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用角蛋白酶降解马来穿山甲鳞片以提取生殖甾体激素。

Degradation of Temminck's pangolin () scales with a keratinase for extraction of reproductive steroid hormones.

作者信息

Blecher Arantxa S, Scheun Juan, Ganswindt André

机构信息

Mammal Research Institute, Department of Zoology and Entomology, Faculty of Natural and Agricultural Sciences, University of Pretoria, South Africa.

Biodiversity Research Services, National Zoological Gardens, South Africa.

出版信息

MethodsX. 2021 Jan 13;8:101229. doi: 10.1016/j.mex.2021.101229. eCollection 2021.

Abstract

Hormone monitoring in keratinous tissues has become increasingly popular. The insoluble keratin materials are generally pulverised before hormone extraction; however, this is difficult for thicker keratin structures like baleen plates or hooves. A new method, involving the use of keratinase, allows enzymatic digestion of keratin and hormone analysis in the resulting suspension. Pangolins are unique mammals covered in keratinous scales, which are one of the reasons these animals are extensively trafficked. This study aimed to investigate the suitability of Temminck's pangolin scales as hormone matrix for quantifying reproductive steroids. A protocol was developed to digest scales with a keratinase before measuring hormone concentrations. This method can be used to investigate the reproductive endocrinology of Temminck's pangolins but may also be extended to the other extant pangolin species.•Keratinase digests Temminck's pangolin scales and reproductive steroid metabolite concentrations are measurable in the resulting suspension.•Isopropanol is an ideal washing solvent for scales to remove surface contaminants and scale sample mass should be standardised to allow comparisons.•Any section of a scale and scales from any pangolin body region can be used as samples for hormone quantification.

摘要

角质组织中的激素监测越来越受到关注。在提取激素之前,通常会将不溶性角蛋白材料粉碎;然而,对于像须鲸板或蹄子这样较厚的角蛋白结构来说,这很困难。一种涉及使用角蛋白酶的新方法,可以对角蛋白进行酶解,并对所得悬浮液中的激素进行分析。穿山甲是一种独特的哺乳动物,全身覆盖着角质鳞片,这也是这些动物被大量非法交易的原因之一。本研究旨在探讨马来穿山甲鳞片作为定量生殖类固醇激素基质的适用性。在测量激素浓度之前,制定了一个用角蛋白酶消化鳞片的方案。该方法可用于研究马来穿山甲的生殖内分泌学,也可能适用于其他现存的穿山甲物种。

  • 角蛋白酶可消化马来穿山甲鳞片,所得悬浮液中可测量出生殖类固醇代谢物浓度。

  • 异丙醇是鳞片去除表面污染物的理想洗涤溶剂,鳞片样品质量应标准化以便进行比较。

  • 鳞片的任何部分以及来自穿山甲身体任何部位的鳞片都可作为激素定量的样本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1044/8374191/50a3a6784f03/fx1.jpg

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