Title: Rbpj-κ mediated Notch signaling plays a critical role in development of hypothalamic Kisspeptin neurons.

Authors: Biehl, Matthew J; Raetzman, Lori T

Published In Dev Biol, (2015 Oct 15)

Abstract: The mammalian arcuate nucleus (ARC) houses neurons critical for energy homeostasis and sexual maturation. Proopiomelanocortin (POMC) and Neuropeptide Y (NPY) neurons function to balance energy intake and Kisspeptin neurons are critical for the onset of puberty and reproductive function. While the physiological roles of these neurons have been well established, their development remains unclear. We have previously shown that Notch signaling plays an important role in cell fate within the ARC of mice. Active Notch signaling prevented neural progenitors from differentiating into feeding circuit neurons, whereas conditional loss of Notch signaling lead to a premature differentiation of these neurons. Presently, we hypothesized that Kisspeptin neurons would similarly be affected by Notch manipulation. To address this, we utilized mice with a conditional deletion of the Notch signaling co-factor Rbpj-κ (Rbpj cKO), or mice persistently expressing the Notch1 intracellular domain (NICD tg) within Nkx2.1 expressing cells of the developing hypothalamus. Interestingly, we found that in both models, a lack of Kisspeptin neurons are observed. This suggests that Notch signaling must be properly titrated for formation of Kisspeptin neurons. These results led us to hypothesize that Kisspeptin neurons of the ARC may arise from a different lineage of intermediate progenitors than NPY neurons and that Notch was responsible for the fate choice between these neurons. To determine if Kisspeptin neurons of the ARC differentiate similarly through a Pomc intermediate, we utilized a genetic model expressing the tdTomato fluorescent protein in all cells that have ever expressed Pomc. We observed some Kisspeptin expressing neurons labeled with the Pomc reporter similar to NPY neurons, suggesting that these distinct neurons can arise from a common progenitor. Finally, we hypothesized that temporal differences leading to premature depletion of progenitors in cKO mice lead to our observed phenotype. Using a BrdU birthdating paradigm, we determined the percentage of NPY and Kisspeptin neurons born on embryonic days 11.5, 12.5, and 13.5. We found no difference in the timing of differentiation of either neuronal subtype, with a majority occurring at e11.5. Taken together, our findings suggest that active Notch signaling is an important molecular switch involved in instructing subpopulations of progenitor cells to differentiate into Kisspeptin neurons.

PubMed ID: 26318021

MeSH Terms: Animals; Arcuate Nucleus of Hypothalamus/cytology; Arcuate Nucleus of Hypothalamus/embryology*; Bromodeoxyuridine; Gene Expression Regulation, Developmental/genetics; Gene Expression Regulation, Developmental/physiology*; Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics; Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism*; Immunohistochemistry; In Situ Hybridization; Kisspeptins/metabolism*; Mice; Mice, Knockout; Models, Neurological; Neurons/metabolism*; Neurons/physiology*; Pro-Opiomelanocortin/metabolism; Receptors, Notch/metabolism; Signal Transduction/physiology*