The correctness and efficiency of the adapter connection are important factors affecting the quality and yield of the libraries in next-generation sequencing (NGS). Lengthening the sticky ends of the adapters and the targets can effectively improve the efficiency of ligation, however, it may cause a new trouble of adapter self-ligation. In this study, a strategy based on nickase is designed to limit or abolish long sticky-end adapters self-ligation. We firstly explored the enzymatic characteristics of Nt.BsmAI, a typical nickase. Then based on the working properties of Nt.BsmAI to design Y-adapters for NGS and evaluated the efficiency of this strategy for reducing long sticky-end adapter dimers. Moreover, the feasibility of Nb.BtsI (another nickase) was verified using the same strategy. This study provides new application for nickase in NGS library construction, and the possibility of using multi-base sticky end adapters in the future. NGS adapters with long sticky-ends paves a new avenue for enhancing the application performance of NGS, particularly for methylation sequencing, in biomedicine, with a higher efficiency in library preparation.

Coprophagy is an innate behavior of rabbits, and rabbit meat is favored by consumers for its distinctive nutritional characteristics. However, the relationship between coprophagy and meat quality in rabbits remains unclear. Therefore, this study investigated the effects of coprophagy prevention on growth performance and meat quality using a coprophagy-prevention model. The results showed that coprophagy prevention reduced growth performance and several meat quality parameters while increasing muscle fiber density. In addition, coprophagy prevention altered the nutritional composition of skeletal muscle by reducing unsaturated fatty acids (UFAs) and essential amino acids (EAAs), while increasing total lipid content and the proportion of saturated fatty acids (SFAs). Tandem mass tag (TMT)-based quantitative proteomic analysis identified 267 differentially expressed proteins (DEPs) in skeletal muscle following coprophagy prevention, which were mainly enriched in pathways related to muscle development, energy metabolism, fatty acid transport, and lipid metabolism. Further analyses suggested that alterations in lipid deposition and fatty acid composition may be associated with changes in mitochondrial oxidative phosphorylation and ATP production; however, these mechanistic relationships require further validation. Although several cognition-related pathways and reduced serum dopamine (DA) and 5-hydroxytryptamine (5-HT) levels were observed, the functional significance of these findings remains unclear due to the lack of behavioural assessments. Overall, the present study suggests that coprophagy prevention is associated with impaired growth performance and altered meat quality in rabbits. These findings provide preliminary insights into the potential metabolic changes associated with coprophagy prevention and highlight the biological importance of coprophagy in rabbit production.

The experiment aimed to explore the impact of diarrhea on the intestinal microbiota of calves. Four healthy calves and four diarrhea calves with similar weight at three months of age, were selected and divided into a healthy group (group CK) and a diarrhea group (group FX), with four replicates per group and one calf per replicate. Fresh fecal samples were collected for 16S rRNA sequencing to analyze microbial diversity and structural composition differences. The results showed that the richness index and diversity of fecal microbiota in the group FX were significantly or extremely lower than those in the group CK (P<0.05 or P<0.01). Beta diversity analysis showed there were significant differences in the composition of fecal microbiota between the two groups of calves (P<0.05). At the phylum level, the relative abundance of Firmicutes_A and Bacteroidetes in the group FX were lower than that in the group CK, while the relative abundance of Firmicutes_D was higher than that in the group CK. At the genus level, the relative abundance of Escherichia in the group FX was higher than that in the group CK. Mann-Whitney U test and linear discriminant analysis effect size (LEfSe) were used to screen for important microbial groups in the feces of the group CK, including Faecousia, Cryptobacteroides, Paraprevotella, Spirochaetota, Fibrobacterota, while Corynebacterium and Mycobacteriaceae could be potential biomarkers for the group FX. The study shows that there is a significant imbalance in the gut microbiota of diarrheal calves, and regulating the gut microbiota can help prevent and treat calf diarrhea.

Milk fat is a crucial component for evaluating the production performance and nutritional value of goat milk. Previous research indicated that the composition of ruminal microbiota plays a significant role in regulating milk fat percentage in ruminants. Thus, this study aimed to identify key ruminal microorganisms and blood metabolites relevant to milk fat synthesis in dairy goats as a mean to explore their role in regulating milk fat synthesis. Sixty clinically healthy Xinong Saanen dairy goats at mid-lactation and of similar body weight, and similar milk yield were used in a feeding study for 15 days. Based on daily milk yield of dairy goats and the results of milk component determination on the 1st and 8th days, five goats with the highest milk fat content (H group) and five goats with the lowest milk fat content (L group) were selected for further analysis. Before the morning feeding on the 15th day of the experiment, samples of milk, blood and ruminal fluid were collected for analyses of components, volatile fatty acids, microbiota and metabolites. Results revealed that acetate content in the rumen of H group was greater compared with L group. H group had abundant beneficial bacteria including Ruminococcaceae_UCG-005, Saccharofermentans, Ruminococcaceae-UCG-002 and Prevotellaceae_UCG-3, which were important for plant cellulose and hemicellulose degradation and immune regulation. Metabolomics analysis revealed H group had greater relative concentrations of 4-acetamidobutanoic acid and azelaic acid in serum, and had lower relative concentrations of Arginyl-Alanine, SM(d18:1/12:0) and DL-Tryptophan. These altered metabolites are involved in the sphingolipid signaling pathway, arginine and proline metabolism. Overall, this study identified key ruminal microorganisms and serum metabolites associated with milk fat synthesis in dairy goats. These findings offer insights for enhancing the quality of goat milk and contribute to a better understanding of the regulatory mechanisms involved in milk fat synthesis in dairy goats.

In recent years, consumers have shown a growing interest in purchasing welfare-friendly livestock products. Environmental enrichment has emerged as an effective approach to enhancing animal welfare. Based on this, this study aims to investigate the effects of welfare housing systems incorporating environmental enrichment on growth performance, meat quality, and intestinal health in rabbits. Rabbits were assigned into three groups: Cage group (Cage), Pen group (Pen), and Pen + environmental enrichment material group (PE). The Pen group exhibited a lower average daily weight gain and a higher feed conversion ratio. However, the meat color and nutrient content of the leg muscles were improved in both the Pen and PE groups to varying extents. Furthermore, fatty acid β-oxidation in the leg muscles was elevated in the Pen group, resulting in reduced lipid deposition. Additionally, both the Pen and PE groups displayed altered cecal microbiota composition and increased short-chain fatty acids content, contributing to enhanced intestinal health through the activation of G protein-coupled receptors. In conclusion, these findings suggest that the two welfare housing systems can improve meat quality and intestinal health. This study offers a theoretical foundation for welfare-oriented rabbit farming and the production of healthier rabbit meat to meet consumer demands.

Meat quality is a complex trait affected by genetic and environmental factors. Intramuscular fat (IMF) is one of the most important factors affecting meat quality. However, there is little research focusing on IMF deposition in rabbits. In this study, we compared the meat quality and nutritional composition of New Zealand White rabbits (NZWRs) and Yufeng Yellow rabbits (YFYRs). The results showed that the drip loss of longissimus dorsi muscle (LD muscle) of YFYR was significantly lower than that of NZWR (p < 0.05), and the pH45 of YFYR was significantly higher than that of NZWR (p < 0.001). Compared with NZWR, the content of IMF, C22:6 n3, C16:0, C20:3 n3, and n-3 PUFA in the LD muscle of YFYR was significantly increased (p < 0.05), while the content of C15:0 was significantly decreased (p < 0.05). RNA-Seq and miRNA-Seq of the LD muscle tissue were performed to measure the mRNA and miRNA expressions of both NZWR and YFYR. A total of 1175 differentially expressed genes (DEGs) were identified between the two rabbit species, including 340 upregulated and 835 downregulated mRNAs in NZWR. A total of 138 differentially expressed miRNAs (DE-miRNAs) were identified, including 31 upregulated and 107 downregulated miRNAs in NZWR. Functional enrichment analysis revealed that the target genes of differential genes and miRNAs were enriched in lipid binding, long-chain fatty acid binding, lipid metabolic process, and fatty acid metabolism, as well as the PPAR signaling pathway, glycerol phospholipid metabolism, and other pathways related to lipid deposition. MiR-973-y, ocu-miR-124-3p, ocu-miR-2355-5p, miR-197-y, and ocu-miR-301b-3p are key miRNAs regulating IMF deposition, and GPX7, GPD2, BDH1, PCYT1B, and ACER2 are key target genes related to lipid deposition and metabolism. In addition, correlation analysis suggested that the IMF content was positively correlated with PAFAH2 and HSD11B2 genes (p < 0.05), while negatively correlated with PRKAG2 (p < 0.01). Taken together, this study suggests that YFYR has relatively better meat quality and nutritional value relative to NZWR; the identified DEGs and target miRNAs provide strong evidence for further studying the regulatory mechanism of IMF deposition in rabbits.

The gut microbiota plays a crucial role in host homeostasis, influencing digestion, metabolism, and immune regulation. Neonatal birth weight is pivotal for early development and long-term productivity. The study identified distinct maternal gut microbiota and metabolomic profiles associated with neonatal calf birth weight, providing preliminary biomarkers for future microbiota-driven reproductive strategies. A cohort of periparturient Holstein dairy cows was strategically selected to establish two distinct experimental groups based on neonatal birth weight stratification: a high birth weight cohort (WU) and a low birth weight cohort (WF). WU cows exhibited enrichment of Burkholderiaceae_A, Turicibacter, Romboutsia_B, Blautia_A, Barnesiella, Clostridium_T, and Paraclostridium, while WF cows had higher levels of Elusimicrobiota, Ruminiclostridium_E, Campylobacter_B, and Odoribacter. Metabolomic profiling identified 85 differentially abundant metabolites. WU cows showed higher levels of 4-imidazoleacrylic acid, 1,2,3,4-tetrahydroacridin-9-ol, 8S-hydroxy-4Z,6E,10Z-hexadecatrienoic acid, 7-hydroxy methotrexate, sorbitol 6-phosphate, and captopril. WF cows exhibited elevated d-proline, l-hydroxy arginine, alanine betaine, sn-glycerol 3-phosphoethanolamine, and anthranilic acid. Correlation analysis further revealed strong associations between microbial taxa and key metabolic pathways. In the WU group, the genera Romboutsia_B, Paraclostridium, Clostridium_T, Turicibacter, and Blautia_A were positively correlated with metabolites such as neoabietic acid, 4-imidazoleacrylic acid, 8 s-hydroxy-4Z,6E,10Z-hexadecatrienoic acid, n-acetylcytidine, 7-hydroxymethotrexate, and O-succinyl-l-homoserine. Conversely, these genera were negatively correlated with alanine betaine, l-hydroxy arginine, d-proline, anthranilic acid (Vitamin L1), sn-glycerol 3-phosphoethanolamine, and 1-deoxy-1-(methylamino)-d-galactitol. In the WF group, Campylobacter_B was positively correlated with 1-myristoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine, sn-glycerol 3-phosphoethanolamine, and anthranilic acid (Vitamin L1), and negatively correlated with 4-imidazole acrylic acid and 7-hydroxy methotrexate. Additionally, Ruminiclostridium_E and Odoribacter negatively correlated with 7-hydroxy methotrexate and captopril. These findings suggest a coordinated microbiota-metabolite interplay influencing fetal development. Identifying maternal specific microbial taxa and metabolic pathways associated with higher-birth-weight calves offers potential biomarkers and intervention targets for optimizing neonatal growth and perinatal health management in dairy cattle.

This study systematically evaluated the carcass characteristics and meat quality of the Nanyang yellow cattle (NYC), Pinan cattle (Piedmontese × NYC; PNC) and Denan cattle (German yellow ×NYC; DNC). DNC and PNC exhibited significantly higher carcass weight, lean meat percentage, dressing percentage, and meat-to-bone ratio, while NYC demonstrated superior tenderness and water-holding capacity. NYC had higher saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA), whereas PNC contained more polyunsaturated fatty acids (PUFA) and potassium (K) content. The Mantel correlation analysis demonstrated that essential amino acids (EAA) and flavor amino acids (FAA) levels were closely associated with crude protein content. Furthermore, SFA, PUFA and MUFA exhibited significant relationships with lean meat yield, whereas K and phosphorus (P) were correlated with carcass weight and meat pH. Collectively, Nanyang yellow crossbred cattle could enhance production performance and slaughter performance, providing valuable insights for conservation and genetic improvement of Chinese indigenous cattle.

The pervasive presence and bioaccumulation of lead (Pb2 +) in living organisms pose a significant risk to human health. Extensive research has been conducted on the neurotoxicity and reproductive toxicity of lead, but the impact of maternal lead exposure on subsequent generations still needs to be addressed. Therefore, we aimed to explore the adverse effects and mechanisms of lead exposure in pregnant mice on the next generation. The present study indicated that lead exposure decreased growth performance and impaired the function of the colon, spleen, testes, and kidneys in the next generation of mice. Furthermore, we found that lead exposure activated autophagy by regulating the AMPK/ULK1 pathway and reduced the relative expression levels of tight junction proteins in a dose-dependent manner. Moreover, the composition of the gut microbiota showed significant alterations compared to the control group. These changes were characterized by a marked reduction in the relative abundance of beneficial bacteria, including Variovorax, Harryflintia, and Romboutsi, while the abundance of pathogenic bacteria such as Mucispirillum and Klebsiella was significantly increased. Such shifts in microbial composition indicated a disruption in microbial homeostasis, contributing to health impairments. In summary, our study indicated that lead exposure in pregnant mice not only decreased the growth performance and impaired multiple organs in the next generation of mice but also disrupted gut microbial homeostasis and induced autophagy via activating the AMPK/ULK1 pathway, resulting in adverse health outcomes in offspring. Additionally, the current study provides a solid experimental foundation for environmental agencies worldwide to implement effective strategies to mitigate lead contamination.

Supplementation of sodium butyrate (SB) has been proved to be beneficial for improving the growth performance and health of animals, but its effects on rabbits have not been well understood. Therefore, this study aimed to investigate the effects of SB supplementation on growth performance, meat quality, and intestinal health of rabbits by evaluating feed intake and efficiency, diarrhea index, blood and cecum metabolites, cecal pH and short-chain fatty acids (SCFAs), histological staining, nutritional composition of meat, and gene expression profile of cecum. A total of 14 weaned male New Zealand White rabbits (35 +/- 2 days, 1.70 +/- 0.09 kg) were randomly divided into two groups: rabbits in Control group were fed normally with the basal diet, and rabbits in Butyrate group were supplemented with 0.75 g/kg SB on the basis of the basal diet. After a 1-week acclimatization period, the formal trial lasted for 10 weeks. Results suggest that dietary SB reduced the feed conversion ratio and diarrhea index of rabbits (p < 0.05). SB treatment significantly increased serum glutamyl transpeptidase, total bile acid, total cholesterol, triglyceride, and glucose levels and decreased tCO(2) content relative to the Control group (p < 0.05). Meanwhile, crude ash content in rabbits' meat was significantly increased by SB treatment (p < 0.05). Furthermore, SB treatment significantly increased the integrity of epithelial villi and number of goblet cells in the cecum and decreased cecal pH compared with the Control group. Dietary SB also upregulated levels of Na+-K+ ATPase, Ca2+-Mg2+ ATPase, total superoxide dismutase, and glutathione peroxidase in cecum. SCFAs analysis revealed that dietary supplementation with SB increases butyric acid in the cecum contents. Furthermore, SB supplementation is demonstrated to stimulate the proliferation and migration of cecal epithelial cells through activating Wnt/beta-catenin pathways in rabbits both in vitro and in vivo by using transcriptome sequencing and gene function verification technologies. Taken together, our findings provide a theoretical basis for the application of SB as an alternative to antibiotics in livestock production.

More sensitive evaluation of the off-target effects of gene editing nucleases is crucial for human gene therapy. Here we report chromogenic assays designed for sensitive evaluation of gene editing activities using CRISPR/Cas9 test system. Based on beta-galactosidase alpha complementation, qualitative and quantitative evaluations of the target and off-target effects of CRISPR/Cas9 were well established through the color alteration of the E.coli colonies. In addition to target effect analysis, these new assays provide extremely sensitive and efficient tool to profile the off-target effects with one or more bases mismatched between the targets and the gRNAs. Moreover, these assays allow the identification of gene editing effects for off-targets with one base mismatched PAM sites.

As bacteria synthesize nutrients primarily in the cecum, coprophagy is indispensable for supplying rabbits with essential nutrients. Recent research has demonstrated its pivotal role in maintaining intestinal microbiota homeostasis and immune regulation in rabbits, although the specific mechanism remains unknown. Here, we used coprophagy prevention (CP) to investigate the effects of coprophagy on the cecum homeostasis and microbiota in New Zealand white rabbits. Furthermore, whether supplementation of Clostridium butyricum (C. butyricum) may alleviate the cecum inflammation and apoptosis caused by CP was also explored. Four groups were randomly assigned: control (Con), sham-coprophagy prevention (SCP), coprophagy prevention (CP), and CP and C. butyricum addition (CPCB). Compared to Con and SCP, CP augmented cecum inflammation and apoptosis, as well as bacterial adhesion to the cecal epithelial mucosa, while decreasing the expression of tight junction proteins (ZO-1, occluding, and claudin-1). The relative abundance of short-chain fatty acids (SCFAs)-producing bacteria was significantly decreased in the CP group. Inversely, there was an increase in the Firmicutes/Bacteroidetes ratio and the relative abundance of Christensenellaceae_R-7_group. Additionally, CP increased the levels of Flagellin, IFN-γ, TNF-a, and IL-1β in cecum contents and promoted the expression of TLR5/MyD88/NF-κB pathway in cecum tissues. However, the CPCB group showed significant improvements in all parameters compared to the CP group. Dietary C. butyricum supplementation significantly increased the production of SCFAs, particularly butyric acid, triggering anti-inflammatory, tissue repairing, and barrier-protective responses. Notably, CPCB effectively mitigated CP-induced apoptosis and inflammation. In summary, CP disrupts the cecum epithelial barrier and induces inflammation in New Zealand white rabbits, but these effects can be alleviated by C. butyricum supplementation. This process appears to be largely associated with the TLR5/MyD88/NF-κB signaling pathway.

Alternative splicing is a ubiquitous regulatory mechanism in gene expression that allows a single gene to generate multiple messenger RNAs (mRNAs). Adipocyte development is regulated by many processes, and recent studies have found that splicing factors also play an important role in adipogenic development. In the present study, we further investigated the differences in selective shearing during different periods of adipocyte differentiation. We identified five alternative splicing types including skipped exon, mutually exclusive exon, Alternative 5′ splice site, Alternative 3′ splice site, and Retained intron, with skipped exons being the most abundant type of selective shearing. In total, 641 differentially expressed selective shearing genes were obtained, enriched in 279 pathways, from which we selected and verified the accuracy of the sequencing results. Overall, RNA-seq revealed changes in the splicing and expression levels of these new candidate genes between precursor adipocytes and adipocytes, suggesting that they may be involved in adipocyte generation and differentiation.

Long non-coding RNAs (lncRNAs) have been shown to be involved in the regulation of skeletal muscle development through multiple mechanisms. The present study revealed that the lncRNA SOX6 AU (SRY-box transcription factor 6 antisense upstream) is reverse transcribed from upstream of the bovine sex-determining region Y (SRY)-related high-mobility-group box 6 (SOX6) gene. SOX6 AU was significantly differentially expressed in muscle tissue among different developmental stages in Xianan cattle. Subsequently, knockdown and overexpression experiments discovered that SOX6 AU promoted primary skeletal muscle cells proliferation, apoptosis, and differentiation in bovine. The overexpression of SOX6 AU in bovine primary skeletal muscle cells resulted in 483 differentially expressed genes (DEGs), including 224 upregulated DEGs and 259 downregulated DEGs. GO functional annotation analysis showed that muscle development-related biological processes such as muscle structure development and muscle cell proliferation were significantly enriched. KEGG pathway analysis revealed that the PI3K/AKT and MAPK signaling pathways were important pathways for DEG enrichment. Notably, we found that SOX6 AU inhibited the mRNA and protein expression levels of the SOX6 gene. Moreover, knockdown of the SOX6 gene promoted the proliferation and apoptosis of bovine primary skeletal muscle cells. Finally, we showed that SOX6 AU promoted the proliferation and apoptosis of bovine primary skeletal muscle cells by cis-modulation of SOX6 in cattle. This work illustrates our discovery of the molecular mechanisms underlying the regulation of SOX6 AU in the development of beef.

Intestinal microbial community plays an important part in maintaining health and skeletal muscle development in livestock. This study is the first of its kind in the world. In order to better understand the relationship between gut microbiota and gene expression in skeletal muscle of rabbits, caecum contents and longissimus dorsi tissues of rabbits at 0 d (S1), 35 d (S2) and 70d (S3) were collected and subjected for 16S rRNA sequencing and transcriptome sequencing. Our results showed that, among three groups of rabbits, Firmicutes and Bacteroidetes were the dominant phyla at the phylum level, while Akmansia, Bacteroides and Ruminobacter were the dominant genera at the genus level, and the relative abundance of Akmansia and Bacteroides increased firstly and then decreased from 0 d to 70 d. By analyzing the transcriptome sequencing data, we identified 2866, 2446 and 4541 differentially expressed genes (DEGs) in S1 vs S2, S2 vs S3 and S1 vs S3 groups, respectively. Finally, we performed correlation analysis between gut microbiota and the expression levels of muscle development-related genes of rabbits at 0 d and 70 d. Compared with 0 day old rabbits, in 70 day old rabbits Acinetobacter and Cronbacter with decreased abundance, and Ruminococcaceae_UCG-014 and Ruminococcus_1 with increase abundance is beneficial to caecum health in rabbits. These results will lay a foundation for further re-searches about the relationship between caecum microflora and muscle development in rabbits.

Previous researches revealed a copy number variation (CNV) region in the bovine fibroblast growth factor 13 (FGF13) gene. However, its effects remain unknown. This study detected the various copy number types in seven Chinese cattle breeds and analysed their population genetic characteristics and effects on growth traits and transcription levels. Copy number Loss was more frequent in Caoyuan Red cattle and Xianan cattle than in the other breeds. Association analysis between CNV and growth traits of Qinchuan indicated that the CNV was significantly related to chest depth, hip width and hucklebone width (P < 0.05). Additionally, the growth traits of individuals with copy number Loss were significantly inferior to those with copy number Gain or Median (P < 0.05). Besides, we found two splicing isoforms, AS1 and AS2, in FGF13 gene, which resulted from alternative 5′ splicing sites of intron 1. These isoforms showed varied expression levels in various tissues. Moreover, CNV was significantly and negatively associated with the mRNA expression of AS1 (r = −0.525, P < 0.05). The CNVs in bovine FGF13 gene negatively regulated growth traits and gene transcription. These observations provide new insights into bovine FGF13 gene, delivering potentially useful information for future Chinese cattle breeding programs.

The Hippo-YAP signaling pathway plays a central role in many biological processes such as regulating cell fate, organ size, and tissue growth, and its key components are spatiotemporally expressed and posttranslationally modified during these processes. Neddylation is a posttranslational modification that involves the covalent attachment of NEDD8 to target proteins by NEDD8-specific E1-E2-E3 enzymes. Whether neddylation is involved in Hippo-YAP signaling remains poorly understood. Here, we provide evidence supporting the critical role of NEDD8 in facilitating the Hippo-YAP signaling pathway by mediating neddylation of the transcriptional coactivator yes-associated protein 1 (YAP1). Overexpression of NEDD8 induces YAP1 neddylation and enhances YAP1 transactivity, but inhibition of neddylation suppresses YAP1 transactivity and attenuates YAP1 nuclear accumulation. Furthermore, inhibition of YAP1 signaling promotes MLN4924-induced ovarian granulosa cells apoptosis and disruption of nedd8 in zebrafish results in downregulation of yap1-activated genes and upregulation of yap1-repressed genes. Further assays show that the xiap ligase promotes nedd8 conjugates to yap1 and that yap1 neddylation. In addition, we identify lysine 159 as a major neddylation site on YAP1. These findings reveal a novel mechanism for neddylation in the regulation of Hippo-YAP signaling.

Background Neddylation, an important post-translational modification (PTM) of proteins, plays a crucial role in follicular development. MLN4924 is a small-molecule inhibitor of the neddylation-activating enzyme (NAE) that regulates various biological processes. However, the regulatory mechanisms of neddylation in rabbit ovarian cells have not been emphasized. Here, the transcriptome and metabolome profiles in granulosa cells (GCs) treated with MLN4924 were utilized to identify differentially expressed genes, followed by pathway analysis to precisely define the altered metabolisms. Results The results showed that 563 upregulated and 910 downregulated differentially expressed genes (DEGs) were mainly enriched in pathways related to cancer, cell cycle, PI3K-AKT, progesterone-mediated oocyte maturation, and PPAR signaling pathway. Furthermore, we characterized that MLN4924 inhibits PPAR-mediated lipid metabolism, and disrupts the cell cycle by promoting the apoptosis and proliferation of GCs. Importantly, we found the reduction of several metabolites in the MLN4924 treated GCs, including glycerophosphocholine, arachidic acid, and palmitic acid, which was consistent with the deregulation of PPAR signaling pathways. Furthermore, the increased metabolites included 6-Deoxy-6-sulfo-D-glucono-1,5-lactone and N-Acetyl-D-glucosaminyldiphosphodolichol. Combined with transcriptome data analyses, we identified genes that strongly correlate with metabolic dysregulation, particularly those related to glucose and lipid metabolism. Therefore, neddylation inhibition may disrupt the energy metabolism of GCs. Conclusions These results provide a foundation for in-depth research into the role and molecular mechanism of neddylation in ovary development.

Research on adipogenesis will help to improve the meat quality of livestock. Long noncoding RNAs (lncRNAs) are involved in mammalian adipogenesis as epigenetic modulators. In this study, we analyzed lncRNA expression during bovine adipogenesis and detected 195 differentially expressed lncRNAs, including lncRNA BlncAD1, which was significantly upregulated in mature bovine adipocytes. Gain- and loss-of-function experiments confirmed that BlncAD1 promoted the proliferation, apoptosis, and differentiation of bovine preadipocytes. RNA pull-down revealed that the nonmuscle myosin 10 (MYH10) is a potential binding protein of BlncAD1. Then, we elucidated that loss of BlncAD1 caused increased ubiquitination of MYH10, which confirmed that BlncAD1 regulates adipogenesis by enhancing the stability of the MYH10 protein. Western blotting was used to demonstrate that BlncAD1 activated the PI3K/Akt signaling pathway. Bioinformatic analysis and dual-luciferase reporter assays indicated that BlncAD1 competitively absorbed miR-27a-5p. The overexpression and interference of miR-27a-5p in bovine preadipocytes displayed that miR-27a-5p inhibited proliferation, apoptosis, and differentiation. Further results suggested that miR-27a-5p targeted the CDK6 gene and that BlncAD1 controlled the proliferation of bovine preadipocytes by modulating the miR-27a-5p/CDK6 axis. This study revealed the complex mechanisms of BlncAD1 underlying bovine adipogenesis for the first time, which would provide useful information for genetics and breeding improvement of Chinese beef cattle.

Acetic acid, which is one of the most abundant short-chain fatty acids (SCFA) in rabbits' cecum, has been reported to play an important function during various physiological metabolic processes. The present study was conducted to elucidate the effects of sodium acetate on growth performance and intestinal health by evaluating feed intake and efficiency, diarrhea score, serum and cecum metabolites, cecal pH and SCFA, histological staining, nutritional composition of meat and gene expression profile of cecum in rabbits. As a result of sodium acetate supplement, the feed conversion ratio, diarrhea score, and diameter of muscle fiber were significantly decreased (P < 0.05). Additionally, dietary sodium acetate significantly increased in total area of muscle fibers and content of crude ash (P < 0.05). Dietary sodium acetate significantly increased serum glucose, total bile acid, and total cholesterol levels and decreased amylase, lipase, and tCO(2) content (P < 0.05). Further examination suggested that sodium acetate supplementation enhanced the micro-environment of cecum, evidenced by significantly increased levels of total antioxidant capacity, total superoxide dismutase, and glutathione peroxidase, and decreased pH and amylase levels (P < 0.05). According to transcriptome sequencing of cecal tissues, differentially expressed genes were predominantly enriched in cell cycle, ABC transporters, and chemokine signaling pathways. Sodium acetate was further suggested to stimulate the proliferation and migration of rabbits' cecum epithelial cells by activating Wnt/beta-catenin pathway both in vivo and in vitro. In conclusion, dietary sodium acetate supplementation improved growth performance and intestinal health in rabbits.