Electrophysiological properties and thermosensitivity of mouse preoptic and anterior hypothalamic neurons in culture

Neuroscience. 2005;135(2):433-49. doi: 10.1016/j.neuroscience.2005.06.053.

Abstract

Responses of mouse preoptic and anterior hypothalamic neurons to variations of temperature are key elements in regulating the setpoint of homeotherms. The goal of the present work was to assess the relevance of culture preparations for investigating the cellular mechanisms underlying thermosensitivity in hypothalamic cells. Our working hypothesis was that some of the main properties of preoptic/anterior hypothalamic neurons in culture are similar to those reported by other authors in slice preparations. Indeed, cultured preoptic/anterior hypothalamic neurons share many of the physiological and morphological properties of neurons in hypothalamic slices. They display heterogenous dendritic arbors and somatic shapes. Most of them are GABAergic and their activity is synaptically driven by the activation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate receptors. Active membrane properties include a depolarizing "sag" in response to hyperpolarization, and a low threshold spike, which is present in a majority of cells and is generated by T-type Ca2+ channels. In a fraction of the cells, the low threshold spike repeats rhythmically, either spontaneously, or in response to depolarization. The background synaptic noise in cultured neurons is characterized by the presence of numerous postsynaptic potentials which can be easily distinguished from the baseline, thus providing an opportunity for assessing their possible roles in thermosensitivity. An unexpected finding was that GABA-A receptors can generate both hyper- and depolarizing postsynaptic potentials in the same neuron. About 20% of the spontaneously firing preoptic/anterior hypothalamic neurons are warm-sensitive. Warming (32-41 degrees C) depolarizes some cells, a phenomenon which is Na+-dependent and tetrodotoxin-insensitive. The increased firing rate of warm-sensitive cells in response to warming can be prepotential and/or synaptically driven. Overall, our data suggest that a warm-sensitive phenotype is already developed in cultured cells. Therefore, and despite obvious differences in their networks, cultured and slice preparations of hypothalamic neurons can complement each other for further studies of warm-sensitivity at the cellular and molecular level.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 6-Cyano-7-nitroquinoxaline-2,3-dione / pharmacology
  • Action Potentials / physiology
  • Action Potentials / radiation effects
  • Animals
  • Bicuculline / pharmacology
  • Calcium Channel Blockers / pharmacology
  • Cells, Cultured
  • Electric Impedance
  • Electric Stimulation / methods
  • Embryo, Mammalian
  • Excitatory Amino Acid Antagonists / pharmacology
  • GABA Agonists / pharmacology
  • GABA Antagonists / pharmacology
  • Glutamate Decarboxylase / metabolism
  • Hypothalamus, Anterior / cytology*
  • Immunohistochemistry / methods
  • Isoenzymes / metabolism
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology*
  • Membrane Potentials / radiation effects
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Muscimol / pharmacology
  • Neural Inhibition / drug effects
  • Neural Inhibition / physiology
  • Neural Inhibition / radiation effects
  • Neurons / physiology
  • Neurons / radiation effects*
  • Nimodipine / pharmacology
  • Patch-Clamp Techniques / methods
  • Pyrimidines / pharmacology
  • Receptors, AMPA / metabolism
  • Sodium / pharmacology
  • Sodium Channel Blockers / pharmacology
  • Temperature*
  • Tetrodotoxin / pharmacology
  • Valine / analogs & derivatives
  • Valine / pharmacology
  • gamma-Aminobutyric Acid / pharmacology

Substances

  • Calcium Channel Blockers
  • Excitatory Amino Acid Antagonists
  • GABA Agonists
  • GABA Antagonists
  • Isoenzymes
  • MAP2 protein, rat
  • Microtubule-Associated Proteins
  • Pyrimidines
  • Receptors, AMPA
  • Sodium Channel Blockers
  • ICI D2788
  • Muscimol
  • Tetrodotoxin
  • gamma-Aminobutyric Acid
  • Nimodipine
  • 6-Cyano-7-nitroquinoxaline-2,3-dione
  • 2-amino-5-phosphopentanoic acid
  • Sodium
  • Glutamate Decarboxylase
  • glutamate decarboxylase 1
  • Valine
  • Bicuculline