Egg Development After In Vitro Insemination in Japanese Quail (Coturnix japonica).

IF 1.8 4区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Journal of Poultry Science Pub Date : 2023-01-25 DOI:10.2141/jpsa.2023001
Yoshinobu Ichikawa, Shusei Mizushima, Noritaka Hirohashi, Tomohiro Sasanami
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Abstract

In vitro fertilization has been widely used to produce offspring in several mammalian species. We previously successfully produced Japanese quail chicks using intracytoplasmic sperm injection (ICSI), whereas in vitro insemination was not successful. This may be due to the difficulties associated with mimicking the sperm-egg fusion process and subsequent events in physiological polyspermic fertilization in vitro. In the present study, we observed egg development after in vitro insemination and investigated the inactivation of metaphase-promoting factor (MPF) and cytostatic factor (CSF), which are downstream of the Ca2+ signaling pathway in the egg, due to fertilizing sperm. We found a sperm number-dependent increase in hole formation caused by sperm penetration of the perivitelline membrane, the extracellular coat surrounding the egg. Egg development was observed following in vitro insemination; however, the developmental rate and stages after 24-h culture were inferior to those of ICSI eggs, even when insemination was performed with a high number of sperm (2 × 104). We also noted the downregulation of inositol 1,4,5-trisphosphate receptor-1, ryanodine receptor-3, cyclin B1, and c-MOS, which are important regulatory components of MPF and CSF in the egg, which was dependent on the number of sperm used for insemination. However, the decreases observed in these components did not reach the levels observed in the ICSI eggs. Collectively, the present results suggest that a sperm number higher than 2 × 104 is required for the progression of the Ca2+ signaling pathway, which initiates subsequent egg development in Japanese quail.

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日本鹌鹑体外受精后卵子发育的研究。
体外受精已被广泛应用于多种哺乳动物的子代生产。我们以前成功地用卵胞浆内单精子注射(ICSI)生产了日本鹌鹑雏鸡,而体外人工授精没有成功。这可能是由于在体外模拟精卵融合过程和生理多精受精的后续事件相关的困难。在本研究中,我们观察了体外受精后卵子的发育情况,并研究了卵子中Ca2+信号通路下游的中期促进因子(MPF)和细胞抑制因子(CSF)因受精精子而失活的情况。我们发现精子数量的增加是由于精子穿透卵泡外膜(卵子周围的细胞外膜)造成的。体外授精后观察卵子发育;然而,培养24小时后的发育速度和阶段低于ICSI卵子,即使在大量精子的情况下(2 × 104)。我们还注意到肌醇1,4,5-三磷酸受体-1,ryanodine受体-3,细胞周期蛋白B1和c-MOS的下调,它们是卵子中MPF和CSF的重要调节成分,这取决于用于受精的精子数量。然而,在这些成分中观察到的减少并没有达到在ICSI卵子中观察到的水平。总的来说,目前的研究结果表明,精子数量高于2 × 104是Ca2+信号通路的进展所必需的,Ca2+信号通路启动了日本鹌鹑随后的卵子发育。
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来源期刊
Journal of Poultry Science
Journal of Poultry Science AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
2.80
自引率
13.30%
发文量
26
审稿时长
12 months
期刊介绍: The Journal of Poultry Science will publish original reports and reviews which either make an original contribution to fundamental science or are of obvious application to the industry. Subjects which are covered include: breeding and genetics, nutrition and feeds, physiology, reproduction, immunology, behavior, environmental science, management and housing welfare, processing and products, and health in poultry. Submission of original articles to the Journal is open to all poultry researchers. The review articles are invited papers written by international outstanding researchers. Articles will be published in English, American style.
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