Immunohistochemical Studies Of Heat Shock Proteins In Cynomolgus Monkeys (Macaca Fascicularis) Testes During Postnatal Development

Objective:To study the relationship between the age-related expression of Heat Shock Proteins 60,70,90α(HSP60,70,90α)and testicular development and spermatogenesis in cynomolgus monkey testes during postnatal development.Materials and Methods:30 male cynomolgus monkeys(Macaca Fascicularis)were divided into 5 developmental stages according to maturational morphological parameters,neonatal(n=5),early infantile(n=5), late infantile(n=6),pubertal(n=6),adult(n=8).Testicular tissues of the monkeys were fixed in Bouin's solution for 4-12 hours and embeded in paraffin wax.Sections were cut into 6-μM thick slices,which were then mounted on slides coated with Poly-L-Lysine.Deparaffinized sections were stained by haematoxylin and eosin.HSP60,HSP70 and HSP90αwere dectected by immunohistochemistry and computer image analysis system.SPSS 13.0 for windows statistical software package was used to assess the difference of ratio of positive cells in postnatal developmental stages.Results:HSP60,70,90αwere all shown to be expressed in cynomolgus monkey testes during postnatal development.Immunoreactive HSP60 was mainly located in the cytoplasm of spermatogonia and Sertoli cells,occasionally was labeled in Leydig cells in each developmental stage.In puberty and adult, HSP60 was also located in the cytoplasm of primary spermatocytes.There was significant difference(P＜0.01)in the ratio of HSP60-positive cells in 5 developmental stages,which was highest in late infantile period and lowest in adult.In neonatal,early infantile and late infantile periods,immunoreactive HSP70 was located in the cytoplasm of spermatogonia.In puberty, immunoreactive HSP70 was located in the cytoplasm of primary spermatocytes. In adult,immunoreactive HSP70 was located in the cytoplasm of spermatocytes and spermatids.There was significant difference(P＜0.01)in the ratio of HSP70-positive spermatogonia before puberty,which was lowest in neonatal and highest in late infantile.During all developmental stages,immunoreactive HSP90αwas only located in the cytoplasm of spermatogonia.There was significant difference(P＜0.01)in the ratio of HSP90α-positive cells in 5 developmental stages,which was lowest in neonatal and highest in late infantile.Conclusion:The variation of HSP60,70,90αexpression in cynomolgus monkey testes during postnatal development suggests they may play an important role in morphological development of testis and spermatogenesis.