GABAA receptor subunit mRNA expression in cultured embryonic and adult human dorsal root ganglion neurons.
Previous studies have demonstrated significant pharmacological differences between the GABA(A) receptors expressed by neurons cultured from embryonic and adult human dorsal root ganglia (DRG). GABA(A) receptors of both embryonic and adult neurons are potentiated by diazepam and low concentrations of pentobarbital, and are activated by high concentrations of pentobarbital. However, in contrast to the GABA responses of embryonic neurons, the GABA responses of adult neurons are insensitive to both bicuculline and picrotoxin. We performed RT-PCR using subunit specific primer pairs, followed by Southern blot analysis with a third specific primer, to determine the pattern of subunit mRNA expression in cultures of embryonic and adult human DRG neurons. alpha2 and beta3 mRNA were expressed in all embryonic and adult cultures, while beta2 mRNA was present in all adult cultures but none of the embryonic cultures. Transcripts expressed by at least half of both embryonic and adult cultures were alpha3, alpha5, gamma2S, gamma3, theta, and rho1. Transcripts for gamma1 and delta were expressed in most adult cultures, but only a single embryonic culture. alpha4 mRNA was expressed by a single embryonic culture and pi mRNA was expressed by a single adult culture. We found no evidence for expression of alpha1, alpha6, beta1, gamma2L or rho2 transcripts. Changes in receptor subunit composition may underlie the novel pharmacological properties of GABA(A) receptor responses in adult cells. However, post-translational modification of a known subunit or the expression of a novel subunit may also contribute to the unique pharmacology of these neurons.
Maddox
Floyd N
FN
Department of Molecular and Cellular Pharmacology, R-189, Miami Project to Cure Paralysis, University of Miami School of Medicine, Miami, FL 33101, USA.
Valeyev
Alexander Y
AY
Poth
Kevin
K
Holohean
Alice M
AM
Wood
Patrick M
PM
Davidoff
Robert A
RA
Hackman
John C
JC
Luetje
Charles W
CW
D000375:Aging / Q000378:metabolism; D001640:Bicuculline / Q000494:pharmacology; D015139:Blotting, Southern / Q000379:methods; D002478:Cells, Cultured; D000697:Central Nervous System Stimulants; D004347:Drug Interactions; D004622:Embryo, Mammalian; D018756:GABA Antagonists / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects / Q000196:embryology*; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques / Q000379:methods; D010852:Picrotoxin / Q000494:pharmacology; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011963:Receptors, GABA-A / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D001640:Bicuculline; D000697:Central Nervous System Stimulants; D018756:GABA Antagonists; D010852:Picrotoxin; D021122:Protein Subunits; D012333:RNA, Messenger; D011963:Receptors, GABA-A; D005680:gamma-Aminobutyric Acid