Quantification of Urinary Exosome Sex Hormone Binding Globulin Levels during Estrus and Diestrus Stages of Buffaloes

Manasa Varra, V. Girish Kumar, B.M. Chandranaik, M.T. Sanjeeva Kumar, N.R. Sundaresan, Veerasamy Sejian, G. Sudha
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Abstract

Background: The efficiency of reproduction in buffaloes can be increased by being able to spot buffaloes in estrus, as silent heat is a severe problem in this species. Proteins of urinary exosomes are identified to be better physiological biomarkers when compared to cell free proteins as these extracellular vesicles (EVs) are tiny in size, have additional protection from degradation by endogenous protease activity and are involved in intercellular communication. Hence, the present study was aimed to quantify the urinary exosome sex hormone binding globulin (SHBG) levels during estrus and diestrus stages of buffaloes to determine its suitability as a marker for identifying buffaloes in estrus. Methods: The grouping of animals as mid-diestrus (Group-I/G-I/Control group), regular estrus (Group-II/G-II) and silent estrus (Group-III/G-III) buffaloes was done using a combinatorial approach. The levels of SHBG in urinary exosomes of three groups of animals was quantified using the bovine SHBG ELISA kit. Result: The present study’s findings showed significantly higher urinary exosome SHBG concentration (p less than 0.05) in G-I in comparison to G-II and G-III. The present study revealing for decreased concentration of urinary exosome SHBG in G-II and G-III animals when compared to G-I animals probably construes the physiological role of SHBG in the female reproductive tract at the estrus stage by mediating the action of estradiol 17 â (Eα) at the target site. However, for urinary exosome SHBG to be considered as a biomarker, there is need for additional research on a bigger population.
水牛发情期和发情期尿外泌体性激素结合球蛋白水平的定量测定
背景:能够发现发情期的水牛,可以提高水牛的繁殖效率,因为在这个物种中,无热是一个严重的问题。与游离细胞蛋白相比,尿外泌体的蛋白质被认为是更好的生理生物标志物,因为这些细胞外囊泡(ev)体积很小,具有内源性蛋白酶活性降解的额外保护,并参与细胞间通讯。因此,本研究旨在定量测定水牛发情期和发情期尿外泌体性激素结合球蛋白(SHBG)水平,以确定其作为鉴定发情期水牛的标志物的适用性。方法:采用组合法将动物分为中期发情组(i组/G-I组/对照组)、正常发情组(ii组/G-II组)和沉默发情组(iii组/G-III组)。采用牛SHBG酶联免疫吸附测定试剂盒,定量测定三组动物尿外泌体中SHBG的含量。结果:G-I组尿外泌体SHBG浓度明显高于G-II和G-III组(p < 0.05)。本研究显示,与G-I动物相比,G-II和G-III动物尿外泌体SHBG浓度降低,这可能解释了SHBG在发情期通过介导雌二醇17 (Eα)在靶部位的作用在雌性生殖道中的生理作用。然而,为了将尿外泌体SHBG作为一种生物标志物,还需要在更大的人群中进行进一步的研究。
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