The Journal of neuroscience : the official journal of the Society for Neuroscience
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Channels from KCNQ2 and KCNQ3 genes have been suggested to underlie the neuronal M-type K(+) current. The M current is modulated by muscarinic agonists via G-proteins and an unidentified diffusible cytoplasmic messenger. Using whole-cell clamp, we studied tsA-201 cells in which cloned KCNQ2/KCNQ3 channels were coexpressed with M(1) muscarinic receptors. ⋯ Finally, when KCNQ2 subunits were overexpressed by intranuclear DNA injection in sympathetic neurons, total M current was fully modulated by the endogenous neuronal muscarinic signaling mechanism. Our data further rule out Ca(2+) as the diffusible messenger. The reconstitution of muscarinic modulation of the M current that uses cloned components should facilitate the elucidation of the muscarinic signaling mechanism.
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Intracellular recording and extracellular field potential (FP) recordings were obtained from spinal cord dorsal horn neurons (laminae I-IV) in a rat transverse slice preparation with attached dorsal roots. To study changes in synaptic inputs after neuroma formation, the sciatic nerve was sectioned and ligated 3 weeks before in vitro electrophysiological analysis. Horseradish peroxidase labeling of dorsal root axons indicated that Abeta fibers sprouted into laminae I-II from deeper laminae after sciatic nerve section. ⋯ The majority of low-threshold EPSPs in lamina II neurons after axotomy displayed properties similar to low-threshold EPSPs in lamina III of control slices. These results indicate that reoccupation of lamina II synapses by sprouting Abeta fibers normally terminating in lamina III occurs after sciatic nerve neuroma formation. Furthermore, these observations indicate that the lamina II neurons receive inappropriate sensory information from low-threshold mechanoreceptor after sciatic nerve neuroma formation.
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The processing of signals by integrative neurons in the retina and CNS relies strongly on inhibitory synaptic inputs, principally from GABAergic and glycinergic neurons that serve primarily to hyperpolarize postsynaptic neurons. Recent evidence indicates that the neuron-specific K-Cl cotransporter 2 (KCC2) is the major chloride extrusion system permitting hyperpolarizing inhibitory responses. It has been hypothesized that depolarizing GABA responses observed in immature neurons are converted to hyperpolarizing responses in large part by the expression of KCC2 during the second week of postnatal development. ⋯ Developmentally, KCC2 expression in the retina increased gradually from postnatal day 1 (P1) until P14 in the inner retina, whereas expression was delayed in the outer plexiform layer until P7 but reached its adult level by P14. These data support the hypothesis that the function of KCC2 is intimately involved in GABAergic synaptic processing. Furthermore, the delayed temporal expression of KCC2 in the outer plexiform layer indicates that GABAergic function may be differentially regulated in retina during postnatal development and that GABA may produce depolarizing responses in the outer plexiform layer at times when it generates hyperpolarizing responses in the inner plexiform layer.
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This study examined the antihyperalgesic and antinociceptive effects of opioid receptor agonists microinjected in the rostral ventromedial medulla (RVM) of rats 4 hr, 4 d, and 2 weeks after the induction of an inflammatory injury by injection of complete Freund's adjuvant (CFA) in one hindpaw. Nociceptive sensitivity of the ipsilateral, inflamed and the contralateral, uninflamed hindpaws was determined by the radiant-heat paw withdrawal test. The antihyperalgesic potency of the mu opioid receptor agonist [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]enkephalin (DAMGO), determined for the inflamed hindpaw, was enhanced 4 d and 2 weeks after injury. ⋯ The antihyperalgesic and antinociceptive effects of the delta opioid receptor agonist [D-Ala(2),Glu(4)]deltorphin were also increased 2 weeks after injury. These results indicate that peripheral inflammatory injury alters the pharmacology of excitatory and inhibitory inputs that modulate the activity of RVM neurons in such a manner as to enhance the effects of opioid agonists in this region. These changes have ramifications not only for the alleviation of hyperalgesia at the site of injury but also for opioid-induced antinociception at sites remote to the injury as revealed by increases in the potency of opioid agonists to suppress nociceptive responses of the contralateral, uninflamed hindpaw.
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We investigated the cellular mechanisms underlying the Ca(2+)-dependent release of glutamate from cultured astrocytes isolated from rat hippocampus. Using Ca(2+) imaging and electrophysiological techniques, we analyzed the effects of disrupting astrocytic vesicle proteins on the ability of astrocytes to release glutamate and to cause neuronal electrophysiological responses, i.e., a slow inward current (SIC) and/or an increase in the frequency of miniature synaptic currents. ⋯ Injection of astrocytes with the light chain of the neurotoxin Botulinum B that selectively cleaves the vesicle-associated SNARE protein synaptobrevin inhibited the astrocyte-induced glutamate response in neurons. Therefore, the Ca(2+)-dependent glutamate release from astrocytes is a SNARE protein-dependent process that requires the presence of functional vesicle-associated proteins, suggesting that astrocytes store glutamate in vesicles and that it is released through an exocytotic pathway.