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Gene Expression in Forebrain of Male Goldfish Following Exposure to Waterborne Sex Pheromones

ABSTRACT: Sex pheromones rapidly affect endocrine physiology and behaviour, but little is known about their effects on gene expression in the neuroendocrine tissues that mediate olfactory processing. In this study we exposed male goldfish for 6 h to waterborne 17,20betaP (4.3 nM) and PGF2α (3 nM), the main pre-ovulatory and post-ovulatory pheromones, respectively. Both treatments elevated milt volume (P = 0.001). Microarray hybridizations were used to male telencephalon samples following PGF2α treatment and identified 71 unique transcripts that were differentially expressed (q < 5%; 67 up, 4 down). Functional annotation of these regulated genes indicates that PGF2α pheromone exposure affects diverse biological processes including nervous system functions, energy metabolism, cholesterol/lipoprotein transport, translational regulation, transcription and chromatin remodelling, protein processing, cytoskeletal organization, and signalling. By using real-time RT-PCR, we further validated three candidate genes, ependymin-II, calmodulin-A and aldolase C, which exhibited 3 - 5 fold increase in expression following PGF2α exposure. Expression levels of some other genes that are thought to be important for reproduction were also determined using real-time RT-PCR. Expression of sGnRH was increased by PGF2α, but not 17,20betaP, whereas cGnRH expression was increased by 17,20betaP but not PGF2α. In contrast, both pheromones increase the expression of glutamate (GluR2a, NR2A) and gamma-aminobutyric acid (GABAA gamma2) receptor subunit mRNAs. This gene expression data link milt release and rapid modulation of neuronal transcription as part of the response to female sex pheromones.

ORGANISM(S): Carassius auratus Cyprinus carpio

PROVIDER: GSE41918 | GEO | 2013/05/01

SECONDARY ACCESSION(S): PRJNA179219

REPOSITORIES: GEO

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Project description:Sex pheromones rapidly affect endocrine physiology and behaviour, but little is known about their effects on gene expression in the neuroendocrine tissues that mediate olfactory processing. In this study we exposed male goldfish for 6 h to waterborne 17,20betaP (4.3 nM) and PGF2α (3 nM), the main pre-ovulatory and post-ovulatory pheromones, respectively. Both treatments elevated milt volume (P = 0.001). Microarray hybridizations were used to male telencephalon samples following PGF2α treatment and identified 71 unique transcripts that were differentially expressed (q < 5%; 67 up, 4 down). Functional annotation of these regulated genes indicates that PGF2α pheromone exposure affects diverse biological processes including nervous system functions, energy metabolism, cholesterol/lipoprotein transport, translational regulation, transcription and chromatin remodelling, protein processing, cytoskeletal organization, and signalling. By using real-time RT-PCR, we further validated three candidate genes, ependymin-II, calmodulin-A and aldolase C, which exhibited 3 - 5 fold increase in expression following PGF2α exposure. Expression levels of some other genes that are thought to be important for reproduction were also determined using real-time RT-PCR. Expression of sGnRH was increased by PGF2α, but not 17,20betaP, whereas cGnRH expression was increased by 17,20betaP but not PGF2α. In contrast, both pheromones increase the expression of glutamate (GluR2a, NR2A) and gamma-aminobutyric acid (GABAA gamma2) receptor subunit mRNAs. This gene expression data link milt release and rapid modulation of neuronal transcription as part of the response to female sex pheromones. A total of four microarray slides were hybridized to study the effects of PGF2α on gene expression in the telencephalon. Control samples (N = 4) were pooled to form our reference or technical replicates while treatment samples (N = 4), which formed our biological replicates, were ran separately. In addition, two dye swaps were performed for our biological samples for the treatments.

Project description:Preterm birth is the leading cause of infant morbidity and mortality. There has been interest in developing prostaglandin F2α (PGF2α) antagonists as a new treatment for preterm birth although much of the rationale for their use is based on studies in rodents where PGF2α initiates labour by regressing the corpus luteum and reducing systemic progesterone concentrations. How PGF2α antagonism would act in humans that do not have a fall in systemic progesterone remains unclear. One possibility, in addition to an acute stimulation of contractions, is a direct alteration of the myometrial smooth muscle cell state towards a pro-labour phenotype. In this study we developed an immortalised myometrial cell line, MYLA, derived from myometrial tissue obtained from a pregnant, non‐labouring patient as well as a novel class of PGF2α receptor (FP) antagonist. We verified the functionality of the cell line by stimulation with PGF2α, which resulted in Gαq specific coupling and Ca2+ release that were inhibited by FP antagonism. Compared to four published FP receptor antagonists, the novel FP antagonist N582707 was the most potent compound (Fmax 7.67 ± 0.63 (IC50 21.26 nM), AUC 7.30 ± 0.32 (IC50 50.43 nM) and frequency of Ca2+ oscillations 7.66 ± 0.41 (IC50 22.15 nM)). RNA‐sequencing of the MYLA cell line at 1, 3, 6, 12, 24 & 48 hours post PGF2α treatment revealed a transforming phenotype from a fibroblastic to smooth muscle mRNA profile. PGF2α treatment increased expression of MYLK, CALD1 and CNN1 as well as the pro-labour genes OXTR, IL6 and IL11 which were inhibited by FP antagonism. Concomitant with inhibition of a smooth muscle, pro-labour transition, FP antagonism increased the expression of the fibroblast marker genes DCN, FBLN1 and PDGFRA. Our findings suggest that in addition to the well described acute contractile effect, PGF2α transforms myometrial smooth muscle cells from a myofibroblast to smooth muscle, pro-labour like state, and that the novel compound N582707 has potential for prophylactic use in preterm labour management beyond its use as an acute tocolytic drug.

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Gene Expression in Forebrain of Male Goldfish Following Exposure to Waterborne Sex Pheromones

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