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대한생식의학회지 제26권 제1호 2010년
비정상적 정자형성 환자의 정소에서 Heat Shock Protein A2 (hspA2) mRNA발현의 감소
대진의료재단 분당 제생병원 불임 및 생식의학연구소1, 산부인과2,비뇨기과3, 서강대학교 이공대학 생명과학과4.
손원영, 황서하, 한징택, 이재호, 김석중, 김영찬,
Repression of HspA2 mRNA Expression in Human Testes with Abnormal Spermatogenesis
Son wy, Hwang sh, Han ct, Lee jh, Kim sj, Kim yc
Objective: Heat shock protein 70.2 (Hsp70.2) gene knockout mice are found to have premeiotic arrest at the primary spermatocyte stage with a complete absence of spermatids and spermatozoa. This observation led to the hypothesis that hspA2 may be disrupted in human testes with abnormal spermatogenesis. To test this hypothesis, we studied the mRNA expression of hspA2 in infertile men with azoospermia. Design: The mRNA expression were analyzed by competitive RT-PCR among testes with normal spermatogenesis, pachytene spermatocyte arrest, and sertoli-cell only syndrome. Materials and methods: Testicular biopsy was performed in men with azoospermia (n=15). Specimens were subdivided into three groups: (group1) normal spermatogenesis (n=5), (group 2) spermatocyte arrest (n=5), (group 3) Sertoli-cell only syndrome (n=5). Total RNA was extracted by Trizol reagent. Total extracted RNA was reverse transcribed into cDNA and amplified by PCR using specific primers for hspA2 target cDNAs. A competitive cDNA fragment was constructed by deleting a defined fragment from the target cDNA sequence, and then coamplified with the target cDNA for competitive PCR. Internal control was used glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Results: On Competitive RT-PCR analyses for hspA2 mRNA, significant hspA2 expression was observed in group 1, whereas a constitutively low level of hspA2 was expressed in groups 2 and 3. Conclusion(s): The study demonstrates that the hspA2 gene expression is down-regulated in the human testes with abnormal spermatogenesis, which in turn suggests that hspA2 gene may play a specific role during meiosis in human testes.
키워드 : Spermatogenic arrest, testes, hspA2 gene, Sertoli-cell only syndrome, and competitive RT-PCR.
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