title: [Expression of GDNF in rat Sertoli cells cultured in vitro at different temperatures].
abstract: OBJECTIVE
  To explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures.
  
  
  METHODS
  Using combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot.
  
  
  RESULTS
  Sertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01).
  
  
  CONCLUSIONS
  The proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.
authors: Department of Urology, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Ji-Cun
authors: JC
authors: Zhao
authors: Department of Cryomedicine, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Zai-Li
authors: ZL
authors: Sun
authors: Department of Cryomedicine, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Xiao-Peng
authors: XP
authors: Li
authors: Department of Cryomedicine, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Ting-Shuai
authors: TS
authors: Cao
authors: Department of Cryomedicine, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Jian-Ting
authors: JT
authors: Hu
authors: Department of Cryomedicine, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Hong-Qiang
authors: HQ
authors: Wang
authors: Central Laboratory, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Xiang-Ping
authors: XP
authors: Liu
authors: Central Laboratory, The Affiliated Hospital of Qingdao University Medical College, Qingdao, Shandong 266003, China.
authors: Pei-Tao
authors: PT
authors: Wang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012708:Sertoli Cells / Q000166:cytology / Q000378:metabolism*; D013696:Temperature; D013737:Testis / Q000166:cytology
chemical_list: C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor