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早期牛胚胎中透明质酸结合蛋白(RHAMM/IHABP)表达的发育调控

Developmental regulation of hyaluronan-binding protein (RHAMM/IHABP) expression in early bovine embryos.

作者信息

Stojkovic Miodrag, Krebs Ottheinz, Kölle Sabine, Prelle Katja, Assmann Volker, Zakhartchenko Valeri, Sinowatz Fred, Wolf Eckhard

机构信息

Department of Molecular Animal Breeding and Biotechnology, LMU, D-85764 Oberschleissheim, Germany.

出版信息

Biol Reprod. 2003 Jan;68(1):60-6. doi: 10.1095/biolreprod.102.007716.

Abstract

Hyaluronan or hyaluronic acid (HA) is a normal component of mammalian follicular, oviduct, and uterine fluids. Granulosa and expanding cumulus cells secrete large amounts of HA, and when HA is added in maturation and culture media, it improves the developmental potential of oocytes and embryos. HA regulates gene expression, signaling, proliferation, motility, adhesion, and morphogenesis. Many of these biological activities of HA are mediated through binding to the receptor for HA-mediated motility/intracellular HA-binding protein (RHAMM/IHABP). We evaluated the presence and dynamics of RHAMM/IHABP mRNA and protein expression in different stages of in vitro-produced bovine embryos using quantitative reverse transcriptase-real time-polymerase chain reaction and immunohistochemistry. We also analyzed the effects of different culture systems on the relative abundance of RHAMM/IHABP transcripts. RHAMM/IHABP mRNA levels decreased from the 2-cell to the 16-cell stage, increased again at the morula stage, and reached their highest level at the expanded blastocyst stage. RHAMM/IHABP mRNA abundance was significantly (P < 0.05) lower in embryos recovered in serum-containing medium than in embryos from serum-free media. Immunohistochemistry revealed the presence of RHAMM/IHABP first in 8-cell stages. Whereas RHAMM staining in 8-cell and morula stages was intense, it was weaker in blastocysts. Embryonic secretion of HA increased from the 2-cell stage until the 8-cell stage and then decreased in 16-cell embryos. After this, HA secretion increased in expanded and hatched blastocyst stages. These data suggest that the positive effects of HA on in vitro-produced bovine embryos may be mediated at least in part by RHAMM/IHABP.

摘要

透明质酸(HA)是哺乳动物卵泡液、输卵管液和子宫液的正常成分。颗粒细胞和扩展的卵丘细胞会分泌大量的HA,当在成熟和培养基中添加HA时,它能提高卵母细胞和胚胎的发育潜能。HA可调节基因表达、信号传导、增殖、运动、黏附和形态发生。HA的许多这些生物学活性是通过与HA介导的运动性受体/细胞内HA结合蛋白(RHAMM/IHABP)结合来介导的。我们使用定量逆转录实时聚合酶链反应和免疫组织化学方法,评估了体外生产的牛胚胎不同阶段中RHAMM/IHABP mRNA和蛋白表达的存在情况及动态变化。我们还分析了不同培养系统对RHAMM/IHABP转录本相对丰度的影响。RHAMM/IHABP mRNA水平从2细胞阶段到16细胞阶段下降,在桑椹胚阶段再次升高,并在扩张囊胚阶段达到最高水平。在含血清培养基中回收的胚胎中,RHAMM/IHABP mRNA丰度显著(P < 0.05)低于无血清培养基中的胚胎。免疫组织化学显示,RHAMM/IHABP最早在8细胞阶段出现。8细胞和桑椹胚阶段的RHAMM染色强烈,而在囊胚中则较弱。HA的胚胎分泌从2细胞阶段直到8细胞阶段增加,然后在16细胞胚胎中减少。在此之后,HA分泌在扩张和孵化的囊胚阶段增加。这些数据表明,HA对体外生产的牛胚胎的积极作用可能至少部分由RHAMM/IHABP介导。

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