Proteomic analysis of amniotic and allantoic fluid from buffaloes during foetal development

Abstract

The objective of this study was to describe the dynamic changes in protein composition and protein abundance in amniotic and allantoic fluids from buffaloes during gestation. Amniotic and allantoic fluids were collected during the first, second and third trimesters of gestation. The foetuses were measured and weighed. Fluid samples were centrifuged at 800 g for 10 min and then at 10,000 g for 60 min at 4°C. The supernatant was collected to determine the total protein concentration. Based on total protein concentration, an aliquot (50 μg) was used for in‐solution tryptic digestion, and mass spectrometry analysis (nano‐LC‐MS/MS) was performed. A multivariate statistical analysis of the proteomic data was conducted. Across the different stages of buffalo gestation, fifty‐one proteins were found in the amniotic fluid, and twenty‐one were found in the allantoic fluid. A total of twelve proteins were common among the stages, and four presented significant differences (VIP score α > 1). Fibronectin and alpha‐1‐antiproteinase were more abundant in the amniotic fluid than in the allantoic fluid. Alpha‐2‐macroglobulin and alpha‐2‐HS‐glycoprotein were more abundant in the allantoic fluid than in the amniotic fluid. Alpha‐2‐macroglobulin participates in remodelling and growth of the uterus at beginning of the gestation (first trimester), and these findings indicate that can serve as a potential tool for the early diagnosis of pregnancy in buffaloes.

Evaluation of Biochemical Composition of Amniotic and Allantoic Fluids at Different Stages of Pregnancy in Queens

“…A similar diagnosis can be observed in mares, where the up- or down-regulation of some components of fetal fluids can be indicative of a pathological condition [ 20 ]. Results from a recent study with buffalo indicated that some proteins can be in greater concentrations during the first stages of gestation, indicating that evaluations of the concentrations of these proteins can be used as a diagnostic approach for an early diagnosis of pregnancy [ 21 ].…”