Chk1和Chk2蛋白在大鼠睾丸发育过程中的表达与定位
|
摘要
目的检测细胞周期检测点激酶1(Checkpoint kinase 1,Chk1)和细胞周期检测点激酶2(Checkpoint kinase 2,Chk2)蛋白在大鼠睾丸不同发育阶段的表达与定位,探讨2种蛋白在大鼠精子发生过程中的功能。方法选择4日龄、28日龄、56日龄、84日龄的大鼠睾丸和附睾组织为实验材料,采用免疫组化方法检测Chk1和Chk2蛋白在大鼠睾丸中的表达与定位。结果在精原细胞中,Chk1和Chk2都仅有微弱的表达; Chk2在细线期初级精母细胞中出现了强表达,而在粗线期初级精母细胞、早期精子细胞中未见表达,但在晚期精子细胞中再次出现强表达; Chk1蛋白在细线期初级精母细胞中未见表达,而在粗线期初级精母细胞中出现强表达,在早期精子细胞中表达不明显,在晚期精子细胞中出现强表达。Chk1在Leydig细胞中始终保持较强的表达,Chk2则是大鼠进入成年期后在Leydig细胞中表达出现增强。Chk1和Chk2在Sertoli细胞中始终维持微弱表达或不表达。结论通过对不同发育阶段大鼠睾丸细胞上Chk1和Chk2的表达与定位的研究发现,Chk1和Chk2可能分别在精子发生的不同阶段参于精子发育的调节,并共同在精子发生过程中发挥作用。
Objective The roles of Checkpoint kinase 1( Chk1) and Checkpoint kinase 2( Chk2) in sper-matogenesis were evaluated by detecting the expression and localization of Chk1 and Chk2 in different developmental stages of the rat testis. Methods Developmental expression and localization of Chk1 and Chk2 were investigated on days 4,28,56,and 84 of age in the male rat testis by immunohistochemistry( IHC). Results No significant staining of Chk1 and Chk2 were observed in spermatogonia. Chk2 predominated in leptotene spermatocytes and older spermatids,but was absent in pachytene spermatocytes and younger spermatids. Whereas Chk1 predominated in pachytene spermatocytes and older spermatids,but was absent or faint in leptotene spermatocytes and younger spermatids. Additionally,Intense staining for Chk1 was maintained in Leydig cells. When adult stage comes,Chk2 was detectable in the Leydig cells. Faint to absent immunoreactivity of Chk1 and Chk2 were detected in Sertoli cells throughout testicular development. Conclusions By investigating the expression and localization of Chk1 and Chk2 in testicular cells at different developmental stages. It is founded that Chk1 may display a different profile of stage-specificity compared to Chk2 and both are considered playing a role in the spermatogenesis.
Objective The roles of Checkpoint kinase 1( Chk1) and Checkpoint kinase 2( Chk2) in sper-matogenesis were evaluated by detecting the expression and localization of Chk1 and Chk2 in different developmental stages of the rat testis. Methods Developmental expression and localization of Chk1 and Chk2 were investigated on days 4,28,56,and 84 of age in the male rat testis by immunohistochemistry( IHC). Results No significant staining of Chk1 and Chk2 were observed in spermatogonia. Chk2 predominated in leptotene spermatocytes and older spermatids,but was absent in pachytene spermatocytes and younger spermatids. Whereas Chk1 predominated in pachytene spermatocytes and older spermatids,but was absent or faint in leptotene spermatocytes and younger spermatids. Additionally,Intense staining for Chk1 was maintained in Leydig cells. When adult stage comes,Chk2 was detectable in the Leydig cells. Faint to absent immunoreactivity of Chk1 and Chk2 were detected in Sertoli cells throughout testicular development. Conclusions By investigating the expression and localization of Chk1 and Chk2 in testicular cells at different developmental stages. It is founded that Chk1 may display a different profile of stage-specificity compared to Chk2 and both are considered playing a role in the spermatogenesis.
引文
[1] Hull MG,Glazener CM,Kelly NJ,et al. Population study of causes,treatment,and outcome of infertility[J]. Br Med J(Clin Res Ed),1985,291(6510):1693-1697.
[2] Ahn J,Urist M,Prives C. The Chk2 protein kinase[J]. DNA Repair(Amst),2004,3(8-9):1039-1047.
[3] Bartek J,Lukas J. Chk1 and Chk2 kinases in checkpoint control and cancer[J]. Cancer Cell,2003,3(5):421-429.
[4] Kastan MB,Bartek J. Cell-cycle checkpoints and cancer[J]. Nature,2004,432(7015):316-323.
[5] Mcneely S,Conti C,Sheikh T,et al. Chk1 inhibition after replicative stress activates a double strand break response mediated by ATM and DNA-dependent protein kinase[J]. Cell Cycle,2010,9(5):995-1004.
[6]刘德风,李斌,姜辉,等. Chk1基因在人类精子中的表达及其意义[J].中华男科学杂志,2010 16(9):811-815.
[7] Flaggs G,Plug AW,Dunks KM,et al. Atm-dependent interactions of a mammalian chk1 homolog with meiotic chromosomes[J]. Curr Biol,1997,7(12):977-986.
[8] Falck J,Mailand N,Syljuasen RG,et al. The ATM-Chk2-Cdc25A checkpoint pathway guards against radioresistant DNA synthesis[J].Nature,2001,410(6830):842-847.
[9] Chen CC,Kennedy RD,Sidi S,et al. CHK1 inhibition as a strategy for targeting fanconi Anemia(FA)DNA repair pathway deficient tumors[J]. Mol Cancer,2009,8(1):8-24.
[2] Ahn J,Urist M,Prives C. The Chk2 protein kinase[J]. DNA Repair(Amst),2004,3(8-9):1039-1047.
[3] Bartek J,Lukas J. Chk1 and Chk2 kinases in checkpoint control and cancer[J]. Cancer Cell,2003,3(5):421-429.
[4] Kastan MB,Bartek J. Cell-cycle checkpoints and cancer[J]. Nature,2004,432(7015):316-323.
[5] Mcneely S,Conti C,Sheikh T,et al. Chk1 inhibition after replicative stress activates a double strand break response mediated by ATM and DNA-dependent protein kinase[J]. Cell Cycle,2010,9(5):995-1004.
[6]刘德风,李斌,姜辉,等. Chk1基因在人类精子中的表达及其意义[J].中华男科学杂志,2010 16(9):811-815.
[7] Flaggs G,Plug AW,Dunks KM,et al. Atm-dependent interactions of a mammalian chk1 homolog with meiotic chromosomes[J]. Curr Biol,1997,7(12):977-986.
[8] Falck J,Mailand N,Syljuasen RG,et al. The ATM-Chk2-Cdc25A checkpoint pathway guards against radioresistant DNA synthesis[J].Nature,2001,410(6830):842-847.
[9] Chen CC,Kennedy RD,Sidi S,et al. CHK1 inhibition as a strategy for targeting fanconi Anemia(FA)DNA repair pathway deficient tumors[J]. Mol Cancer,2009,8(1):8-24.