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Applicability of Anti-Human Estrogen Receptor β Antibody PPZ0506 for the Immunodetection of Rodent Estrogen Receptor β Proteins
Open AccessArticle

Neurochemical Characterization of Neurons Expressing Estrogen Receptor β in the Hypothalamic Nuclei of Rats Using in Situ Hybridization and Immunofluorescence

Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan
*
Author to whom correspondence should be addressed.
Current addresses: Department of Neurophysiology, School of Medicine, Tokyo Women’s Medical University, 8–1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan; Japan Society for the Promotion of Science, Tokyo 102-8472, Japan.
Int. J. Mol. Sci. 2020, 21(1), 115; https://doi.org/10.3390/ijms21010115
Received: 31 October 2019 / Revised: 14 December 2019 / Accepted: 18 December 2019 / Published: 23 December 2019
(This article belongs to the Special Issue Steroids and Lipophilic Hormones, and Their Actions)
Estrogens play an essential role in multiple physiological functions in the brain, including reproductive neuroendocrine, learning and memory, and anxiety-related behaviors. To determine these estrogen functions, many studies have tried to characterize neurons expressing estrogen receptors known as ERα and ERβ. However, the characteristics of ERβ-expressing neurons in the rat brain still remain poorly understood compared to that of ERα-expressing neurons. The main aim of this study is to determine the neurochemical characteristics of ERβ-expressing neurons in the rat hypothalamus using RNAscope in situ hybridization (ISH) combined with immunofluorescence. Strong Esr2 signals were observed especially in the anteroventral periventricular nucleus (AVPV), bed nucleus of stria terminalis, hypothalamic paraventricular nucleus (PVN), supraoptic nucleus, and medial amygdala, as previously reported. RNAscope ISH with immunofluorescence revealed that more than half of kisspeptin neurons in female AVPV expressed Esr2, whereas few kisspeptin neurons were found to co-express Esr2 in the arcuate nucleus. In the PVN, we observed a high ratio of Esr2 co-expression in arginine-vasopressin neurons and a low ratio in oxytocin and corticotropin-releasing factor neurons. The detailed neurochemical characteristics of ERβ-expressing neurons identified in the current study can be very essential to understand the estrogen signaling via ERβ. View Full-Text
Keywords: estrogen receptor β; Esr2; RNAscope; kisspeptin; anteroventral periventricular nucleus; arcuate nucleus; vasopressin; oxytocin; corticotropin-releasing factor; paraventricular nucleus estrogen receptor β; Esr2; RNAscope; kisspeptin; anteroventral periventricular nucleus; arcuate nucleus; vasopressin; oxytocin; corticotropin-releasing factor; paraventricular nucleus
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Kanaya, M.; Higo, S.; Ozawa, H. Neurochemical Characterization of Neurons Expressing Estrogen Receptor β in the Hypothalamic Nuclei of Rats Using in Situ Hybridization and Immunofluorescence. Int. J. Mol. Sci. 2020, 21, 115.

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