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Effect of thermal stress on expression profile of apoptosis related genes in peripheral blood mononuclear cells of transition Sahiwal cow.

ABSTRACT: The study was conducted to evaluate the effect of thermal stress on expression profile of genes related to apoptosis in peripartum Sahiwal cows. For this, twelve pregnant dry Sahiwal cows were selected from Livestock Research Centre at National Dairy Research Institute, Karnal. The cows were divided into two groups consisting of six Sahiwal cows each. Cows of group I calved during thermoneutral temperature conditions (THI=67.3) and cows of group II calved in summer season (THI=79.9). Blood samples were collected on -15, 0 and +15 days with respect to calving where day '0' represents the day of calving. The peripheral blood mononuclear cells (PBMC) were separated and total RNA was isolated for the BCL-2 (B-Cell Lymphoma-2), BAX (BCL-2 antagonist killer-1), BAK (Bcl-2-associated X protein), CASP-3 (cysteine-aspartic proteases-3) and P53 (tumour protien-53) mRNAs expression. It was found that there was up regulation of CASP-3 on the day of calving during both temperature conditions. Comparison between the two temperature conditions showed that expression of CASP-3, BCL-2, BAK, P53 and ratio of BAX/BCL-2 in PBMC increased during summer as compared to thermoneutral condition suggesting the susceptibility of these cells to apoptosis. Based on the above findings it can be concluded that during calving PBMC are more susceptible to apoptosis, and summer being more stressful potentiates the apoptosis of PBMC in Sahiwal cows.

SUBMITTER: Somal A

PROVIDER: S-EPMC4827677 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Effect of thermal stress on expression profile of apoptosis related genes in peripheral blood mononuclear cells of transition Sahiwal cow.

Somal A A Aggarwal A A Upadhyay R C RC

Iranian journal of veterinary research 20150101 2

The study was conducted to evaluate the effect of thermal stress on expression profile of genes related to apoptosis in peripartum Sahiwal cows. For this, twelve pregnant dry Sahiwal cows were selected from Livestock Research Centre at National Dairy Research Institute, Karnal. The cows were divided into two groups consisting of six Sahiwal cows each. Cows of group I calved during thermoneutral temperature conditions (THI=67.3) and cows of group II calved in summer season (THI=79.9). Blood sampl ...[more]

PMID: 27175165

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Project description:The objectives of the present study were to identify key genes and biological pathways associated with thermal stress in Chinese Holstein dairy cattle. Hence, we constructed a cell-model, applied various molecular biology experimental techniques and bioinformatics analysis. A total of 55 candidate genes were screened from published literature and the IPA database to examine its regulation under cold (25°C) or heat (42°C) stress in PBMCs. We identified 29 (3 up-regulated and 26 down-regulated) and 41 (15 up-regulated and 26 down-regulated) significantly differentially expressed genes (DEGs) (fold change ≥ 1.2-fold and P < 0.05) after cold and heat stress treatments, respectively. Furthermore, bioinformatics analyses confirmed that major biological processes and pathways associated with thermal stress include protein folding and refolding, protein phosphorylation, transcription factor binding, immune effector process, negative regulation of cell proliferation, autophagy, apoptosis, protein processing in endoplasmic reticulum, estrogen signaling pathway, pathways related to cancer, PI3K- Akt signaling pathway, and MAPK signaling pathway. Based on validation at the cellular and individual levels, the mRNA expression of the HIF1A gene showed upregulation during cold stress and the EIF2A, HSPA1A, HSP90AA1, and HSF1 genes showed downregulation after heat exposure. The RT-qPCR and western blot results revealed that the HIF1A after cold stress and the EIF2A, HSPA1A, HSP90AA1, and HSF1 after heat stress had consistent trend changes at the cellular transcription and translation levels, suggesting as key genes associated with thermal stress response in Holstein dairy cattle. The cellular model established in this study with PBMCs provides a suitable platform to improve our understanding of thermal stress in dairy cattle. Moreover, this study provides an opportunity to develop simultaneously both high-yielding and thermotolerant Chinese Holstein cattle through marker-assisted selection.

Project description:Trypanosoma cruzi (Tc) infection causes chagasic cardiomyopathy; however, why 30-40% of the patients develop clinical disease is not known. To discover the pathomechanisms in disease progression, we obtained the proteome signature of peripheral blood mononuclear cells (PBMCs) of normal healthy controls (N/H, n = 30) and subjects that were seropositive for Tc-specific antibodies, but were clinically asymptomatic (C/A, n = 25) or clinically symptomatic (C/S, n = 28) with cardiac involvement and left ventricular dysfunction. Protein samples were labeled with BODIPY FL-maleimide (dynamic range: > 4 orders of magnitude, detection limit: 5 f-mol) and resolved by two-dimensional gel electrophoresis (2D-GE). After normalizing the gel images, protein spots that exhibited differential abundance in any of the two groups were analyzed by mass spectrometry, and searched against UniProt human database for protein identification. We found 213 and 199 protein spots (fold change: |≥ 1.5|, p< 0.05) were differentially abundant in C/A and C/S individuals, respectively, with respect to N/H controls. Ingenuity Pathway Analysis (IPA) of PBMCs proteome dataset identified an increase in disorganization of cytoskeletal assembly and recruitment/activation and migration of immune cells in all chagasic subjects, though the invasion capacity of cells was decreased in C/S individuals. IPA predicted with high probability a decline in cell survival and free radical scavenging capacity in C/S (but not C/A) subjects. The MYC/SP1 transcription factors that regulate hypoxia and oxidative/inflammatory stress were predicted to be key targets in the context of control of Chagas disease severity. Further, MARS-modeling identified a panel of proteins that had >93% prediction success in classifying infected individuals with no disease and those with cardiac involvement and LV dysfunction. In conclusion, we have identified molecular pathways and a panel of proteins that could aid in detecting seropositive individuals at risk of developing cardiomyopathy.

Dataset Information

Effect of thermal stress on expression profile of apoptosis related genes in peripheral blood mononuclear cells of transition Sahiwal cow.

Publications

Effect of thermal stress on expression profile of apoptosis related genes in peripheral blood mononuclear cells of transition Sahiwal cow.

Similar Datasets

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