Role of GABAB receptors in intracellular Ca2+ homeostasis and possible interaction between GABAA and GABAB receptors in regulation of transmitter release in cerebellar granule neurons

J. Kardos, L. Elster, I. Damgaard, P. Krogsgaard‐Larsen, A. Schousboe

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The expression of GABAB receptors in cultured mouse cerebellar granule cells was investigated in binding experiments using [3H](S, R)‐baclofen as well as in functional assessment of the ability of (R)‐baclofen to interact with depolarization (15–40 mM KCI) coupled changes in intracellular Ca2+ homeostasis and neurotransmitter release. In the latter case a possible functional coupling between GABAA and GABAB receptors was investigated. The binding studies showed that the granule cells express specific binding sites for (R)‐baclofen. The number of binding sites could be increased by exposure of the cells to the GABAA receptor agonist THIP (4,5,6,7‐tetrahy‐droisoxazolo[5,4‐c]pyridin‐3‐ol) during the culture period. Pretreatment of the neurons with pertussis toxin showed that the GABAB receptors are coupled to G‐proteins. This coupling was, however, less pronounced when the cells had been cultured in the presence of THIP. When 45Ca2+ uptake was measured or the intracellular Ca2+ concentration ([Ca2+]i) determined using the fluorescent Ca2+ chelator Fluo‐3 it could be demonstrated that culturing the neurons in THIP influences intracellular Ca2+ homeostasis. Moreover, this homeostasis was found to be functionally coupled to the GABAB receptors as (R)‐baclofen inhibited depolarization‐induced increases in 45Ca2+ uptake and [Ca2+]i. (R)‐Baclofen also inhibited K+‐induced transmitter release from the neurons as monitored by the use of [3H]D‐aspartate which labels the neurotransmitter pool of glutamate. Using the selective GABAA receptor agonist isoguvacine it could be demonstrated that the GABAB receptors are functionally coupled to GABAA receptors in the neurons leading to a disinhibitory action of GABAB receptor agonists. © 1994 Wiley‐Liss, Inc.

Original languageEnglish
Pages (from-to)646-655
Number of pages10
JournalJournal of Neuroscience Research
Volume39
Issue number6
DOIs
Publication statusPublished - Dec 15 1994

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Keywords

  • G‐proteins
  • THIP
  • baclofen
  • cerebellum
  • disinhibition
  • granule neurons
  • intracellular Ca
  • pertussis toxin

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

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