Proteome profiling of chicken ovarian follicles immediately before and after cyclic recruitment

Abstract

A shotgun proteomics study using isobaric tags for relative and absolute quantification labeling was conducted to characterize proteins in chicken ovarian follicles immediately before and after cyclic recruitment. Granulosa cell (GC) layers from the most recently recruited follicle (GC9) and from each of the four largest prerecruitment follicles (GC1-4) plus theca tissue (TH) from the most recently recruited (TH9) and largest prerecruitment (TH1) follicles were compared. Of 1535 proteins identified, none were determined to be differentially expressed between TH9 and TH1. A pairwise comparison between GC9 and GC1, GC2, GC3, or GC4 resulted in one, five, five, and six differentially expressed proteins, respectively, including yolk and cholesterol transport proteins (vitellogenin 1-3 and apolipoprotein B). In addition, transforming growth factor-beta 1 and microRNA-21 pathways were predicted to be activated at recruitment. We also report, for the first time, the expression of the neuropeptide, RELAXIN-3 (RLN3), in GC. Quantitative polymerase chain reaction determined RLN3 expression to be highest in GC9 and GC1, but its receptors, RXFP1 and RXFP3, were highest in TH and ovarian stroma, respectively. Overall, cyclic recruitment is associated with changes in protein expression predominantly within follicle GC, and a potential role for RLN3 in follicle recruitment and the initiation of GC differentiation warrants further investigation.

Identification and functional analysis of ovarian lncRNAs during different egg laying periods in Taihe Black-Bone Chickens

“…RLN3 is significantly enriched in this pathway and is highly expressed during the peak egg-laying period. Relaxin-like peptides, such as RLN3 , are produced in granulosa cells post-ovulation and play a role in promoting egg laying, impacting the oviduct and ovary ( Ghanem and Johnson, 2021 ). Additionally, in the protein-protein interaction network, we observed an interaction between RLN3 and SCG2 ( Figure 6B ).…”

Characterization of relaxin 3 and its receptors in chicken: Evidence for relaxin 3 acting as a novel pituitary hormone

“…In another transcriptome analysis between the two divergent chicken breeds, which are in the laying phase and the brooding phase, the expression of RLN3 mRNA shows downregulation in brooding chickens, thus pointing out its role actively involved in reproductive biology ( Shen et al, 2016 ). The report that RLN3 is actively involved in female reproductive biology also comes from the study by Ghanem and Johnson (2021 ) in a shotgun proteomics study using isobaric tags to characterize the proteins in chicken ovarian follicles immediately before and after cyclic recruitment. RLN3 is revealed to be distinctively expressed in a total of 1,535 proteins.…”

Transcriptome and Proteome Analysis Revealed Key Pathways Regulating Final Stage of Oocyte Maturation of the Turkey (Meleagris gallopavo)

“…In our study, we indicated that eIF2 signaling and the regulation of eIF4 and p70S6K signaling pathways-both involved in protein synthesisare the most significant canonical pathways in the transcriptome, as well as in the proteome, of mature egg coats. The eIF2 signaling pathway has recently been identified in chicken follicles after cyclic recruitment, and eIF2 activation reflects the demand for enhanced protein synthesis during follicle development [28]. In bird oogenesis, primary follicles aprox.…”

“…Interestingly, we also observed the presence of a membrane estrogen receptor, GPER1, which, together with ESRs, may be involved in estrogen signaling in mature egg coats. In birds, GPER signaling has been described in the GCs of chicken ovarian follicles before and after cyclic recruitment [28], and its role in GC differentiation and promoting follicle survival has been reported [71]. Estrogen binding to GPER activates a stimulatory G protein, resulting in the stimulation of adenylyl cyclase activity and cAMP levels, which are involved in maintaining the estrogen-induced meiotic arrest of oocytes before fertilization in vertebrates [72].…”

“…Candidate microarray analysis of the germinal disc region of chicken oocytes has revealed genes potentially involved in "laying" and "meat" line hen fertility [25], as well as a few genes involved in oocyte maturation and early embryo development in chicken hens [13]. Transcriptomic and proteomic analyses have been used to determine the mechanisms involved in chicken follicle selection [26][27][28], and proteomic analysis has been widely used for the analysis of the laid chicken egg vitelline membrane [29,30], and egg yolk [31] and egg white proteome [32]. However, there is a lack of similarly comprehensive studies for the turkey hen reproductive tract.…”