Glycosylphosphatidylinositol-anchored sperm hyaluronidases have long been believed to assist in sperm penetration through the cumulus-oocyte complex(COC);however,their role in mammalian fertilization remains unclear.P...Glycosylphosphatidylinositol-anchored sperm hyaluronidases have long been believed to assist in sperm penetration through the cumulus-oocyte complex(COC);however,their role in mammalian fertilization remains unclear.Previously,we have shown that hyaluronidase 5(Hyal5)/Hyal7 double-knockout(dKO)mice produce significantly fewer offspring than their wild-type(WT)counterparts because of defective COC dispersal.Male infertility is mainly caused by a low sperm count.It can be further exacerbated by the deficiency of sperm hyaluronidase,which disperses the cumulus cells of the outer layer of the COC.In the current study,we evaluated the effects of a low count of Hyal-deficient sperm and conditions of ovulated oocytes on the fertilization rate using a mouse model.Our results demonstrated that a low sperm count further decreases the in vitro fertilization(IVF)rate of Hyal-deficient dKO spermatozoa.In addition,the dKO spermatozoa resulted in a fertilization rate of 12.5%upon fertilizing COCs with a thick cumulus layer,whereas the IVF rate was comparable to that of WT spermatozoa when oocytes with a thin or no cumulus layer were fertilized.Finally,we proved that the IVF rate of dKO spermatozoa could be recovered by adding rat spermatozoa as a source of sperm hyal.Our results suggest that a deficiency of proteins involved in fertilization,such as sperm hyal,has a vital role in fertilization.展开更多
Changes of ovarian tPA,uPA and PA inhibitor activities were examined in PMSG-and hCG-treatedimmature mice during periovulatory periods.The results show that 15% of the gonadotropin-treatedanimals ovulated 8 hrs after ...Changes of ovarian tPA,uPA and PA inhibitor activities were examined in PMSG-and hCG-treatedimmature mice during periovulatory periods.The results show that 15% of the gonadotropin-treatedanimals ovulated 8 hrs after hCG administration,about 6-8 hrs earlier than in rat.It is also shownthat not only tPA activity,but also uPA activity,was regulated by gonadotropins in ovarianhomogenates and granulosa cells,and they reached maximum prior to ovulation.No measurableamount of PAI-1 activity could be detected in mouse granulosa cell conditioned medium andfollicular fluid,but considerable amount of α_2-antiplasmin,a specific inhibitor for plasmin,wasfound in follicular fluid.Cumulus-oocyte complexes contain mainly tPA.Since the ovulated eggsstill have high tPA activity,it is thought that the enzyme in the oocyte may play an important rolein implantation.展开更多
Raising Min pigs(MPs)in northern China is challenging,because of harsh environmental conditions in winter.There is a hypothesis that MPs can tolerate cold stress better than Large White pigs(LWPs)and their oocyte comp...Raising Min pigs(MPs)in northern China is challenging,because of harsh environmental conditions in winter.There is a hypothesis that MPs can tolerate cold stress better than Large White pigs(LWPs)and their oocyte competence is particularly protected.Over a period of 3 years,multiple cohorts of MPs and LWPs under different environmental conditions were studied in this paper.Follistatin(FST)serum level was used as a marker for swine oocyte competence.Oocytes were collected from the swine and their maturation rate,cleavage rate and blastocyst rate were measured.Lastly,mRNA expression in ovary and longissimus dorsi for the genes FST,luteinizing hormone receptor(LHR)and cold-inducible RNA-binding protein(CIRP)was measured.Under cold stress,serum follistatin concentrations of both LWPs and MPs were markedly increased,and they were more in LWPs than in MPs.However,oocyte competence was found to be unaffected by cold stress in MPs.It was found that in a separate group of LWPs,oocyte competence was lower in samples collected in winter than that in other seasons.Interestingly,follistatin levels in the follicular fluid of MPs showed no change with cold stress,and serum follistatin levels were increased in sows.Gene expression analysis showed a significant reduction in FST,LHR and CIRP expressions under cold stress in longissimus dorsi,but not in the ovaries.It was found that MPs were better able to maintain oocyte competence under cold stress,which might be due to improved regulation of follistatin in follicular fluid,and MPs appeared likely an adaptation to the harsh conditions that they faced under domestication in northern China.展开更多
基金supported by the KRIBB Research Initiative Program(KGM4252122)the National Research Foundation(2018M2A9Hl078340)the National Research Foundation of Korea Grant funded by the Korean Government(NRF-2020R111A3072358).
文摘Glycosylphosphatidylinositol-anchored sperm hyaluronidases have long been believed to assist in sperm penetration through the cumulus-oocyte complex(COC);however,their role in mammalian fertilization remains unclear.Previously,we have shown that hyaluronidase 5(Hyal5)/Hyal7 double-knockout(dKO)mice produce significantly fewer offspring than their wild-type(WT)counterparts because of defective COC dispersal.Male infertility is mainly caused by a low sperm count.It can be further exacerbated by the deficiency of sperm hyaluronidase,which disperses the cumulus cells of the outer layer of the COC.In the current study,we evaluated the effects of a low count of Hyal-deficient sperm and conditions of ovulated oocytes on the fertilization rate using a mouse model.Our results demonstrated that a low sperm count further decreases the in vitro fertilization(IVF)rate of Hyal-deficient dKO spermatozoa.In addition,the dKO spermatozoa resulted in a fertilization rate of 12.5%upon fertilizing COCs with a thick cumulus layer,whereas the IVF rate was comparable to that of WT spermatozoa when oocytes with a thin or no cumulus layer were fertilized.Finally,we proved that the IVF rate of dKO spermatozoa could be recovered by adding rat spermatozoa as a source of sperm hyal.Our results suggest that a deficiency of proteins involved in fertilization,such as sperm hyal,has a vital role in fertilization.
基金This work was supported by National Natural Science Foundation,the"Eighth Five"Scientific Development Program,research fund for State Key Laboratories and Rockefeller Foundation,New York,USA.
文摘Changes of ovarian tPA,uPA and PA inhibitor activities were examined in PMSG-and hCG-treatedimmature mice during periovulatory periods.The results show that 15% of the gonadotropin-treatedanimals ovulated 8 hrs after hCG administration,about 6-8 hrs earlier than in rat.It is also shownthat not only tPA activity,but also uPA activity,was regulated by gonadotropins in ovarianhomogenates and granulosa cells,and they reached maximum prior to ovulation.No measurableamount of PAI-1 activity could be detected in mouse granulosa cell conditioned medium andfollicular fluid,but considerable amount of α_2-antiplasmin,a specific inhibitor for plasmin,wasfound in follicular fluid.Cumulus-oocyte complexes contain mainly tPA.Since the ovulated eggsstill have high tPA activity,it is thought that the enzyme in the oocyte may play an important rolein implantation.
基金Supported by the National Natural Science Foundation of China(32172696,U20A2052,31872980)Heilongjiang Provincial Scientific Research Business Fund Project(CZKYF2020A004,CZKYF2021-2-C025)Heilongjiang Provincial Natural Science Foundation(LH2019C064)。
文摘Raising Min pigs(MPs)in northern China is challenging,because of harsh environmental conditions in winter.There is a hypothesis that MPs can tolerate cold stress better than Large White pigs(LWPs)and their oocyte competence is particularly protected.Over a period of 3 years,multiple cohorts of MPs and LWPs under different environmental conditions were studied in this paper.Follistatin(FST)serum level was used as a marker for swine oocyte competence.Oocytes were collected from the swine and their maturation rate,cleavage rate and blastocyst rate were measured.Lastly,mRNA expression in ovary and longissimus dorsi for the genes FST,luteinizing hormone receptor(LHR)and cold-inducible RNA-binding protein(CIRP)was measured.Under cold stress,serum follistatin concentrations of both LWPs and MPs were markedly increased,and they were more in LWPs than in MPs.However,oocyte competence was found to be unaffected by cold stress in MPs.It was found that in a separate group of LWPs,oocyte competence was lower in samples collected in winter than that in other seasons.Interestingly,follistatin levels in the follicular fluid of MPs showed no change with cold stress,and serum follistatin levels were increased in sows.Gene expression analysis showed a significant reduction in FST,LHR and CIRP expressions under cold stress in longissimus dorsi,but not in the ovaries.It was found that MPs were better able to maintain oocyte competence under cold stress,which might be due to improved regulation of follistatin in follicular fluid,and MPs appeared likely an adaptation to the harsh conditions that they faced under domestication in northern China.
