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Fluorescence imaging of GABAA receptor-mediated intracellular [Cl-] in P19-N cells reveals unique pharmacological properties.

Publication ,  Journal Article
Chistina Grobin, A; Inglefield, JR; Schwartz-Bloom, RD; Devaud, LL; Morrow, AL
Published in: Brain Res
May 8, 1999

This study describes the pharmacological properties of GABAA receptors expressed in P19-N cells using fluorescence imaging of intracellular chloride with 6-methoxy-N-ethylquinolinium iodide (MEQ). We show that application of the GABA agonist, muscimol (10-200 microM), produces time- and concentration-dependent increases in intracellular [Cl-] that are blocked by bicuculline. Diazepam (10 microM) and pentobarbital (1 mM) potentiate muscimol-stimulation. These receptors exhibit novel pharmacological properties. The neurosteroid, 3alpha-hydroxy-5alpha-pregnane-20-one (1-10 microM) exhibited weak potency in enhancement of muscimol-stimulation. Ethanol (50 and 100 mM) exhibited high efficacy on muscimol responses, a 4- to 5-fold potentiation, respectively, of muscimol (10 microM) alone. GABA and muscimol allosterically modulated specific binding of [3H]flunitrazepam to differentiated P19 cells. Modulation of GABAA receptor mediated increases in intracellular [Cl-] demonstrated stability in response magnitude from 7 to 15 days following removal of retinoic acid. In concert, GABAA receptor subunit mRNA and protein expression patterns in these neuron-like cells were stable over the same period. Using RT-PCR we determined that differentiated P19 cells lack gamma1, gamma2L, alpha6 and delta subunit mRNAs while expressing alpha1, alpha2, alpha3, alpha4, alpha5, beta1, beta2, beta3, gamma2S and gamma3. Furthermore, subunit specific antibody immunocytochemical labeling of cells with a neuronal morphology indicated the presence of alpha1, alpha2, alpha4, and gamma2 subunits (the only subunits tested). Therefore, P19-N cells should prove useful to researchers in need of a model cell culture system in which to study function and regulation of neuronal GABAA receptors.

Duke Scholars

Published In

Brain Res

DOI

ISSN

0006-8993

Publication Date

May 8, 1999

Volume

827

Issue

1-2

Start / End Page

1 / 11

Location

Netherlands

Related Subject Headings

  • Tritium
  • Reverse Transcriptase Polymerase Chain Reaction
  • Receptors, GABA-A
  • RNA, Messenger
  • Quinolinium Compounds
  • Pregnanolone
  • Pentobarbital
  • Neurology & Neurosurgery
  • Neuroglia
  • Neoplastic Stem Cells
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Chistina Grobin, A., Inglefield, J. R., Schwartz-Bloom, R. D., Devaud, L. L., & Morrow, A. L. (1999). Fluorescence imaging of GABAA receptor-mediated intracellular [Cl-] in P19-N cells reveals unique pharmacological properties. Brain Res, 827(1–2), 1–11. https://doi.org/10.1016/s0006-8993(99)01223-8
Chistina Grobin, A., J. R. Inglefield, R. D. Schwartz-Bloom, L. L. Devaud, and A. L. Morrow. “Fluorescence imaging of GABAA receptor-mediated intracellular [Cl-] in P19-N cells reveals unique pharmacological properties.Brain Res 827, no. 1–2 (May 8, 1999): 1–11. https://doi.org/10.1016/s0006-8993(99)01223-8.
Chistina Grobin A, Inglefield JR, Schwartz-Bloom RD, Devaud LL, Morrow AL. Fluorescence imaging of GABAA receptor-mediated intracellular [Cl-] in P19-N cells reveals unique pharmacological properties. Brain Res. 1999 May 8;827(1–2):1–11.
Chistina Grobin, A., et al. “Fluorescence imaging of GABAA receptor-mediated intracellular [Cl-] in P19-N cells reveals unique pharmacological properties.Brain Res, vol. 827, no. 1–2, May 1999, pp. 1–11. Pubmed, doi:10.1016/s0006-8993(99)01223-8.
Chistina Grobin A, Inglefield JR, Schwartz-Bloom RD, Devaud LL, Morrow AL. Fluorescence imaging of GABAA receptor-mediated intracellular [Cl-] in P19-N cells reveals unique pharmacological properties. Brain Res. 1999 May 8;827(1–2):1–11.
Journal cover image

Published In

Brain Res

DOI

ISSN

0006-8993

Publication Date

May 8, 1999

Volume

827

Issue

1-2

Start / End Page

1 / 11

Location

Netherlands

Related Subject Headings

  • Tritium
  • Reverse Transcriptase Polymerase Chain Reaction
  • Receptors, GABA-A
  • RNA, Messenger
  • Quinolinium Compounds
  • Pregnanolone
  • Pentobarbital
  • Neurology & Neurosurgery
  • Neuroglia
  • Neoplastic Stem Cells