Complement C3a Not C4a Activates Osteoclasts By Regulating the PI3K/PDK1/SGK3 Pathway in Patients with Multiple Myeloma

Objective: Myeloma bone disease (MBD) is the most common complication of multiple myeloma (MM). We found that the serum levels of C3/C4 in MM patients were significantly positive correlated with the severity of bone disease in our previously study. Thus, we conduct detailed studies to explore the effect and potential mechanism of C3a/C4a on osteoclasts in patients with MM. Methods: By adding C3a/C4a to culture system of osteoclasts induced from mononuclear cells in vitro, and the expression of related gene, number and function of osteoclasts were detected. RNA-Seq analysis was used to detect the differentially expressed genes on osteoclasts between the complement group and control group, and the possible signal pathways were analyzed. Quantitative real-time PCR (qRT-PCR), Western blot and pathway inhibitors were used for further validation. R esults: In vitro, the osteoclasts area per view induced by C3a 1μg/ml (52.794±13.027 %) and 10μg/ml (51.797±12.464 %) was significantly increased than control group (0μg/ml) (33.668±8.427 %) (P<0.001; P<0.001), the mRNA relative expression of osteoclasts related genes OSCAR/TRAP/RANKL/Cathepsin K induced by C3a 1μg/ml (median 5.041; 3.726; 1.638; 4.752) and 10μg/ml (median 5.140; 3.702; 2.250; 5.172) was significantly increased than control group (0μg/ml) (median 3.137; 2.004; 0.573; 2.257) (1μg/ml P=0.001; P=0.003; P<0.001; P=0.008; 1μg/ml P<0.001; P=0.019; P<0.001; P=0.002), and the absorption area of osteoclast resorption pit per view induced by C3a 1μg/ml (51.464±11.983 %) and 10μg/ml (50.219±12.067 %) was also significantly increased than control group (0μg/ml) (33.845±8.331 %) (P<0.001; P<0.001) in NDMM patients. There was no difference among the osteoclasts area, relative expression of osteoclasts related genes and absorption area of osteoclast resorption pit between C4a (1μg/ml and 10μg/ml) group and control group (0μg/ml). RNA-Seq was performed on total RNA of osteoclasts induced by C3a in 1μg/ml group and 0μg/ml group of 4 patients with NDMM. There were 184 differentially expressed genes that were detected by RNA-Seq analysis. KEGG Pathway enrichment bubble chart shows C3a may through Phosphoinositide 3-kinase (PI3K) signaling pathways (including PI3K-Akt pathway and AKT-independent signaling pathway) promotes the proliferation of osteoclast. Upregulated differentially expressed genes in this pathway among at least 3 patients with sequencing were validated by qRT-PCR and Western Blot. It was found that the relative expression level of Phosphoinositide dependent kinase-1 (PDK1) / Serum and glucocorticoid inducible protein kinases (SGK3) genes (median 2.078; 4.428) in C3a group (1μg/ml) was significantly higher than control group (0μg/ml) (median 1.336; 1.714) (P<0.001; P=0.001). The relative grayscale levels of PDK1/ P-SGK3 protein (1.785±0.323; 2.190±0.274) in C3a group (1μg/ml) was significantly stronger than control group (0μg/ml) (0.8653±0.588; 0.176±0.152) (P=0.034; P<0.001). Under the action of C3a in patients with NDMM, osteoclasts area per view in SGK inhibitor (EMD638683) 1μM group (39.244±9.089 %) and 10μM group (39.299±9.587 %) significantly reduced than control group (0μM) (54.884±12.837 %) (P<0.001; P<0.001), the relative expression of osteoclast related genes OSCAR/RANKL/TRAP/Cathepsin K in EMD638683 1μM group (median 0.869; 1.097; 0.902; 1.328) and 10μM group (median 0.703; 1.391; 0.843; 1.418) significantly decreased than control group (0μM) (median 2.270; 3.024; 2.208; 3.237) (1μM P=0.015; P=0.002; P=0.003; P=0.015; 10μM P=0.012; P=0.006; P<0.001; P=0.017), and the absorption area per view of osteoclast resorption pit in EMD638683 1μM (35.383±7.794 %) group and 10μM group (32.886±8.993 %) significantly reduced than control group (0μM) (49.358±11.856 %) (P < 0.001; P < 0.001). Conclusions:ComplementC3a activates osteoclasts by regulating the PI3K/PDK1/SGK3 pathway in patients with MM, thus leading to the occurrence of bone diseases. SGK inhibitor has a significant inhibitory effect on osteoclasts in patients with MM. This study provide important evidences for the search for new therapeutic targets and strategies for myeloma bone disease patients. Disclosures No relevant conflicts of interest to declare.

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RNA-Seq Analysis Identifies Differentially Expressed Genes in Subcutaneous Adipose Tissue in Qaidaford Cattle, Cattle-Yak, and Angus Cattle

Animals ◽

10.3390/ani9121077 ◽

2019 ◽

Vol 9 (12) ◽

pp. 1077 ◽

Cited By ~ 2

Author(s):

Chengchuang Song ◽

Yongzhen Huang ◽

Zhaoxin Yang ◽

Yulin Ma ◽

Buren Chaogetu ◽

...

Keyword(s):

Adipose Tissue ◽

Meat Quality ◽

Differentially Expressed Genes ◽

Transcriptome Analysis ◽

Cell Metabolism ◽

Differentially Expressed ◽

Receptor Interaction ◽

Control Group ◽

Rna Seq ◽

Angus Cattle

In the beef industry, fat tissue is closely related to meat quality. In this study, high-throughput RNA sequencing was utilized for adipose tissue transcriptome analysis between cattle-yak, Qaidamford cattle, and Angus cattle. The screening and identification of differentially expressed genes (DEGs) between different breeds of cattle would facilitate cattle breeding. Compared to Angus cattle adipose tissue, a total of 4167 DEGs were identified in cattle-yak adipose tissue and 3269 DEGs were identified in Qaidamford cattle adipose tissue. Considering cattle-yak as a control group, 154 DEGs were identified in Qaidamford cattle adipose tissue. GO analysis indicated the significant enrichment of some DEGs related to lipid metabolism. The KEGG pathway database was also used to map DEGs and revealed that most annotated genes were involved in ECM-receptor interaction and the PI3K-Akt signal pathway, which are closely related to cell metabolism. Eight selected DEGs related to adipose tissue development or metabolism were verified by RT-qPCR, indicating the reliability of the RNA-seq data. The results of this comparative transcriptome analysis of adipose tissue and screening DEGs suggest several candidates for further investigations of meat quality in different cattle breeds.

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Effect of hyperoside on cervical cancer cells and transcriptome analysis of differentially expressed genes

Cancer Cell International ◽

10.1186/s12935-019-0953-4 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 4

Author(s):

Weikang Guo ◽

Hui Yu ◽

Lu Zhang ◽

Xiuwei Chen ◽

Yunduo Liu ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Differentially Expressed Genes ◽

Interaction Network ◽

Differentially Expressed ◽

Control Group ◽

Rna Seq ◽

Network Construction ◽

Target Network ◽

Key Genes

Abstract Background Hyperoside (Hy) is a plant-derived quercetin 3-d-galactoside that exhibits inhibitory activities on various tumor types. The objective of the current study was to explore Hy effects on cervical cancer cell proliferation, and to perform a transcriptome analysis of differentially expressed genes. Methods Cervical cancer HeLa and C-33A cells were cultured and the effect of Hy treatment was determined using the Cell Counting Kit-8 (CCK-8) assay. After calculating the IC50 of Hy in HeLa and C-33A cells, the more sensitive to Hy treatment cell type was selected for RNA-Seq. Differentially expressed genes (DEGs) were identified by comparing gene expression between the Hy and control groups. Candidate genes were determined through DEG analysis, protein interaction network (PPI) construction, PPI module analysis, transcription factor (TF) prediction, TF-target network construction, and survival analysis. Finally, the key candidate genes were verified by RT-qPCR and western blot. Results Hy inhibited HeLa and C33A cell proliferation in a dose- and time-dependent manner, as determined by the CCK-8 assay. Treatment of C-33A cells with 2 mM Hy was selected for the subsequent experiments. Compared with the control group, 754 upregulated and 509 downregulated genes were identified after RNA-Seq. After functional enrichment, 74 gene ontology biological processes and 43 Kyoto Encyclopedia of Genes and Genomes pathways were obtained. According to the protein interaction network (PPI), PPI module analysis, TF-target network construction, and survival analysis, the key genes MYC, CNKN1A, PAX2, TFRC, ACOX2, UNC5B, APBA1, PRKACA, PEAR1, COL12A1, CACNA1G, PEAR1, and CCNA2 were detected. RT-qPCR was performed on the key genes, and Western blot was used to verify C-MYC and TFRC. C-MYC and TFRC expressions were lower and higher than the corresponding values in the control group, respectively, in accordance with the results from the RNA-Seq analysis. Conclusion Hy inhibited HeLa and C-33A cell proliferation through C-MYC gene expression reduction in C-33A cells and TFRC regulation. The results of the current study provide a theoretical basis for Hy treatment of cervical cancer.

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Study on the Effect of Macrophages on Vascular Endothelium in Mice With Different TCM Syndromes of Dyslipidemia and its Biological Basis Based on RNA-Seq Technology

Frontiers in Pharmacology ◽

10.3389/fphar.2021.665635 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jing Chen ◽

Chao Ye ◽

Zheng Yang ◽

Tieshan Wang ◽

Bing Xu ◽

...

Keyword(s):

Quality Assessment ◽

Differentially Expressed Genes ◽

Normal Control ◽

Normal Control Group ◽

Differentially Expressed ◽

Control Group ◽

Biological Processes ◽

Rna Seq ◽

Pathogenic Factors ◽

Protective Processes

Background: “Treating the same disease with different methods” is a Traditional Chinese medicine (TCM) therapeutic concept suggesting that, while patients may be diagnosed with the same disease, they may also have different syndromes that require distinct drug administrations.Objective: This study aimed to identify the differentially expressed genes and related biological processes in dyslipidemia in relation to phlegm–dampness retention (PDR) syndrome and spleen and kidney Yang deficiency (SKYD) syndrome using transcriptomic analysis.Methods: Ten ApoE−/− mice were used for the establishment of dyslipidemic disease–syndrome models via multifactor-hybrid modeling, with five in the PDR group and five in the SKYD group. Additionally, five C57BL/6J mice were employed as a normal control group. Test model-quality aortic endothelial macrophages in mice were screened using flow cytometry. Transcriptomic analysis was performed for macrophages using RNA-Seq.Results: A quality assessment of the disease–syndrome model showed that levels of lipids significantly increased in the PDR and SKYD groups, compared to the normal control group, p < 0.05. Applying, in addition, hematoxylin and eosin staining of aorta, the disease model was also successfully established. A quality assessment of the syndrome models showed that mice in the PDR group presented with typical manifestations of PDR syndrome, and mice in the SKYD group had related manifestations of SKYD syndrome, indicating that the syndrome models were successfully constructed as well. After comparing the differentially expressed gene expressions in macrophages of the dyslipidemic mice with different syndromes, 4,142 genes were identified with statistical significance, p < 0.05. Gene ontology analysis for the differentially expressed genes showed that the biological process of difference between the PDR group and the SKYD group included both adverse and protective processes.Conclusion: The differentially expressed genes between PDR syndrome and SKYD syndrome indicate different biological mechanisms between the onsets of the two syndromes. They have distinctive biological processes, including adverse and protective processes that correspond to the invasion of pathogenic factors into the body and the fight of healthy Qi against pathogenic factors, respectively, according to TCM theory. Our results provide biological evidence for the TCM principle of “treating the same disease with different treatments.”

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Dorsal Striatum Transcriptome Profile Profound Shift in Repeated Aggression Mouse Model Converged to Networks of 12 Transcription Factors after Fighting Deprivation

Genes ◽

10.3390/genes13010021 ◽

2021 ◽

Vol 13 (1) ◽

pp. 21

Author(s):

Vladimir Babenko ◽

Olga Redina ◽

Dmitry Smagin ◽

Irina Kovalenko ◽

Anna Galyamina ◽

...

Keyword(s):

Differentially Expressed Genes ◽

Expression Profile ◽

Transcriptional Repression ◽

Gene Networks ◽

Dorsal Striatum ◽

Differentially Expressed ◽

Control Group ◽

Rna Seq ◽

Withdrawal Effect ◽

Transcriptome Expression

Both aggressive and aggression-deprived (AD) species represent pathologic cases intensely addressed in psychiatry and substance abuse disciplines. Previously, we reported that AD mice displayed a higher aggressive behavior score than the aggressive group, implying the manifestation of a withdrawal effect. We employed an animal model of chronic social conflicts, curated in our lab for more than 30 years. In the study, we pursued the task of evaluating key events in the dorsal striatum transcriptome of aggression experienced mice and AD species compared to controls using RNA-Seq profiling. Aggressive species were subjected to repeated social conflict encounters (fights) with regular positive (winners) experience in the course of 20 consecutive days (A20 group). This led to a profoundly shifted transcriptome expression profile relative to the control group, outlined by more than 1000 differentially expressed genes (DEGs). RNA-Seq cluster analysis revealed that elevated cyclic AMP (cAMP) signaling cascade and associated genes comprising 170 differentially expressed genes (DEGs) in aggressive (A20) species were accompanied by a downturn in the majority of other metabolic/signaling gene networks (839 DEGs) via the activation of transcriptional repressor DEGs. Fourteen days of a consecutive fighting deprivation period (AD group) featured the basic restoration of the normal (control) transcriptome expression profile yielding only 62 DEGs against the control. Notably, we observed a network of 12 coordinated DEG Transcription Factor (TF) activators from 62 DEGs in total that were distinctly altered in AD compared to control group, underlining the distinct transcription programs featuring AD group, partly retained from the aggressive encounters and not restored to normal in 14 days. We found circadian clock TFs among them, reported previously as a withdrawal effect factor. We conclude that the aggressive phenotype selection with positive reward effect (winning) manifests an addiction model featuring a distinct opioid-related withdrawal effect in AD group. Along with reporting profound transcriptome alteration in A20 group and gaining some insight on its specifics, we outline specific TF activator gene networks associated with transcriptional repression in affected species compared to controls, outlining Nr1d1 as a primary candidate, thus offering putative therapeutic targets in opioid-induced withdrawal treatment.

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Transcriptome analysis of fasting caecotrophy on hepatic lipid metabolism in New Zealand rabbits

10.1101/493957 ◽

2018 ◽

Author(s):

yadong wang ◽

Huifen Xu ◽

Guirong Sun ◽

Mingming Xue ◽

Shuaijie Sun ◽

...

Keyword(s):

Growth Rate ◽

Lipid Metabolism ◽

Growth Performance ◽

Differentially Expressed Genes ◽

Molecular Mechanisms ◽

Effective Means ◽

Differentially Expressed ◽

Control Group ◽

Rna Seq ◽

Final Body Weight

Background: Rabbit produce two kinds of feces: hard and soft feces, and they have a preference for consuming the latter. Although this habit of rabbits has been reported for many years, little is known on whether this behavior will impact growth performance and metabolism. The RNA-Seq technology is an effective means of analyzing transcript groups to clarify molecular mechanisms. The aim of the present study was to investigate the effects of fasting caecotrophy on growth performance and lipid metabolism in rabbits. Results: Our results indicated that, compared with the control group, the final body weight, weight gain, liver weight, specific growth rate and feed conversion ratio were all decreased in the experimental group (P<0.05). Oil red staining of the liver tissue indicated that fasting caecotrophy resulted in decrease of lipid droplet accumulation. RNA sequencing (RNA-seq) analysis revealed a total of 301.2 million raw reads approximately 45.06 Gb of high-quality clean data. The data were mapped to the rabbit genome (http://www.ensembl.org/Oryctolagus_cuniculus). After a five step filtering process, 14964 genes were identified, including 444 differentially expressed genes (P<0.05, foldchange≥1). Especially for remarkable changes of genes related to lipid metabolism, RT-PCR further validated the remarkable decrease of these genes in fasting caecotrophy group, including CYP7A1, PPARG, ABCA1, ABCB1, ABCG1, GPAM, SREBP, etc. KEGG annotation of the differentially expressed genes indicated that the main pathways affected were retinol metabolism, pentose and glucuronide interactions, starch and sucrose metabolism, fatty acid degradation, steroid hormone biosynthesis. Conclusion: In conclusion, the present study revealed that banning caecotrophy reduced growth rate and altered lipid metabolism, our results laid instructive basis for rabbit feeding and production. These data also provides a reference for studying the effects of soft feces on other small herbivores.

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The bacteriostatic effect and mechanism of berberine on Methicillin resistant Staphylococcus aureus in vitro

10.21203/rs.2.9640/v1 ◽

2019 ◽

Author(s):

Lei Wang ◽

Fangfang Zhou ◽

Minyi Xu ◽

Wei Gong ◽

Shuiying Ji

Keyword(s):

Cell Wall ◽

Rna Sequencing ◽

Differentially Expressed Genes ◽

Differentially Expressed ◽

Control Group ◽

High Concentration ◽

Bacteriostatic Effect ◽

Low Concentration ◽

Kinetics Of

Abstract Background: To observe the bacteriostatic effect of berberine on MRSA, while also exploring the bacteriostatic mechanism of BBR on MRSA. Methods: The MIC of BBR, gentamicin, levofloxacin，amikacin was determined by broth microdilution, while the MICs of BBR combined with gentamicin, levofloxacin，amikacin against MRSA were determined using microdilution checkerboard. Time-killing test were used to determine the kinetics of BBR combined with antibiotics for MRSA. We used conductivity to assess the changes in membrane permeability in response to BBR on MRSA, while also investigating the changes in MRSA morphology by TEM. RNA-sequencing was used to analyze the expression of differentially expressed genes in USA300 after its treatment with BBR. Results: The MICs range of BBR on MRSA was 32-256 µg/mL. The range of FICIs of BBR combined with gentamicin, levofloxacin，amikacin were 0.53-1.06, 0.62-1.5, 0.16-1.25. After co-culturing MRSA with BBR at 512 ug/mL, 64 ug/mL,8 ug/mL, respectively, the conductivity of these group increased by 8.14%,13.08% and 12.01%, respectively. Using TEM, we found that low-concentration of BBR had no significant effect on MRSA structure, medium-concentration of BBR thinned the cell wall of MRSA, while high-concentration of BBR destroyed cell wall, leading to bacterial lysis. RNA-sequencing results showed that there were 754 differentially expressed genes in the high-concentration group compared with the control group, of which 561 genes were up-regulated and 193 genes were down-regulated. Compared with the low-concentration group, there were 590 differentially expressed genes, of which 402 genes were up-regulated and 188 genes were down-regulated. Compared with the control group, 19 genes were differentially expressed in the low-concentration group, of which 11 genes were up-regulated,8 genes were down-regulated. Conclusions: BBR displayed an excellent bacteriostatic effect on MRSA. BBR combined with antibiotics significantly enhanced the bacteriostatic effect on MRSA. BBR inhibited bacteria by destroying the structure of cell wall. RNA-sequencing results demonstrated that the expression of cell wall hydrolysis genes and virulence factor were significantly differentially expressed on MRSA.

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RNA-Seq profiling of microdissected glomeruli identifies potential biomarkers for human IgA nephropathy

AJP Renal Physiology ◽

10.1152/ajprenal.00037.2020 ◽

2020 ◽

Vol 319 (5) ◽

pp. F809-F821

Author(s):

Sehoon Park ◽

Seung Hee Yang ◽

Chang Wook Jeong ◽

Kyung Chul Moon ◽

Dong Ki Kim ◽

...

Keyword(s):

Gene Ontology ◽

Iga Nephropathy ◽

Differentially Expressed Genes ◽

Mesangial Cells ◽

Principal Component ◽

Differentially Expressed ◽

Rna Seq ◽

Ontology Term ◽

Transcriptomic Profiling

Few studies have examined gene expression changes occurring in the glomeruli of IgA nephropathy (IgAN) using a sensitive transcriptomic profiling method such as RNA sequencing (RNA-Seq). We collected glomeruli from biopsy specimens from patients with IgAN with relatively preserved kidney function (estimated glomerular filtration rate ≥ 60 mL·min−1·1.73 m−2 and urine protein-to-creatinine ratio < 3 g/g) and from normal kidney cortexes by hand microdissection and performed RNA-Seq. Differentially expressed genes were identified, and gene ontology term annotation and pathway analysis were performed. Immunohistochemical labeling and primary mesangial cell cultures were performed to confirm the findings of RNA-Seq analysis. Fourteen patients with IgAN and ten controls were included in this study. Glomerulus-specific genes were highly abundant. Principal component analysis showed clear separation between the IgAN and control groups. There were 2,497 differentially expressed genes, of which 1,380 were upregulated and 1,117 were downregulated (false discovery rate < 0.01). The enriched gene ontology terms included motility/migration, protein/vesicle transport, and immune system, and kinase binding was the molecular function overrepresented in IgAN. B cell signaling, chemokine signal transduction, and Fcγ receptor-mediated phagocytosis were the canonical pathways overrepresented. In vitro experiments confirmed that spleen tyrosine kinase (SYK), reported as upregulated in the IgAN transcriptome, was also upregulated in glomeruli from an independent set of patients with IgAN and that treatment with patient-derived IgA1 increased the expression of SYK in mesangial cells. In conclusion, transcriptomic profiling of the IgAN glomerulus provides insights in the intraglomerular pathophysiology of IgAN before it reaches profound kidney dysfunction. SYK may have a pathogenetic role in IgAN.

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Comparative Analysis of Mouse Decidualization Models at the Molecular Level

Genes ◽

10.3390/genes11080935 ◽

2020 ◽

Vol 11 (8) ◽

pp. 935 ◽

Cited By ~ 1

Author(s):

Chong Wang ◽

Miao Zhao ◽

Wen-Qian Zhang ◽

Ming-Yu Huang ◽

Can Zhu ◽

...

Keyword(s):

Comparative Analysis ◽

Mouse Models ◽

Differentially Expressed Genes ◽

Molecular Level ◽

Differentially Expressed ◽

Rna Seq ◽

Golden Standard ◽

Transcriptomic Level ◽

Similarity And Difference

The mouse is widely used to study decidualization and there are three well-established mouse models of decidualization, namely natural pregnancy decidualization (NPD), artificial decidualization (AD), and in vitro decidualization (IVD). However, the extent of similarity and difference between these models at the molecular level remains largely unknown. Here, we performed a comparative analysis using the RNA-seq approach. In the NPD model, which is thought to be the golden standard of mouse decidualization, we found a total of 5277 differentially expressed genes, with 3158 genes being up-regulated and 2119 genes being down-regulated. A total of 4294 differentially expressed genes were identified in the AD model: 1127 up-regulated genes and 3167 down-regulated genes. In comparison to NPD, 1977 genes were consistently expressed, whereas only 217 genes were inconsistently expressed, indicating that AD is a reliable model for mouse decidualization. In the IVD model, RNA-seq analysis revealed that 513 genes were up-regulated and 988 genes were down-regulated. Compared to NPD, 310 genes were consistently expressed, whereas 456 genes were inconsistently expressed. Moreover, although the decidualization marker Prl8a2 (prolactin family 8 subfamily a member 2) was up-regulated, the widely-used marker Alpl (alkaline phosphatase liver/bone/kidney) was down-regulated in the IVD model. Therefore, we suggest that the IVD model should be optimized to mimic NPD at the transcriptomic level. Our study contributes to an increase in the knowledge about mouse models of decidualization.

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Exon-level estimates improve the detection of differentially expressed genes in RNA-seq studies

RNA Biology ◽

10.1080/15476286.2020.1868151 ◽

2021 ◽

pp. 1-8

Author(s):

Arfa Mehmood ◽

Asta Laiho ◽

Laura L. Elo

Keyword(s):

Differentially Expressed Genes ◽

Differentially Expressed ◽

Rna Seq ◽

Exon Level

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PSVII-2 Differentially expressed genes and their biological function in skeletal muscle of calves born from cows with or without protein supplementation during mid-gestation

Journal of Animal Science ◽

10.1093/jas/skaa054.293 ◽

2020 ◽

Vol 98 (Supplement_3) ◽

pp. 165-166

Author(s):

Elisa B Carvalho ◽

Letícia P Sanglard ◽

Karolina B Nascimento ◽

Javier M Meneses ◽

Daniel R Casagrande ◽

...

Keyword(s):

Skeletal Muscle ◽

Differentially Expressed Genes ◽

Muscle Development ◽

Negative Binomial ◽

Muscle Protein ◽

Basal Diet ◽

Protein Supplementation ◽

Differentially Expressed ◽

Control Group ◽

Q Value

Abstract Gestating cows have an increased nutrient demand to meet the needs of developing the fetus and the mid-gestation is a critical period for the fetal skeletal muscle development. The aim of this study was to evaluate the skeletal muscle transcriptome in the progeny as a function of the maternal protein nutrition during mid-gestation. Eleven Tabapuã cows and their male calves were used in this study. In the first third of gestation (0 to 100 days of gestation; dg), all cows were kept on pasture. From 100 to 200 dg, the control group (CTRL; 7 animals) received a basal diet achieving 5.5% crude protein (CP), whereas the supplemented group (SUPPL; 4 animals) received a basal diet plus protein supplementation (40% CP). After 200 dg, all animals received the same diet. Weaning was performed at 205 ± 7.5 days of age and animals were kept on pasture until reaching 240 days of age, when they were transferred to a feedlot. Muscle samples were collected at 260 days of age and RNA was extracted for RNA-seq analysis. Gene expression data was analyzed with a negative binomial model to identify (q-value ≤ 0.05) differentially expressed genes (DEG) between treatments. A total of 716 DEG were identified (289 DEG up-regulated and 427 down-regulated in SUPPL group; q-value ≤ 0.05). From the 10 most significant down-regulated DEG in the SUPPL group, two genes associated with apoptotic process were identified: MAPK8IP1 and GRINA, with log2 Fold-Changes (log2FC) of 1.04 and 0.49, respectively. From the 10 most significant up-regulated DEG in the SUPPL group, mTOR was identified, with log2FC=0.31. This is a well-known gene involved in muscle protein synthesis. In conclusion, maternal protein supplementation during mid-gestation affects the expression of genes related to energy metabolism and muscle development, which can lead to long-term impacts on production efficiency.

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