DOI: /j.issn ,44(09): ChineseVeterinaryScience :S :A : (2014) 袁莉刚, 曲亚玲, 谷来凤, 闫振龙,

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1 DOI: /j.issn ,44(09): ChineseVeterinaryScience :S :A : (2014) 袁莉刚, 曲亚玲, 谷来凤, 闫振龙, 田旦增 ( ) : 为了探索蛋白基因产物 9.5(proteingeneproduct9.5,PGP9.5) 在不同海拔地区黄牛睾丸组织中的分布特征及其与生精功能的关系, 采用免疫组织化学 SP 法观察了 PGP9.5 在平原地区和高原地区黄牛睾丸组织中的分布特点结果显示,PGP9.5 显著分布于平原黄牛和高原黄牛睾丸 Leydig 细胞中, 间质血管周围或血管壁内为阴性, 生精小管内初级精母细胞为阴性, 其余生精细胞为阳性 ;PGP9.5 在平原黄牛 Sertoli 细胞为强阳性表达, 而高原黄牛阳性信号有所减弱, 仅在 Sertoli 细胞顶部呈弱阳性表达 ;PGP9.5 阳性信号高原黄牛和平原黄牛表达均在睾丸头和睾丸尾较强, 睾丸体最弱结论 :PGP9.5 在不同海拔地区黄牛睾丸生精小管及 Leydig 细胞中均有分布, 表明 PGP9.5 影响精子发生和激素合成等生殖机能活动 ; PGP9.5 在高原黄牛 Sertoli 细胞中的分布明显较平原黄牛弱, 提示高原环境对睾丸生精功能有一定影响 : 蛋白基因产物 9.5; 免疫组织化学 SP 法 ; 睾丸 ; 黄牛 ; 高原 ; 平原 Comparisonofdistributionofproteingeneproduct9.5inbovine testisfromplateauwiththatfromplain YUAN Li-gang,QU Ya-ling,GU Lai-feng,YANZhen-long,TIAN Dan-zeng (Colegeof Veterinary Medicine,Gansu Agricultural University,Lanzhou730070,China) Abstract:Thisexperimentwasconductedtoinvestigatethedistributionofproteingeneproduct9.5 tributioncharacteristicsofpgp9.5inbovinetestiswereanalyzed.intheresults,thetesticularparenchyma (PGP9.5)inbovinetestisfromplainandplateau.ByusingtheimmunohistochemicalSPtechnique,thedis- exhibitedarichinnervationbypgp9.5-positivefibers,mainlyassociatedwithleydigcelnestsandsemi- niferoustubules.pgp9.5 was mainly expressedin spermatogonia ofcatlefrom diferentaltitudes, however,noexpressionofpgp9.5 wasobserved withintheprimaryspermatocytes.theexpressionof PGP9.5incaputandcaudaofthetestis were muchstrongerthancorpusoftestis.inconclusion,the PGP9.5-positivefibersofthecatlefromdiferentaltitudesmayserveasneuronalregulatorforbovinetes- ticularfunctionsandregulatethespermatogenesisandandrogensynthesis,andtheplateauenvironmentis aninevitableefectoronthefunctionsoftesticular. Keywords:PGP9.5;immunohistochemicalSPtechnique;testis;catle;plateau;plain, - (GnRH) PGP9.5(proteingeneproduct9.5), : ;: : ( ); (1010RJZA210); (0902B-03) : (1974-),,,,,,Tel: , yuan2918@126.com

2 [1],PGP9.5 SP PGP9.5 ;30g/L H 2O 2 10 min, 15min ; [2] PGP μL PGP9.5,37 2h,PBS [3] ;PGP9.5 50μL IgG, 50μL (HRP) [4] PGP9.5,37 DAB,, PBS, LeydigPGP9.5 Olympus [5] Wrobel [6],PGP Leydig, 5, , 400 DP 5, Image-Pro Plus6.0 Suburo [10] [7] :-,+, [8-9], ; ;,, 2 PGP9.5, 2.1, toli, 2~3 7, 3~6,,2~3 6, 1~3 1.2 PGP9.5 3,3- (DAB) SP ZYMED 1.3 2~3,Leydig ; 样品固定和切片制备 Sertoli, 7, ;,, ( 1C D) 20h 24h,, 2.2 PGP9.5, HE 染色及免疫组织化学染色 5μm,HE Leydig,,, Ser-, ( 1A B), 5~8, ; PGP9.5 Leydig ; Leydig

9 : 9.5 983 ;PGP9.5 PGP9.5, PGP9.5, Sertoli, Leydig ; PGP9.5 ;, Sertoli PGP9.

1 Comparisonofbovinetestisbetweenplateaucatleandplaincatle A B: 分别是高原黄牛睾丸组织 ;C D:

A,B:Plateaubovinetestis,respectively;C,D:Plainbovinetestis,respectively;

5inthebovinetestisfromplateauandplain Bovine origins Plateaucatle Plaincatle

3 9 : ;PGP9.5 PGP9.5, PGP9.5, Sertoli, Leydig ; PGP9.5 ;, Sertoli PGP9.5 (, Sertoli 1 2A~D) ( 1 2E~I) 1 HE Fig.1 Comparisonofbovinetestisbetweenplateaucatleandplaincatle A B: 分别是高原黄牛睾丸组织 ;C D: 分别是平原黄牛睾丸组织 ; : 间质细胞 ;ST: 生精小管 A,B:Plateaubovinetestis,respectively;C,D:Plainbovinetestis,respectively; :Leydigcels;ST:Seminiferoustubule 1 PGP9.5 Table1 ThedensityofPGP9.5inthebovinetestisfromplateauandplain Bovine origins Plateaucatle Plaincatle Sertoli Sertolicels Spermatogonia Primary oftestis spermatocytes Spermatid Leydig Leydigcelsof caputand caudatestis Leydig Leydigcels ofcorpustestis Myoid peritubular cels Vascelum wal ,,, [2-6] Wrobel [11]

5inbovinetestisfromplateauandplain A B C: 分别是 PGP9.

5 在高原黄牛睾丸尾的表达情况, 标尺示 10μm;E G H: 分别是 PGP9.

5 在平原黄牛睾丸头尾的表达情况, 标尺示 10 μm; : 间质细胞 ; : 间质 PGP9.

4 PGP9.5 Fig.2 ImmunohistochemicalstainingofPGP9.5inbovinetestisfromplateauandplain A B C: 分别是 PGP9.5 在高原黄牛睾丸头体尾的表达情况, 标尺示 20μm;D:PGP9.5 在高原黄牛睾丸尾的表达情况, 标尺示 10μm;E G H: 分别是 PGP9.5 在平原黄牛睾丸头体尾的表达情况, 标尺示 20μm;F I: 分别是 PGP9.5 在平原黄牛睾丸头尾的表达情况, 标尺示 10 μm; : 间质细胞 ; : 间质 PGP9.5 阳性神经纤维 ; :Sertoli 细胞 ; : 肌样细胞 ; : 初级精母细胞 ;ST: 生精小管 ;V: 血管 A,B,C:PGP9.5expressionincaput,corpusandcaudaofplateaubovinetestis,respectively,Bar=20μm;D:PGP9.5expressioninplateaubovine caudatestis,bar=10μm;e,g,h:pgp9.5expressionincaput,corpusandcaudaofplainbovinetestis,respectively,bar=20μm;f,i:pgp9.5 expressionincaputandcaudaofplainbovinetestis,respectively,bar=10μm; :Leydigcels; :Nervebundle; :Sertolicels; :Myoid cels; :Primaryspermatocytes;ST:Seminiferoustubule;V:Vascelum,, PGP9.5,,PGP9.5 PGP9.5,, Saleh [5] [4], PGP9.5 PGP9.5 35,,PGP9.5 [12-13], Chow [14], PGP9.5, Leydig,,,PGP9.5 [2,15] PGP9.5,, 1 PGP9.5 [16-17], A, PGP9.5 [4-5] [18], [19] [20]

5 9 : [21] [22],, PGP9.5,, PGP9.5,, (References) [1] TAINIO H.Peptidergicinnervationofthehumantestisand epididymis[j].acta Histochem,1994,96(4): [2] GONG Y G,FENG M M,HU X N,etal.Peptidergicnot monoaminergicfibers profusely innervate the young adult [3] WROBEL K H,BRANDLB.Theautonomousinnervationof theporcinetestisintheperiodfrom birthtoadulthood[j]. [4] WROBEL K H,MOUSTAFA M N.Ontheinnervationofthe [6] WROBEL K H,G RTLER A.Thenervedistributioninthe [7] BELKIN V S H,KAIUMOV A K.Vascularization ofthe testesofthewhiteratinthehighaltitudeconditionsofthe PamirandAntarctica[J].Arkh AnatGistolEmbriol,1986,91,. Sertoli humantestis[j].j Anat,2009,214(3): ,, PGP9.5 Sertoli Anatomischer Anzeiger,1998,180(2): , [18] Zeng [22] donkeytestis[j].ann Anat,2000,182(1):13-22.,PGP9.5 Sertoli,, darius)[j].ann Anat,2002,184(3): [8-9] PGP9.5 testisofthecat[j].ann Anat,2001,183(4): Sertoli,,, Leydig - (9): Leydig [8], Davidof [23] [J].,2007,13(6): Leydig, Gong [2], PGP9.5 Leydig tion,2010,139(6): Leydig PGP9.5 Leydig testisofthecat[j].j Androl,2002,23(1): Leydig, thebovinetestis[j].acta Anat,1997,160(1):1-14., Neuropeptides,2011,45(1):55-61., Leydig Sertoli [15,24] Miyake [24],, Ser- toli, PGP9.5 Prince [15] [5] SALEH A M,ALAMELDIN M A,ABDELMONIEM M E,et al.immunohistochemicalinvestigations of the autonomous nervedistributioninthetestisofthecamel(camelusdrome- LIAO Wei-gong,GAO Yu-qi,CAI Ming-chun,etal.Hypoxia promotesapoptosisofgerm celsinrattestes[j].national Journalof Andrology,2007,13(6): (inChinese) [9] LIAO W,CAI M,CHENJ,etal.Hypobarichypoxiacauses deleteriousefectsonspermatogenesisinrats[j].reproduc- [10] SUBURO A M,CHIOCCHIOSR,SOLER M,etal.Peptider- gicinnervationofbloodvesselsandinterstitialcelsinthe [11] WROBEL K H,ABU-GHALIN.Autonomicinnervationof [12] ALLEN CD,WASERB,K RNER M,etal.NeuropeptideY actswithintherattestistoinhibittestosteronesecretion[j]. [13] GRUENEWALD D A,NAAIM A,MARCKBT,etal.Age- relateddecreaseinneuropeptide-ygeneexpressioninthear- cuatenucleusofthemaleratbrainisindependentoftesticular feedback[j].endocrinology,1994,134(6): [14] CHOW S H,GIGLIO W,ANESETTIR,etal.Theefectsof testiculardenervation on spermatogenesisin the Sprague- Dawleyrat[J].Neuroendocrinology,2000,72(1): [15] PRINCEFP.Ultrastructuralevidenceofadrenergic,aswel ascholinergic,nervevaricositiesinrelationtothelaminapro- priaofthehumanseminiferoustubulesduringchildhood[j].

6 TissueCel,1996,28(5): [16] BANAFSHEH H,MARYAM R A,MOHAMMAD M A,et al.isolation,identification,andcultureofgoatspermatogonial stemcelsusingc-kitandpgp9.5 markers[j].j AssistRe- prod Genet,2012,29(10): [17] HEIDARIB,GIFANI M,SHIRAZIA,etal.Enrichmentof undiferentiatedtypeaspermatogoniafromgoattestisusing discontinuouspercoldensitygradientanddiferentialplating [J].Avicenna J Med Biotechnol,2014,6(2): [18] KON Y,ENDOH D,IWANAGA T.Expressionofprotein geneproduct9.5,aneuronalubiquitinc-terminalhydrolase, anditsdevelopingchangeinsertolicelsofmousetestis[j]. MolReprod Dev,1999,54(4): [19] RODRIGUEZ-SOSAJR,DOBSON H,HAHNEL A.Isola- tionandtransplantationofspermatogoniainsheep[j].the- riogenology,2006,66(9): [20] ZHANGZ,HILLJ,HOLLAND M,etal.Bovinesertolicels colonizeandform tubulesin murinehostsfolowingtrans- plantationandgrafting procedures[j].j Androl,2008,29 (4): [21] VON KOPYLOW K,KIRCHHOFF C,JEZEK D,etal. Screeningforbiomarkersofspermatogoniawithinthehuman testis:awholegenomeapproach[j].hum Reprod,2010,25 (5): [22] ZENG W,SNEDAKER A K,MEGEES,etal.Preservation andtransplantationofporcinetestistissue[j].reprod Fer- tile Dev,2009,21(3): [23] DAVIDOFF M S,SCHULZE W,MIDDENDORFFR,etal. TheLeydigcelofthehumantestis-anew memberofthe difuseneuroendocrinesystem[j].cel Tissue Res,1993,271 (3): [24] MIYAKE K,YAMAMOTO M,MITSUYA H.Pharmacologi- calandhistologicalevidenceforadrenergicinnervationofthe myoidcelsintheratseminiferoustubule[j].tohoku J Exp Med,1986,149(1): ( )

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