Proteomic Profiles of the Embryonic Chorioamnion and Uterine Caruncles in Buffaloes (Bubalus bubalis) with Normal and Retarded Embryonic Development1

Abstract

The aim of this study was to compare the proteome profiles of the chorioamnion and corresponding caruncle for buffalo embryos that had either normal or retarded development on Day 25 after artificial insemination (AI). In experiment 1, embryos that were to subsequently undergo late embryonic mortality had a smaller width on Day 25 after AI than embryos associated with pregnancy on Day 45 after AI. In experiment 2, 25 Italian Mediterranean buffaloes underwent transrectal ultrasonography on Day 25 after AI, and pregnant animals were categorized as one of two groups based on embryonic width: normal embryos (embryonic width > 2.7 mm) and retarded embryos (embryonic width < 2.7 mm). Three buffaloes of each group were slaughtered on Day 27 after AI to collect chorioamnion and caruncle tissues for subsequent proteomic analyses. Two-dimensional difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight mass spectrometer analysis were used to ascertain the proteomic profiles. To confirm 2D-DIGE-results, three selected proteins were analyzed by Western blot. The proteomic profiles of the chorioamnion of retarded embryos and the corresponding caruncles showed differences in the expression of several proteins compared to normal embryos. In particular, a down-regulation was observed for proteins involved in protein folding (HSP 90-alpha, calreticulin), calcium binding (annexin A1, annexin A2), and coagulation (fibrinogen alpha-chain) (P < 0.05), whereas proteins involved in protease inhibition (alpha-1-antiproteinase, serpin H1, serpin A3-8), DNA and RNA binding (heterogeneous nuclear ribonucleoproteins A2/B1 and K), chromosome segregation (serine/threonine-protein phosphatase 2A), cytoskeletal organization (ezrin), cell redox homeostasis (amine oxidase-A), and hemoglobin binding (haptoglobin) were up-regulated (P < 0.05).

Influences of different space allowance on reproductive performances in buffalo

“…The results obtained in this study demonstrated that reproductive status (pregnant, not pregnant and LEM) and progesterone blood levels were not affected by the different space allowance conditions. On the contrary, P4 blood levels were significantly higher in pregnant buffaloes, compared to not pregnant and buffaloes underwent LEM, in agreement with other studies (Vecchio et al, 2012;Balestrieri et al, 2013). These findings support the hypothesis that LEM in buffalo is primarily due to a reduced P4 secretion by the ] LDG HDG corpus luteum.…”

“…These findings support the hypothesis that LEM in buffalo is primarily due to a reduced P4 secretion by the ] LDG HDG corpus luteum. Adequate luteal function and threshold circulating concentrations of P4 in circulation are required to guarantee an uterine environment that promotes ongoing elongation of the embryo during the critical peri-implantation period of pregnancy (Balestrieri et al, 2013). In our study no differences were found on blood metabolities and redox status between buffaloes maintained in high and low density.…”

Global Transcriptome Profiles of Italian Mediterranean Buffalo Embryos with Normal and Retarded Growth

“…Differences in proteome profiles were also reported for the caruncles adjacent to the chorioamnions of embryos with retarded and normal growth. Differentially expressed proteins were related to antioxidant protection, protease inhibition and protein folding [13]. These findings supported other evidence that a disruption of conceptus-uterus interaction, as a result of reduced progesterone, is the major cause of late embryonic mortality in buffaloes [1], [2].…”

“…The latter covers the period during which there is maternal recognition of pregnancy. It was proposed that late embryonic mortality in buffaloes results from the failure of embryonic attachment [2], [13] which is a consequence, at least in part, of reduced concentrations of P4 in circulation from around Day 10 after mating [3].…”

“…Balestrieri et al [13] recently reported that the chorioamnion of embryos with retarded growth (embryo width<2.7 mm) had a different proteomic profile to that of embryos with normal growth (embryo width>2.7 mm). Differences in proteome profiles were also reported for the caruncles adjacent to the chorioamnions of embryos with retarded and normal growth.…”