Puerarin has the potential of regulating the differentiation of preadipocytes, but its mechanism of action has not yet been elucidated. Adipocytes found in adipose tissue, the main endocrine organ, are the main sites of lipid deposition, and are widely used as a cell model in the study of in vitro fat deposition. This study aimed to investigate the effects of puerarin on adipogenesis in vitro. Puerarin was added to the culture medium during the process of adipogenesis. The proliferation and differentiation of bovine preadipocytes was measured through cell viability and staining with Oil Red O. The content of triacylglycerol (TG) was measured using a triglyceride assay kit. The mRNA and protein expression levels of adipogenic genes, peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer-binding protein-α (C/EBPα), were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, respectively. The addition of puerarin significantly increased adipogenesis of bovine preadipocytes and enhanced the mRNA and protein level expression of PPARγ (p<0.01). The expression of P-Akt increased after adipogenic hormonal induction, whereas puerarin significantly increased PPARγ expression by promoting the Akt signaling component, P-Akt. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473, which may activate the downstream signaling of the Akt pathway. Puerarin was able to promote the differentiation of preadipocytes and improve fat deposition in cattle. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473.

Histone deacetylase 9 (HDAC9) is a histone deacetylase (HDAC) subtype IIa protein that deacetylates histone 3 (H3), histone 4 (H4), and nonhistone proteins in vivo to alter chromosomal shape and regulate gene transcription. There have been few studies on the regulatory influence of the HDAC9 gene on the differentiation of chicken embryonic stem cells (cESCs) into male germ cells, and the significance of HDAC9 is still unknown. Therefore, we explored the specific role of HDAC9 during differentiation of the cESCs of Jilin Luhua chickens through inhibition or overexpression. In medium supplemented with 10^-5 mol/L retinoic acid (RA), cESCs were stimulated to develop into germ cells. HDAC9 and germline marker gene mRNA and protein levels were measured using qRT‒PCR and western blotting. During the differentiation of cESCs into male germ cells, overexpression of the HDAC9 gene greatly increased the mRNA and protein expression levels of the germline marker genes Stra8, Dazl, c-kit, and integrin ɑ6. The HDAC9 inhibitor TMP195 significantly decreased the mRNA and protein expression levels of the above markers. In summary, HDAC9 positively regulates the differentiation of cESCs.

Intramuscular fat (IMF) deposition is the primary economic driver of premium beef production, yet conventional phenotypic selection requires costly rearing to slaughter age. Although whole-genome sequencing has repeatedly flagged the KIRREL3 region on BTA29 as a candidate interval for meat quality in beef cattle, whether intronic polymorphisms within this gene are directly associated with carcass or meat-quality traits--and whether such marker effects remain stable across physiological stages--has not been validated. To address these gaps, we genotyped two intronic SNPs in KIRREL3 (g.30324329C>T, 63-bp site; g.30324473C>T, 203-bp site) in two age cohorts of Woking heifers (n = 226 at 36 months; n = 206 at 48 months), together with an independent validation cohort (n = 41) for fatty-acid profiling, meat-quality phenotyping, and multi-tissue expression analysis. Both loci were moderately polymorphic (PIC = 0.37) and in Hardy-Weinberg equilibrium. At 36 months, the TT genotype was associated with significantly higher dressing percentage, rib thickness, and meat grade than CC at both loci. By 48 months, associations at the 63-bp site persisted and the T allele showed complete dominance (CT and TT both exceeded CC in grade and rib thickness), whereas associations at the 203-bp site dissipated with age, showing no significant effect on intermuscular fat thickness or grade. In the validation cohort, palmitic acid (C16:0), linolenic acid (C18:3n3), and arachidonic acid (C20:4) followed a CC > TC >= TT trend (P < 0.05). Notably, CC animals displayed higher IMF and muscle KIRREL3 mRNA levels than TT animals, yet received lower meat grades--a pattern that awaits functional clarification. These findings identify KIRREL3 g.30324329C>T as a robust age-stable marker, providing a concrete target for early marker-assisted selection in Woking cattle.

Intramuscular fat (IMF) deposition drives premium beef value, but phenotypic selection is costly and slow. Although genome-wide studies have linked the KIRREL3 region to meat quality, whether intronic variants directly affect carcass traits and retain predictive power across physiological stages remains unproven. We genotyped two intronic SNPs (g.29756010C>T and g.29756149C>T) in Woking cattle, a high-quality beef breed developed in Jilin Province, China (n = 473; 226 and 206 heifers at 36 and 48 months, respectively, plus an independent validation cohort of n = 41), assessed for fatty acid composition, meat quality, and multi-tissue gene expression. Both loci were polymorphic and in Hardy–Weinberg equilibrium. At 36 months, TT individuals showed higher dressing percentage, rib thickness, and meat grade than CC at both loci. By 48 months, the 63-bp locus retained broader associations with carcass traits, whereas the 203-bp locus effects weakened. In the validation group, CC animals had higher IMF and muscle KIRREL3 expression than TT, yet received lower meat grades. Five fatty acids differed significantly among genotypes after Benjamini–Hochberg correction (q < 0.05), whereas the dominant fatty acids (C16:0, C18:1n9c, C18:0, C18:2n6c) showed no proportional differences. KIRREL3 was most highly expressed in the kidney among eight tissues examined. These findings identify the 63-bp site as a candidate marker for meat quality in local Chinese cattle, with potential for marker-assisted selection pending confirmation in larger cohorts and functional studies.