The genome tagging project(GTP)plays a pivotal role in addressing a critical gap in the understanding of protein functions.Within this framework,we successfully generated a human influenza hemagglutinin-tagged sperm-s...The genome tagging project(GTP)plays a pivotal role in addressing a critical gap in the understanding of protein functions.Within this framework,we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411(HA-tagged Ssp411)mouse model.This model is instrumental in probing the expression and function of Ssp411.Our research revealed that Ssp411 is expressed in the round spermatids,elongating spermatids,elongated spermatids,and epididymal spermatozoa.The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis.Nevertheless,rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions.Ssp411 is not detectable in metaphase Ⅱ(MⅡ)oocytes,zygotes,or 2-cell stage embryos,highlighting its intricate role in early embryonic development.These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP,fostering groundbreaking advancements in the f ields of spermiogenesis and reproductive biology.展开更多
Lactate dehydrogenase C4 (LDHC4) is a key enzyme for sperm metabolism. It is distributed specifically in testis and is highly immunogenic. In this study, two DNA vaccines pVAX1-hLDHC and pVAX1-mLDHC were constructed b...Lactate dehydrogenase C4 (LDHC4) is a key enzyme for sperm metabolism. It is distributed specifically in testis and is highly immunogenic. In this study, two DNA vaccines pVAX1-hLDHC and pVAX1-mLDHC were constructed by inserting coding sequences of human and mice LDHC4 into the eukaryotic ex-pression vector pVAX1. The production of LDHC4 specific antibodies was induced in the sera of vac-cinated mice and the reproductive tract secretions of vaccinated female mice through immunization by mucosal surface instillation. Furthermore, the antibody titer increased with the times of immunization. In the mating experiment, the number of newborns of the vaccinated mice reduced significantly and some immunized female mice even lost the ability to bear any offsprings, suggesting that the difference between the immunized and control mice was statistically significant. Sperm agglutination analysis indicated that both the antisera from immunized mice and the reproductive tract secretions of vacci-nated female mice could agglutinate normal sperms. Results of immunohistochemistry showed that the antibodies present in the sera of immunized mice and the reproductive tract secretions of vaccinated female mice could specifically react with LDHC4 antigen, which mainly locates in the cytoplasm, acrosome membrane externa and acrosome capsule of the sperm. Taken together, our results indicated that the constructed contraceptive DNA vaccines did yield immunocontraceptive effects on mice and this would enable clinical trials in near future.展开更多
基金the support from the National Natural Science Foundation of China(No.32070849)The Foundation of Science and Technology Commission of Shanghai Municipality(No.22DX1900400)+1 种基金Science and Technology Commission of Shanghai Municipality(No.23JC1403803)Shanghai Municipal Science and Technology Commission Targeted Funding Project(No.22DX1900400).
文摘The genome tagging project(GTP)plays a pivotal role in addressing a critical gap in the understanding of protein functions.Within this framework,we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411(HA-tagged Ssp411)mouse model.This model is instrumental in probing the expression and function of Ssp411.Our research revealed that Ssp411 is expressed in the round spermatids,elongating spermatids,elongated spermatids,and epididymal spermatozoa.The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis.Nevertheless,rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions.Ssp411 is not detectable in metaphase Ⅱ(MⅡ)oocytes,zygotes,or 2-cell stage embryos,highlighting its intricate role in early embryonic development.These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP,fostering groundbreaking advancements in the f ields of spermiogenesis and reproductive biology.
基金Grants from the Key Research Projects of Science and Technology of Fujian Province,China (Grant No. 2002Y032)Chinese PLA Foundation of Medical Sciences during the Tenth Five–Year Period (Grant No. 01MA032)
文摘Lactate dehydrogenase C4 (LDHC4) is a key enzyme for sperm metabolism. It is distributed specifically in testis and is highly immunogenic. In this study, two DNA vaccines pVAX1-hLDHC and pVAX1-mLDHC were constructed by inserting coding sequences of human and mice LDHC4 into the eukaryotic ex-pression vector pVAX1. The production of LDHC4 specific antibodies was induced in the sera of vac-cinated mice and the reproductive tract secretions of vaccinated female mice through immunization by mucosal surface instillation. Furthermore, the antibody titer increased with the times of immunization. In the mating experiment, the number of newborns of the vaccinated mice reduced significantly and some immunized female mice even lost the ability to bear any offsprings, suggesting that the difference between the immunized and control mice was statistically significant. Sperm agglutination analysis indicated that both the antisera from immunized mice and the reproductive tract secretions of vacci-nated female mice could agglutinate normal sperms. Results of immunohistochemistry showed that the antibodies present in the sera of immunized mice and the reproductive tract secretions of vaccinated female mice could specifically react with LDHC4 antigen, which mainly locates in the cytoplasm, acrosome membrane externa and acrosome capsule of the sperm. Taken together, our results indicated that the constructed contraceptive DNA vaccines did yield immunocontraceptive effects on mice and this would enable clinical trials in near future.
