初步评价精浆蛋白和神经生长因子的免疫反应性作为牙魔排卵诱导因子的指示性。

D. Bergfelt, J. Blum, Jill R. Ratner, M. Ratto, J. O'Brien, T. Robeck
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引用次数: 0

摘要

在被称为诱导排卵(如骆驼)和自发排卵(如牛)的陆生哺乳动物的精浆中,已经发现了一种具有类似于β-神经生长因子(β-NGF)的蛋白质结构的排卵诱导因子(OIF)。鲸目动物精浆中OIF/NGF的检测将在生殖生物学和保护管理计划中具有基础和应用意义。对水族馆中的白鲸、太平洋白边海豚和宽吻海豚精液浆蛋白的分布和丰度进行了初步评估。初始采用SDS-PAGE,总蛋白50 μg进行分离;随后,采用Western免疫印迹法检测抗ngf。除齿蟾精浆外,还采用羊驼精浆纯化部分(蛋白含量100 ng)和小鼠脑提取物(总蛋白含量20 μg)作为NGF阳性对照。在两只白鲸体内,目视检查显示蛋白质带的分布和强度相似。然而,在白鲸、太平洋白颊海豚和宽吻海豚之间,精浆蛋白的分布和丰度的多样性大于相似性。虽然NGF的免疫反应性在骆驼和小鼠阳性对照中非常明显,但在任何牙髓样本中都没有视觉反应性。这些初步结果提供了新的信息,表明有齿螈精浆蛋白内部有更多的同质性和异质性。虽然NGF没有被免疫检测到,但未来的研究需要解决NGF免疫检测的明显局限性,特别是如果翻译后形式的β-NGF在白鲸、太平洋白边海豚和宽吻海豚的精浆中丰度很低。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PRELIMINARY EVALUATION OF SEMINAL PLASMA PROTEINS AND IMMUNOREACTIVITY OF NERVE GROWTH FACTOR AS INDICATIVE OF AN OVULATION INDUCING FACTOR IN ODONTOCETES.
In the seminal plasma of terrestrial mammalian species known as induced (e.g., camels) and spontaneous (e.g., cattle) ovulators, an ovulation-inducing factor (OIF) with a protein structure similar to beta-nerve growth factor (β-NGF) has been identified. Detection of an OIF/NGF in the seminal plasma of cetaceans would have both basic and applied implications in reproductive biology and conservation management programs. A preliminary evaluation was conducted to characterize the distribution and abundance of seminal plasma proteins in aquarium-based belugas and a Pacific white-sided and bottlenose dolphin. Initially, SDS-PAGE was used with 50 μg of total protein for separation; thereafter, Western immunoblot was used with anti-NGF. In addition to odontocete seminal plasma, a purified fraction of llama seminal plasma (100 ng protein) and an extract of mouse brain (20 μg total protein) were included as positive controls for NGF. Within the two belugas, visual inspection of the protein bands indicated similar distribution and intensity. However, among the belugas and Pacific white-sided and bottlenose dolphins there was more diversity than similarity in the distribution and abundance of seminal plasma proteins. While immunoreactivity of NGF was distinctly evident in the llama and mouse positive controls, there was no visual reactivity in any of the odontocete samples. These preliminary results provide novel information indicating more homogeneity within and heterogeneity among seminal plasma proteins of ondentocetes. Although NGF was not immunologically detected, future studies are required to address the apparent limitations of immuno-detection of NGF, especially if the post-translational form of β-NGF is in low abundance in the seminal plasma of belugas and Pacific white-sided and bottlenose dolphins.
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