The DDK strain (Mus musculus domesticus) of inbred mouse has a unique peculiarity known as DDK syndrome. The DDK females are mostly infertile when crossed with males of other inbred strains, while DDK males exhibit ...The DDK strain (Mus musculus domesticus) of inbred mouse has a unique peculiarity known as DDK syndrome. The DDK females are mostly infertile when crossed with males of other inbred strains, while DDK males exhibit normal fertility in the reciprocal crosses, as intrastrain matings. This DDK syndrome has been demonstrated to be caused by an incompatibility system between DDK ooplasmic factor and the sperm gene of other strains owing to the ovum mutant (Ore) locus on mouse Chromosome 11. Recently, it was reported that DDK females are fully fertile when crossed to males of MOM (M. m. molossinus) and CASP (M. m. castaneus) strains, indicating that no incompatibilities exist between DDK ooplasmic factor and sperm gene of MOM or CASP males. In the present study, DDK females were found to be also fully fertile when crossed to the males of PWK wild-derived inbred strain (originated from Czech Republic wild mice, M. m. musculus). The crosses of DDK females × F1 (DDK9 × PWK♀) males also resulted in normal fertility. Furthermore, the transmission ratios of Om alleles from these Fl males to their backcross N2 offspring are 50%:50% as genotyped by microsatellite markers closely linked to Om locus. Moreover, it was demonstrated that PWK females are also fully fertile when crossed to DDK males. All above results indicated that no incompatibility exists between ooplasmic factor and sperm gene in the intersubspecific crosses with DDK and PWK strains. PWK strain would also be useful for further investigations on the DDK syndrome, and DDK strain can be used more widely for various studies in the mouse.展开更多
Aim: To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic (Cres) gene in mouse testis and epididymis during postnatal development. Methods: The QuantiGene assay and i...Aim: To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic (Cres) gene in mouse testis and epididymis during postnatal development. Methods: The QuantiGene assay and indirect immunofluorescence technique were used to examine the Cres mRNA and Cres protein level in mouse testis and epididymis on postnatal days 14, 20, 22, 28, 35, 49, 70 and 420. Results: (1) In both the testis and epididymis, Cres mRNA was fast detected on day 20, then it increased gradually from day 20 to day 70, and the high expression level maintained till day 420. (2) In the testis, the Cres protein was exclusively localized to the elongating spermatids and was first detected on day 22. The number of Cres-positive spermatids increased progressively till day 49. From day 49 to day 420, the number of Cres-positive cells was almost stable. (3) The Cres protein was first detected on day 20 in the proximal caput epididymal epithelium. By day 35, the expression level of the Cres protein increased dramatically and the high level was maintained till day 420. Moreover, the luminal fluid of the midcaput epididymis was also stained Cres-positive from day 35 on. No Cres-positive staining was observed in distal caput, corpus and cauda epididymis throughout. Conclusion: The Cres gene displays a specific age-dependent expression pattern in mouse testis and epididymis on both the mRNA and protein level.展开更多
Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( ...Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( SMGT). Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.展开更多
Use of germ cells as vectors for transgenesis in mammals has been well developed and offers exciting prospects for experimental and applied biology, agricultural and medical sciences. Such approach is referred to as e...Use of germ cells as vectors for transgenesis in mammals has been well developed and offers exciting prospects for experimental and applied biology, agricultural and medical sciences. Such approach is referred to as either male germ cell mediated gene transfer (MGCMGT) or female germ cell mediated gene transfer (FGCMGT) technique. Sperm-mediated gene transfer (SMGT), including its alternative method, testis-mediated gene transfer (TMGT), becomes an established and reliable method for transgenesis. They have been extensively used for producing transgenic animals. The newly developed approach of FGCMGT, ovary-mediated gene transfer (OMGT) is also a novel and useful tool for efficient transgenesis. This review highlights an overview of the recent progress in germ cell mediated gene transfer techniques, methods developed and mechanisms of nucleic acid uptake by germ cells.展开更多
Aim: To investigate the possible functions of human sperm membrane protein (hSMP-1) in the process of fertilization. Methods: A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was ...Aim: To investigate the possible functions of human sperm membrane protein (hSMP-1) in the process of fertilization. Methods: A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 (anti-rhSMP-1) antibodies. Sperm acrosome reaction and spermzona pellucida (ZP) binding assay were carried out in 10-week-old BALB/c mice. Results: Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 (P 〈 0.05). Conclusion: The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.展开更多
An average human ejaculate contains over 100 million sperm, but only a few succeed in accomplishing the journey to an egg by migration through the female reproductive tract. Among these few sperm, only one participate...An average human ejaculate contains over 100 million sperm, but only a few succeed in accomplishing the journey to an egg by migration through the female reproductive tract. Among these few sperm, only one participates in fertilization. There might be an ingenious molecular mechanism to ensure that the very best sperm fertilize an egg. However, recent gene disruption experiments in mice have revealed that many factors previously described as important for fertilization are largely dispensable. One could argue that the fertilization mechanism is made robust against gene disruptions. However, this is not likely, as there are already six different gene-disrupted mouse lines (Calmegin, Adam Ia, Adam2, Adam3, Ace and Pgapl), all of which result in male sterility. The sperm from these animals are known to have defective zona-binding ability and at the same time lose oviduct-migrating ability. Concerning spermzona binding, the widely accepted involvement of sugar moiety on zona pellucida 3 (ZP3) is indicated to be dispensable by gene disruption experiments. Thus, the landscape of the mechanism of fertilization is revolving considerably. In the sperm-egg fusion process, CD9 on egg and IZUMO1 on sperm have emerged as essential factors. This review focuses on the mechanism of fertilization elucidated by gene-manipulated animals.展开更多
Diabetes Mellitus is a chronic disease that affects important body organs in a very serious manner. The consequences of this disease turn out to be a significant problem for the patient, who tries to cope with the new...Diabetes Mellitus is a chronic disease that affects important body organs in a very serious manner. The consequences of this disease turn out to be a significant problem for the patient, who tries to cope with the new condition his organism has been placed in. The most common effect of the disease, hyperglycaemia, leads over time to serious damage to various body systems, such as nerves and blood vessels. What is not widely known among the population is that diabetes may have harmful effects on the reproductive system of the men suffering from diabetes type 1 and 2 and that such a parameter could lead to or might be the reason for infertility problems for couples, for example, miscarriage or embryonic failure. AGEs is a number of products which are believed to play an important role, because their presence has been detected in increased level in diabetic men. This implies that those glycation products might play a key role in diabetic complications. Their receptor, RAGE, member of the immunoglobulin superfamily has been detected in the reproductive tract of diabetic men. Reactive oxygen species (ROS), a possible product of AGEs appear in high levels in seminal plasma and are believed to be the cause of DNA fragmentation. The objective of this review was to gather all the available material i.e. studies on diabetes mellitus in one article, to study the research which has already been conducted and the conclusions that have been drawn, in order to offer, if possible, new pathways and perspectives to the scientists, who focus on fertility problems, sometimes intractable.展开更多
HASPIN is a serine/threonine protein kinase predominantly expressed during spermatogenesis and localized in the nucleus. The HASPIN gene is conserved from yeast to mammals and plants. To investigate any possible assoc...HASPIN is a serine/threonine protein kinase predominantly expressed during spermatogenesis and localized in the nucleus. The HASPIN gene is conserved from yeast to mammals and plants. To investigate any possible associations between HASPIN polymorphisms and impaired spermatogenesis in Japanese males, we screened for mutations in the HASPIN coding sequence (CDS) using DNA from 282 sterile male patients and 262 fertile male volunteers. Polymorphisms were found at 10 positions within the HASPIN CDS. Among these 10 polymorphisms, 5 were found only in the infertile group, 3 of which were nonsynonymous. These polymorphisms found only in the infertile patients may be a cause of male infertility and thus valuable candidates for further study of this condition.展开更多
This paper aims to determine the possible role of estrogen receptor-β (ERβ) gene Rsal polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited: in vitr...This paper aims to determine the possible role of estrogen receptor-β (ERβ) gene Rsal polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited: in vitro fertilization (IVF) group, including 374 couples who underwent conventional IVF; intracytoplasmic sperm injection (ICSI) group, including 294 couples who underwent an ICSI procedure using ejaculated sperm; and azoospermic group, consisting of 197 couples who underwent ICSI using either testis or epididymis sperm. Rsal polymorphism in the ERβ gene was detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism technique; fertilization and high-quality embryo rates were evaluated for each group. In each group, no significant differences were found in the overall rates of fertilization and high-quality embryos among GG, AG and AA genotypes. However, the proportion of cycles possessing a satisfactory high-quality embryo rate with the AA genotype was significantly lower than that in the wild-type GG genotype from each group. These results demonstrated that sperm possessing the ERβ RsalA genotype may have reduced fertilization ability and decreased early embryonic developmental potential, which could directly or indirectly contribute to the low fertilization rate and early embryonic developmental arrest in some cases.展开更多
In the present study, a model is suggested to describe hormone control in male blue gourami (<i><span style="font-family:Verdana;">Trichogaster trichopterus</span></i><span style=&...In the present study, a model is suggested to describe hormone control in male blue gourami (<i><span style="font-family:Verdana;">Trichogaster trichopterus</span></i><span style="font-family:Verdana;">) along the gonadotropic brain</span><span style="font-size:10pt;font-family:Verdana;">-</span><span style="font-size:10pt;font-family:Verdana;">pituitary</span><span style="font-size:10pt;font-family:Verdana;">- </span><span style="font-size:10pt;font-family:Verdana;">gonad axis (BPG axis) and the hypothalamic-pituitary-somatotropic axis (HPS axis). This model is based on the cloning</span><span style="font-size:10.0pt;font-family:""><span style="font-family:Verdana;"> and transcription of genes encoding hormones of the two axes involved in spermatogenesis during blue gourami reproduction. Gene transcription is affected by environmental, biological, </span><span style="font-family:Verdana;">and behavioral factors. Mature males were examined in two different stages—nonreproductive in high-density habitats and reproductive in low-density </span><span style="font-family:Verdana;">habitats. Based on gene transcription, gonadotropin-releasing hormone 1 (GnRH1) was involved in controlling spermatogenesis (spermatogonia to spermatids) via the BPG axis in nonreproductive and reproductive stages by controlling follicle-stimulating hormone (FSH), 11-ketotestosterone (11KT) and 17</span><i><span style="font-family:Verdana;">β</span></i><span style="font-family:Verdana;">-estradiol (E</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">). However, GnRH3 had a larger effect during the reproductive stage via the BPG axis (spermatids to sperm) on luteinizing hormone (LH), 11KT, and 17</span><i><span style="font-family:Verdana;">α</span></i></span><span style="font-size:10pt;font-family:Verdana;">-</span><span style="font-size:10pt;font-family:Verdana;">hydroxyprogesterone (17P). At the same time, the HPS axis was involved in spermatogenesis via pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide PRP (formerly known as GHRH-like peptide) in the brain, and growth hormone (GH) in the pituitary affected synthesis of insulin-like growth factor 1 (IGF1) in the liver.</span>展开更多
基金supported by the National Natural Science Foundation of China(No.30770321)
文摘The DDK strain (Mus musculus domesticus) of inbred mouse has a unique peculiarity known as DDK syndrome. The DDK females are mostly infertile when crossed with males of other inbred strains, while DDK males exhibit normal fertility in the reciprocal crosses, as intrastrain matings. This DDK syndrome has been demonstrated to be caused by an incompatibility system between DDK ooplasmic factor and the sperm gene of other strains owing to the ovum mutant (Ore) locus on mouse Chromosome 11. Recently, it was reported that DDK females are fully fertile when crossed to males of MOM (M. m. molossinus) and CASP (M. m. castaneus) strains, indicating that no incompatibilities exist between DDK ooplasmic factor and sperm gene of MOM or CASP males. In the present study, DDK females were found to be also fully fertile when crossed to the males of PWK wild-derived inbred strain (originated from Czech Republic wild mice, M. m. musculus). The crosses of DDK females × F1 (DDK9 × PWK♀) males also resulted in normal fertility. Furthermore, the transmission ratios of Om alleles from these Fl males to their backcross N2 offspring are 50%:50% as genotyped by microsatellite markers closely linked to Om locus. Moreover, it was demonstrated that PWK females are also fully fertile when crossed to DDK males. All above results indicated that no incompatibility exists between ooplasmic factor and sperm gene in the intersubspecific crosses with DDK and PWK strains. PWK strain would also be useful for further investigations on the DDK syndrome, and DDK strain can be used more widely for various studies in the mouse.
文摘Aim: To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic (Cres) gene in mouse testis and epididymis during postnatal development. Methods: The QuantiGene assay and indirect immunofluorescence technique were used to examine the Cres mRNA and Cres protein level in mouse testis and epididymis on postnatal days 14, 20, 22, 28, 35, 49, 70 and 420. Results: (1) In both the testis and epididymis, Cres mRNA was fast detected on day 20, then it increased gradually from day 20 to day 70, and the high expression level maintained till day 420. (2) In the testis, the Cres protein was exclusively localized to the elongating spermatids and was first detected on day 22. The number of Cres-positive spermatids increased progressively till day 49. From day 49 to day 420, the number of Cres-positive cells was almost stable. (3) The Cres protein was first detected on day 20 in the proximal caput epididymal epithelium. By day 35, the expression level of the Cres protein increased dramatically and the high level was maintained till day 420. Moreover, the luminal fluid of the midcaput epididymis was also stained Cres-positive from day 35 on. No Cres-positive staining was observed in distal caput, corpus and cauda epididymis throughout. Conclusion: The Cres gene displays a specific age-dependent expression pattern in mouse testis and epididymis on both the mRNA and protein level.
基金Scientific Research Project for High Schools of the Educational Department of Liaoning Province,China(No.2008643)
文摘Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( SMGT). Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.
基金supported by the Peking University People’s Hospital Research and Development Foundation(No.RDB2007-03)
文摘Use of germ cells as vectors for transgenesis in mammals has been well developed and offers exciting prospects for experimental and applied biology, agricultural and medical sciences. Such approach is referred to as either male germ cell mediated gene transfer (MGCMGT) or female germ cell mediated gene transfer (FGCMGT) technique. Sperm-mediated gene transfer (SMGT), including its alternative method, testis-mediated gene transfer (TMGT), becomes an established and reliable method for transgenesis. They have been extensively used for producing transgenic animals. The newly developed approach of FGCMGT, ovary-mediated gene transfer (OMGT) is also a novel and useful tool for efficient transgenesis. This review highlights an overview of the recent progress in germ cell mediated gene transfer techniques, methods developed and mechanisms of nucleic acid uptake by germ cells.
文摘Aim: To investigate the possible functions of human sperm membrane protein (hSMP-1) in the process of fertilization. Methods: A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 (anti-rhSMP-1) antibodies. Sperm acrosome reaction and spermzona pellucida (ZP) binding assay were carried out in 10-week-old BALB/c mice. Results: Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 (P 〈 0.05). Conclusion: The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.
文摘An average human ejaculate contains over 100 million sperm, but only a few succeed in accomplishing the journey to an egg by migration through the female reproductive tract. Among these few sperm, only one participates in fertilization. There might be an ingenious molecular mechanism to ensure that the very best sperm fertilize an egg. However, recent gene disruption experiments in mice have revealed that many factors previously described as important for fertilization are largely dispensable. One could argue that the fertilization mechanism is made robust against gene disruptions. However, this is not likely, as there are already six different gene-disrupted mouse lines (Calmegin, Adam Ia, Adam2, Adam3, Ace and Pgapl), all of which result in male sterility. The sperm from these animals are known to have defective zona-binding ability and at the same time lose oviduct-migrating ability. Concerning spermzona binding, the widely accepted involvement of sugar moiety on zona pellucida 3 (ZP3) is indicated to be dispensable by gene disruption experiments. Thus, the landscape of the mechanism of fertilization is revolving considerably. In the sperm-egg fusion process, CD9 on egg and IZUMO1 on sperm have emerged as essential factors. This review focuses on the mechanism of fertilization elucidated by gene-manipulated animals.
文摘Diabetes Mellitus is a chronic disease that affects important body organs in a very serious manner. The consequences of this disease turn out to be a significant problem for the patient, who tries to cope with the new condition his organism has been placed in. The most common effect of the disease, hyperglycaemia, leads over time to serious damage to various body systems, such as nerves and blood vessels. What is not widely known among the population is that diabetes may have harmful effects on the reproductive system of the men suffering from diabetes type 1 and 2 and that such a parameter could lead to or might be the reason for infertility problems for couples, for example, miscarriage or embryonic failure. AGEs is a number of products which are believed to play an important role, because their presence has been detected in increased level in diabetic men. This implies that those glycation products might play a key role in diabetic complications. Their receptor, RAGE, member of the immunoglobulin superfamily has been detected in the reproductive tract of diabetic men. Reactive oxygen species (ROS), a possible product of AGEs appear in high levels in seminal plasma and are believed to be the cause of DNA fragmentation. The objective of this review was to gather all the available material i.e. studies on diabetes mellitus in one article, to study the research which has already been conducted and the conclusions that have been drawn, in order to offer, if possible, new pathways and perspectives to the scientists, who focus on fertility problems, sometimes intractable.
文摘HASPIN is a serine/threonine protein kinase predominantly expressed during spermatogenesis and localized in the nucleus. The HASPIN gene is conserved from yeast to mammals and plants. To investigate any possible associations between HASPIN polymorphisms and impaired spermatogenesis in Japanese males, we screened for mutations in the HASPIN coding sequence (CDS) using DNA from 282 sterile male patients and 262 fertile male volunteers. Polymorphisms were found at 10 positions within the HASPIN CDS. Among these 10 polymorphisms, 5 were found only in the infertile group, 3 of which were nonsynonymous. These polymorphisms found only in the infertile patients may be a cause of male infertility and thus valuable candidates for further study of this condition.
文摘This paper aims to determine the possible role of estrogen receptor-β (ERβ) gene Rsal polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited: in vitro fertilization (IVF) group, including 374 couples who underwent conventional IVF; intracytoplasmic sperm injection (ICSI) group, including 294 couples who underwent an ICSI procedure using ejaculated sperm; and azoospermic group, consisting of 197 couples who underwent ICSI using either testis or epididymis sperm. Rsal polymorphism in the ERβ gene was detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism technique; fertilization and high-quality embryo rates were evaluated for each group. In each group, no significant differences were found in the overall rates of fertilization and high-quality embryos among GG, AG and AA genotypes. However, the proportion of cycles possessing a satisfactory high-quality embryo rate with the AA genotype was significantly lower than that in the wild-type GG genotype from each group. These results demonstrated that sperm possessing the ERβ RsalA genotype may have reduced fertilization ability and decreased early embryonic developmental potential, which could directly or indirectly contribute to the low fertilization rate and early embryonic developmental arrest in some cases.
文摘In the present study, a model is suggested to describe hormone control in male blue gourami (<i><span style="font-family:Verdana;">Trichogaster trichopterus</span></i><span style="font-family:Verdana;">) along the gonadotropic brain</span><span style="font-size:10pt;font-family:Verdana;">-</span><span style="font-size:10pt;font-family:Verdana;">pituitary</span><span style="font-size:10pt;font-family:Verdana;">- </span><span style="font-size:10pt;font-family:Verdana;">gonad axis (BPG axis) and the hypothalamic-pituitary-somatotropic axis (HPS axis). This model is based on the cloning</span><span style="font-size:10.0pt;font-family:""><span style="font-family:Verdana;"> and transcription of genes encoding hormones of the two axes involved in spermatogenesis during blue gourami reproduction. Gene transcription is affected by environmental, biological, </span><span style="font-family:Verdana;">and behavioral factors. Mature males were examined in two different stages—nonreproductive in high-density habitats and reproductive in low-density </span><span style="font-family:Verdana;">habitats. Based on gene transcription, gonadotropin-releasing hormone 1 (GnRH1) was involved in controlling spermatogenesis (spermatogonia to spermatids) via the BPG axis in nonreproductive and reproductive stages by controlling follicle-stimulating hormone (FSH), 11-ketotestosterone (11KT) and 17</span><i><span style="font-family:Verdana;">β</span></i><span style="font-family:Verdana;">-estradiol (E</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">). However, GnRH3 had a larger effect during the reproductive stage via the BPG axis (spermatids to sperm) on luteinizing hormone (LH), 11KT, and 17</span><i><span style="font-family:Verdana;">α</span></i></span><span style="font-size:10pt;font-family:Verdana;">-</span><span style="font-size:10pt;font-family:Verdana;">hydroxyprogesterone (17P). At the same time, the HPS axis was involved in spermatogenesis via pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide PRP (formerly known as GHRH-like peptide) in the brain, and growth hormone (GH) in the pituitary affected synthesis of insulin-like growth factor 1 (IGF1) in the liver.</span>