{"title":"Progesterone increases the success of in vitro fertilization via enhanced sperm hyperactivation in mice.","authors":"Risa Suzuki, Masakatsu Fujinoki","doi":"10.1262/jrd.2022-114","DOIUrl":"https://doi.org/10.1262/jrd.2022-114","url":null,"abstract":"<p><p>Progesterone (P) enhances spermatozoal hyperactivation, a capacitation event. Hyperactivation is associated with successful in vitro fertilization (IVF). In this study, we examined the effects of P on hyperactivation and IVF in mice. P enhanced spermatozoal hyperactivation and increased IVF success rate in a dose-dependent manner. Moreover, P affected spermatozoal hyperactivation and IVF through the membrane progesterone receptor of the spermatozoal head. These results show that P regulates spermatozoal capacitation and fertilization in mice. The concentration of P changes during the estrous cycle, indicating that spermatozoa are capacitated in response to the oviductal environment and subsequently fertilize the oocyte.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/87/20/jrd-69-147.PMC10267584.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9991849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Lysophosphatidic acid stimulates rat uterine contraction in vitro.","authors":"Satoshi Nagashima, Takuma Kimura, Ryota Terashima, Makoto Sugiyama, Keiichiro Kizaki, Mitsumori Kawaminami, Shiro Kurusu","doi":"10.1262/jrd.2023-011","DOIUrl":"https://doi.org/10.1262/jrd.2023-011","url":null,"abstract":"<p><p>Lysophosphatidic acid (LPA) has been implicated in the uterine endometrial functions of implantation and decidualization; however, not much is known about its myometrial contractile function. Herein we characterized the uterotonic effects of LPA in non-pregnant (estrus) and peri-parturient rats in vitro. LPA dose-dependently (0.01-10 μM) stimulated the amplitude and integral, but not the frequency, of the uterine strip contraction of estrous rats. The stimulatory effect of LPA was enhanced 1 day before parturition but was lost 1 day postpartum. LPA did not cause the de novo synthesis of prostaglandin (PG) F<sub>2</sub>α but stimulated contractions cooperatively with the PG. LPA-induced contractions were significantly inhibited by an LPA1/2/3 antagonist in the uteri of estrous rats but not in term rats. This study characterized the uterotonic effect of a natural LPA that occurs at physiological concentrations, changes with reproductive states, and is independent of mediation by the newly synthesized PG.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/2e/85/jrd-69-163.PMC10267583.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9992339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Comparative characteristics between calyculin A-induced and thimerosal-induced hyperactivation of cryopreserved bovine spermatozoa.","authors":"Natsuko Miyamoto, Akihiro Ohya, Mitsuhiro Sakase, Hiroshi Harayama","doi":"10.1262/jrd.2023-007","DOIUrl":"https://doi.org/10.1262/jrd.2023-007","url":null,"abstract":"<p><p>This study aimed to characterize calyculin A (CL-A)-induced and thimerosal-induced hyperactivation of cryopreserved bovine spermatozoa. Hyperactivation was effectively induced by treating with 10 nM CL-A for 60 min in the presence of cyclic AMP analogs, extracellular Ca<sup>2+</sup>, and albumin or with 12.5 µM thimerosal briefly in the absence of these capacitation-supporting factors. Majority of the spermatozoa exhibiting CL-A-induced hyperactivation were characterized by the 3-dimensional helical movement with head rotation, higher degree of flagellar curvature, and faster beating of the flagella than those exhibiting thimerosal-induced hyperactivation of the 2-dimensional planar movement without head rotation. The CL-A-induced hyperactivation was linked to the activation of cAMP/protein phosphorylation-dependent signaling cascades and to the decreased activity of glycogen synthase kinase-3α (GSK-3α). In contrast, the thimerosal-induced hyperactivation was suppressed by pretreatment with CL-A and cyclic AMP analogs in the absence of CaCl<sub>2</sub> to activate cAMP/protein phosphorylation-dependent signaling cascades. Additionally, the intracellular Ca<sup>2+</sup> level in live sperm flagella was significantly higher in the CL-A-treated samples than in the thimerosal-treated samples. These results indicate that CL-A-induced hyperactivation of cryopreserved bovine spermatozoa is an extracellular Ca<sup>2+</sup>-dependent type with the 3-dimensional helical movement, which can be regulated not only by the activation of cAMP/protein phosphorylation-dependent signaling cascades, leading to a large enhancement of the intracellular Ca<sup>2+</sup> level, but also by the reduction in GSK-3α activity. Considering the different characteristics of thimerosal-induced hyperactivation, our results suggest that the diversity of sperm hyperactivation arises from different combinations of flagellar bending and head rotation.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/b1/94/jrd-69-170.PMC10267582.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9627691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"First Kiso pony foal produced via transfer of long-distance shipped fresh embryo to Hokkaido native pony.","authors":"Reza Rajabi-Toustani, Munkhtuul Tsogtgerel, Yuanzhi Gao, Canbo Li, Miou Sakato, Shingo Haneda, Soon Hon Cheong, Yasuo Nambo","doi":"10.1262/jrd.2022-125","DOIUrl":"https://doi.org/10.1262/jrd.2022-125","url":null,"abstract":"<p><p>Japanese native horses, which consists of 8 breeds, are threatened with extinction. Embryo transfer (ET) is used to reproduce endangered animals in various mammalian species. We aimed to perform ET using native ponies from Kiso and Hokkaido as donors and recipients, respectively. ET operation included long-distance transport of non-cryopreserved embryos from Nagano Prefecture to Hokkaido. Embryos were transported 1500 km over 9 h in a container maintained at 22°C. After transferring two embryos to two recipients, one mare delivered a healthy live foal. These results demonstrated that reciprocal ET with long-distance transportation of fresh embryos between the isolated breeds may allow for the proliferation of Japanese native horses.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/64/b6/jrd-69-125.PMC10085766.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9293286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Effects of short-term dietary supplementation on the number of ovarian follicles, quantity and quality of oocytes, and in vitro embryo production in Japanese Black cows.","authors":"Kensuke Tomita, Toshimichi Ishii, Natsumi Endo, Tomomi Tanaka","doi":"10.1262/jrd.2022-103","DOIUrl":"https://doi.org/10.1262/jrd.2022-103","url":null,"abstract":"<p><p>This study aimed to test the hypothesis that short-term supplementation with a high-energy diet promotes embryo production following ovum pick-up (OPU) in Japanese Black cows. After a period of adaptation to the maintenance diet, a 200% maintenance diet was fed to the high-energy diet group (HD group, n = 6) for four weeks, and a maintenance diet was fed to the other group (MD group, n = 6). OPU-in vitro fertilization (IVF) procedures were performed on days 14, 21, and 28; follicles and oocytes were counted and morphologically graded, and cultivable oocytes were cultured for in vitro maturation, fertilization, and culture. The mean plasma insulin concentrations on days 14 and 21 were significantly higher in the HD group than in the MD group (P < 0.05). The number of follicles observed at OPU, recovered oocytes, cultivable (Grades 1 to 4) oocytes, and the rate of degenerated (Grade 6) oocytes in the HD group were significantly higher than those in the MD group (P < 0.05). The proportion of cleaved oocytes was lower in the HD group than in the MD group (P < 0.05); consequently, there was no significant difference in the number of blastocysts obtained between the HD and MD groups. The present findings suggest that high-energy diets can promote follicular growth in parallel with an increase in plasma concentrations of insulin, but have a detrimental effect on the quality of oocytes with the OPU-IVF procedure in Japanese Black cows.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c2/62/jrd-69-065.PMC10085769.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9286488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Eri Furukawa, Yojiro Yanagawa, Akira Matsuzaki, Heejin Kim, Hanako Bai, Masashi Takahashi, Seiji Katagiri, Shogo Higaki
{"title":"Analysis of sequential ruminal temperature sensor data from dairy cows to identify cow subgroups by clustering and predict calving through supervised machine learning.","authors":"Eri Furukawa, Yojiro Yanagawa, Akira Matsuzaki, Heejin Kim, Hanako Bai, Masashi Takahashi, Seiji Katagiri, Shogo Higaki","doi":"10.1262/jrd.2022-107","DOIUrl":"https://doi.org/10.1262/jrd.2022-107","url":null,"abstract":"<p><p>The present study investigated the applicability of a calving prediction model based on supervised machine learning of ruminal temperature (RT) data in dairy cows. The existence of cow subgroups for prepartum RT changes was also examined, and the predictive performance of the model was compared among these subgroups. RT data were collected from 24 Holstein cows at 10 min intervals using an RT sensor system. The average hourly RT was calculated and data were expressed as residual RTs (rRT = actual RT - mean RT for the same time on the previous three days). The mean rRT decreased beginning at approximately 48 h before calving to a low of -0.5°C at 5 h before calving. However, two cow subgroups were identified: cows with a late and small rRT decrease (Cluster 1, n = 9) and those with an early and large rRT decrease (Cluster 2, n = 15). A calving prediction model was developed using five features extracted from the sensor data (indicative of prepartum rRT changes) through a support vector machine. Cross-validation showed that calving within 24 h was predicted with a sensitivity of 87.5% (21/24) and precision of 77.8% (21/27). A significant difference in sensitivity was observed between Clusters 1 and 2 (66.7 vs. 100%, respectively), while none was observed for precision. Therefore, the model based on RT data with supervised machine learning has the potential to efficiently predict calving, although improvements for specific cow subgroups are required.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/0a/c9/jrd-69-103.PMC10085770.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9657002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Development of cryopreservation media for the slow-freezing of cultured primordial germ cells in chicken.","authors":"Natsuko Hamai, Chihiro Koide, Yuki Tansho, Yukino Ooka, Mayo Hirano, Effrosyni Fatira, Masaoki Tsudzuki, Yoshiaki Nakamura","doi":"10.1262/jrd.2022-123","DOIUrl":"https://doi.org/10.1262/jrd.2022-123","url":null,"abstract":"<p><p>Conservation of chicken germplasm is crucial in supporting commercial breeds for sustainable egg and meat production and preserving the genetic diversity of indigenous breeds for future breeding. Cryopreservation of chicken fertilized eggs or embryos is not feasible, owing to the large yolk-laden structure of the eggs. Primordial germ cells (PGCs), the embryonic precursors of gametes, are the best candidates for the cryobanking of chicken germplasm. Effective cryobanking of chicken PGCs requires an optimal cryopreservation protocol. Cryomedia containing dimethyl sulfoxide (DMSO) or DMSO combined with serum have been widely used for the cryopreservation of chicken PGCs. However, as cryoprotectants are yet to be optimized for chicken PGCs, the efficacy of cryomedia can be further improved. Here, we investigated the cryoprotective effects of propylene glycol (PG), an alternative to DMSO, on chicken PGCs. We found that the addition of non-permeable cryoprotectants, such as trehalose or chicken serum, to DMSO or PG cryomedia improved the recovery and survival rates of post-thawed PGCs. We further investigated the cryoprotective effects of trehalose and chicken serum and found that these additives have different cryoprotective actions. Based on these findings, we designed two different cryomedia: DTS, including 5% DMSO, 0.3 M trehalose, and 1% chicken serum, and PTS, including 7.5% PG, 0.1 M trehalose, and 5% chicken serum. Among the different PGC lines and freshly isolated PGCs, the cryomedia showed similar post-thaw recovery rates. Following transplantation, post-thawed male PGCs can colonize gonads and differentiate into functional sperm. We successfully revived the offspring of Kurokashiwa, a rare chicken breed in Japan, with cryopreserved PGCs. In conclusion, we developed two different cryomedia that achieved > 50% recovery of viable PGCs after thawing while maintaining germline competency.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/65/e4/jrd-69-109.PMC10085767.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9281868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A more accurate analysis of maternal effect genes by siRNA electroporation into mouse oocytes.","authors":"Takuto Yamamoto, Shinnosuke Honda, Issei Ideguchi, Motoki Suematsu, Shuntaro Ikeda, Naojiro Minami","doi":"10.1262/jrd.2022-122","DOIUrl":"https://doi.org/10.1262/jrd.2022-122","url":null,"abstract":"<p><p>Maternal RNA and proteins accumulate in mouse oocytes and regulate initial developmental stages. Sperm DNA combines with protamine, which is exchanged after fertilization with maternal histones, including H3.3; however, the effect of H3.3 on development post-fertilization remains unclear. Herein, we established an electroporation method to introduce H3.3 siRNA into germinal vesicle (GV)-stage oocytes without removing cumulus cells. Oocyte-attached cumulus cells need to be removed during the traditional microinjection method; however, we confirmed that artificially removing cumulus cells from oocytes reduced fertilization rates, and oocytes originally free of cumulus cells had reduced developmental competence. On introducing H3.3 siRNA at the GV stage, H3.3 was maintained in the maternal pronucleus and second polar body but not in the paternal pronucleus, resulting in embryonic lethality after fertilization. These findings indicate that H3.3 protein was not incorporated into the paternal pronucleus, as it was repeatedly translated and degraded over a relatively short period. Conversely, H3.3 protein incorporated into the maternal genome in the GV stage escaped degradation and remained in the maternal pronucleus after fertilization. This new method of electroporation into GV-stage oocytes without cumulus cell removal is not skill-intensive and is essential for the accurate analysis of maternal effect genes.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/b6/6c/jrd-69-118.PMC10085774.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9281872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Effects of the temperature-humidity index on conception rates in Holstein heifers and cows receiving in vitro-produced Japanese Black cattle embryos.","authors":"Takamasa Nishisozu, Jaswant Singh, Akinori Abe, Kazuo Okamura, Osamu Dochi","doi":"10.1262/jrd.2022-112","DOIUrl":"https://doi.org/10.1262/jrd.2022-112","url":null,"abstract":"<p><p>We investigated the effect of the temperature-humidity index (THI) on the conception rate (CR) in Holstein heifers and cows receiving in vitro-produced (IVP) Japanese Black cattle fresh embryos. IVP embryos were transferred to Holstein heifers (n = 1,407) and cows (n = 3,189) on 245 commercial farms. The monthly average ambient temperature (AT) and THI ranged from 4.7 to 29°C and 41 to 81, respectively; both were the highest in August. The monthly CR ranged from 16.3% to 46.7% in cows and 23.8% to 74.1% in heifers. The CR of heifers was unaffected by THI, AT, or the month of embryo transfer. However, these parameters affected the CR of cows. The CR at THI values of 61-65 and 71-75 was greater than that at THI > 75, whereas other THI values had no effect. The CR at temperatures > 25°C was lower (P = 0.008) than that at temperatures of 15-20°C and 20-25°C. Moreover, the CR was lowest (P = 0.003) in July. THI and parity (P = 0.057 and P = 0.001, respectively) and AT and parity (P = 0.019 and P = 0.001, respectively) showed significant effects on CR; however, there was no interaction between these two factors. In conclusion, AT > 25°C and THI > 75 adversely affect the CR outcome in cows but not in heifers.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/42/4c/jrd-69-072.PMC10085771.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9293287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yuto Taniuchi, Kazutaka Hiraide, Rilige Su, Kazune Ijuin, XingQiang Wei, Takuro Horii, Izuho Hatada, Jibak Lee
{"title":"Analysis of absolute amounts of two meiotic cohesin subunits, RAD21L and REC8, in mouse spermatocytes.","authors":"Yuto Taniuchi, Kazutaka Hiraide, Rilige Su, Kazune Ijuin, XingQiang Wei, Takuro Horii, Izuho Hatada, Jibak Lee","doi":"10.1262/jrd.2022-075","DOIUrl":"10.1262/jrd.2022-075","url":null,"abstract":"<p><p>RAD2lL and REC8, meiosis-specific paralogs of the canonical cohesin subunit RAD21, are essential for proper formation of axial/lateral elements of the synaptonemal complex, synapsis of homologous chromosomes, and crossover recombination in mammalian meiosis. However, how many meiotic cohesins are present in germ cells has not been investigated because of the lack of an appropriate method of analysis. In the present study, to examine the intracellular amount of meiotic cohesins, we generated two strains of knock-in (KI) mice that expressed a 3×FLAG-tag at the C-terminus of RAD21L or REC8 protein using the CRISPR/Cas9 genome editing system. Both KI mice were fertile. Western blot analyses and immunocytochemical studies revealed that expression levels and localization patterns of both RAD21L-3×FLAG and REC8-3×FLAG in KI mice were similar to those in wild-type mice. After confirming that tagging of endogenous RAD21L and REC8 with 3×FLAG did not affect their expression profiles, we evaluated the levels of RAD21L-3×FLAG and REC8-3×FLAG in the testes of 2-week-old mice in which only RAD21L and REC8 but little RAD21 are expressed in the meiocytes. By comparing the band intensities of testicular RAD21L-3×FLAG and REC8-3×FLAG with 3×FLAG-tagged recombinant proteins of known concentrations in western blot analysis, we found that there were approximately 413,000 RAD21L and 453,000 REC8 molecules per spermatocyte in the early stages of prophase I. These findings provide new insights into the role played by cohesins in the process of meiotic chromosome organization in mammalian germ cells.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e0/38/jrd-69-078.PMC10085773.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9293294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}