季节性减重（SWL）是热带和地中海地区畜牧生产最重要的限制因素，制约生产者的收入和农村社区的营养状况。制定抵抗SWL不利影响的生产策略是至关重要的。经过选择，在恶劣的气候中演变出的品种对SWL具有耐受性。决定这种能力的因素大部分与受SWL影响的生化途径变化有关。

在本研究中，基于凝胶的蛋白质组学策略(BN：蓝色-聚丙烯酰胺凝胶电泳和2DE：双向凝胶电泳)描述山羊乳腺分泌组织线粒体蛋白质组。此外，研究人员还对两个具有不同营养应激适应性水平的山羊品种：Majorera(适应)和Palmera (敏感)，进行了减重的影响因素的调查。本研究采用Majorera和Palmera奶山羊，分为4组，每2组为不同品种：营养不良组饲喂小麦秸秆(限制饲粮，所以他们的体重在实验结束时将减少15-20%)，对照组饲喂能量平衡饲粮。在实验期结束后(22天)，所有的实验组都进行乳腺活体检查。线粒体的蛋白质组分析鉴别了总共277种蛋白质，其中148(53%)种由飞行质谱/质谱法确定。一些蛋白质被确定为来自线粒体的谷氨酸脱氢酶复合物的亚基和呼吸复合物I、II、IV、V，许多其他蛋白质功能：新陈代谢、发育、定位、细胞组织和合成、生物调节、对刺激的反应，都在BN和2DE凝胶中得到测定。比较蛋白质组学分析实现了几种蛋白质的鉴定：NADH泛醌氧化还原酶75 kDa的亚基和核纤层蛋白B1线粒体（在Palmera品种中上调），鸟嘌呤核苷酸结合蛋白G（I）/ G（S）/G（T）亚基β2（在Majorera品种中上调）以及细胞色素b- c1复合物亚基1，线粒体和链D，ATP合成酶牛F1-C8亚复合物（在Majorera品种中下调），作为一种减重的影响因素。

原文摘要如下：

The Goat (Capra hircus) Mammary Gland Mitochondrial Proteome: A Study on the Effect of Weight Loss Using Blue-Native PAGE and Two-Dimensional Gel Electrophoresis

Cugno G^1,2, Parreira JR^3,4, Ferlizza E^1,5, Hernández-Castellano LE^2,6, Carneiro M¹, Renaut J⁷, Castro N², Arguello A², Capote J⁸, Campos AM¹, Almeida AM^3,4,9

Abstract：Seasonal weight loss (SWL) is the most important limitation to animal production in the Tropical and Mediterranean regions, conditioning producer's incomes and the nutritional status of rural communities. It is of importance to produce strategies to oppose adverse effects of SWL. Breeds that have evolved in harsh climates have acquired tolerance to SWL through selection. Most of the factors determining such ability are related to changes in biochemical pathways as affected by SWL. In this study, a gel based proteomics strategy (BN: Blue-Native Page and 2DE: Two-dimensional gel electrophoresis) was used to characterize the mitochondrial proteome of the secretory tissue of the goat mammary gland. In addition, we have conducted an investigation of the effects of weight loss in two goat breeds with different levels of adaptation to nutritional stress: Majorera (tolerant) and Palmera (susceptible). The study used Majorera and Palmera dairy goats, divided in 4 sets, 2 for each breed: underfed group fed on wheat straw (restricted diet, so their body weight would be 15-20% reduced by the end of experiment), and a control group fed with an energy-balanced diet. At the end of the experimental period (22 days), mammary gland biopsies were obtained for all experimental groups. The proteomic analysis of the mitochondria enabled the resolution of a total of 277 proteins, and 148 (53%) were identified by MALDI-TOF/TOF mass spectrometry. Some of the proteins were identified as subunits of the glutamate dehydrogenase complex and the respiratory complexes I, II, IV, V from mitochondria, as well as numerous other proteins with functions in: metabolism, development, localization, cellular organization and biogenesis, biological regulation, response to stimulus, among others, that were mapped in both BN and 2DE gels. The comparative proteomics analysis enabled the identification of several proteins: NADH-ubiquinone oxidoreductase 75 kDa subunit and lamin B1 mitochondrial (up-regulated in the Palmera breed), Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2 (up-regulated in the Majorera breed) and cytochrome b-c1 complex subunit 1, mitochondrial and Chain D, Bovine F1-C8 Sub-Complex Of Atp Synthase (down-regulated in the Majorera breed) as a consequence of weight loss.

Keywords: Seasonal weight loss (SWL), Mitochondrial proteome, Mammary gland, Blue-Native Page（BN）, Two-dimensional gel electrophoresis（2DE）

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