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斑马鱼(Danio rerio)卵母细胞生长和卵黄蛋白原基因表达的时间动态变化

Temporal dynamics of oocyte growth and vitellogenin gene expression in zebrafish (Danio rerio).

作者信息

Connolly Michelle H, Dutkosky Rachel M, Heah Tze P, Sayler Gary S, Henry Theodore B

机构信息

1 Center for Environmental Biotechnology, University of Tennessee , Knoxville, Tennessee.

出版信息

Zebrafish. 2014 Apr;11(2):107-14. doi: 10.1089/zeb.2013.0938. Epub 2014 Mar 10.

Abstract

Little is known about how hepatic vitellogenin gene (vtg) expression relates to oogenesis in fish, especially among fractional spawners. The objective of this study was to relate hepatic vtg 1A/B expression to stage-specific oocyte development in zebrafish (Danio rerio), an asynchronous spawning fish. Liver samples were collected at seven time points postspawning (1-32 days) and fish were preserved for subsequent histological analyses. Relative vtg 1A/B expression among liver samples was quantified by reverse transcription-quantitative PCR and oogenesis was evaluated following standard hematoxylin and eosin staining of serial ovarian sections. Histological analyses indicate that a subset of previtellogenic oocytes (stages 1-2) transitioned into postvitellogenic oocytes (stages 3-4) within 4 days (96 h) postspawning. By 8 days postspawning (192 h), the majority of the ovary was occupied by mature (stage 4) oocytes, a trend that continued through 32 days postspawning. Hepatic vtg 1A/B gene expression was upregulated 3.89-fold 1-h postspawning relative to the average gene expression across all time points, but was not correlated to stage-specific oogenesis. Follicular atresia among fish sampled 32 days postspawning highlights the importance of regular spawning in zebrafish and suggests that the event of spawning itself may be integral to the regulation of oocyte development.

摘要

关于肝脏卵黄蛋白原基因(vtg)表达与鱼类卵子发生的关系,人们了解甚少,尤其是在分批产卵的鱼类中。本研究的目的是将斑马鱼(Danio rerio)——一种异步产卵鱼类的肝脏vtg 1A/B表达与特定阶段的卵母细胞发育联系起来。在产卵后7个时间点(1 - 32天)采集肝脏样本,并保存鱼体用于后续的组织学分析。通过逆转录定量PCR对肝脏样本中的相对vtg 1A/B表达进行定量,并对卵巢连续切片进行标准苏木精和伊红染色后评估卵子发生情况。组织学分析表明,一部分卵黄生成前的卵母细胞(1 - 2期)在产卵后4天(96小时)内转变为卵黄生成后的卵母细胞(3 - 4期)。到产卵后8天(192小时),卵巢的大部分被成熟(4期)卵母细胞占据,这一趋势持续到产卵后32天。肝脏vtg 1A/B基因表达在产卵后1小时相对于所有时间点的平均基因表达上调了3.89倍,但与特定阶段的卵子发生无关。产卵后32天采集的鱼中卵泡闭锁现象突出了斑马鱼定期产卵的重要性,并表明产卵事件本身可能是卵母细胞发育调控的一个组成部分。

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