Phenotyping of nNOS neurons in the postnatal and adult female mouse hypothalamus

J Comp Neurol. 2017 Oct 15;525(15):3177-3189. doi: 10.1002/cne.24257. Epub 2017 Jun 19.

Abstract

Neurons expressing nitric oxide (NO) synthase (nNOS) and thus capable of synthesizing NO play major roles in many aspects of brain function. While the heterogeneity of nNOS-expressing neurons has been studied in various brain regions, their phenotype in the hypothalamus remains largely unknown. Here we examined the distribution of cells expressing nNOS in the postnatal and adult female mouse hypothalamus using immunohistochemistry. In both adults and neonates, nNOS was largely restricted to regions of the hypothalamus involved in the control of bodily functions, such as energy balance and reproduction. Labeled cells were found in the paraventricular, ventromedial, and dorsomedial nuclei as well as in the lateral area of the hypothalamus. Intriguingly, nNOS was seen only after the second week of life in the arcuate nucleus of the hypothalamus (ARH). The most dense and heavily labeled population of cells was found in the organum vasculosum laminae terminalis (OV) and the median preoptic nucleus (MEPO), where most of the somata of the neuroendocrine neurons releasing GnRH and controlling reproduction are located. A great proportion of nNOS-immunoreactive neurons in the OV/MEPO and ARH were seen to express estrogen receptor (ER) α. Notably, almost all ERα-immunoreactive cells of the OV/MEPO also expressed nNOS. Moreover, the use of EYFPVglut2 , EYFPVgat , and GFPGad67 transgenic mouse lines revealed that, like GnRH neurons, most hypothalamic nNOS neurons have a glutamatergic phenotype, except for nNOS neurons of the ARH, which are GABAergic. Altogether, these observations are consistent with the proposed role of nNOS neurons in physiological processes.

Keywords: GABA; GnRH neurons; Nitric oxide; RRID:AB_2340375; RRID:AB_2534017; RRID:AB_2535865; RRID:AB_2651133; RRID:AB_300798; RRID:AB_310305; RRID:IMSR_JAX:006148; RRID:IMSR_JAX:007677; RRID:IMSR_JAX:016962; RRID:IMSR_JAX:016963; RRID:SCR_014199; development; estrogen receptor; glutamate; hypothalamus; immunofluorescence.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Estrogen Receptor alpha / metabolism
  • Female
  • Glutamate Decarboxylase / genetics
  • Glutamate Decarboxylase / metabolism
  • Hypothalamus / cytology
  • Hypothalamus / growth & development*
  • Hypothalamus / metabolism*
  • Immunohistochemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Neurons / cytology*
  • Neurons / metabolism*
  • Nitric Oxide Synthase Type I / metabolism*
  • Vesicular Glutamate Transport Protein 2 / genetics
  • Vesicular Glutamate Transport Protein 2 / metabolism
  • Vesicular Inhibitory Amino Acid Transport Proteins / genetics
  • Vesicular Inhibitory Amino Acid Transport Proteins / metabolism

Substances

  • Bacterial Proteins
  • Estrogen Receptor alpha
  • Luminescent Proteins
  • Slc17a6 protein, mouse
  • Vesicular Glutamate Transport Protein 2
  • Vesicular Inhibitory Amino Acid Transport Proteins
  • Viaat protein, mouse
  • yellow fluorescent protein, Bacteria
  • Nitric Oxide Synthase Type I
  • Nos1 protein, mouse
  • Glutamate Decarboxylase
  • glutamate decarboxylase 1