ABSTRACT
Phosphoribosyl-pyrophosphate synthetase 2 (PRPS2) is a rate-limiting enzyme and plays an important role in purine and pyrimidine nucleotide synthesis. Recent studies report that PRPS2 is involved in male infertility. However, the role of PRPS2 in hypospermatogenesis is unknown. In this study, the relationship of PRPS2 with hypospermatogenesis and spermatogenic cell apoptosis was investigated. The results showed that PRPS2 depletion increased the number of apoptotic spermatogenic cells in vitro. PRPS2 was downregulated in a mouse model of hypospermatogenesis. When PRPS2 expression was knocked down in mouse testes, hypospermatogenesis and accelerated apoptosis of spermatogenic cells were noted. E2F transcription factor 1 (E2F1) was confirmed as the target gene of PRPS2 and played a key role in cell apoptosis by regulating the P53/Bcl-xl/Bcl-2/Caspase 6/Caspase 9 apoptosis pathway. Therefore, these data indicate that PRPS2 depletion contributes to the apoptosis of spermatogenic cells and is associated with hypospermatogenesis, which may be helpful for the diagnosis of male infertility.
ABSTRACT
<p><b>Objective</b>To study the expression of CLAUDIN-11 in the testis tissue of non-obstructive azoospermia (NOA) patients with different severities and investigate its clinical significance.</p><p><b>METHODS</b>Sixty-two NOA patients were divided into a hypospermatogenesis (HS) group (n = 30) and a Sertoli cell only syndrome (SCO) group (n =32). The expression of CLAUDIN-11 in the testicular tissue of the patients was detected by immunohistochemistry, that of CLAUDIN-11 mRNA determined by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the levels of serum reproductive hormones measured by chemiluminescent immunoassay.</p><p><b>RESULTS</b>Immunohistochemistry showed that the expression of CLAUDIN-11 was mainly in the cytoplasm of the Sertoli cells around the seminiferous tubule wall in the HS group, but diffusely distributed in the membrane of the Sertoli cells in the SCO group. RT-qPCR revealed a significantly lower expression of CLAUDIN-11 mRNA in the HS than in the SCO group (0.008 ± 0.001 vs 0.013 ± 0.002, t = 10.616, P<0.01). The level of serum luteotropic hormone (LH) was also markedly lower in the HS than in the SCO group (［3.62 ± 1.34］ vs ［4.96 ± 3.10］ IU/L, P<0.05) and so was that of follicle-stimulating hormone (FSH) (［5.36 ± 2.80］ vs ［10.65 ± 9.18］ IU/L, P<0.05).</p><p><b>CONCLUSIONS</b>The up-regulated expression of CLAUDIN-11 in Sertoli cells may play an important role in the development and progression of spermatogenic dysfunction in NOA patients.</p>
Subject(s)
Humans , Male , Azoospermia , Genetics , Metabolism , Claudins , Metabolism , Follicle Stimulating Hormone , Metabolism , Oligospermia , Genetics , Metabolism , RNA, Messenger , Metabolism , Seminiferous Tubules , Metabolism , Sertoli Cell-Only Syndrome , Genetics , Metabolism , Sertoli Cells , Metabolism , Spermatogenesis , Testis , MetabolismABSTRACT
PURPOSE: This study was performed to assess the fertilization, embryonic development and pregnancy rate using fresh and frozen-thawed testicular sperm from hypospermatogenic patients. MATERIALS AND METHODS: A total of 84 cycles of ICSI were performed with fresh or frozen-thawed testicular sperm frm hypospermatogenesis patients. Of these, 55 cycles(65.5%) were performed with fresh sperm, and 29 cycles(34.5%) were performed with frozen-thawed sperm. Testicular tissue was frozen with the programmed cell freezer(Cryomagic I, Miraebiotech, Seoul, Korea). Fertilization was checked 18-20 hrs after ICSI. Embryo grade was assessed based on the day 2 and day 3 embryo morphology. Embryo grade was scored five groups, and good embryos were classified as grade I to grade II. RESULTS: The total percentage of fertilized embryos was 62.4%, the development of good embryos was 58.8%(290/493), and pregnancy rate was 39.2%(29/74). For fresh sperm, the percentage of fertilized embryos was 65.7%, the percentage of good embryos was 57.6%(204/354) and the pregnancy rate was 34.8%(16/46). For thawed sperm, the percentage of fertilized embryos was 54.4%, the percentage of good embryos was 61.9%(86/139), and pregnancy rate was 46.4%(13/28). Thawed sperm showed a higher percentage of good embryos and higher pregnancy rate than fresh sperm. CONCLUSIONS: In this study we obtained acceptable rates of fertilization and good embryos after ICSI with testicular sperm from hypospermatogenesis patients. Frozen sperm showed higher rates of good embryos and pregnancy than fresh sperm. Therefore both fresh and frozen testicular sperm are effective in ICSI for embryonic development and pregnancy in patients with hypospermatogenesis.
Subject(s)
Female , Humans , Male , Pregnancy , Pregnancy , Cryopreservation , Embryonic Development , Embryonic Structures , Fertilization , Oligospermia , Pregnancy Rate , Seoul , Sperm Injections, Intracytoplasmic , SpermatozoaABSTRACT
PURPOSE: This study was performed to assess the fertilization, embryonic development and pregnancy rate using fresh and frozen-thawed testicular sperm from hypospermatogenic patients. MATERIALS AND METHODS: A total of 84 cycles of ICSI were performed with fresh or frozen-thawed testicular sperm frm hypospermatogenesis patients. Of these, 55 cycles(65.5%) were performed with fresh sperm, and 29 cycles(34.5%) were performed with frozen-thawed sperm. Testicular tissue was frozen with the programmed cell freezer(Cryomagic I, Miraebiotech, Seoul, Korea). Fertilization was checked 18-20 hrs after ICSI. Embryo grade was assessed based on the day 2 and day 3 embryo morphology. Embryo grade was scored five groups, and good embryos were classified as grade I to grade II. RESULTS: The total percentage of fertilized embryos was 62.4%, the development of good embryos was 58.8%(290/493), and pregnancy rate was 39.2%(29/74). For fresh sperm, the percentage of fertilized embryos was 65.7%, the percentage of good embryos was 57.6%(204/354) and the pregnancy rate was 34.8%(16/46). For thawed sperm, the percentage of fertilized embryos was 54.4%, the percentage of good embryos was 61.9%(86/139), and pregnancy rate was 46.4%(13/28). Thawed sperm showed a higher percentage of good embryos and higher pregnancy rate than fresh sperm. CONCLUSIONS: In this study we obtained acceptable rates of fertilization and good embryos after ICSI with testicular sperm from hypospermatogenesis patients. Frozen sperm showed higher rates of good embryos and pregnancy than fresh sperm. Therefore both fresh and frozen testicular sperm are effective in ICSI for embryonic development and pregnancy in patients with hypospermatogenesis.