The Ca 2؉ channel ␣1A-subunit is a voltage-gated, pore-forming membrane protein positioned at the intersection of two important lines of research: one exploring the diversity of Ca 2؉ channels and their physiological roles, and the other pursuing mechanisms of ataxia, dystonia, epilepsy, and migraine. ␣1A-Subunits are thought to support both P-and Q-type Ca 2؉ channel currents, but the most direct test, a null mutant, has not been described, nor is it known which changes in neurotransmission might arise from elimination of the predominant Ca 2؉ delivery system at excitatory nerve terminals. We generated ␣1A-deficient mice (␣1A ؊͞؊ ) and found that they developed a rapidly progressive neurological deficit with specific characteristics of ataxia and dystonia before dying Ϸ3-4 weeks after birth. P-type currents in Purkinje neurons and P-and Q-type currents in cerebellar granule cells were eliminated completely whereas other Ca 2؉ channel types, including those involved in triggering transmitter release, also underwent concomitant changes in density. Synaptic transmission in ␣1A ؊͞؊ hippocampal slices persisted despite the lack of P͞Q-type channels but showed enhanced reliance on N-type and R-type Ca 2؉ entry. The ␣1A ؊͞؊ mice provide a starting point for unraveling neuropathological mechanisms of human diseases generated by mutations in ␣1A. The ␣ 1A -subunit, the most abundant ␣ 1 -subunit in vertebrate brain (1), mediates Ca 2ϩ influx across presynaptic and somatodendritic membranes, thereby triggering fast neurotransmitter release and other key neuronal responses (2-5). Because of its high expression levels in the brain, the ␣ 1A -subunit was the first representative of its subclass to be isolated by cDNA cloning (1, 6). This predominantly neuronal subclass also includes ␣ 1B (N-type Ca 2ϩ channel) and ␣ 1E [possibly R-type Ca 2ϩ channel (7-9)] and is referred to as ABE or Ca V 2. There is no information to date on the behavioral or electrophysiological consequences of deleting a member of the ABE subfamily.␣ 1A Transcripts are widely distributed in rat (10) and human brain (11), most prominently in cell body layers in cerebellum and hippocampus. At the subcellular level, ␣ 1A immunoreactivity has been found in cell bodies, dendrites, and presynaptic terminals (12). Less clear has been the role of ␣ 1A in supporting Ca 2ϩ channel components defined by biophysical and pharmacological criteria. In either Xenopus oocytes (13, 14) or HEK293 cells (15), expression of ␣ 1A -subunits along with ancillary ␣ 2 ͞␦-and ␤-subunits generated currents with properties closely resembling the Q-type current found in cerebellar granule cells (8) and much less the P-type current first described in cerebellar Purkinje neurons by Llinás and colleagues (16,17). Unlike native P-type channels (18), the expressed currents showed pronounced inactivation during sustained depolarizations and responded to -agatoxin IVA (-Aga-IVA) at half-blocking doses of Ϸ100 nM, not Ϸ1 nM (13). Various explanations for the discrepancies have been advanced...

SUMMARY TRPV1 receptors feature prominently in nociception of spinal primary afferents but are also expressed in unmyelinated cranial visceral primary afferents linked to homeostatic regulation. Cranial visceral afferents enter the brain at the solitary tract nucleus (NTS) to control the heart, lungs and other vital organs. Here we identify a novel role for central TRPV1 in the activity-dependent facilitation of glutamatergic transmission from solitary tract (ST) afferents. Fast, synchronous ST-NTS transmission from capsaicin sensitive (TRPV1+) and insensitive (TRPV1−) afferents was similar. However, afferent activation triggered long lasting asynchronous glutamate release only from TRPV1+ synapses. Asynchronous release was proportional to synchronous EPSC amplitude, activity, and calcium entry. TRPV1 antagonists and low temperature blocked asynchronous release but not evoked EPSCs. At physiological afferent frequencies, asynchronous release strongly potentiated the duration of postsynaptic spiking. This activity dependent TPRV1-mediated facilitation is a novel form of synaptic plasticity that brings a unique central integrative feature to the CNS and autonomic regulation.

Increased extravascular lung water is a feature of early acute respiratory distress syndrome and predicts survival. Indexing extravascular lung water to predicted body weight, instead of actual body weight, improves the predictive value of extravascular lung water for survival and correlation with markers of disease severity.

The hypothalamus coordinates autonomic responses in part through arginine vasopressin (AVP) released in medial nucleus tractus solitarius (NTS).

Spontaneous release of glutamate is important for maintaining synaptic strength and controlling spike timing in the brain. Mechanisms regulating spontaneous exocytosis remain poorly understood. Extracellular calcium concentration ([Ca2+]o) regulates Ca2+ entry through voltage-activated calcium channels (VACCs) and consequently is a pivotal determinant of action potential-evoked vesicle fusion. Extracellular Ca2+ also enhances spontaneous release, but via unknown mechanisms. Here we report that external Ca2+ triggers spontaneous glutamate release more weakly than evoked release in mouse neocortical neurons. Blockade of VACCs has no effect on the spontaneous release rate or its dependence on [Ca2+]o. Intracellular [Ca2+] slowly increases in a minority of neurons following increases in [Ca2+]o. Furthermore, the enhancement of spontaneous release by extracellular calcium is insensitive to chelation of intracellular calcium by BAPTA. Activation of the calcium-sensing receptor (CaSR), a G-protein coupled receptor present in nerve terminals, by several specific agonists increased spontaneous glutamate release. The frequency of spontaneous synaptic transmission was decreased in CaSR mutant neurons. The concentration effect relationship for extracellular calcium regulation of spontaneous release was well described by a combination of CaSR-dependent and CaSR-independent mechanisms. Overall these results indicate that extracellular Ca2+ does not trigger spontaneous glutamate release by simply increasing calcium influx but stimulates CaSR and thereby promotes resting spontaneous glutamate release.

Several modes of synaptic vesicle release, retrieval and recycling have been identified. In a well-established mode of exocytosis, termed ‘full-collapse fusion’, vesicles empty their neurotransmitter content fully into the synaptic cleft by flattening out and becoming part of the presynaptic membrane. The fused vesicle membrane is then reinternalized via a slow and clathrin-dependent mode of compensatory endocytosis that takes several seconds. A more fleeting mode of vesicle fusion, termed ‘kiss-and-run’ exocytosis or ‘flicker-fusion’, indicates that during synaptic transmission some vesicles are only briefly connected to the presynaptic membrane by a transient fusion pore. Finally, a mode that retrieves a large amount of membrane, equivalent to that of several fused vesicles, termed ‘bulk endocytosis’, has been found after prolonged exocytosis. We are of the opinion that both fast and slow modes of endocytosis co-exist at central nervous system nerve terminals and that one mode can predominate depending on stimulus strength, temperature and synaptic maturation.

Synaptic activity causes reductions in cleft [Ca(2+)] that may impact subsequent synaptic efficacy. Using modified patch-clamp techniques to record from single neocortical nerve terminals, we report that physiologically relevant reductions of extracellular [Ca(2+)] ([Ca(2+)](o)) activate voltage-dependent outward currents. These outward currents are carried by a novel nonselective cation (NSC) channel that is indirectly inhibited by various extracellular agents (rank order potency, Gd(3+) > spermidine > Ca(2+) > Mg(2+), typical for [Ca(2+)](o) receptors). The identification of a Ca(2+) sensor-NSC channel pathway establishes the existence of a mechanism by which presynaptic terminals can detect and respond to reductions in cleft [Ca(2+)]. Activation of NSC channels by falls in [Ca(2+)](o) would be expected during periods of high activity in the neocortex and may modulate the excitability of the presynaptic terminal.

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