Umezu Kohei, Yajima Risa, Hiradate Yuuki, Yanai Rin, Numabe Takashi, Hara Kenshiro, Oikawa Toshinori, Tanemura Kentaro
Laboratory of Animal Reproduction and Development, Graduate School of Agricultural Science, Tohoku University, Miyagi 980-0845, Japan.
Miyagi Prefectural Livestock Experiment Station, Miyagi 989-6445, Japan.
J Reprod Dev. 2019 Apr 12;65(2):147-153. doi: 10.1262/jrd.2018-147. Epub 2019 Jan 19.
Previously, we reported that neurotensin (NT), which is expressed in the uterus and oviduct, enhanced bovine sperm capacitation and acrosome reactions. As NT mRNA expression in bovine oviducts increases dramatically in the follicular phase, we hypothesized that NT modulates fertilization and subsequent conception in cattle. The objective of this study was to evaluate the effect of NT on embryo development and blastocyst quality. The rate of embryo cleavage was significantly increased by the addition of NT to the fertilization medium. Furthermore, the total number of cells and numbers of cells in the inner cell mass of blastocysts were significantly increased by NT during in vitro fertilization (IVF). These results suggested that NT enhanced the efficiency of early bovine embryo development and blastocyst quality. The expression of NT receptors (NTRs) in sperm, testes, oocytes, and cumulus cells was evaluated to determine whether NT acted via NTRs in sperm alone or in both male and female reproductive cells during IVF. Immunocytochemistry and reverse transcription polymerase chain reaction revealed that NTR1 and NTR2 were expressed in sperm and testes, but not in oocytes and cumulus cells. We propose that NT selectively acts upon sperm via NTR1 and NTR2 during IVF to improve the cleavage rate and quality of blastocysts, which are important determinants of sperm quality for successful conception. This research supports our hypothesis that NT acts as a key modulator of fertilization and conception in cattle. Further studies are necessary to apply our findings to the industrial framework of bovine reproduction.
此前,我们报道过在子宫和输卵管中表达的神经降压素(NT)可增强牛精子的获能和顶体反应。由于牛输卵管中NT mRNA的表达在卵泡期会显著增加,我们推测NT可调节牛的受精及后续受孕过程。本研究的目的是评估NT对胚胎发育和囊胚质量的影响。向受精培养基中添加NT可显著提高胚胎的分裂率。此外,在体外受精(IVF)过程中,NT可显著增加囊胚的总细胞数和内细胞团的细胞数。这些结果表明,NT可提高牛早期胚胎发育的效率和囊胚质量。对精子、睾丸、卵母细胞和卵丘细胞中NT受体(NTRs)的表达进行评估,以确定NT在IVF过程中是仅通过精子中的NTRs起作用,还是在雄性和雌性生殖细胞中都起作用。免疫细胞化学和逆转录聚合酶链反应显示,NTR1和NTR2在精子和睾丸中表达,但在卵母细胞和卵丘细胞中不表达。我们提出,在IVF过程中,NT通过NTR1和NTR2选择性地作用于精子,以提高囊胚的分裂率和质量,而囊胚的分裂率和质量是成功受孕的精子质量的重要决定因素。本研究支持了我们的假设,即NT是牛受精和受孕的关键调节因子。有必要进一步开展研究,将我们的研究结果应用于牛繁殖的产业框架中。