GABA excites immature CA3 pyramidal cells through bicuculline-sensitive and -insensitive chloride-dependent receptors.
Intracellular and patch clamp recording techniques were used to investigate the role of GABA in immature CA3 hippocampal neurons. During the first postnatal week spontaneous GABA release was detected as spontaneous ongoing synaptic potentials (SPSPs) or giant depolarizing potentials (GDPs). GDPs were generated at regular intervals and regulated by ionotropic glutamate receptors (GluRs), whereas SPSPs occurred randomly and were unaffected by ionotropic GluRs. Both GDPs and SPSPs were positively modulated by metabotropic GluRs through cyclic AMP-dependent protein kinase. Moreover GABA controlled its own release through GABAA and GABAB receptors, probably localized on GABAergic nerve terminals. At this developmental stage, GABA depolarized CA3 pyramidal cells through two distinct classes of chloride-permeable receptors: bicuculline sensitive and insensitive, respectively. The bicuculline-insensitive responses were blocked by picrotoxin in a noncompetitive way. Whole-cell GABA currents, recorded in the presence of bicuculline, had a slower desensitization rate and faster recovery from desensitization. In excised outside-out patches, in the presence of bicuculline, GABA activated single-channel currents with conductances of 14, 22, and 31 pS. These values were similar to those obtained when GABA was applied in the absence of bicuculline. Interestingly, GABA responses obtained in the absence of bicuculline, were sensitive to the blocking effect of zinc, whereas bicuculline-resistant responses were almost unaffected by this divalent cation. Expression of different subunits in native receptors (particularly of the alpha and rho type) may account for the functional differences observed in the present experiments. Activation of bicuculline-insensitive receptors would strengthen and prolong the depolarizing action of GABA, thus favoring the entry of calcium through voltage-dependent calcium channels. This calcium signal may be essential in promoting stabilization of synaptic contacts during a critical period of postnatal development.
Cherubini
E
E
Biophysics Laboratory, International School for Advanced Studies (SISSA), Trieste, Italy.
Martina
M
M
Sciancalepore
M
M
Strata
F
F
D000818:Animals; D000831:Animals, Newborn; D001418:Baclofen / Q000494:pharmacology; D001640:Bicuculline / Q000494:pharmacology*; D018118:Chloride Channels / Q000187:drug effects* / Q000378:metabolism; D046508:Culture Techniques; D005071:Evoked Potentials / Q000187:drug effects; D007736:Kynurenic Acid / Q000494:pharmacology; D009557:Nipecotic Acids / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010852:Picrotoxin / Q000494:pharmacology; D011392:Proline / Q000031:analogs & derivatives*; D017966:Pyramidal Cells / Q000187:drug effects* / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D011963:Receptors, GABA-A / Q000187:drug effects* / Q000378:metabolism; D018080:Receptors, GABA-B / Q000187:drug effects* / Q000378:metabolism; D015032:Zinc / Q000502:physiology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism / Q000502:physiology*
D001418:Baclofen; D001640:Bicuculline; D018118:Chloride Channels; D007736:Kynurenic Acid; D009557:Nipecotic Acids; D010852:Picrotoxin; D011392:Proline; D011963:Receptors, GABA-A; D018080:Receptors, GABA-B; D015032:Zinc; D005680:gamma-Aminobutyric Acid; C030278:nipecotic acid; C010983:homoproline