Neuropeptide profiling of the bovine hypothalamus: Thermal stabilization is an effective tool in inhibiting post-mortem degradation
Neuropeptide profiling of the bovine hypothalamus: Thermal stabilization is an effective tool in inhibiting post-mortem degradation
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DOI：
10.1002/pmic.201000423
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发表时间：
2011-04-01
期刊：
影响因子：
3.4
通讯作者：
Wijffels, Gene
中科院分区：
文献类型：
--
作者：
Colgrave, Michelle L.;Xi, Li;Wijffels, Gene
下丘脑是大脑的中央调节区域，通过垂体将神经系统与内分泌系统连接起来。它合成并分泌神经肽激素，神经肽激素反过来又刺激或抑制垂体激素的分泌。我们通过采用一种新颖的热稳定方法，对牛下丘脑中存在的肽进行了详细的质谱研究，该方法改进了肽的发现，以指导我们对牛繁殖所涉及的分子机制的研究。未经处理的样品含有大量蛋白质降解产物，干扰神经肽的分析。在热稳定样品中，我们能够识别出更多已知在牛下丘脑中表达的神经肽。此外，我们还表征了一系列翻译后修饰，表明稳定的组织样本中存在经过加工的完整成熟神经肽，而我们在未处理的组织样本中检测到了许多由死后降解产生的修剪或截短的肽。总之，通过优化的工作流程，我们能够鉴定出 140 种候选神经肽。我们还提名了源自 proSAAS、secretogranin-2 和 proTRH 的六种新候选神经肽。
The hypothalamus is the central regulatory region of the brain that links the nervous system to the endocrine system via the pituitary gland. It synthesizes and secretes neuropeptide hormones, which in turn act to stimulate or inhibit the secretion of pituitary hormones. We have undertaken a detailed MS investigation of the peptides present in the bovine hypothalamus by adapting a novel heat stabilization methodology, which improved peptide discovery to direct our studies into the molecular mechanisms involved in bovine reproduction. The untreated samples contained large numbers of protein degradation products that interfered with the analysis of the neuropeptides. In the thermally stabilized samples, we were able to identify many more neuropeptides that are known to be expressed in the bovine hypothalamus. Furthermore, we have characterized a range of post-translational modifications that indicate the presence of processed intact mature neuropeptides in the stabilized tissue samples, whereas we detected many trimmed or truncated peptides resulting from post-mortem degradation in the untreated tissue samples. Altogether, using an optimized workflow, we were able to identify 140 candidate neuropeptides. We also nominate six new candidate neuropeptides derived from proSAAS, secretogranin-2 and proTRH.