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Preparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid Pathology
Published on: March 23, 2011
Age-dependent changes in the functional expression of two nicotinic receptor subtypes in CA1 stratum radiatum
Manickavasagom Alkondon1, Edna F R Pereira, Edson X Albuquerque
1Department of Pharmacology and Experimental Therapeutics, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Abstract:
Protein density measurements and mRNA analysis have provided valuable information on age-dependent changes in the distribution of different nicotinic receptor (nAChR) subtypes in various areas of the rat brain, including the hippocampus. However, very little is known regarding the functional expression of nAChRs in individual neuron types at various ages. Likewise, there is paucity of information regarding the functional and pharmacological profile of nAChRs in the mature rat hippocampus. To address these issues, we used the whole-cell patch-clamp technique to record nicotinic responses from CA1 stratum radiatum (SR) interneurons in hippocampal slices from rat pups (5-19 days old) and adult rats (2-5 months old). As previously observed in the hippocampus of rat pups, CA1 SR interneurons in the hippocampus of adult rats responded to choline (10mM, 12s) with whole-cell currents that decayed to the baseline within the agonist pulse, were sensitive to inhibition by methyllycaconitine (10nM) or alpha-bungarotoxin (50 nM), and were, therefore, mediated by alpha7*(1)[1] nAChRs. Likewise, as previously observed in the hippocampus of young rats, in the adult rat hippocampus excitatory postsynaptic currents (EPSCs) were recorded from SR interneurons in response to a pulse of ACh (0.1 mM, 12s) applied in the presence of the GABA(A) receptor antagonist bicuculline. ACh-triggered EPSCs were inhibited by mecamylamine (1 microM) or choline (1 mM) and were, therefore, likely to have resulted from activation of alpha3beta4beta2* nAChR. The magnitude of alpha7* nAChR-mediated responses increased with the age of the animals. In contrast, the magnitude of alpha3beta4beta2* nAChR-mediated responses was highest at the second postnatal week. The distinct age dependency of functional expression of alpha7* and alpha3beta4beta2* nAChRs strongly suggests that the excitability of CA1 SR interneurons is differentially regulated by the nicotinic cholinergic system in the hippocampus of rat pups and adult rats.
