Study on the transcriptome for breast muscle of chickens and the function of key gene RAC2 on fibroblasts proliferation

Abstract

Background Growth performance is significant in broiler production. In the growth process of broilers, gene expression varies at different growth stages. However, limited research has been conducted on the molecular mechanisms of muscle growth and development in yellow-feathered male chickens. Results In the study, we used RNA-seq to study the transcriptome of the breast muscle of male Jinghai yellow chickens at 4 (M4F), 8 (M8F) and 12 weeks (M12F) of age. The results showed that 4608 differentially expressed genes (DEGs) were obtained by comparison in pairs of the three groups with Fold Change (FC) ≥ 2 and False Discovery Rate (FDR) ≤ 0.05, and 83, 3445 and 3903 DEGs were obtained separately from M4FvsM8F, M4FvsM12F and M8FvsM12F. Six genes were found as co-differentially expressed in the three age groups, namely SNCG, MYH1A, ARHGDIB, ENSGALG00000031598, ENSGALG00000035660 and ENSGALG00000030559. The GO analysis showed that 0, 304 and 408 biological process (BP) were significantly enriched in M4FvsM8F, M4FvsM12F and M8FvsM12F groups, respectively. KEGG pathway enrichment showed that 1, 2, 4 and 4 pathways were significantly enriched in M4FvsM8F, M4FvsM12F, M8FvsM12F and all DEGs, respectively. They were steroid biosynthesis, carbon metabolism, focal adhesion, cytokine-cytokine receptor interaction, biosynthesis of amino acids and salmonella infection. We constructed short hairpin RNA (shRNA) to interfere the differentially expressed gene RAC2 in DF-1 cells and detected mRNA and protein expression of the downstream genes PAK1 and MAPK8. Results of qPCR showed that RAC2, PAK1 and MAPK8 mRNA expression significantly decreased in the shRAC2–2 group compared with the negative control (NC) group. Western Blot (WB) results showed that the proteins of RAC2, PAK1 and MAPK8 also decreased in the shRAC2–2 group. Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2′-deoxyuridine (EdU) assay both showed that the proliferation of DF-1 cells was significantly inhibited after transfection of shRAC2–2. Conclusions The results of RNA-seq revealed genes, BP terms and KEGG pathways related to growth and development of male Jinghai yellow chickens, and they would have important guiding significance to our production practice. Further research suggested that RAC2 might regulate cell proliferation by regulating PAKs/MAPK8 pathway and affect growth of chickens.

Delivery of a tissue derived extracellular matrix gel modulates early fibro-adipogenic cell behavior and improves recovery following both acute and chronic atrophy muscle injury

“…LGALS3 which aids in the fusion and differentiation of myoblasts during muscle repair and regeneration were increased by 4.6, 4.8, and 3 folds in 7DPI transcripts, 3 DPI and 14DPI proteins abundance respectively 40,41 . Similar increases were observed for RAC2 which is essential in myotube formation and regulation of hypertrophy 42,43 . Comparably, FABP5, regulating the insulin-mediated glucose uptake through activation of AKT and AMPK and promoting cell proliferation, also increased 44,45 .…”

Transcriptome analysis of embryonic muscle development in Chengkou Mountain Chicken

“…At the same time, the ECM–receptor interaction and focal adhesion were significantly enriched in KEGG pathway analysis with differentially expressed genes [ 12 ]. In addition, transcriptome sequencing of breast muscle of Jinghai Huang chicken at different times (M4F, M8Fand M12F) was conducted to screen out 3,903 DEGs, and it was analyzed that RAC2 may affect the growth of chickens by regulating the PAKs/MAPK8 pathway[ 20 ]. In the later stage of embryonic development, the number of differentially expressed genes decreased sharply.…”

Unveiling Comparative Genomic Trajectories of Selection and Key Candidate Genes in Egg-Type Russian White and Meat-Type White Cornish Chickens

“…Within the same genomic region, there are three other genes, BTG1 (44,429,339 … 44,433,309 bp) , KITLG (43,015,486 … 43,066,975 bp), and MGAT4C (42,251,047 … 42,358,204 bp). BTG anti-proliferation factor 1 ( BTG1 ) is a candidate gene expressed during early chick development [ 82 ], being related to muscle structure growth and development at early stages [ 83 ]. The KITLG (ligand of tyrosine–kinase receptor) gene was cytogenetically mapped to GGA1 [ 84 ] and more recently ascertained as a gene related to pigmentation traits, having been under selection before chicken domestication and overlapping a selective sweep [ 40 ].…”

“…Initially, the chicken CTSL (now CTSV ) gene that has an increased expression during oviduct regression [ 110 ] was mapped to the Z chromosome [ 111 ], while LOC420160 is considered as its paralog on GGA28. MYDGF (myeloid-derived growth factor; 5,034,853 … 5,039,077 bp) is a candidate gene related to growth and development at early stages [ 83 ]. The PLIN3 (perilipin 3; 4,952,051 … 4,961,161 bp) gene is associated with immunity and enhances growth performance in broilers [ 112 ].…”

“…. 5,039,077 bp) is a candidate gene related to growth and development at early stages [83]. The PLIN3 (perilipin 3; 4,952,051 .…”