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白鲈(Morone americana)卵黄形成机制及其对早期生活史的影响。

Mechanisms of Egg Yolk Formation and Implications on Early Life History of White Perch (Morone americana).

作者信息

Schilling Justin, Loziuk Philip L, Muddiman David C, Daniels Harry V, Reading Benjamin J

机构信息

Department of Applied Ecology, North Carolina State University, Raleigh, North Carolina, United States of America.

W. M. Keck FTMS Laboratory for Human Health Research, Department of Chemistry, North Carolina State University, Raleigh, North Carolina, United States of America.

出版信息

PLoS One. 2015 Nov 18;10(11):e0143225. doi: 10.1371/journal.pone.0143225. eCollection 2015.

Abstract

The three white perch (Morone americana) vitellogenins (VtgAa, VtgAb, VtgC) were quantified accurately and precisely in the liver, plasma, and ovary during pre-, early-, mid-, and post-vitellogenic oocyte growth using protein cleavage-isotope dilution mass spectrometry (PC-IDMS). Western blotting generally mirrored the PC-IDMS results. By PC-IDMS, VtgC was quantifiable in pre-vitellogenic ovary tissues and VtgAb was quantifiable in pre-vitellogenic liver tissues however, neither protein was detected by western blotting in these respective tissues at this time point. Immunohistochemistry indicated that VtgC was present within pre-vitellogenic oocytes and localized to lipid droplets within vitellogenic oocytes. Affinity purification coupled to tandem mass spectrometry using highly purified VtgC as a bait protein revealed a single specific interacting protein (Y-box binding protein 2a-like [Ybx2a-like]) that eluted with suramin buffer and confirmed that VtgC does not bind the ovary vitellogenin receptors (LR8 and Lrp13). Western blotting for LR8 and Lrp13 showed that both receptors were expressed during vitellogenesis with LR8 and Lrp13 expression highest in early- and mid-vitellogenesis, respectively. The VtgAa within the ovary peaked during post-vitellogenesis, while VtgAb peaked during early-vitellogenesis in both white perch and the closely related striped bass (M. saxatilis). The VtgC was steadily accumulated by oocytes beginning during pre-vitellogenesis and continued until post-vitellogenesis and its composition varies widely between striped bass and white perch. In striped bass, the VtgC accounted for 26% of the vitellogenin-derived egg yolk, however in the white perch it comprised only 4%. Striped bass larvae have an extended developmental window and these larvae have yolk stores that may enable them to survive in the absence of food for twice as long as white perch after hatch. Thus, the VtgC may play an integral role in providing nutrients to late stage fish larvae prior to the onset of exogenous feeding and its composition in the egg yolk may relate to different early life histories among this diverse group of animals.

摘要

利用蛋白质裂解-同位素稀释质谱法(PC-IDMS),在卵黄发生前、早期、中期和后期卵母细胞生长过程中,对三条白鲈(美洲鲈,Morone americana)的三种卵黄蛋白原(VtgAa、VtgAb、VtgC)在肝脏、血浆和卵巢中的含量进行了准确而精确的定量分析。蛋白质印迹法的结果总体上与PC-IDMS的结果一致。通过PC-IDMS分析,在卵黄发生前的卵巢组织中可检测到VtgC,在卵黄发生前的肝脏组织中可检测到VtgAb;然而,在这个时间点,通过蛋白质印迹法在这些相应组织中均未检测到这两种蛋白质。免疫组织化学分析表明,VtgC存在于卵黄发生前的卵母细胞中,并定位于卵黄发生期卵母细胞内的脂滴上。以高度纯化的VtgC作为诱饵蛋白,进行亲和纯化并结合串联质谱分析,发现了一种单一的特异性相互作用蛋白(Y盒结合蛋白2a样蛋白[Ybx2a样蛋白]),该蛋白在苏拉明缓冲液中洗脱,并证实VtgC不与卵巢卵黄蛋白原受体(LR8和Lrp13)结合。对LR8和Lrp13进行蛋白质印迹分析表明,这两种受体在卵黄发生过程中均有表达,其中LR8和Lrp13的表达分别在卵黄发生早期和中期达到最高。卵巢中的VtgAa在卵黄发生后期达到峰值,而VtgAb在白鲈和近缘种条纹鲈(M. saxatilis)的卵黄发生早期达到峰值。VtgC从卵黄发生前开始由卵母细胞稳步积累,一直持续到卵黄发生后期,其组成在条纹鲈和白鲈之间差异很大。在条纹鲈中,VtgC占卵黄蛋白原衍生卵黄的26%,而在白鲈中仅占4%。条纹鲈幼体有一个延长的发育窗口,这些幼体的卵黄储备可能使它们在孵化后在没有食物的情况下存活的时间是白鲈的两倍。因此,VtgC可能在为外源摄食开始前的晚期鱼类幼体提供营养方面发挥不可或缺的作用,其在卵黄中的组成可能与这一多样化动物群体中不同的早期生活史有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2cd8/4651544/a9bb222d0be4/pone.0143225.g001.jpg

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