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Transcriptomic signature of in vitro heat stress challenged porcine granulosa cells

ABSTRACT: Exploration of heat stress induced genes in granulosa cells is critical to characterization of candidate genes for thermal stress research in pigs. This study aimed to gain insight into the differentially expressed genes (DEGs) and signalling pathways regulating porcine granulosa cell adaptation to in vitro heat stress challenge at 42OC for 6 hours. Transcriptome profiling of heat stressed (treated) and non-heat stressed (control) granulosa cells was conducted using Illumina NextSeq2000 sequencing platform. The significant DEGs were selected using NOISeq R package with cut-offs, probability value ≥ 0.95 and absolute log2(fold change) ≥1. Bioinformatics analysis of DEGs were conducted to explore functional annotations enrichment, protein-protein interaction network and hub genes regulating the cellular homeostasis and survivability during heat stress challenge.

ORGANISM(S): Sus scrofa

PROVIDER: GSE243234 | GEO | 2024/10/22

REPOSITORIES: GEO

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Project description:Direct exposure to physiologically-relevant elevated temperatures during the first half of in vitro maturation heightens cumulus-derived progesterone production, suppresses matrix metallopeptidase 9 secretion, and alters cumulus transcriptome. We hypothesized that heat-induced perturbations in cumulus cells enveloping maturing oocyte may extend to the mural granulosa of the periovulatory follicle in the heat-stressed cow to subsequently follicular fluid proteome. Lactating Holsteins were pharmacologically stimulated to produce a dominant follicle then given GnRH to induce an LH surge. Following GnRH administration, cows were maintained at ~67 temperature humidity index (THI; thermoneutral conditions) or exposed to conditions simulating an acute heat stress event (71 to 86 THI; heat stress for ~12 h). Fluid was aspirated from periovulatory follicle ~16 h after GnRH. Follicular fluid proteome from thermoneutral (n = 5) and hyperthermic (n = 5) cows was evaluated by quantitative tandem mass spectrometry (nano LC-MS/MS). We identified 35 differentially-abundant proteins. Functional annotation revealed numerous immune-related proteins. Subsequent efforts revealed an increase in levels of the proinflammatory mediator bradykinin in follicular fluid (P = 0.0456) but not in serum (P = 0.9319) of hyperthermic cows. Intrafollicular increases in transferrin (negative acute phase protein) in hyperthermic cows (P = 0.0181) coincided with a tendency for levels to be increased in the circulation (P = 0.0683). Nine out of 15 cytokines evaluated were detected in follicular fluid. Heat stress increased intrafollicular IL6 levels (P = 0.0160). Whether hyperthermia-induced changes in the heat-stressed cow’s follicular fluid milieu reflect changes in mural granulosa, cumulus, other cell types secretions, and/or transudative changes from circulation remains unclear. Regardless of origin, heat stress/hyperthermia related changes in the follicular fluid milieu may have an impact on components important for ovulation and competence of the cumulus-oocyte complex contained within the periovulatory follicle

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Transcriptomic signature of in vitro heat stress challenged porcine granulosa cells

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