V!l 'F;7DS!BA79:F!=I!7!18C$ 'B8!D:HH!D=H=D7HBT:C!PB;>!#%g 'B8!B:!(X0XR!18C$W!#%gW!7BDSW!MRO! rA!F;:GVR!\>B;:!FD7H:! @78!8:G8:F:<;F!NM!rAR!&8;>=9=<7H!EB:P!;>8=?9>!;>:!ABCCH:!=I!;>:!D:HH!D=:!8:C!788=P!FB9:!Q!GH7<:W!98:::!6!GH7<:W!7:!T!GH7<:R!U1V!0B:! D>78;!C:GBD;B<9!;>:!A:7:!(X0X!;>7;!D=H=D7HBT:!PB;>! #%gR! (!1!Y Z X !!eM!020406080Treatment Time Mean maximum [LH] (ng/ml) puberty adulthood ME trt: control > AraC Control AraC 0100002000030000Treatment Time Area Under Curve (ng/ml/hr) puberty adulthood Control AraC !!!!!!!!!!'''''+0JCD:'O. XQ'4A@?NB@RA'@E'?:GG'0@A'BCD0AJ'FCM:D>H'@D'9@@B'D:=CG>='0A'< 'B:G0@A'@E' >9:'(I'=CDJ:.' 'U(V!(874!;8:7;A:<;!C?8B<9! G?@:8;6!7 ==C!8:C?D: C!;>:!+/!F?89: R!)=;;:C! E:8;BD7H!HB<:F!B;F!=IIR! U4V!(8:7!$7;!(874!;8:7;A:<;! C?8B<9!G?@:8;6!7==C!8:C?D:C!;>:!+/!F?89: !@6! Ndq!7:8:!(874 ';8:7;:C! 77C!7!H=P:8!A7QBA?A!i+/j!D=AG78:C!;=!D=<;8=H!7F!8:G8:F:<;!A:7D:<>K:A>['K@D:'?:GG'0@A'0A'>9:'!6;6'0='<==@?0<>:B'R0>9' 90J9:D'?@A?:A>D<>0@A'@E'(I'0A'>9:'MG@@B'E@GG@R0AJ'<'9@D K@A: L0ABC?:B'(I'=CDJ:.' ',:!:C! F:D;B=:!(X0X !8:E:7HF!7!HB<:78!D=88:H7;B==8A=<:!G8BAB<9!7:!:!(X0X !U#!p!MRMOeW! %L!p!MRLdW!FH=G:!p! MRM c!x!MRM `VR!!-=HBC!HB<:!BD:<>K:A>'D:BC?:='>9:'ACKM:D'@E'"DB2 L0D'?:GG='0A'>9:'!6;6 '@'>9:'!6;6'BCD0AJ'FCM:D>H'>99@@B.' 'U(V!%:G8:F:<;7;BE:!G>=;=ABD8=987G>F!F>=PB<9!18C$ 'B8!D:HHF!B:!(X0X!=I!I:A7H:!87;F!;8:7;:C! PB;>!E:>BDH: !U18C$!B==CR! ,A79:F!;7S:!(874 ';8:7;:C!77EB<9!H:FF!18C$ 'B8!D:HHF!;>77;!^?F;!9=;!18C$ !U37NWLNV!p!NcRcaeW! #!MRMMN VW!7:8:!78:! A=8:!18C$ 'B8!D:HHF!B :!(X0X !=I!77;!P:8:! 9BE:7;!P:8:!9BE:==C! U37NWLNV!p!cRcedW! #!p!MRMNd VR!*87G>!8:G8:F:<;F!A:7 AraC ME trt time: puberty > adulthood 0200400600800Mean # BrdU-ir cells in AVPV puberty adulthood Treatment Time ME trt: control > AraC ME trt time: puberty > adulthood 1!!!e`!Y Z X BrdU NeuN Merge BrdU GFAP Merge Y Z X Y Z X Pubertal Control 51 ± 3% BrdU only 39 ± 1% BrdU/GFAP 10 ± 3% BrdU/NeuN Avg Total BrdU = 105 Adult Control 59 ± 1% BrdU only 28 ± 2% BrdU/GFAP 13 ± 1% BrdU/NeuN Avg Total BrdU = 164 Pubertal AraC 43 ± 7% BrdU only 52 ± 2% BrdU/GFAP 5 ± 5% BrdU/NeuN Avg Total BrdU = 41 Adult AraC 67 ± 5% BrdU only 31 ± 7% BrdU/GFAP 2 ± 2% BrdU/NeuN Avg Total Brd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l'F;7DS!BA79:F!=I!7!18C$ 'B8!D:HH!D=H=D7 HBT:C!PB;>!5=F 'B8!B:!(X0XR!18C$W!5=FW!7BDSW!MRO!rA!F;:GV R!\>B;:!FD7H:! @78!8:G8:F:<;F!NM!rAR! &8;>=9=<7H!EB:P !;>8=?9>!;>:!ABCCH:!=I!;>:!D:HH!D=:!8:C!788=P!FB9:!Q!GH7<:W!98:::!6!GH7<:W!7:!T!GH7<:R! U1V!0B:! D>78;F!C:GBD;B<9!;>:!G :8D:<; 79: !=I!G?@:8;7HH6!=8!7C?H; '@=8< !U18C$ 'B8V !D:HHF!B:!(X0X!;>7;! 7HF=! :QG8:FF!5=F 'B8! C?8B<9!;>:!;BA:!=I!;>:!+/!F?89: !B7;!P:8:!;8:7;:C!PB;>!E:>BDH:! U18C$!B==C R!)7;7!78:!G8:F:<;:C!7F!A:7@'/:A>D0?CG'D:9:D'0A'>9:'F9:'!,4X!(874!I=8!a!P::SF!7:!7A69C7H7!UH=P!G=P:8 !BA79: !BB9>:8!G=P:8 !BA79:! 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