ABSTRACT
INTRODUCTION: Despite the known negative health outcomes of concussions in minor level boys' hockey, there has been significant resistance to creating a safer game with less body checking. METHODS: To better understand cultural barriers that prevent making the sport safer for youth and adolescents, semistructured interviews, with 20 ice hockey stakeholders, were conducted and analysed using thematic analysis. RESULTS: Through this analysis, two primary concepts arose from respondents. The first concept is that body checking, despite the harm it can cause, should be done in a respectful sportsmanlike fashion. The second concept is the contradiction that the game of ice hockey is both dynamic and unchangeable. DISCUSSION: Using structural functionalist theory, we propose an argument that the unfortunate perpetuation of violence and body checking in youth ice hockey serves to maintain the social order of the game and its culture. Any strategies aimed at modifying and promoting healthy behaviour in the game should take these concepts into account.
Subject(s)
Aggression/psychology , Athletic Injuries/prevention & control , Brain Concussion/prevention & control , Hockey/injuries , Safety Management/methods , Athletic Injuries/complications , Brain Concussion/complications , Canada , Group Processes , Health Knowledge, Attitudes, Practice , Health Policy , Hockey/psychology , Humans , Incidence , Qualitative Research , Stakeholder ParticipationABSTRACT
Reproductive function is coordinated by kisspeptin (Kiss) and GnRH neurons. Phoenixin-20 amide (PNX) is a recently described peptide found to increase GnRH-stimulated LH secretion in the pituitary. However, the effects of PNX in the hypothalamus, the putative signaling pathways, and PNX receptor have yet to be identified. The mHypoA-GnRH/GFP and mHypoA-Kiss/GFP-3 cell lines represent populations of GnRH and Kiss neurons, respectively. PNX increased GnRH and GnRH receptor (GnRH-R) mRNA expression, as well as GnRH secretion, in the mHypoA-GnRH/GFP cell model. In the mHypoA-Kiss/GFP-3 cell line, PNX increased Kiss1 mRNA expression. CCAAT/enhancer-binding protein (C/EBP)-ß, octamer transcription factor-1 (Oct-1), and cAMP response element binding protein (CREB) binding sites are localized to the 5' flanking regions of the GnRH, GnRH-R, and Kiss1 genes. PNX decreased C/EBP-ß mRNA expression in both cell models and increased Oct-1 mRNA expression in the mHypoA-GnRH/GFP neurons. PNX increased CREB phosphorylation in both cell models and phospho-ERK1/2 in the mHypoA-GnRH/GFP cell model, whereas inhibiting the cAMP/protein kinase A pathway prevented PNX induction of GnRH and Kiss1 mRNA expression. Importantly, we determined that the G protein-coupled receptor, GPR173, was strongly expressed in both GnRH and kisspeptin cell models and small interfering RNA knockdown of GPR173 prevented the PNX-mediated up-regulation of GnRH, GnRH-R, and Kiss1 mRNA expression and the down-regulation of C/EBP-ß mRNA expression. PNX also increased GPR173 mRNA expression in the mHypoA-GnRH/GFP cells. Taken together, these studies are the first to implicate that PNX acts through GPR173 to activate the cAMP/protein kinase A pathway through CREB, and potentially C/EBP-ß and/or Oct-1 to increase GnRH, GnRH-R, and Kiss1 gene expression, ultimately having a stimulatory effect on reproductive function.
Subject(s)
Amides/pharmacology , Gonadotropin-Releasing Hormone/metabolism , Kisspeptins/metabolism , Neurons/metabolism , Peptides/pharmacology , Receptors, G-Protein-Coupled/metabolism , Amides/chemistry , Animals , Cell Line , Cells, Cultured , Cyclic AMP Response Element-Binding Protein/metabolism , Hypothalamus/drug effects , Hypothalamus/metabolism , Kisspeptins/genetics , Mice , Neurons/drug effects , Peptides/chemistry , Phosphorylation/drug effects , Receptors, G-Protein-Coupled/genetics , Receptors, LHRH/metabolismABSTRACT
Kisspeptin (Kiss) and G-protein-coupled receptor (Gpr)54 have emerged as key regulators of reproduction. 17ß-estradiol (E2)-mediated regulation of these neurons is nuclei specific, where anteroventral periventricular (AVPV) Kiss neurons are positively regulated by E2, whereas arcuate nucleus (ARC) neurons are inhibited. We have generated immortalized Kiss cell lines from male and female adult-derived murine hypothalamic primary culture, as well as cell lines from microdissected AVPV and ARC from female Kiss-green fluorescent protein (GFP) mice. All exhibit endogenous Kiss-1 expression, estrogen receptors (ER)s (ERα, ERß, and Gpr30), as well as known markers of AVPV Kiss neurons in the mHypoA-50 and mHypoA-Kiss/GFP-4, vs markers of ARC Kiss neurons in the mHypoA-55 and the mHypoA-Kiss/GFP-3 lines. There was an increase in Kiss-1 mRNA expression at 24 hours in the AVPV lines and a repression of Kiss-1 mRNA at 4 hours in the ARC lines. An E2-mediated decrease in ERα mRNA expression at 24 hours in the AVPV cell lines was detected, and a significant decrease in Gpr30, ERα, and ERß mRNA levels at 4 hours in the ARC cell lines was evident. ER agonists and antagonists determined the specific ERs responsible for mediating changes in gene expression. In the AVPV, ERα is required but not ERß or GPR30, vs the ARC Kiss-expressing cell lines that require GPR30, and either ERα and/or ERß. We determined cAMP response element-binding protein 1 was necessary for the down-regulation of Kiss-1 mRNA expression using small interfering RNA knockdown in the ARC cell model. These studies elucidate some of the molecular events involved in the differential E2-mediated regulation of unique and specific Kiss neuronal models.
