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Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia

Hypocalcemia is closely associated with inflammatory diseases in dairy cows. Recent research has underscored the key role of calcium in the adaptations of the innate immune system during this period. The main objective in the present study was to compare the transcriptome profiles and analyze differ...

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Autores principales: Zhang, Bingbing, Ma, Xinru, Huang, Baoyin, Jiang, Qianming, Loor, Juan J., Lv, Xinquan, Zhang, Wei, Li, Ming, Wen, Jianan, Yin, Yufeng, Wang, Jingjing, Yang, Wei, Xu, Chuang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9514324/
https://www.ncbi.nlm.nih.gov/pubmed/36176696
http://dx.doi.org/10.3389/fvets.2022.959831
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author Zhang, Bingbing
Ma, Xinru
Huang, Baoyin
Jiang, Qianming
Loor, Juan J.
Lv, Xinquan
Zhang, Wei
Li, Ming
Wen, Jianan
Yin, Yufeng
Wang, Jingjing
Yang, Wei
Xu, Chuang
author_facet Zhang, Bingbing
Ma, Xinru
Huang, Baoyin
Jiang, Qianming
Loor, Juan J.
Lv, Xinquan
Zhang, Wei
Li, Ming
Wen, Jianan
Yin, Yufeng
Wang, Jingjing
Yang, Wei
Xu, Chuang
author_sort Zhang, Bingbing
collection PubMed
description Hypocalcemia is closely associated with inflammatory diseases in dairy cows. Recent research has underscored the key role of calcium in the adaptations of the innate immune system during this period. The main objective in the present study was to compare the transcriptome profiles and analyze differences in the expression of neutrophil (PMNL) immune function-related genes and calcium binding-related genes in hypocalcemic cows. At 2 days postpartum, a concentration >2.10 mmol Ca(2+)/L was used to classify cows as controls (CON), and a concentration <2.00 mmol Ca(2+)/L used to classify cows as low-calcium (LCAL) (n = 8 in each group). A routine medical examination was conducted by the attending veterinarian to ensure there were no other complications and that the blood β-hydroxybutyrate was <1.2 mmol/L. Blood was collected from the tail vein (20 mL) to isolate PMNL, and 5 cows in each group were used for RNA sequencing and statistical analysis of gene expression differences. Transcriptome RNA-seq sequencing analysis was via omicsstudio using the R package edgeR. GO and KEGG enrichment analysis were used for bioinformatics. The remaining 3 cows in each group were used for validation of RNA sequencing data via quantitative PCR, which confirmed the observed responses. Compared with CON, 158 genes in LCAL were significantly up-regulated and 296 genes were down-regulated. The downregulation of Interleukin-12 (CXCL12), Tubulin beta chain (TUBB1), L1 cell adhesion molecule (L1CAM), and Myeloperoxidase (MPO) indicated a decrease in immune function of PMNL in LCAL cows. The decreased expression of calcium-binding pathway-related genes in PMNL of LCAL cows indicated a decrease in immune function of PMNL likely related to calcium ions. For example, cartilage acid protein 1 (CRTAC1) and calcium/calmodulin-dependent kinase 4 (CAMK4) were significantly reduced in LCAL cows. The upregulation of Cyclin dependent kinase inhibitor 1A (CDKN1A), Perforin 1 (PRF1), and Homeodomain interacting protein kinase 3 (HIPK3) indicated that LCAL led to greater cell apoptosis and senescence. Overall, the analyses indicated that the reduction in PMNL immune function during hypocalcemia is associated with downregulation of intracellular Ca(2+) related genes and upregulation of genes controlling apoptosis and senescence. Together, these alterations contribute to an immunosuppressive state during the transition period.
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spelling pubmed-95143242022-09-28 Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia Zhang, Bingbing Ma, Xinru Huang, Baoyin Jiang, Qianming Loor, Juan J. Lv, Xinquan Zhang, Wei Li, Ming Wen, Jianan Yin, Yufeng Wang, Jingjing Yang, Wei Xu, Chuang Front Vet Sci Veterinary Science Hypocalcemia is closely associated with inflammatory diseases in dairy cows. Recent research has underscored the key role of calcium in the adaptations of the innate immune system during this period. The main objective in the present study was to compare the transcriptome profiles and analyze differences in the expression of neutrophil (PMNL) immune function-related genes and calcium binding-related genes in hypocalcemic cows. At 2 days postpartum, a concentration >2.10 mmol Ca(2+)/L was used to classify cows as controls (CON), and a concentration <2.00 mmol Ca(2+)/L used to classify cows as low-calcium (LCAL) (n = 8 in each group). A routine medical examination was conducted by the attending veterinarian to ensure there were no other complications and that the blood β-hydroxybutyrate was <1.2 mmol/L. Blood was collected from the tail vein (20 mL) to isolate PMNL, and 5 cows in each group were used for RNA sequencing and statistical analysis of gene expression differences. Transcriptome RNA-seq sequencing analysis was via omicsstudio using the R package edgeR. GO and KEGG enrichment analysis were used for bioinformatics. The remaining 3 cows in each group were used for validation of RNA sequencing data via quantitative PCR, which confirmed the observed responses. Compared with CON, 158 genes in LCAL were significantly up-regulated and 296 genes were down-regulated. The downregulation of Interleukin-12 (CXCL12), Tubulin beta chain (TUBB1), L1 cell adhesion molecule (L1CAM), and Myeloperoxidase (MPO) indicated a decrease in immune function of PMNL in LCAL cows. The decreased expression of calcium-binding pathway-related genes in PMNL of LCAL cows indicated a decrease in immune function of PMNL likely related to calcium ions. For example, cartilage acid protein 1 (CRTAC1) and calcium/calmodulin-dependent kinase 4 (CAMK4) were significantly reduced in LCAL cows. The upregulation of Cyclin dependent kinase inhibitor 1A (CDKN1A), Perforin 1 (PRF1), and Homeodomain interacting protein kinase 3 (HIPK3) indicated that LCAL led to greater cell apoptosis and senescence. Overall, the analyses indicated that the reduction in PMNL immune function during hypocalcemia is associated with downregulation of intracellular Ca(2+) related genes and upregulation of genes controlling apoptosis and senescence. Together, these alterations contribute to an immunosuppressive state during the transition period. Frontiers Media S.A. 2022-09-13 /pmc/articles/PMC9514324/ /pubmed/36176696 http://dx.doi.org/10.3389/fvets.2022.959831 Text en Copyright © 2022 Zhang, Ma, Huang, Jiang, Loor, Lv, Zhang, Li, Wen, Yin, Wang, Yang and Xu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Zhang, Bingbing
Ma, Xinru
Huang, Baoyin
Jiang, Qianming
Loor, Juan J.
Lv, Xinquan
Zhang, Wei
Li, Ming
Wen, Jianan
Yin, Yufeng
Wang, Jingjing
Yang, Wei
Xu, Chuang
Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
title Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
title_full Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
title_fullStr Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
title_full_unstemmed Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
title_short Transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
title_sort transcriptomics of circulating neutrophils in dairy cows with subclinical hypocalcemia
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9514324/
https://www.ncbi.nlm.nih.gov/pubmed/36176696
http://dx.doi.org/10.3389/fvets.2022.959831
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