To date, a practical method for inducing superovulation in the Mongolian gerbil has not been defined; therefore, in this study, we attempted to develop a flexible superovulation protocol for this species. Superovulation can be induced in the Mongolian gerbils by using PMSG with or without hCG. The injection schedule for PMSG (and hCG) has a high degree of flexibility, but the best protocol for embryo collection for reproductive biology is 20 IU PMSG followed by 20 IU hCG 54 hours later.
{"title":"Interval between PMSG Priming and hCG Injection in Superovulation of the Mongolian Gerbil","authors":"Y. Kameyama, Kaori Arai, Y. Ishijima","doi":"10.1274/JMOR.21.105","DOIUrl":"https://doi.org/10.1274/JMOR.21.105","url":null,"abstract":"To date, a practical method for inducing superovulation in the Mongolian gerbil has not been defined; therefore, in this study, we attempted to develop a flexible superovulation protocol for this species. Superovulation can be induced in the Mongolian gerbils by using PMSG with or without hCG. The injection schedule for PMSG (and hCG) has a high degree of flexibility, but the best protocol for embryo collection for reproductive biology is 20 IU PMSG followed by 20 IU hCG 54 hours later.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"56 1","pages":"105-109"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77863339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Egashira, M. Motoishi, M. Sugioka, Emiko Nagafuti, Kanako Ohtsu, Akemi Nishigaki, Naomi Yoshioka, T. Kuramoto
The cycles were divided into the following five groups (100-500, 500-1,000, 1,000-1,500, 1,500-2,000, ≥2,000 × 104/ml) by the concentration of motile sperm and compared the rates of normal fertilization and cycle from which normal fertilization was not obtained. In motile sperm concentration from 100 to <500 × 104/ml, the normal fertilization rate was significantly lower (48.8%) than other groups. The accordance with the aging and declining sperm counts, the normal fertilization rate has declined and ration of the cycle, which could not obtain normal fertilization has risen. This study suggested that the cycle in motile sperm concentration from 100 to <500 × 104/ml is not be suitable for IVF. Moreover, the cycle in motile sperm concentration from 500 to <1,000 × 104/ml, we have to consider the number of oocyte, female age and number of treatment cycle.
{"title":"Evaluation of Sperm Fertilizing Ability by Using Motility Sperm Concentration in In-vitro Fertilization Programs","authors":"A. Egashira, M. Motoishi, M. Sugioka, Emiko Nagafuti, Kanako Ohtsu, Akemi Nishigaki, Naomi Yoshioka, T. Kuramoto","doi":"10.1274/JMOR.21.214","DOIUrl":"https://doi.org/10.1274/JMOR.21.214","url":null,"abstract":"The cycles were divided into the following five groups (100-500, 500-1,000, 1,000-1,500, 1,500-2,000, ≥2,000 × 104/ml) by the concentration of motile sperm and compared the rates of normal fertilization and cycle from which normal fertilization was not obtained. In motile sperm concentration from 100 to <500 × 104/ml, the normal fertilization rate was significantly lower (48.8%) than other groups. The accordance with the aging and declining sperm counts, the normal fertilization rate has declined and ration of the cycle, which could not obtain normal fertilization has risen. This study suggested that the cycle in motile sperm concentration from 100 to <500 × 104/ml is not be suitable for IVF. Moreover, the cycle in motile sperm concentration from 500 to <1,000 × 104/ml, we have to consider the number of oocyte, female age and number of treatment cycle.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"10 1","pages":"214-219"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86574499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Amemiya, Y. Iwanami, Toshihiro Kobayashi, T. Terao, Y. Fukui, H. Ishikawa, S. Ohsumi, M. Hirabayashi, S. Hochi
The presence or absence of sperm-borne oocyte-activating factor (SOAF) in the Antarctic minke whale haploid spermatogenic cells was determined by assessing the meiosis resumption of microinseminated mouse oocytes. The relative capacity of mature spermatozoa from mouse, cattle and whale to resume the meiosis of BDF1 mouse oocytes was, respectively, 90.5, 84.6 and 76.5%, while nuclear changes in non-treated or buffer-injected oocytes did not occur after 90-min culture. In the whales, the late-stage elongating spermatids as well as the testicular spermatozoa triggered the meiosis resumption of mouse oocytes at similar rates (oocyte activation rates; 68.0 and 62.5%, respectively). The oocyte activating capacity of the early-stage elongating spermatids was significantly lower (25.0%), and the round spermatids did not activate mouse oocytes at all. This result suggests that the SOAF activity in the Antarctic minke whales is acquired during the early phase of spermiogenesis.
{"title":"Acquirement of Oocyte-activating Factor in Antarctic Minke Whale (Balaenoptera bonaerensis) Spermatogenic Cells, Assessed by Meiosis Resumption of Microinseminated Mouse Oocytes","authors":"K. Amemiya, Y. Iwanami, Toshihiro Kobayashi, T. Terao, Y. Fukui, H. Ishikawa, S. Ohsumi, M. Hirabayashi, S. Hochi","doi":"10.1274/JMOR.21.149","DOIUrl":"https://doi.org/10.1274/JMOR.21.149","url":null,"abstract":"The presence or absence of sperm-borne oocyte-activating factor (SOAF) in the Antarctic minke whale haploid spermatogenic cells was determined by assessing the meiosis resumption of microinseminated mouse oocytes. The relative capacity of mature spermatozoa from mouse, cattle and whale to resume the meiosis of BDF1 mouse oocytes was, respectively, 90.5, 84.6 and 76.5%, while nuclear changes in non-treated or buffer-injected oocytes did not occur after 90-min culture. In the whales, the late-stage elongating spermatids as well as the testicular spermatozoa triggered the meiosis resumption of mouse oocytes at similar rates (oocyte activation rates; 68.0 and 62.5%, respectively). The oocyte activating capacity of the early-stage elongating spermatids was significantly lower (25.0%), and the round spermatids did not activate mouse oocytes at all. This result suggests that the SOAF activity in the Antarctic minke whales is acquired during the early phase of spermiogenesis.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"84 1","pages":"149-156"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82417402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To demonstrate delivery by using re-vitrified blastocysts derived from supernumerary embryos in the event of an unsuccessful pregnancy attempt. Nine early stage cleaved embryos were frozen by vitrification, and subsequently thawed. Four of the nine embryos developed to the blastocyst stage. Two of the four blastocysts were transferred, and two supernumerary good embryos remained unexpectedly, and they were refrozen by vitrification under consent. Subsequently, one of the re-vitrified embryos developed to 38 weeks and was delivered by cesarean section (a health female; 46, XX). The transfer of re-vitrified supernumerary blastocysts resulted in a successful pregnancy and delivery outcome. This study suggests that re-vitrification is one rescue procedure which allows the re-use of supernumerary embryos in patients who failed to have a pregnancy after frozen embryo transfer.
{"title":"Successfully healthy baby delivery from human refrozen blastocyst embryos by vitrification","authors":"Tomomi Takahashi, Y. Araki","doi":"10.1274/JMOR.21.162","DOIUrl":"https://doi.org/10.1274/JMOR.21.162","url":null,"abstract":"To demonstrate delivery by using re-vitrified blastocysts derived from supernumerary embryos in the event of an unsuccessful pregnancy attempt. Nine early stage cleaved embryos were frozen by vitrification, and subsequently thawed. Four of the nine embryos developed to the blastocyst stage. Two of the four blastocysts were transferred, and two supernumerary good embryos remained unexpectedly, and they were refrozen by vitrification under consent. Subsequently, one of the re-vitrified embryos developed to 38 weeks and was delivered by cesarean section (a health female; 46, XX). The transfer of re-vitrified supernumerary blastocysts resulted in a successful pregnancy and delivery outcome. This study suggests that re-vitrification is one rescue procedure which allows the re-use of supernumerary embryos in patients who failed to have a pregnancy after frozen embryo transfer.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"40 1","pages":"162-165"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73475262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chromosomal abnormalities were evaluated in mouse embryos derived from in vitro fertilization using oocytes from old (12 months) BALB/c (12-BALB) and ICR (12-ICR) females and young (3 months; 3-BALB and 3-ICR) female mice. Chromosomal analysis was carried out at two developmental stages, 12-BALB and 12-ICR vs 3-BALB and 3-ICR at the first cleavage division and 12-BALB vs 3-ICR at the second division. Although all young females ovulated, this was not so in the old females; additionally, significantly fewer oocytes were ovulated by the old females and the fertilization rate was significantly lower in the 12-ICR group, but not in the 12-BALB group. Significantly fewer of the embryos in the old group had a metaphase figure at the first cleavage division and this correlated with significantly more of the embryos in this group being still at the pronuclear stage. The incidence of polyploidy, probably due to polyspermy, was found in both the 12-BALB and 12-ICR groups at the first cleavage division, but this was not seen in embryos at the second division. These results indicate that the aging of females impairs reproductive success, as evidenced by anovulation, polyspermy, and delays in fertilization/asynchronous development in resulting embryos.
{"title":"Chromosomal Analysis of Embryos Derived from In Vitro Fertilization of Oocytes from Old and Young Female Mice","authors":"Kaori Yoshizawa, M. Yoshizawa, Shino Sasaki","doi":"10.1274/JMOR.21.192","DOIUrl":"https://doi.org/10.1274/JMOR.21.192","url":null,"abstract":"Chromosomal abnormalities were evaluated in mouse embryos derived from in vitro fertilization using oocytes from old (12 months) BALB/c (12-BALB) and ICR (12-ICR) females and young (3 months; 3-BALB and 3-ICR) female mice. Chromosomal analysis was carried out at two developmental stages, 12-BALB and 12-ICR vs 3-BALB and 3-ICR at the first cleavage division and 12-BALB vs 3-ICR at the second division. Although all young females ovulated, this was not so in the old females; additionally, significantly fewer oocytes were ovulated by the old females and the fertilization rate was significantly lower in the 12-ICR group, but not in the 12-BALB group. Significantly fewer of the embryos in the old group had a metaphase figure at the first cleavage division and this correlated with significantly more of the embryos in this group being still at the pronuclear stage. The incidence of polyploidy, probably due to polyspermy, was found in both the 12-BALB and 12-ICR groups at the first cleavage division, but this was not seen in embryos at the second division. These results indicate that the aging of females impairs reproductive success, as evidenced by anovulation, polyspermy, and delays in fertilization/asynchronous development in resulting embryos.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"18 1","pages":"192-199"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72713133","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Katayose, A. Shinohara, M. Chiba, H. Yamada, K. Tominaga, A. Sato, K. Yanagida
This study aimed to investigate the effects of metals on semen profiles. The concentrations of 50 elements in seminal plasma collected from 128 infertile men were measured. Eleven (Na, Mg, P, K, Ca, Fe, Cu, Zn, Se, Rb, Sr) elements were positively detected in all samples. Another eight elements (V, Mn, Co, As, Mo, Cd, Sn, Ba) were detected in over 75% of the samples. In these 19 elements, significant correlations were observed only between copper concentration and sperm motility. The presence of cadmium and zinc in seminal plasma was associated with a low total sperm number (p=0.067) and low sperm motility (p=0.052), respectively. Higher concentrations of cadmium were observed in the Brinkmann index under 100 than in that over 100 (p=0.055). Recovery of sperm motility after EDTA treatment was observed with in vitro exposure to 300 μg/ml of zinc sulfate. Declines in sperm motility after exposure to 50 μg/ml of copper sulfate were irreversible, even with EDTA treatment. It was suggested that excess copper and zinc in seminal plasma was detrimental for male reproductive capacity by reducing sperm motility. It also appeared that cadmium may exert toxic effects on spermatogenesis, after long-term exposure, as occurs with cigarette smoking.
{"title":"Effects of Various Elements in Seminal Plasma on Semen Profiles","authors":"H. Katayose, A. Shinohara, M. Chiba, H. Yamada, K. Tominaga, A. Sato, K. Yanagida","doi":"10.1274/JMOR.21.141","DOIUrl":"https://doi.org/10.1274/JMOR.21.141","url":null,"abstract":"This study aimed to investigate the effects of metals on semen profiles. The concentrations of 50 elements in seminal plasma collected from 128 infertile men were measured. Eleven (Na, Mg, P, K, Ca, Fe, Cu, Zn, Se, Rb, Sr) elements were positively detected in all samples. Another eight elements (V, Mn, Co, As, Mo, Cd, Sn, Ba) were detected in over 75% of the samples. In these 19 elements, significant correlations were observed only between copper concentration and sperm motility. The presence of cadmium and zinc in seminal plasma was associated with a low total sperm number (p=0.067) and low sperm motility (p=0.052), respectively. Higher concentrations of cadmium were observed in the Brinkmann index under 100 than in that over 100 (p=0.055). Recovery of sperm motility after EDTA treatment was observed with in vitro exposure to 300 μg/ml of zinc sulfate. Declines in sperm motility after exposure to 50 μg/ml of copper sulfate were irreversible, even with EDTA treatment. It was suggested that excess copper and zinc in seminal plasma was detrimental for male reproductive capacity by reducing sperm motility. It also appeared that cadmium may exert toxic effects on spermatogenesis, after long-term exposure, as occurs with cigarette smoking.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"28 1","pages":"141-148"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78185945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Principles of the modern bioethics and the ethical status of the fetus are interpreted in this article. The modern bioethics is based on the three basic ethical principles reported in Belmont report. The principles consist of respect for persons, beneficence and justice. Respect for persons was considered to be the most important principle and have the priority among the principles. However some ethicist dissent from the priority of respect for persons. When we face a case in which the principles conflict each other, the top priority needs to be granted to one of the principles. How we select one of the principles for the top priority remains an unsolved issue. Considering the ethical status of the fetus, the most important thing is when the embryo or fetus becomes a person. Roman Catholic declared in 1974 that the embryo just after fertilization is considered as a person. However many bio-ethicists believe that only an autonomous person who is an individual capable of deliberation about personal goals, and of acting under the direction of such deliberation has the right of living. This opinion permits us not only to abort fetus artificially but also to kill infants. In the latter half of the article personal opinion on the ethical status of the fetus is explained.
{"title":"Ethical Base of Artificial Reproductive Medicine","authors":"S. Yamano","doi":"10.1274/JMOR.21.177","DOIUrl":"https://doi.org/10.1274/JMOR.21.177","url":null,"abstract":"Principles of the modern bioethics and the ethical status of the fetus are interpreted in this article. The modern bioethics is based on the three basic ethical principles reported in Belmont report. The principles consist of respect for persons, beneficence and justice. Respect for persons was considered to be the most important principle and have the priority among the principles. However some ethicist dissent from the priority of respect for persons. When we face a case in which the principles conflict each other, the top priority needs to be granted to one of the principles. How we select one of the principles for the top priority remains an unsolved issue. Considering the ethical status of the fetus, the most important thing is when the embryo or fetus becomes a person. Roman Catholic declared in 1974 that the embryo just after fertilization is considered as a person. However many bio-ethicists believe that only an autonomous person who is an individual capable of deliberation about personal goals, and of acting under the direction of such deliberation has the right of living. This opinion permits us not only to abort fetus artificially but also to kill infants. In the latter half of the article personal opinion on the ethical status of the fetus is explained.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"45 1","pages":"177-184"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79095504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The proliferation and differentiation of most cells are regulated by cytokine signaling, but the mechanism that regulates pre-implantation development remains unclear. Recently, it has been shown that Jak2, which mediates various cytokine signaling pathways, is expressed in pre-implantation mouse embryos. In this study, we investigated the expression of the cytokine receptors that activate Jak2, i.e., the receptors for prolactin (PrlR), growth hormone (GHR), tumor necrosis factor (TNFR), interleukin-3 (IL-3R), interleukin-5 (IL-5R), and granulocyte-macrophage colony stimulating factor (GM-CSFR). RT-PCR analysis revealed that PrlR was expressed in MII stage oocytes at a relatively high level, and that the level of expression decreased between the 2-cell and 4-cell stages. The expression levels of GHR, TNFR, IL-3R, IL-5R and GM-CSFR were relatively low before the morula stage, but they increased thereafter until the hatched blastocyst stage. These results suggest that various cytokine signaling pathways mediated by Jak2 activation are involved in the regulation of pre-implantation development.
{"title":"Expression of Cytokine Receptors in Pre-implantation Mouse Embryos","authors":"M. Nakasato, M. Nagata, F. Aoki","doi":"10.1274/JMOR.21.128","DOIUrl":"https://doi.org/10.1274/JMOR.21.128","url":null,"abstract":"The proliferation and differentiation of most cells are regulated by cytokine signaling, but the mechanism that regulates pre-implantation development remains unclear. Recently, it has been shown that Jak2, which mediates various cytokine signaling pathways, is expressed in pre-implantation mouse embryos. In this study, we investigated the expression of the cytokine receptors that activate Jak2, i.e., the receptors for prolactin (PrlR), growth hormone (GHR), tumor necrosis factor (TNFR), interleukin-3 (IL-3R), interleukin-5 (IL-5R), and granulocyte-macrophage colony stimulating factor (GM-CSFR). RT-PCR analysis revealed that PrlR was expressed in MII stage oocytes at a relatively high level, and that the level of expression decreased between the 2-cell and 4-cell stages. The expression levels of GHR, TNFR, IL-3R, IL-5R and GM-CSFR were relatively low before the morula stage, but they increased thereafter until the hatched blastocyst stage. These results suggest that various cytokine signaling pathways mediated by Jak2 activation are involved in the regulation of pre-implantation development.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"192 1","pages":"128-133"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78746109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Hashiba, Hiroyoshi Watanabe, T. Maeda, Hiroto Tajima, N. Kuji, K. Sueoka, Y. Yoshimura
Duchenne muscular dystrophy (DMD), caused by mutations of the dystrophin gene, is a severe X-linked recessive neuromuscular disorder. Preimplantation diagnosis of DMD includes three approaches. The first approach is gender determination of embryos by either polymerase chain reaction (PCR) or the fluorescence in situ hybridization (FISH)-based method. While each method is well established, the FISH method has some advantages over PCR in gender determination. The second approach is diagnosis of specific gene mutation. The partial deletions are diagnosed by the PCR with primers constructed to amplify the deletion exons. The partial duplication cannot be detected by now available strategies. The small mutations can be diagnosed by the specific PCR based assay. The third approach is linkage analysis by means of linked markers. CA repeats have been shown to be highly polymorphic and to be of great diagnostic utility because they can be easily assayed by PCR.
{"title":"Preimplantation Diagnosis of Duchenne Muscular Dystrophy","authors":"T. Hashiba, Hiroyoshi Watanabe, T. Maeda, Hiroto Tajima, N. Kuji, K. Sueoka, Y. Yoshimura","doi":"10.1274/JMOR.21.7","DOIUrl":"https://doi.org/10.1274/JMOR.21.7","url":null,"abstract":"Duchenne muscular dystrophy (DMD), caused by mutations of the dystrophin gene, is a severe X-linked recessive neuromuscular disorder. Preimplantation diagnosis of DMD includes three approaches. The first approach is gender determination of embryos by either polymerase chain reaction (PCR) or the fluorescence in situ hybridization (FISH)-based method. While each method is well established, the FISH method has some advantages over PCR in gender determination. The second approach is diagnosis of specific gene mutation. The partial deletions are diagnosed by the PCR with primers constructed to amplify the deletion exons. The partial duplication cannot be detected by now available strategies. The small mutations can be diagnosed by the specific PCR based assay. The third approach is linkage analysis by means of linked markers. CA repeats have been shown to be highly polymorphic and to be of great diagnostic utility because they can be easily assayed by PCR.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"45 1","pages":"7-12"},"PeriodicalIF":0.0,"publicationDate":"2004-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88168273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In this study, to investigate the time dependent changes in progesterone receptor (PR) expression in cumulus cells during meiotic resumption of porcine oocytes, each amount of PR-A and PR-B mRNA was analyzed by RT-PCR with primer sets for the PR-B region and the PR-A/B common region. The results showed that the levels of both PR-A/B and PR-B mRNA were very low in cumulus cells immediately recovered from their follicles. The cultivation with FSH and LH significantly increased the level of both PR-A/B and PR- B mRNA in cumulus cell of COCs, whereas the level of PR-B mRNA significantly decreased at 12-hr cultivation. Nevertheless, the higher level of PR-A/B mRNA was maintained up to 20-hr cultivation, suggesting that PR-A was mainly expressed in cumulus cells during cultivation from 12 hr to 20 hr. When COCs were cultured for 10 hr and then further cultured with RU486 for 10 hr, the proportion of oocytes undergoing GVBD significantly decreased in a dose dependent fashion. These results suggest that the high ratios of PR-A to PR-B in cumulus cells of COCs during 12-hr to 20-hr cultivation, are required for meiotic resumption of porcine cumulus-
为了研究猪卵母细胞减数分裂恢复过程中卵丘细胞中孕酮受体(PR)表达的时间依赖性变化，采用RT-PCR方法分析了PR-B区和PR- a /B共同区各引物的PR- a和PR-B mRNA表达量。结果表明，在刚从卵泡中恢复的积云细胞中，PR-A/B和PR-B mRNA的水平都很低。FSH和LH培养可显著提高COCs积云细胞PR- a /B和PR-B mRNA水平，培养12小时后PR-B mRNA水平显著降低。然而，PR-A/B mRNA在培养20小时后仍保持较高水平，表明PR-A主要在培养12 - 20小时的积云细胞中表达。COCs培养10小时后，再与RU486一起培养10小时，发生GVBD的卵母细胞比例呈剂量依赖性显著降低。这些结果表明，在培养12 ~ 20小时的COCs积云细胞中PR-A与PR-B的高比率是猪积云-恢复减数分裂所必需的
{"title":"Time Dependent Changes in Progesterone Receptor Expression in Cumulus Cells During Meiotic Resumption of Porcine Oocytes","authors":"M. Shimada, M. Nishibori","doi":"10.1274/JMOR.20.113","DOIUrl":"https://doi.org/10.1274/JMOR.20.113","url":null,"abstract":"In this study, to investigate the time dependent changes in progesterone receptor (PR) expression in cumulus cells during meiotic resumption of porcine oocytes, each amount of PR-A and PR-B mRNA was analyzed by RT-PCR with primer sets for the PR-B region and the PR-A/B common region. The results showed that the levels of both PR-A/B and PR-B mRNA were very low in cumulus cells immediately recovered from their follicles. The cultivation with FSH and LH significantly increased the level of both PR-A/B and PR- B mRNA in cumulus cell of COCs, whereas the level of PR-B mRNA significantly decreased at 12-hr cultivation. Nevertheless, the higher level of PR-A/B mRNA was maintained up to 20-hr cultivation, suggesting that PR-A was mainly expressed in cumulus cells during cultivation from 12 hr to 20 hr. When COCs were cultured for 10 hr and then further cultured with RU486 for 10 hr, the proportion of oocytes undergoing GVBD significantly decreased in a dose dependent fashion. These results suggest that the high ratios of PR-A to PR-B in cumulus cells of COCs during 12-hr to 20-hr cultivation, are required for meiotic resumption of porcine cumulus-","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"30 1","pages":"113-117"},"PeriodicalIF":0.0,"publicationDate":"2003-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81371633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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