RESUMEN
Transmembrane protein 8C (Tmem8C) is a muscle-specific membrane protein that controls myoblast fusion, which is essential for the formation of multinucleated muscle fibres. As most of the birds can fly, they have enormous requirement for the muscle, but there are only a few studies of Tmem8C in birds. In this study, we obtained the coding sequence (CDS) of Tmem8C in goose, predicted miRNAs that can act on the 3'UTR, analysed expression profiles of this gene in breast and leg muscles (BM and LM) during the embryonic period and neonatal stages, and identified miRNAs that might affect the targeted gene. The results revealed a high homology between Tmem8C in goose and other animals (indicated by sequence comparisons and phylogenetic trees), some conservative characteristics (e.g., six transmembrane domains and two E-boxes in the 5'UTR might be the potential binding sites of muscle regulatory factors (MRFs)), and the dN/dS ratio indicated purifying selection acting on this gene, facilitating conservatism in vertebrates. Q-PCR indicated Tmem8C had a peak expression pattern, reaching its highest expression levels in stage E15 in LM and E19 in BM, and then dropping transiently in E23 (P < 0.05). We examined 13 candidate miRNAs, and negative relationships were detected both in BM and LM (mir-125b-5p, mir-15a, mir-16-1 and mir-n23). Notably, mir-16-1 significantly decreased luciferase activity in dual luciferase reporter gene (LRG) assay, suggesting that it can be identified as potential factors affecting Tmem8C. This study investigated Tmem8C in water bird for the first time, and provided useful information about this gene and its candidate miRNAs in goose.
Asunto(s)
Gansos/genética , Regulación de la Expresión Génica , MicroARNs/genética , Desarrollo de Músculos/genética , Animales , Línea Celular , Gansos/clasificación , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genes Reporteros , Interacciones Hidrofóbicas e Hidrofílicas , MicroARNs/química , Mioblastos/metabolismo , Conformación de Ácido Nucleico , Filogenia , Interferencia de ARN , ARN Mensajero/química , ARN Mensajero/genética , Análisis de Secuencia de ADNRESUMEN
To analyze the gene structure and the evolutionary roadmap of Fulica atra, the complete mitogenome is sequenced. It is composed of 37 genes and 1 control region, and the structure and arrangement of all genes are identical to other Rallidae. The comparative mitogenome revealed that the start codon and stop codon varied in Rallidae, and the gene lengths are different in ND2, COX1, ND3, ND5 and CYTB due to incompleteness of stop codon, frameshift mutation and various numbers of amino acids. We analyzed the correlation between phylogeny and gene characteristic in Rallidae with respect to the usage of start/stop codon and gene length, but no correlation was found. It indicates these discrepancies might happen independently. This work can afford an in-depth insight of phyletic evolution in Rallidae.