The Effect of FATP1 on Adipocyte Differentiation in Qinchuan Beef Cattle

FATP1 plays an important role in the regulation of fatty acid metabolism and lipid accumulation. In this study, we investigated the patterns of FATP1 expression in various tissues obtained from calf and adult Qinchuan cattle, and in differentiating adipocytes. Next, we investigated the effect of FATP1 expression on preadipocyte differentiation in Qinchuan cattle using overexpression and interference assays. We also identified the differentially expressed genes (DEGs) and pathways associated with FATP1 overexpression/interference. Our results reveal that FATP1 was broadly expressed in heart, kidney, muscle, small intestine, large intestine, and perirenal fat tissues. While FATP1 overexpression promoted preadipocyte differentiation, fat deposition, and the expression of several genes involved in fat metabolism, FATP1 interference had the opposite effects on adipocyte differentiation. Following FATP1 overexpression and FATP1 interference in adipocytes, RNA-seq analysis was performed to identify DEGs related to fat metabolism. The DEGs identified include SLPI, STC1, SEMA6A, TNFRSF19, SLN, PTGS2, ADCYP1, FADS2, and SCD. Pathway analysis revealed that the DEGs were enriched in the PPAR signaling pathway, AMPK signal pathway, and Insulin signaling pathway. Our results provide an in-depth understanding of the function and regulation mechanism of FAPT1 in fat metabolism.

Association of SNPs in AKIRIN2, TTN, EDG1 and MYBPC1 genes with growth and carcass traits in Qinchuan cattle

Growth and carcass traits are the main breeding objectives in beef cattle. The aim of this study was to confirm genetic effects of the c.*188G>A SNP of AKIRIN2, the g.231054C>T SNP of TTN, the g.1471620G>T SNP of EDG1 and the g.70014208A>G SNP of MYBPC1 on growth and carcass traits in Chinese Qinchuan (QC) cattle, as well as to compare the frequencies of the well-characterized alleles of these SNPs among six Chinese cattle populations, three Japanese cattle populations, two European cattle populations and one Korean cattle population. In this study, a total of 665 cattle samples were genotyped using MassARRAY and PCR-RFLP. Association analysis explored effects of four SNPs on growth and carcass traits including body length, wither height, hip height, hip width, rump length, chest depth, chest circumference, back fat thickness, ultrasound longissimus muscle area and ultrasound longissimus muscle depth in QC (P<0.05 to P<0.001). The well-characterized A (c.*188G>A), T (g.231054C>T) and T (g.1471620G>T) alleles in Japanese Black cattle were significantly higher than Chinese cattle breeds, on the contrary, the G allele (g.70014208A>G) was markedly higher in Chinese cattle breeds than other cattle breeds. These results suggest that the four SNPs might be useful as a molecular marker for growth-related traits in Chinese QC cattle.

Detection of genetic variation and activity analysis of the promoter region of the cattle tRNA-modified gene <i>TRDMT1</i>

The tRNA modification gene in eukaryotes is relatively conservative. As an important modification gene, the TRDMT1 gene plays an important role in maintaining tRNA structural maintenance and reducing mistranslation of protein translation by methylation of specific tRNA subpopulations. Mouse and zebrafish TRDMT1 knockout experiments indicate that it may mediate growth and development through tRNA modification. However, there are no systematic reports on the function of tRNA-modified genes in livestock. In this study, Qinchuan cattle DNA pool sequencing technology was used. A G>C mutation in the −1223 bp position upstream of the TRDMT1 translation initiator codon was found. At this locus, the dual-luciferase assay indicated that different genotypes cause differences in transcriptional activity (P<0.05). Our experiment detected a natural genetic variation of a tRNA modification gene TRDMT1, which may provide potential natural molecular materials for the study of tRNA modification.

Polymorphism of the PLIN1 gene and its association with body measures and ultrasound carcass traits in Qinchuan beef cattle

The PLIN1 gene produces a phosphorylated protein wrapped in lipid droplets in adipocytes. This phosphorylation assists the mobilization of fat into adipose tissue. The purpose of the experiment was to study the polymorphism of the PLIN1 gene and its relationship with the body and carcass characteristics of Qinchuan cattle to find molecular genetic markers that can be used for breeding. The expression level of the PLIN1 gene was determined in various tissues by qRT-PCR. The results showed that the highest level of PLN1 expression was found in subcutaneous fat, followed by the heart and longissimus muscle, and the lowest level was found in the kidney. Five SNP loci of the PLIN1 gene were identified in 510 Qinchuan cattle, including g.3580T>C (SNP1), g.3898G>A (SNP2), g.8333G>A (SNP3), g.10517T>C (SNP4), and g.10538G>T (SNP5). The results show that SNP1, SNP2, SNP3, and SNP4 were moderately polymorphic (0.25 < PIC < 0.5), while SNP5 was minimally polymorphic (PIC < 0.25). SNP2, SNP3, and SNP5 were within Hardy–Weinberg equilibrium (P > 0.05), but SNP1 and SNP4 were not (P < 0.05). Correlation analysis showed that the five SNPs of the PLIN1 gene were correlated with back-fat depth, intramuscular fat, and chest depth of Qinchuan cattle. The double haplotype H2H4 in Qinchuan beef was associated with body and carcass traits. We conclude that variants mapped within PLIN1 can be used in marker-assisted selection for carcass quality and body traits in breed improvement programs for Qinchuan cattle.

Effect of Expressions and SNPs of Candidate Genes on Intramuscular Fat Content in Qinchuan Cattle

Marbling is characterized by the amount and distribution of intramuscular fat (IMF). The AKIRIN2, TTN, EDG1, and MYBPC1 genes are well-known marbling-related genes, which were first identified in Japanese Black beef cattle. The objectives of this study were to analyze the correlation of the expression levels of these genes in the longissimus muscle (LM) with IMF content, and the associations between the single nucleotide polymorphisms (SNPs) in these genes and IMF content in Chinese Qinchuan cattle (n = 350). The association analyses showed that the g.42041062G>T SNP in the EDG1 gene was significantly associated with IMF content in Qinchuan (p < 0.05). Further, the expressions of the EDG1 and MYBPC1 were up-regulated (p < 0.05) in LM of Qinchuan cattle group with low IMF content. Down-regulations of the AKIRIN2 and TTN genes (p < 0.05 and p < 0.01, respectively) were observed in the Qinchuan cattle group with high IMF content. These results suggest possible effects of the expression levels of selected genes on IMF content in the LM, and the g.42041062G>T SNP in the EDG1 gene might be useful as a molecular marker for IMF content in Qinchuan.