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Comparative transcriptome analysis of Qinchuan and Wagyu cattle reveals lnc11599 as a negative regulator of intramuscular fat deposition.

BACKGROUND: Intramuscular fat (IMF) content is a critical factor determining beef quality, influenced by various factors including breed and age. However, the regulatory role of long non-coding RNAs (lncRNAs) in IMF deposition remains unclear. METHODS: This study investigated IMF deposition in the longissimus dorsi muscle of one- and two-year-old Qinchuan and Wagyu cattle through histological examination and fat content measurement. Based on transcriptome sequencing data of intramuscular fat tissue, differential expression analysis and weighted gene co-expression network analysis (WGCNA) were performed to identify lncRNAs associated with IMF deposition. The effects of a key candidate lncRNA on the adipogenic differentiation of cattle intramuscular preadipocytes were further examined. RESULTS: Results showed that Wagyu cattle exhibited stronger IMF deposition capacity than Qinchuan cattle across all age groups, with IMF content increasing with age in both breeds. We identified 7,910 lncRNAs from intramuscular fat tissue transcriptome data, including 6,455 novel lncRNAs. Through integrated differential expression analysis and WGCNA, 88 lncRNAs closely associated with IMF deposition were screened from two-year-old Qinchuan and Wagyu cattle. Notably, lnc11599 was significantly upregulated in Qinchuan cattle intramuscular fat tissue, but its expression decreased during intramuscular preadipocyte differentiation. Functional experiments demonstrated that lnc11599 knockdown enhanced adipogenic differentiation capacity, manifested as a highly significant increase in lipid accumulation, upregulation of key adipogenic genes at the mRNA level, together with increases in total fatty acid content and unsaturated fatty acid proportion. CONCLUSIONS: This study established the lncRNA expression profiles in intramuscular fat tissue of Qinchuan and Wagyu cattle across different developmental stages, and demonstrated that lnc11599 acts as a negative regulator of intramuscular fat deposition. These findings provide new directions for elucidating the mechanisms of cattle IMF deposition and offer potential targets for genetic improvement of beef quality.

Animals↗

DGAT1, a new positional and functional candidate gene for intramuscular fat deposition in cattle.

Intramuscular fat content, also assessed as marbling of meat, represents an important beef quality trait. Recent work has mapped a quantitative trait locus (QTL) with an effect on marbling to the centromeric region of bovine chromosome 14, with the gene encoding thyroglobulin (TG) being proposed as a positional and functional candidate gene for this QTL. Recently, the gene encoding diacylglycerol O-acyltransferase (DGAT1), which also has been mapped within the region of the marbling QTL, has been demonstrated to affect the fat content of milk. In the present study, the effects of a 5'-polymorphism of TG and of a lysine/alanine polymorphism of DGAT1 on the fat content of musculus (m.) semitendinosus and m. longissimus dorsi in 55 bovine animals (28 German Holstein and 27 Charolais) has been investigated. Significant effects were found for both candidate genes in both the breeds. These effects seem to be independent of one another because the alleles of the two polymorphisms showed no statistically significant disequilibrium. The DGAT1 effect is mainly on the m. semitendinosus. The TG polymorphism only affects m. longissimus dorsi. However, both intramuscular fat enhancing effects seem to be recessive. The possibility of two linked loci, acting recessively on intramuscular fat content, will require special strategies when selecting for higher marbling scores.

Acyltransferases↗

Functional characterization of lncIMF_17214 in regulating intramuscular fat deposition of yellow-feathered broilers.

Intramuscular fat (IMF) content and lipid composition are key determinants of both the nutritional value and sensory attributes of poultry meat, yet the underlying regulatory mechanisms remain insufficiently elucidated. In this study, triglyceride (TG) content was employed as a quantitative phenotypic proxy to dissect the molecular basis of IMF deposition in yellow-feathered broilers. By integrating TG phenotypic data from 315 individuals with transcriptomic profiles and whole-genome resequencing datasets, a TG-associated long noncoding RNA (lncRNA), lncIMF_17214, was identified. Functional characterization revealed that lncIMF_17214 functions as a negative regulator of lipid deposition. Specifically, its knockdown led to significant increases in TG and total cholesterol concentrations, promoted lipid droplet accumulation, and decreased shear force in breast muscle, whereas its overexpression elicited the opposite effects. Mechanistically, lncIMF_17214 interacts with the RNA-binding protein CNBP, forming a regulatory complex that inhibits lipid accumulation. Furthermore, liver-directed overexpression increased the abundance of lncIMF_17214 in plasma exosomes, while liver-directed manipulation was associated with changes in hepatic and breast-muscle lipid deposition; direct exosome-mediated transfer to intramuscular adipocytes remains to be established. Transcriptomic profiling coupled with pathway enrichment analyses demonstrated that lncIMF_17214 predominantly influences steroid biosynthesis, unsaturated fatty acid metabolism, and peroxisome proliferator-activated receptor (PPAR) signaling pathways. This suggests that it may be involved in the regulation of these pathways, although the underlying molecular mechanisms remain to be further elucidated. Collectively, these findings define a lncIMF_17214-centered regulatory axis linking intracellular and systemic lipid metabolism and provide a robust molecular framework for the targeted improvement of meat quality traits in yellow-feathered broilers.

Breast muscle↗

Relationship among GeneSTAR marbling marker, intramuscular fat deposition, and expected progeny differences in early weaned Simmental steers.

Research has demonstrated that triiodothyronine and thyroxin are correlated with marbling (MARB) deposition in Wagyu cattle. Polymorphisms in the 5' region of the thyroglobulin gene have been associated with an improvement in overall fattening and could be used as a gene marker for MARB. The commercially available GeneSTAR MARB test measures the specific thyroglobulin gene polymorphism and identifies cattle as having 0, 1, or 2 copies of the allele; these are identified as 0-STAR, 1-STAR, or 2-STAR, respectfully. Early weaned Simmental steers (n = 192) of known genetics were individually fed over a repeated 4-yr trial period to determine the correlations between GeneSTAR MARB test [Genetic Solutions/Bovigen Pty. Ltd. (Australia) in conjunction with Frontier Beef Systems, LLC (Louisville, CO)] results and intramuscular fat (IMF) deposition. Yearling weight, MARB, percent retail cuts, and carcass weight EPD were calculated for each steer. Steers were weaned at 88.0 +/- 1.1 d, pen-fed a high-concentrate diet for 84.5 +/- 0.4 d before allotment, and subsequently individually fed a 90% concentrate diet composed primarily of cracked corn and corn silage for 249.7 +/- 0.7 d. Steers were slaughtered at 423.3 +/- 1.4 d. Deoxyribonucleic acid samples were used by Genetic Solutions/Bovigen (Australia) for GeneSTAR MARB analysis. Steers with allele types of 0-STAR (n = 47), 1-STAR (n = 95), and 2-STAR (n = 33) had no effect (P > 0.10) on MARB score, chemically determined IMF percentage, quality grade, or percent low Choice and better. There were no differences (P > 0.10) in performance or other carcass parameters among the allele types. GeneSTAR results were not associated with MARB (P > 0.10). Conversely, MARB EPD was correlated (P < 0.01) with MARB score (r = 0.44) and IMF percentage (r = 0.27). Thus, in this management system, MARB EPD is an accurate predictor of IMF deposition. These data suggest that the GeneSTAR MARB marker was not an efficacious predictor of IMF deposition in early weaned Simmental steers fed a high-energy diet.

Adipose Tissue↗

Effects of different grazing and feeding periods on performance and carcass traits of beef steers.

Over three consecutive years, 180 (60/yr) fall-born steer calves were weaned in May (average initial BW = 238 kg, SD = 36.2 kg) and allocated to one of three groups: 1) calf-fed steers that entered the feedlot at weaning; 2) short yearlings that grazed irrigated pasture for another 4 mo and entered the feedlot in September; and 3) long yearlings that grazed with short yearlings during the summer, remained on annual California foothills range through the fall, winter, and spring, and entered the feedlot the following May. All steers were fed until the average group backfat (BF), determined by ultrasound, reached 11 to 12 mm. On pasture, short- and long-yearling steers gained weight in the summer; long yearlings then slightly lost weight in the fall and winter, and then gained weight again the following spring. Average days in the feedlot were 188, 158, and 94 (P < 0.10) for calves, short yearlings, and long yearlings, respectively. Feedlot DMI increased with age (and weight) at feedlot entry, with no difference among groups in gain:feed ratio. The gain of BF was nil on pasture, even when animals were gaining weight, and then increased rapidly when animals were placed on a high-energy diet. Final body weights were heaviest (P < 0.10) in long yearlings, followed by short yearlings and then calves, indicating that a prolonged growing period increases the apparent mature size of the animal. Moreover, total carcass fat contents and percentage of Choice or above were all lower (P < 0.10) in cattle that were older at feedlot entry (i.e., long yearlings) compared with the other groups. In conclusion, increasing the backgrounding period decreased time and total concentrate requirements in the feedlot of Angus-Hereford steers. Older cattle reached 10 mm of BF at heavier weights. Grazing animals gained weight without increasing BF; however, BF increased rapidly in the feedlot. Prolonged grazing may decrease quality grade, either by impairing the ability of the animal to deposit intramuscular fat or by decreasing the time during which dietary energy supply is adequate for intramuscular fat deposition to occur.

Adipose Tissue↗

Effects of a dexamethasone implant on deposition of intramuscular fat in genetically identical cattle.

Administration of exogenous glucocorticoids has been shown to enhance deposition of intramuscular fat in cattle. In this study, six pairs of genetically identical (cloned) steers were used to determine the effects of an implant containing 100 mg of dexamethasone on intramuscular fat deposition and on other carcass and meat quality traits. One steer in each pair served as a negative control (CON), whereas the other steer received a dexamethasone implant 30 to 60 d from slaughter (DEX). Treatment with dexamethasone did not (P > .05) enhance intramuscular fat deposition. However, DEX steers had greater (.33 cm; P < .05) thickness of external fat, larger (5.94 cm2; P < .05) longissimus muscle areas, and higher (1.9 percentage points; P < .05) dressing percentages than did CON steers. In addition, treatment with dexamethasone increased (P < .01) serum insulin concentrations. Dexamethasone had no effect (P > .05) on cooked beef palatability traits or on 24-h calpastatin activity. Although the DEX implant tested in this study was not effective for improving carcass quality grade, use of the implant increased external fat thickness and was effective for increasing carcass muscularity and dressing percentage.

Adipose Tissue↗

Exploration of genes showing intramuscular fat deposition-associated expression changes in musculus longissimus muscle.

Marbling, as defined by the amount of intramuscular fat, is an economically important trait in beef cattle. Intramuscular fat deposition is postulated to arise mainly from a series of adipogenic events in intramuscular adipocyte-lineage cells and in the physiological or anatomical milieux surrounding them. This study was designed to investigate gene-expression patterns associated with fat deposition in musculus longissimus muscle, including adipocyte-lineage cells and part of the milieux. Differential-display PCR (ddPCR) was used to examine expression differences between low-marbled and high-marbled steer groups at 8, 10, 12 and 14 months of age, encompassing the time that marbling starts to appear. Seventy-four of 2114 total bands on ddPCR gel-bands were significant (P < 0.05) for the group effect, the interaction effect between group and age, or both the group and the interaction effects. Sequence analysis of 72 of these bands revealed 77 genes, including 35 annotated genes and 42 novel sequences. Among the 35 annotated genes, 6 (BTG2, PDHB, SORBS1, TRDN, TTN and MGP) have been related to changes in intramuscular fat deposition, possibly by exerting effects on adipocyte-lineage cells or on the milieux surrounding them.

Adipose Tissue↗

FOS Regulates Myogenic and Adipogenic Differentiation via Extracellular Matrix Signaling.

In the livestock industry, intramuscular fat deposition is a key factor influencing meat tenderness and flavor. Although FOS (Fos proto-oncogene, AP-1 transcription factor subunit) has been implicated in the regulation of cell proliferation and differentiation, its differential roles in myogenic and adipogenic regulation remain unclear. In this study, we revealed that FOS markedly enhanced myogenic differentiation while inhibiting adipogenic differentiation in muscle stem cells, indicating that it exerts distinct effects on muscle development and intramuscular fat deposition. Mechanistically, FOS modulated extracellular matrix signaling by regulating FAK and PXN phosphorylation, acting as a molecular regulator between the muscle and fat lineages. Furthermore, exon SNPs in FOS were associated with slaughter weight and backfat thickness, and the mutant genotypes weakened its antiadipogenic effect. Collectively, these findings suggest that FOS is an important regulator of myogenic and adipogenic differentiation and is a potential candidate gene for the genetic improvement of meat quality traits.

Animals↗

Fibrolipomatous hamartoma: MR imaging findings.

OBJECTIVE: To analyze the MR imaging features of fibrolipomatous hamartoma (FLH) of nerves. DESIGN AND PATIENTS: MR imaging studies from six patients (three men and three women) were retrospectively reviewed by three musculoskeletal radiologists. In four patients, a biopsy of the nerve lesion was performed. In two patients, biopsy data were unavailable and the diagnosis was based on the clinical history combined with the MR imaging findings. RESULTS AND CONCLUSION: MR imaging demonstrated fusiform nerve enlargement that was caused by fatty proliferation and thickening of nerve bundles. Nerve bundles appeared as serpentine tubular structures, hypointense on both T1- and T2-weighted images. The degree of fatty proliferation varied among patients. In addition, significant variation in the distribution of fat along the course of the nerves was noted. In three patients, FLH followed the branching pattern of the nerves, a characteristic pathologic finding. In two patients, intramuscular fat deposition (biceps and tibialis posterior muscles) was present. MR imaging findings of FLH are typical, allowing a confident diagnosis. The variation of fatty proliferation among patients and involved nerves as well as the tendency of the abnormalities to follow the branching pattern of the nerves is well demonstrated with MR imaging. FLH may present as an isolated nerve lesion, may be associated with intramuscular fat deposition, or may occur as a feature of macrodystrophia lipomatosa (MDL).

Adult↗

A novel nuclear-encoded mitochondrial poly(A) polymerase PAPD1 is a potential candidate gene for the extreme obesity related phenotypes in mammals.

People with obesity, especially extreme obesity, are at risk for many health problems. However, the responsible genes remain unknown in >95% of severe obesity cases. Our previous genome-wide scan of Wagyu x Limousin F2 cattle crosses with extreme phenotypes revealed a molecular marker significantly associated with intramuscular fat deposition. Characterization of this marker showed that it is orthologous to the human gene KIAA1462 located on HSA10p11.23, where a major quantitative trait locus for morbid obesity has been reported. The newly identified mitochondrial poly(A) polymerase associated domain containing 1 (PAPD1) gene, which is located near this marker, is particularly interesting because the polymerase is required for the polyadenylation and stabilization of mammalian mitochondrial mRNAs. In the present study, both cDNA and genomic DNA sequences were annotated for the bovine PAPD1 gene and ten genetic markers were detected in the promoter and exon 1 region. Among seven markers assayed on approximately 250 Wagyu x Limousin F2 animals, two single nucleotide polymorphisms (SNPs) in the promoter region were significantly associated with intramuscular fat (P<0.05). However, there was a significant interaction (P<0.05) between a third SNP, which causes an amino acid change in coding exon 1, and each of these two promoter SNPs on intramuscular fat deposition. In particular, the differences between double heterozygous animals at two polymorphic sites and the slim genotype animals exceeded 2.3 standard deviations for the trait in both cases. Our study provides evidence for a new mechanism--the involvement of compound heterosis in extreme obesity, which warrants further examination.

Adipose Tissue↗

Differential response of obese gene expression from fasting in bovine adipose tissues.

To understand the molecular mechanism for intramuscular fat deposition, the expression of the obese gene was examined in response to fasting. Food deprivation for 48 h induced a decrease in the level of obese mRNA in pooled adipose tissues (abdominal, perirenal, subcutaneous, intermuscular and intramuscular). The expression of obese mRNA was examined for individual adipose tissue from several fat depots. It was highly expressed in perirenal adipose tissue, but fasting did not affect its expression level in this tissue. Moderate levels were detected in subcutaneous and intermuscular adipose tissues, and a fasting-induced decrease in obese mRNA was apparent in these tissues. The expression level of the obese gene in intramuscular adipose tissue was very low and did not respond to fasting.

Adipose Tissue↗

[Comparative study on lipogenic and lipolytic gene expression in intramuscular fat tissue between growing Erhualian and large white pigs].

To investigate the mechanism of difference in intramuscular fat deposition between Erhualian and Large White pigs,single tube relative-quantitative RT-PCR method was used to investigate the development patterns of lipogenic (ACX, LPL, ME) and lipolytic (HSL) gene expression with 18S internal standard control. Sixteen Large White boars and twenty Erhualian boars were selected and raised according to normal nutrition standard respectively. The animals were selected randomly and slaughtered at 15 kg, 40 kg, 60 kg and 90 kg for Large White pigs and at 18 kg, 40 kg, 60 kg, 80 kg and 90 kg for Erhualian pigs respectively; with four animals of each breed at each time. The supraspinatus and semimembranosus muscles were removed for total RNA extraction and longisimus dorsi muscle for intramuscular fat (IMF) analysis using ether extract method. The results showed: (1) The property of IMF between Erhualian and Large White boars was similar during early growing period (before 40 kg) (P > 0.05) ,thereafter, IMF level of Erhualian boars increased dramatically to 4% at 60 kg and over 5% at 90 kg while Large White boars kept steadily at about 2% (P < 0.05); (2) The pattern of lipogenic and lypolytic gene expression was similar between semimembranosus and supraspinatus muscle in each breed; (3) The tendency for LPL and ME mRNA expression coincided with that of IMF development among 20 approximately 60 kg in Erhualian pigs. The results suggest that the development of IMF between 20 kg and 60 kg in Erhualian pigs may play a meaningful role in deposition of IMF,and that the expression of ME and LPL mRNA may contribute to fast sediment of IMF in Erhualian pigs.

Adipose Tissue↗

Growth, carcass characteristics, muscle conjugated linoleic acid (CLA) content, and response to intravenous glucose challenge in high percentage Wagyu, Wagyu x Limousin, and Limousin steers fed sunflower oil-containing diet.

The effect of breed and diet on insulin response to glucose challenge and its relation to intramuscular fat deposition was determined in 36 steers with 12 each of greater than 87% Wagyu (referred to as Wagyu), Wagyu x Limousin, and Limousin breeds. Weaned steers were blocked by weight into heavy, medium, and light calves and placed in six pens with two pens per weight type and with two steers of each breed per pen. Three pens with steers from each weightclass were fed backgrounding and finishing diets for 259 d, while the other three pens were fed the same diets where 6% of the barley grain was replaced with sunflower oil. Prior to initiation of the finishing phase of the study the intravenous glucose tolerance test (VGTIT) was conducted in all steers. Once steers were judged as carrying adequate 12th-rib fat, based on weight and days on feed, they were harvested and graded and samples of the longissimus muscle were procured for determination of fat content and fatty acid composition. Dietary oil improved (P = 0.011; 0.06) ADG and feed conversion efficiency of steers during the latter part of backgrounding and only ADG during early part ofthe finishing period. Generally percent kidney, pelvic, and heart fat was the only adiposity assessment increased (P = 0.003) by dietary oil. The IVGTT results indicated that insulin response to intravenous glucose was lower in Limousin steers than in Wagyu steers. Dietary oil decreased (P = 0.052) fasting plasma insulin concentration in Wagyu steers compared with Limousin steers. The correlation coefficients among the IVGTT measures and intramuscular fat content or marbling score were less than 0.4, and only a negative trend existed between fasting insulin and USDA marbling scores. However, the carcasses of the Wagyu steers graded US Choice, and 66% of the Wagyu carcasses graded US Prime, which were substantially better than the quality grades obtained for the carcasses from the other breed types. Dietary oil did not affect muscle fat content but increased (P = 0.01) conjugated linoleic acid (CLA) concentrations by 339%. Results indicated that IVGTT measures were not appropriate indices of marbling potential in cattle and that dietary oil can enhance CLA content of beef.

Adipose Tissue↗

Molecular characterization and chromosomal mapping of porcine adipose differentiation-related protein (ADRP).

ADRP plays an important role in regulating lipid storage in various cells. We investigated the ADRP gene as a candidate gene for intramuscular fat deposition and marbling traits in pigs. A full-length transcript of porcine ADRP was cloned by RT-PCR and RACE. The porcine ADRP cDNA (1848 bp) contains a 1377-bp open reading frame, encoding a deduced protein of 459 amino acids, which has amino acid sequence identities of 89, 89, 82 and 81% with cattle, human, mouse and rat ADRP genes respectively. The genomic structure and sequence of the porcine ADRP were also analysed using a BAC clone of a Korean native pig. Pig ADRP comprises eight exons spanning approximately 13 kb and is located on chromosome 1 q2.3-q2.7 between microsatellite markers SW2185 and SW974. Several sequence variations were detected from nine different pig breeds. The biological role of this gene and the mapping localization indicated that the porcine ADRP is a possible candidate gene for fat deposition and marbling traits.

Adipose Tissue↗

Feedlot performance, carcass characteristics, hormones, and metabolites in steers actively immunized against growth hormone-releasing factor.

Large-framed Simmental and Charolais steers were actively immunized against growth hormone-releasing factor (GRF) to evaluate the effect on growth, carcass characteristics (especially intramuscular fat deposition), and concentrations of somatotropin (ST) and IGF-I. Primary immunizations of 1.5 mg of GRF-(1-29)-Gly-Gly-Cys-NH2 conjugated to 1.5 mg of human serum albumin (GRFi, n = 12) or 1.5 mg of human serum albumin (HSAi, n = 12) were given at approximately 10 mo of age. Booster immunizations of .5 mg of the appropriate antigen were given at d 49 and 125. Weights of steers administered GRFi were less (P < .05) than those given HSAi at 126 d (34.6 kg) or at 262 d (48.2 kg) after treatment. Carcass weights were 28.2 kg less (P < .01) for GRFi than for HSAi steers. Dry matter intake was not affected by immunization treatment, whereas feed efficiency was reduced in GRFi steers. Marbling scores were higher (P < .05) for HSAi than for GRFi steers but similar percentages (83.3) of both treatments graded Low Choice or higher. Rib sections of GRFi steers contained more fat (31.2 vs 25.0%) and less lean (63.3 vs 68.4%) than those of HSAi steers (P < .05). A breed x treatment interaction was observed for percentage of fat within the trimmed longissimus muscle (P < .05); percentage of fat was similar for Charolais and Simmental steers when immunized against HSAi but was higher for Simmental than for Charolais when immunized against GRFi.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗