Quantitative Proteomic Analysis of buffalo ovaries reveals the regulation of kinase and phosphatase during the Formation and Regression of Corpus Luteum

Abstract

Background: Reproductive characteristics are made up of many complicated physiology procedures that have significant effects on the growth of the buffalo industry and are affected by numerous factors. The ovary is the most important sexual organ of female mammals, and during the oestrous cycles, its corpus luteum (CL) plays a significant part in mammalian reproduction. During the development and regression of corpus luteum, however, the differentially expressed proteins are less defined. In this study, we used a 6-plex tandem mass tag (TMT) strategy for quantitative proteomic comparison of three distinct ovary phases (corpus hemorrhagicum, corpus luteum and corpus hemorrhagicum) in buffalo. Results: A total of 148 differentially expressed proteins were identified, 32 of these proteins were identified as differentially expressed in the group CH (corpus hemorrhagicum) and 116 were identified as differentially expressed in the group CF (corpus fibrosum), with the group CL (corpus luteum) serving as the control group. Notably, we discovered that quite some enzymes such as kinase and phosphatase, are upregulated in the ovary CL phase, and three upregulated enzymes and proteins in the CL phase (PLK1, PGP, and HGS) were verified using Western blotting, quantitative RT-PCR, and immunohistochemistry analysis. The results of these validations were consistent with the quantitative analysis of the TMT-label, which indicated that they could play a crucial role during the CL's reproductive cycle. These buffalo results during the formation and regression of the corpus luteum are also shown to be a substantial reference value for comparable human and mouse studies after analyzing homologous BLAST cross-species. The expression information is accessible with the PXD009957 identifier via ProteomeXchange. Conclusions: Our research gives a deeper understanding of CL formation and regression during the oestrous cycles, which shows kinase and phosphatase regulation, and indicates some potential enzymes and proteins that may influence buffalo fertility. Keywords: corpus luteum, quantitative proteomic, oestrous cycles, ovary, cross-species