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      • SCOPUSKCI등재

        인간 조직에서 Heat Shock Protein A2 (HspA2) 단백질의 발현

        손원영,황서하,한징택,이재호,최윤정,김석중,김영찬,Son, W.Y.,Hwang, S.H.,Han, C.T.,Lee, J.H.,Choi, Y.J.,Kim, S.,Kim, Y.C. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.2

        In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

      • SCOPUSKCI등재

        비정상적 정자형성 환자의 정소에서 Heat Shock Protein A2 (hspA2) mRNA 발현의 감소

        손원영,황서하,한징택,이재호,김석중,김영찬,Son, W.Y.,Hwang, S.H.,Han, C.T.,Lee, J.H.,Kim, S.J.,Kim, Y.C. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.1

        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: (group 1) 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. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control. Results: On Competitive RT-PCR analyses for hspA2 mRNA, significant amount of 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 human testes with abnormal spermatogenesis, which in turn suggests that hspA2 gene may play a specific role during meiosis in human testes.

      • KCI등재

        지주세포증후군 환자에서 생식세포 배양의 유용성

        오종진,홍재엽,임정진,이동률,홍영권 대한비뇨의학회 2009 Investigative and Clinical Urology Vol.50 No.3

        Purpose: We determined the usefulness of in vitro germ cell culture in nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome, no sperm in testicular sperm extraction. Materials and Methods: This study included 44 patients (45 testicular tissues) with nonobstructive azoospermia who were diagnosed with Sertoli cell only syndrome and were found to have no sperm in testicular sperm extraction between January 2006 and July 2008. Among the 45 testicular tissues, 22 tissues were processed for culture. In the in vitro cultures, the testicular tissues were dissociated and plated on gelatin-coated dishes. Patients were divided into 2 groups according to culture success: group I, culture positive (+; n=10); and group II, culture negative (−; n=12). Results: The mean patient ages were 31.73 and 31.68 years for groups I and II, respectively. The mean testicular sizes were 10.19 and 10.42 cc, respectively; the semen volumes were 2.86 and 3.04 cc, respectively; and the mean FSH, LH, and testosterone levels were 18.86 mIU/ml, 5.99 mIU/ml, and 4.46 ng/ml vs. 21.02 mIU/ml, 6.29 mIU/ml, and 4.32 ng/ml for groups I and II, respectively, with no significant differences between the groups (p>0.05). The culture rate of nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome was 45.5% (10/22). Round spermatid injection was done in 2 patients with consent of the patients, but implantation failed. Among the 45 tissues, germ cells were found in 8 tissues after pathologic reexamination. Conclusions: The in vitro culture of germ cells would be useful in the advanced treatment of nonobstructive azoospermic patients. Purpose: We determined the usefulness of in vitro germ cell culture in nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome, no sperm in testicular sperm extraction. Materials and Methods: This study included 44 patients (45 testicular tissues) with nonobstructive azoospermia who were diagnosed with Sertoli cell only syndrome and were found to have no sperm in testicular sperm extraction between January 2006 and July 2008. Among the 45 testicular tissues, 22 tissues were processed for culture. In the in vitro cultures, the testicular tissues were dissociated and plated on gelatin-coated dishes. Patients were divided into 2 groups according to culture success: group I, culture positive (+; n=10); and group II, culture negative (−; n=12). Results: The mean patient ages were 31.73 and 31.68 years for groups I and II, respectively. The mean testicular sizes were 10.19 and 10.42 cc, respectively; the semen volumes were 2.86 and 3.04 cc, respectively; and the mean FSH, LH, and testosterone levels were 18.86 mIU/ml, 5.99 mIU/ml, and 4.46 ng/ml vs. 21.02 mIU/ml, 6.29 mIU/ml, and 4.32 ng/ml for groups I and II, respectively, with no significant differences between the groups (p>0.05). The culture rate of nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome was 45.5% (10/22). Round spermatid injection was done in 2 patients with consent of the patients, but implantation failed. Among the 45 tissues, germ cells were found in 8 tissues after pathologic reexamination. Conclusions: The in vitro culture of germ cells would be useful in the advanced treatment of nonobstructive azoospermic patients.

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