Aim:

To examine the possible effect of heat treatment on expression of heat shock proteins (Hsps) 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways.

Methods:

Western blot analysis, realtime polymerase chain reaction (PCR), and confocal immunohistochemistry were used to analyze mRNA and protein levels of the Hsps in response to 43°C treatment of Sertoli cells isolated from pubertal monkey testes.

Results:

Staining with Hoechst 33342 indicated Sertoli cells did not undergo apoptosis after heat treatment. Hsp105 was expressed in cytoplasm of untreated Sertoli cells. Both Hsp105 mRNA and protein levels were increased approximately 20-fold compared to those of the untreated controls at 12 h after heat treatment. Untreated Sertoli cells did not express Hsp70, but heat stress induced its expression in the cell cytoplasm. The time-course of changes in Hsp70 was similar to that of Hsp105. In contrast to Hsp105 and Hsp70, the change in Hsp60 expression was much less obvious. The protein level between 12 h and 48 h after heat treatment was only approximately 1.5-fold that of the untreated control. Extracellular regulated kinase (ERK) 1/2 inhibitor (U0126) or phosphoinositide kinase-3 (PI3K) inhibitor (LY294002) could partially block the response of Hsp105 and Hsp70 induced by heat treatment.

Conclusion:

These results indicate that the heat-induced expression of the three types of Hsp in monkey Sertoli cells might be regulated by ERK and/or PI3K signal pathways, but the profile of their expression is different, suggesting that they might have different regulatory functions in Sertoli cells.

References

1. Kormano M. Development of the rectum–testis temperature difference in the post-natal rat. J Reprod Fertil 1967; 14: 427–37. | PubMed | ChemPort |
2. Tao SX, Guo J, Zhang XS, Li YC, Hu ZY, Han CS, et al. Germ cell apoptosis induced by experimental cryptorchidism is mediated by multiple molecular pathways in cynomolgus macaque. Front Biosci 2006; 11: 1077–89. | Article | PubMed | ChemPort |
3. Lue YH, Hikim AP, Swerdloff RS, Im P, Taing KS, Bui T, et al. Single exposure to heat induces stage-specific germ cell apoptosis in rats: role of intratesticular testosterone on stage specificity. Endocrinology 1999; 140: 1709–17. | Article | PubMed | ISI | ChemPort |
4. Lue YH, Lasley BL, Laughlin LS, Swerdloff RS, Hikim AP, Leung A, et al. Mild testicular hyperthermia induces profound transitional spermatogenic suppression through increased germ cell apoptosis in adult cynomolgus monkeys (Macaca fascicularis). J Androl 2002; 23: 799–805. | PubMed |
5. Griswold MD. Interactions between germ cells and Sertoli cells in the tesis. Biol Reprod 1995; 52: 211–6. | Article | PubMed | ISI | ChemPort |
6. Huleihel M, Lunenfeld E. Regulation of spermatogenesis by paracrine/autocrine testicular factors. Asian J Androl 2004; 6: 259–68. | PubMed | ChemPort |
7. Lindquist S. The heat-shock response. Annu Rev Biochem 1986; 55: 1151–91. | Article | PubMed | ISI | ChemPort |
8. Morimoto RI. Cells in stress: transcriptional activation of heat shock genes. Science 1993; 259: 1409–10. | Article | PubMed | ISI | ChemPort |
9. Jolly C, Morimoto RI. Role of the heat shock response and molecular chaperones in oncogenesis and cell death. J Natl Cancer Inst 2000; 92: 1564–72. | Article | PubMed | ChemPort |
10. Easton DP, Kaneko Y, Subjeck JR. The hsp110 and Grp170 stress proteins: newly recognized relatives of the Hsp70s. Cell Stress Chaperones 2000; 5: 276–90. | Article | PubMed | ISI | ChemPort |
11. Craig EA, Gambill BD, Nelson RJ. Heat shock proteins: molecular chaperones of protein biogenesis. Microbiol Rev 1993; 57: 402–14. | PubMed | ISI | ChemPort |
12. Nonoguchi K, Itoh K, Xue JH, Tokuchi H, Nishiyama H, Kaneko Y, et al. Cloning of human cDNAs for Apg-1 and Apg-2, members of the Hsp110 family, and chromosomal assignment of their genes. Gene 1999; 237: 21–8. | Article | PubMed | ChemPort |
13. Krawczyk Z, Mali P, Parvinen M. Expression of a testis-specific hsp70 gene-related RNA in defined stages of rat seminiferous epithelium. J Cell Biol 1988; 107: 1317–23. | Article | PubMed | ISI | ChemPort |
14. Dix DJ, Rosario-Herrle M, Gotoh H, Mori C, Goulding EH, Barrett CV, et al. Developmentally regulated expression of Hsp70-2 and a Hsp70-2/lacZ transgene during spermatogenesis. Dev Biol 1996; 174: 310–21. | Article | PubMed | ChemPort |
15. Matsumoto M, Fujimoto H. Cloning of a hsp70–related gene expressed in mouse spermatids. Biochem Biophys Res Commun 1990; 166: 43–9. | Article | PubMed | ISI | ChemPort |
16. Meinhardt A, Parvinen M, Bacher M, Aumuller G, Hakovirta H, Yagi A, et al. Expression of mitochondrial heat shock protein 60 in distinct cell types and defined stages of rat seminiferous epithelium. Biol Reprod 1995; 52: 798–807. | Article | PubMed | ChemPort |
17. Mein hardt A, Seitz J, Arslan M, Aumuller G, Weinbauer GF. Hormonal regulation and germ cell-specific expression of heat shock protein 60 (hsp60) in the testis of macaque monkeys (Macaca mulatta and M. fascicularis). Int J Androl 1998; 21: 301–7. | Article | PubMed | ChemPort |
18. Werner A, Meinhardt A, Seitz J, Bergmann M. Distribution of heat-shock protein 60 immunoreactivity in testes of infertile men. Cell Tissue Res 1997; 288: 539–44. | Article | PubMed | ChemPort |
19. Feng HL, Sandlow JI, Sparks AE. Decreased expression of the heat shock protein hsp70–2 is associated with the pathogen-esis of male infertility. Fertil Steril 2001; 76: 1136–9. | Article | PubMed | ChemPort |
20. Zhang XS, Lue YH, Guo SH, Yuan JX, Hu ZY, Han CS, et al. Expression of HSP105 and HSP60 during germ cell apoptosis in the heat-treated testes of adult cynomolgus monkeys (Macaca fascicularis). Front Biosci 2005; 10: 3110–21. | Article | PubMed | ChemPort |
21. Zhou XC, Han XB, Hu ZY, Zhou RJ, Liu YX. Expression of Hsp70–2 in unilateral cryptorchid testis of rhesus monkey during germ cell apoptosis. Endocrine 2001; 16: 89–95. | Article | PubMed | ChemPort |
22. Guo CX, Ma J, Zhou XC, Liu YX. Expression of HSP70–2 gene during germ cell apoptosis in rat unilateral cryptorchid testes. Arch Androl 2001; 46: 109–15. | Article | PubMed | ChemPort |
23. Widlak W. [The heat shock response and HSP70 gene expression in male germ cells.] Postepy Biochem 2006; 52: 289–95. (In Polish.). | PubMed | ChemPort |
24. Widlak W, Winiarski B, Krawczyk A, Vydra N, Malusecka E, Krawczyk Z. Inducible 70 kDa heat shock protein does not protect spermatogenic cells from damage induced by cryptorchidism. Int J Androl 2007; 30: 80–7. | Article | PubMed | ChemPort |
25. Vydra N, Malusecka E, Jarzab M, Lisowska K, Glowala-Kosinska M, Benedyk K, et al. Spermatocyte-specific expression of constitutively active heat shock factor 1 induces HSP70i-resistant apoptosis in male germ cells. Cell Death Differ 2006; 13: 212–22. | Article | PubMed | ChemPort |
26. Zakeri ZF, Welch WJ, Wolgemuth DJ. Characterization and inducibility of hsp 70 proteins in the male mouse germ line. J Cell Biol 1990; 111: 1785–92. | Article | PubMed | ChemPort |
27. Hayashida N, Inouye S, Fujimoto M, Tanaka Y, Izu H, Takaki E, et al. A novel HSF1–mediated death pathway that is suppressed by heat shock proteins. EMBO J 2006; 25: 4773–83. | Article | PubMed | ChemPort |
28. Kaneko Y, Kimara T, Kishishita M, Noda Y, Fujita J. Cloning of apg-2 encoding a novel member of heat shock protein 110 family. Gene 1997; 189: 19–24. | Article | PubMed | ChemPort |
29. Kaneko Y, Nishiyama H, Nonoguchi K, Higashitsuji H, Kishishita M, Fujita J. A novel hsp110–related gene, apg-1, that is abundantly expressed in the testis responds to a low temperature heat shock rather than the traditional elevated temperatures. J Biol Chem 1997; 272: 2640–5. | Article | PubMed | ISI | ChemPort |
30. Majumdar SS, Winters SJ, Plant TM. Procedures for the isolation and culture of Sertoli cells from the testes of infant, juvenile, and adult rhesus monkeys (Macaca mulatta). Biol Reprod 1998; 58: 633–40. | Article | PubMed | ChemPort |
31. Yu FQ, Han CS, Yang W, Jin X, Hu ZY, Liu YX. Activation of the p38 MAPK pathway by follicle-stimulating hormone regulates steroidogenesis in granulosa cells differentially. J Endocrinol 2005; 186: 85–96. | Article | PubMed | ChemPort |
32. Jin X, Han CS, Zhang XS, Yuan JX, Hu ZY, Liu YX. Signal transduction of stem cell factor in promoting early follicle development. Mol Cell Endocrinol 2005; 229: 3–10. | Article | PubMed | ChemPort |
33. Zhang XS, Zhang ZH, Guo SH, Yang W, Zhang ZQ, Yuan JX, et al. Activation of extracellular signal-related kinases 1 and 2 in Sertoli cells in experimentally cryptorchid rhesus monkeys. Asian J Androl 2006; 8: 265–72. | Article | PubMed | ChemPort |
34. Zhang XS, Zhang ZH, Jin X, Wei P, Hu XQ, Chen M, et al. Dedifferentiation of adult monkey Sertoli cells through activation of extracellularly regulated kinase 1/2 induced by heat treatment. Endocrinology 2006; 147: 1237–45. | Article | PubMed | ChemPort |
35. Sharpe RM, McKinnell C, Kivlin C, Fisher JS. Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. Reproduction 2003; 125: 769–84. | Article | PubMed | ChemPort |
36. Gething MJ, Sambrook JF. Protein folding in the cell. Nature 1992; 355: 33–45. | Article | PubMed | ISI | ChemPort |
37. Parsell DA, Lindquist S. The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins. Annu Rev Genet 1993; 27: 437–96. | Article | PubMed | ISI | ChemPort |
38. Zhao ZG, Ma QZ, Xu CX. Abrogation of heat-shock protein (HSP)70 expression induced cell growth inhibition and apoptosis in human androgen-independent prostate cancer cell line PC-3m. Asian J Androl 2004; 6: 319–24. | PubMed | ChemPort |
39. Nonoguchi K, Tokuchi H, Okuno H, Watanabe H, Egawa H, Saito K, et al. Expression of Apg-1, a member of the Hsp110 family, in the human testis and sperm. Int J Urol 2001; 8: 308–14. | Article | PubMed | ChemPort |
40. Tavaria M, Gabriele T, Kola I, Anderson RL. A hitchhiker's guide to the human Hsp70 family. Cell Stress Chaperones 1996; 1: 23–8. | Article | PubMed | ISI | ChemPort |
41. Widlak W, Vydra N, Malusecka E, Dudaladava V, Winiarski B, Sciegliñska D, et al. Heat shock transcription factor 1 down-regulates spermatocyte-specific 70 kDa heat shock protein expression prior to the induction of apoptosis in mouse testes. Genes Cells 2007; 12: 487–99. | Article | PubMed | ChemPort |
42. Eddy EM. Role of heat shock protein HSP70–2 in spermatogenesis. Rev Reprod 1999; 4: 23–30. | Article | PubMed | ISI | ChemPort |
43. Meinhardt A, Wilhelm B, Seitz J. Expression of mitochondrial marker proteins during spermatogenesis. Hum Reprod Update 1999; 5: 108–19. | Article | PubMed | ChemPort |
44. Rohde M, Daugaard M, Jensen MH, Helin K, Nylandsted J, Jaattela M. Members of the heat-shock protein 70 family promote cancer cell growth by distinct mechanisms. Genes Dev 2005; 19: 570–82. | Article | PubMed | ISI | ChemPort |
45. Mosser DD, Caron AW, Bourget L, Denis-Larose C, Massie B. Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis. Mol Cell Biol 1997; 17: 5317–27. | PubMed | ISI | ChemPort |
46. Buzzard KA, Giaccia AJ, Killender M, Anderson RL. Heat shock protein 72 modulates pathways of stress-induced apoptosis. J Biol Chem 1998; 273: 17147–53. | Article | PubMed | ISI | ChemPort |
47. Wei YQ, Zhao X, Kariya Y, Teshigawara K, Uchida A. Inhibition of proliferation and induction of apoptosis by abrogation of heat-shock protein (HSP) 70 expression in tumor cells. Cancer Immunol Immunother 1995; 40: 73–8. | Article | PubMed | ISI | ChemPort |
48. Nylandsted J, Brand K, Jaattela M. Heat shock protein 70 is required for the survival of cancer cells. Ann NY Acad Sci 2000; 926: 122–5. | PubMed | ChemPort |
49. Allen RL, O'Brien DA, Eddy EM. A novel hsp70-like protein (P70) is present in mouse spermatogenic cells. Mol Cell Biol 1988; 8: 828–32. | PubMed | ISI | ChemPort |
50. Allen RL, O'Brien DA, Jones CC, Rockett DL, Eddy EM. Expression of heat shock proteins by isolated mouse spermatogenic cells. Mol Cell Biol 1988; 8: 3260–6. | PubMed | ISI | ChemPort |
51. Cheng MY, Hartl FU, Martin J, Pollock RA, Kalousek F, Neupert W, et al. Mitochondrial heat shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria. Nature 1989; 337: 620–5. | Article | PubMed | ISI | ChemPort |
52. Kyriakis JM, Avruch J. Mammalian mitogenactivated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 2001; 81: 807–69. | PubMed | ISI | ChemPort |
53. Matsuzawa A, Ichijo H. Molecular mechanisms of the decision between life and death: regulation of apoptosis by apoptosis signal-regulating kinase 1. J Biochem 2001; 130: 1–8. | PubMed | ChemPort |
54. Guo J, Tao SX, Chen M, Shi YQ, Zhang ZQ, Li YC, et al. Heat treatment induces liver receptor homolog-1 expression in monkey and rat Sertoli cells. Endocrinology 2007; 148: 1255–65. | Article | PubMed | ChemPort |

MORE ARTICLES LIKE THIS

These links to content published by NPG are automatically generated.