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1.
Reprod Domest Anim ; 56(4): 586-603, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33460477

ABSTRACT

The present study was conducted to decipher the proteome of in vivo-produced pre-implantation ovine embryos. Ten locally adapted Morana Nova ewes received hormonal treatment and were inseminated 12 hr after ovulation. Six days later, 54 embryos (morula and blastocyst developmental state) were recovered from eight ewes and pooled to obtain sufficient protein for proteomic analysis. Extracted embryo proteins were analysed by LC-MS/MS, followed by identification based on four database searches (PEAKS, Proteome Discoverer software, SearchGUI software, PepExplorer). Identified proteins were analysed for gene ontology terms, protein clusters and interactions. Genes associated with the ovine embryo proteome were screened for miRNA targets using data sets of TargetScan (http://www.targetscan.org) and mIRBase (http://www.mirbase.org) servers. There were 667 proteins identified in the ovine embryos. Biological processes of such proteins were mainly related to cellular process and regulation, and molecular functions, to binding and catalytic activity. Analysis of the embryo proteins revealed 49 enriched functional clusters, linked to energy metabolism (TCA cycle, pyruvate and glycolysis metabolism), zona pellucida (ZP), MAPK signalling pathway, tight junction, binding of sperm to ZP, translation, proteasome, cell cycle and calcium/phospholipid binding. Sixteen miRNAs were related to 25 pre-implantation ovine embryo genes, all conserved in human, bovine and ovine species. The interaction network generated by miRNet showed four key miRNAs (hsa-mir-106b-5p; hsa-mir-30-5p; hsa-mir-103a-5p and hsa-mir-106a-5p) with potential interactions with embryo-expressed genes. Functional analysis of the network indicated that miRNAs modulate genes related to cell cycle, regulation of stem cell and embryonic cell differentiation, among others. Retrieved miRNAs also modulate the expression of genes involved in cell signalling pathways, such as MAPK, Wnt, TGF-beta, p53 and Toll-like receptor. The current study describes the first major proteomic profile of 6-day-old ovine embryos produced in vivo, setting a comprehensive foundation for our understanding of embryo physiology in the ovine species.


Subject(s)
Embryo, Mammalian/chemistry , Proteome/analysis , Sheep, Domestic/embryology , Animals , Female , Insemination, Artificial/veterinary , Male , MicroRNAs/genetics , Proteome/genetics , Sheep, Domestic/genetics , Sheep, Domestic/metabolism
2.
Theriogenology ; 94: 59-63, 2017 May.
Article in English | MEDLINE | ID: mdl-28407861

ABSTRACT

Reproductive tract development during puberty is critical to reproductive performance, and the light is crucial in this process in birds. However, in male quail, there is little information on the effects of types of lamps, more specifically the wavelength emitted. Thus, the objective was to evaluate the effects of types of lamps on the reproductive performance of male Japanese quail (Coturnix coturnix japonica). Two hundred and forty male quail were exposed to six different types of lamp (incandescent, white fluorescent, or blue, white, red or green LED). The experimental design was completely randomized with six treatments and five replicates of one quail. Quail were slaughtered on days 35, 47, 57, 71 and 123 to evaluate the development of testes. On day 117, semen samples were analyzed and fertilized eggs were incubated. Body weight of the quails was influenced (P < 0.01) by lamps only until the 47 days of age. Higher body weight until this age were observed with incandescent, blue and green LED bulbs. Fluorescent and red LED bulbs propitiated (P < 0.05) early testicular development of quails but, at 57 days of age, higher testicular development was obtained (P < 0.01) whit white LED bulbs. Lower testicular development was observed (P < 0.01) at 123 days of age with the red LED. No influence of different types of lamps was observed (P > 0.05) on the quality of semen nor on the fertility rates of quail. It is concluded that lamps can influence the histological reproductive characteristics of male quails, but without influencing the semen quality. Fluorescent bulbs and red LED seem to anticipate the sexual maturity, but the white LED results in higher testicular development at 57 days of age.


Subject(s)
Coturnix/growth & development , Sexual Maturation/radiation effects , Testis/radiation effects , Animals , Body Weight/radiation effects , Fertility/radiation effects , Male , Random Allocation , Semen Analysis/veterinary , Testis/growth & development
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