Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Effect of short term diet restriction on gene expression in the bovine hypothalamus using next generation RNA sequencing technology

ABSTRACT: Background: Negative energy balance (NEB) is an imbalance between energy intake and energy requirements for lactation and body maintenance, affecting high-yielding dairy cows after calving and is of considerable economic importance due to its negative impact on fertility and health in dairy herds. The hypothalamus is central to the neural control of homeostasis. It is anticipated that the cow hypothalamus experience extensive biochemical changes during the early post partum period in an effort to re-establish metabolic homeostasis. There is variation in the tolerance to NEB between individual cows. In order to understand the genomic regulation of ovulation in hypothalamic tissue during NEB transcriptional patterns between tolerant and sensitive animals was examined. A short term dietary restriction heifer model was developed which induced abrupt onset of anoestrus in some animals (Restricted Anovulatory; RA) while others maintained oestrous cyclicity (Restricted Ovulatory; RO). In addition a third control group (C) received a higher level of normal feeding. Heifers were slaughtered and hypothalamic tissue was collected after 18 days of differential feeding. mRNAseq was performed to examine differential gene expression profiles in hypothalamic tissue between groups. Results: A total of 15,295 genes were expressed in hypothalamic tissue. The largest difference between groups was observed in the comparison between RA and RO heifers, with 1094 genes shown to be significantly differentially expressed (SDE). Innatedb pathway analysis showed that these SDE genes were associated with 6 canonical pathways (P < 0.01), of which neuroactive ligand-receptor interaction was the most significant. Within the comparisons the main over-represented pathway functions appear to be immune response including neuroprotection (CXCL10, Q1KLR3, IFIH1, IL1 and IL8; RA v C and RA v RO); energy homeostasis (AgRP and NPY; RA v RO); cell motility (CADH1, DSP and TSP4; RO v C) and prevention of GnRH release (NTSR1 IL1?, IL1?, NPY and PACA; RA v RO). Conclusion: This information assists in understanding the genomic factors regulating the influence of diet restriction on fertility and may assist in to optimise nutritional and management systems that improve reproductive performance. A short term dietary restriction heifer model was developed which induced abrupt onset of anoestrus in some animals (Restricted Anovulatory; RA) while others maintained oestrous cyclicity (Restricted Ovulatory; RO). In addition a third control group (C) received a higher level of normal feeding. Heifers were slaughtered and hypothalamic tissue was collected after 18 days of differential feeding. mRNAseq was performed to examine differential gene expression profiles in hypothalamic tissue between groups.

ORGANISM(S): Bos taurus

SUBMITTER: Anthony Doran

PROVIDER: E-GEOD-49540 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background: Negative energy balance (NEB) is an imbalance between energy intake and energy requirements for lactation and body maintenance, affecting high-yielding dairy cows after calving and is of considerable economic importance due to its negative impact on fertility and health in dairy herds. The hypothalamus is central to the neural control of homeostasis. It is anticipated that the cow hypothalamus experience extensive biochemical changes during the early post partum period in an effort to re-establish metabolic homeostasis. There is variation in the tolerance to NEB between individual cows. In order to understand the genomic regulation of ovulation in hypothalamic tissue during NEB transcriptional patterns between tolerant and sensitive animals was examined. A short term dietary restriction heifer model was developed which induced abrupt onset of anoestrus in some animals (Restricted Anovulatory; RA) while others maintained oestrous cyclicity (Restricted Ovulatory; RO). In addition a third control group (C) received a higher level of normal feeding. Heifers were slaughtered and hypothalamic tissue was collected after 18 days of differential feeding. mRNAseq was performed to examine differential gene expression profiles in hypothalamic tissue between groups. Results: A total of 15,295 genes were expressed in hypothalamic tissue. The largest difference between groups was observed in the comparison between RA and RO heifers, with 1094 genes shown to be significantly differentially expressed (SDE). Innatedb pathway analysis showed that these SDE genes were associated with 6 canonical pathways (P < 0.01), of which neuroactive ligand-receptor interaction was the most significant. Within the comparisons the main over-represented pathway functions appear to be immune response including neuroprotection (CXCL10, Q1KLR3, IFIH1, IL1 and IL8; RA v C and RA v RO); energy homeostasis (AgRP and NPY; RA v RO); cell motility (CADH1, DSP and TSP4; RO v C) and prevention of GnRH release (NTSR1 IL1α, IL1β, NPY and PACA; RA v RO). Conclusion: This information assists in understanding the genomic factors regulating the influence of diet restriction on fertility and may assist in to optimise nutritional and management systems that improve reproductive performance.

Project description:Our objectives were to compare gene expression profiles in neutrophils (PMN) during a Streptococcus uberis mastitis challenge between lactating cows subjected to feed restriction to induce negative energy balance (NEB; n = 5) and cows fed ad libitum to maintain positive energy balance (PEB; n = 5). The NEB cows were feed-restricted to 60% of calculated net energy for lactation requirements for 7 d, whereas PEB cows were fed the same diet for ad libitum intake. After 5 d of feed restriction, one rear mammary quarter of each cow was inoculated with 5,000 cfu of Streptococcus uberis (strain O140J). Blood PMN were isolated at 24 h post-inoculation from all cows for RNA extraction and microarray analysis. NEB resulted in 94 differentially expressed genes compared with PEB. Of these, 51 genes were down-regulated, including genes involved with antigen presentation (HLADRA and HLAA), respiratory burst (SOD1), and the pro-inflammatory response (TNFA and IRAK-1). The most affected genes up-regulated by NEB (n = 43) included IL1R2 and IL6, toll-like receptors (TLR2 and TLR4), and THY1. Network analysis by Ingenuity Pathway Analysis Â® revealed that TNFA was associated with the expression of numerous differentially expressed genes involved with immune response in NEB cows compared with PEB cows. Energy balance alters PMN expression of several genes involved with immune response, which provides new information on transcriptomic mechanisms associated with post-partal NEB and immune response during early lactation. Briefly, 10 multiparous Holstein cows in PEB and past peak lactation were used for this study. The NEB cows were feed-restricted to 60% of calculated NEL requirements for 7 d, whereas PEB cows were fed the same diet for ad libitum intake. After 5 d of feed restriction, one rear mammary quarter of each cow was inoculated with 5,000 cfu of Streptococcus uberis (strain O140J). Blood PMN were isolated at 24 h post-inoculation from all cows. A 13,257-oligonucleotide (70-mers) array was used for transcript profiling. Cy3- and Cy5 labelled cDNA from PMN and a reference standard were used for hybridizations. All samples were hybridized to duplicate slides with reverse labeling (20 microarrays total).

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Effect of short term diet restriction on gene expression in the bovine hypothalamus using next generation RNA sequencing technology

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