The birth of quail chicks after intracytoplasmic sperm injection
The birth of quail chicks after intracytoplasmic sperm injection
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DOI：
10.1242/dev.111765
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发表时间：
2014-10-01
期刊：
影响因子：
4.6
通讯作者：
Sasanami, Tomohiro
中科院分区：
文献类型：
--
作者：
Mizushima, Shusei;Hiyama, Gen;Sasanami, Tomohiro
卵胞浆内单精子注射（ICSI）已成功用于包括人类在内的几种哺乳动物的子代生产。然而，ICSI在鸟类中尚未成功，因为卵子的大小和难以模仿正常受精过程中发生的生理多精现象。显微外科手术将20个或更多的精子注射到卵子中对卵子的存活是有害的。在这里，我们报告了注射单个精子与少量精子提取物（SE）或其成分导致健康鹌鹑雏鸟的发育和出生。SE含有磷脂酶C - zeta （PLCZ）、乌头酸水合酶（AH）和柠檬酸合成酶（CS）三种因子，它们都是卵子完全激活和胚胎发育所必需的。PLCZ诱导恢复减数分裂所需的立即、短暂的Ca2+升高。AH和CS是激活卵子内持久的螺旋状Ca2+振荡所必需的，这是早期胚胎细胞周期进程所必需的。我们还发现，在不使用SE的情况下，共注射编码PLCZ、AH和CS的crna支持icsi生成的受精卵的完全发育。这些发现将有助于我们了解鸟类受精和胚胎发育的机制，并有助于鸟类基因组的操纵和转基因和克隆鸟类的生产。
Intracytoplasmic sperm injection (ICSI) has been successfully used to produce offspring in several mammalian species including humans. However, ICSI has not been successful in birds because of the size of the egg and difficulty in mimicking the physiological polyspermy that takes place during normal fertilization. Microsurgical injection of 20 or more spermatozoa into an egg is detrimental to its survival. Here, we report that injection of a single spermatozoon with a small volume of sperm extract (SE) or its components led to the development and birth of healthy quail chicks. SE contains three factors - phospholipase C zeta (PLCZ), aconitate hydratase (AH) and citrate synthase (CS) - all of which are essential for full egg activation and subsequent embryonic development. PLCZ induces an immediate, transient Ca2+ rise required for the resumption of meiosis. AH and CS are required for long-lasting, spiral-like Ca2+ oscillations within the activated egg, which are essential for cell cycle progression in early embryos. We also found that co-injection of cRNAs encoding PLCZ, AH and CS support the full development of ICSI-generated zygotes without the use of SE. These findings will aid our understanding of the mechanism of avian fertilization and embryo development, as well as assisting in the manipulation of the avian genome and the production of transgenic and cloned birds.