Biomedical subjects

Sterling N Sudweeks

Publications and source records attributed to Sterling N Sudweeks.

3 recordsLinked to original sources

PubMed2006

Connexin-36 gap junctions mediate electrical coupling between ventral tegmental area GABA neurons.

Communication between neurons in the mammalian brain is primarily through chemical synapses; however, evidence is accumulating in support of electrical synaptic transmission between some neuronal types in the mature nervous system. The authors have recently demonstrated that the gap junction (GJ) blocker quinidine suppresses stimulus-induced and dopamine-evoked coupling of gamma amino butyric acid (GABA) neurons in the ventral tegmental area (VTA) of mature rats (Stobbs et al., 2004). The aim of this study was to evaluate the role of connexin-36 (Cx36) GJs in mediating electrical coupling between VTA GABA neurons in P50-80 rats in vivo and P25-50 rats in vitro. Single stimulation of the internal capsule (IC) evoked VTA GABA neuron spike couplets in mature rats when activated antidromically, and multiple poststimulus spike discharges (PSDs) when activated with brief high-frequency stimulation of the IC (ICPSDs). The Cx36 GJ blocker mefloquine (30 mg/kg) suppressed VTA GABA neuron ICPSDs in mature freely behaving rats. VTA GABA neurons recorded via whole-cell patch clamp in the midbrain slice preparation of P25-50 rats showed robust expression of Cx36 transcripts when tested with single-cell quantitative reverse transcription polymerase chain reaction. In P50-80 rats, Cx36 protein immunoreactivity was evident in the VTA and surrounding structures. Dye-coupling between VTA neurons was observed following Neurobiotin labeling of VTA GABA neurons, as well as with the fluorochrome Alexa Fluor 488 using real-time video fluorescent microscopy. Thus, mature VTA GABA neurons appear to be connected by electrical synapses via Cx36 GJs, whose coupling is enhanced by corticotegmental input and by dopamine.

Animals↗

PubMed2002

Serotonin 5-HT(3) receptors in rat CA1 hippocampal interneurons: functional and molecular characterization.

The molecular makeup of the serotonin 5-HT(3) receptor (5-HT(3)R) channel was investigated in rat hippocampal CA1 interneurons in slices using single-cell RT-PCR and patch-clamp recording techniques. We tested for the expression of the 5-HT(3A) (both short and long splice variants) and 5-HT(3B) subunits, as well as the expression of the alpha4 subunit of the neuronal nicotinic ACh receptors (nAChRs), the latter of which has been shown to co-assemble with the 5-HT(3A) subunit in heterologous expression systems. Both the 5-HT(3A)-short and alpha4-nAChR subunits were expressed in these interneurons, but we could not detect any expression of either the 5-HT(3B) or the 5-HT(3A)-long subunits. Furthermore, there was a strong tendency for the 5-HT(3A)-short and alpha4-nAChR subunits to be co-expressed in individual interneurons. To assess whether there was any functional evidence for co-assembly between the 5-HT(3A)-short and alpha4-nAChR subunits, we used the sulphydryl agent 2-aminoethyl methanethiosulphonate (MTSEA), which has previously been shown to modulate expressed 5-HT(3)Rs that contain the alpha4-nAChR subunit. In half of the interneurons examined, MTSEA significantly enhanced the amplitude of the 5-HT(3)R-mediated responses, which is consistent with the notion that the alpha4-nAChR subunit co-assembles with the 5-HT(3A) subunit to form a native heteromeric 5-HT(3)R channel in rat CA1 hippocampal interneurons in vivo. In addition, the single-channel properties of the 5-HT(3)R were investigated in outside-out patches. No resolvable single-channel currents were observed. Using non-stationary fluctuation analysis, we obtained an estimate of the single-channel conductance of 4 pS, which is well below that expected for channels containing both the 5-HT(3A) and 5-HT(3B) subunits.

Action Potentials↗

PubMed2002

Rat nicotinic ACh receptor alpha7 and beta2 subunits co-assemble to form functional heteromeric nicotinic receptor channels.

Rat hippocampal interneurons express diverse subtypes of functional nicotinic acetylcholine receptors (nAChRs), including alpha7-containing receptors that have properties unlike those expected for homomeric alpha7 nAChRs. We previously reported a strong correlation between expression of the alpha7 and of the beta2 subunits in individual neurons. To explore whether co-assembly of the alpha7 and beta2 subunits might occur, these subunits were co-expressed in Xenopus oocytes and the functional properties of heterologously expressed nAChRs were characterized by two-electrode voltage clamp. Co-expression of the beta2 subunit, both wild-type and mutant forms, with the alpha7 subunit significantly slowed the rate of nAChR desensitization and altered the pharmacological properties. Whereas ACh, carbachol and choline were full or near-full agonists for homomeric alpha7 receptor channels, both carbachol and choline were only partial agonists in oocytes expressing both alpha7 and beta2 subunits. In addition the EC(50) values for all three agonists significantly increased when the beta2 subunit was co-expressed with the alpha7 subunit. Co-expression with the beta2 subunit did not result in any significant change in the current-voltage curve. Biochemical evidence for the co-assembly of the alpha7 and beta2 subunits was obtained by co-immunoprecipitation of these subunits from transiently transfected human embryonic kidney (TSA201) cells. These data provide direct biophysical and molecular evidence that the nAChR alpha7 and beta2 subunits co-assemble to form a functional heteromeric nAChR with functional and pharmacological properties different from those of homomeric alpha7 channels. This co-assembly may help to explain nAChR channel diversity in rat hippocampal interneurons, and perhaps in other areas of the nervous system.

Acetylcholine↗