Kisspeptin cell models from the hypothalamus
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1
University of Toronto, Department of Physiology, Canada
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2
University of Toronto, Department of Medicine, Canada
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3
University of Toronto, Obstetrics and Gynecology, Canada
Kisspeptin, a peptide hormone secreted by neurons in the hypothalamus, has emerged as a key regulator of the hypothalamic-pituitary-gonadal axis through its potent stimulation of GnRH release. Although this essential role of kisspeptin in reproductive function is well established, there is a paucity of studies focusing on the regulation of the hypothalamic Kiss1 gene (which encodes kisspeptin) due to the lack of appropriate cell models. In order to address this problem, we have generated immortalized, clonal, murine cell lines derived from adult hypothalamic primary culture. Using RT-PCR, immunocytochemistry and ELISAs, we have established over twenty cell lines that exhibit endogenous kisspeptin synthesis. Importantly, there are two distinct populations of kisspeptin neurons found in the hypothalamus: kisspeptin neurons in the arcuate nucleus (ARC), which are negatively regulated by estrogen, and kisspeptin neurons in the anteroventral periventricular nucleus (AVPV), which are positively regulated by estrogen. Four of our kisspeptin-expressing cell lines, mHypoA-48, 50, 51 and 63 are regulated by estrogen, as demonstrated by real-time RT-PCR: estrogen up-regulates Kiss1 mRNA expression in mHypoA-51 and mHypoA-63 neurons, and down-regulates Kiss1 mRNA expression in mHypoA-48 and mHypoA-50 neurons. These findings suggest that mHypoA-51 and mHypoA-63 neurons are cell models of AVPV kisspeptin neurons, and mHypoA-48 and mHypoA-50 neurons are cell models of ARC kisspeptin neurons. These kisspeptin cell models will be key to studying the cellular events underlying differential estrogen regulation of kisspeptin neurons. Furthermore, there is considerable ambiguity over the location of the mouse Kiss1 promoter; thus, our cell models will enable a precise analysis of the hypothalamus-specific 5’-regulatory region. Overall, these cell lines are invaluable for the mechanistic analysis of endogenous Kiss1 gene regulation and kisspeptin secretion. These studies have not been possible in the whole brain and will lead to a better understanding of how Kiss1 neurons are controlled directly by hormones and neuropeptides.
Funded by: CIHR, CRC, and CFI
Conference:
B.R.A.I.N. platform in Physiology poster day 2009, Toronto, ON, Canada, 16 Dec - 16 Dec, 2009.
Presentation Type:
Poster Presentation
Topic:
Poster presentations
Citation:
Kim
GL,
Gingerich
S,
Centeno
M,
Wang
X,
Chalmers
JA,
Koletar
MM,
Thompson
DR and
Belsham
DD
(2009). Kisspeptin cell models from the hypothalamus.
Front. Neurosci.
Conference Abstract:
B.R.A.I.N. platform in Physiology poster day 2009.
doi: 10.3389/conf.neuro.03.2009.17.026
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Received:
17 Dec 2009;
Published Online:
17 Dec 2009.
*
Correspondence:
Ginah L Kim, University of Toronto, Department of Physiology, Toronto, Canada, ginah.lee.kim@gmail.com