566 research outputs found
SK2 channels are required for function and long-term survival of efferent synapses on mammalian outer hair cells
Cochlear hair cells use SK2 currents to shape responses to cholinergic efferent feedback from the brain. Using SK2-/- mice, we demonstrate that, in addition to their previously defined role in modulating hair cell membrane potentials, SK2 channels are necessary for long-term survival of olivocochlear fibers and synapses. Loss of the SK2 gene also results in loss of electrically driven olivocochlear effects in vivo, and down regulation of ryanodine receptors involved in calcium-induced calcium release, the main inducer of nAChR evoked SK2 activity. Generation of double-null mice lacking both the α10 nAChR gene, loss of which results in hypertrophied olivocochlear terminals, and the SK2 gene, recapitulates the SK2-/- synaptic phenotype and gene expression, and also leads to down regulation of α9 nAChR gene expression. The data suggest a hierarchy of activity necessary to maintain early olivocochlear synapses at their targets, with SK2 serving an epistatic, upstream, role to the nAChRs.Fil: Murthy, Vidya. Tufts University; Estados UnidosFil: Maison, StĂ©phane F.. Massachusetts Eye and Ear Infirmary; Estados Unidos. Harvard Medical School; Estados UnidosFil: Taranda, Julian. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; Argentina. Tufts University; Estados UnidosFil: Haque, Nadeem. University of Notre Dame; Estados UnidosFil: Bond, Chris T.. Oregon Health Sciences University; Estados UnidosFil: Elgoyhen, Ana Belen. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; ArgentinaFil: Adelman, John P.. Oregon Health Sciences University; Estados UnidosFil: Liberman, M. Charles. Massachusetts Eye and Ear Infirmary; Estados Unidos. Harvard Medical School; Estados UnidosFil: Vetter, Douglas E.. Tufts University; Estados Unido
Cooperative interactions among subunits of a voltage-dependent potassium channel. Evidence from expression of concatenated cDNAs
Four copies of the coding sequence for a voltage-dependent potassium channel (RBK1, rat Kv1.1) were ligated contiguously and transcribed in vitro. The resulting RNA encodes four covalently linked subunit domains ([4]RBK1). Injection of this RNA into Xenopus oocytes resulted in the expression of voltage-dependent potassium currents. A single amino acid substitution, Tyr--\u3eVal, located within the outer mouth of the pore, introduced into the equivalent position of any of the four domains, reduced affinity for external tetraethylammonium by approximately the same amount. In constructs containing 0, 1, 2, 3, or 4 Tyr residues the free energy of binding tetraethylammonium was linearly related to the number of Tyr residues. A different amino acid substitution, Leu--\u3eIle, located in the S4 region, was made in the equivalent position of one, two, three, or four domains. The depolarization required for channel activation increased approximately linearly with the number of Ile residues, whereas models of independent gating of each domain predict marked nonlinearity. Expression of this concatenated channel provides direct evidence that voltage-dependent potassium channels have four subunits positioned symmetrically around a central permeation pathway and that these subunits interact cooperatively during channel activation
Evidence for a Deep Pore Activation Gate in Small Conductance Ca2+-activated K+ Channels
Small conductance calcium-gated potassium (SK) channels share an overall topology with voltage-gated potassium (Kv) channels, but are distinct in that they are gated solely by calcium (Ca2+), not voltage. For Kv channels there is strong evidence for an activation gate at the intracellular end of the pore, which was not revealed by substituted cysteine accessibility of the homologous region in SK2 channels. In this study, the divalent ions cadmium (Cd2+) and barium (Ba2+), and 2-aminoethyl methanethiosulfonate (MTSEA) were used to probe three sites in the SK2 channel pore, each intracellular to (on the selectivity filter side of) the region that forms the intracellular activation gate of voltage-gated ion channels. We report that Cd2+ applied to the intracellular side of the membrane can modify a cysteine introduced to a site (V391C) just intracellular to the putative activation gate whether channels are open or closed. Similarly, MTSEA applied to the intracellular side of the membrane can access a cysteine residue (A384C) that, based on homology to potassium (K) channel crystal structures (i.e., the KcsA/MthK model), resides one amino acid intracellular to the glycine gating hinge. Cd2+ and MTSEA modify with similar rates whether the channels are open or closed. In contrast, Ba2+ applied to the intracellular side of the membrane, which is believed to block at the intracellular end of the selectivity filter, blocks open but not closed channels when applied to the cytoplasmic face of rSK2 channels. Moreover, Ba2+ is trapped in SK2 channels when applied to open channels that are subsequently closed. Ba2+ pre-block slows MTSEA modification of A384C in open but not in closed (Ba2+-trapped) channels. The findings suggest that the SK channel activation gate resides deep in the vestibule of the channel, perhaps in the selectivity filter itself
Immunocytochemical localization of small-conductance, calcium-dependent potassium channels in astrocytes of the rat supraoptic nucleus
Supraoptic nucleus (SON) neurons possess a prominent afterhyperpolarization (AHP) that contributes to spike patterning. This AHP is probably underlain by a small-conductance, CA2+-dependent, K+ type 3 (SK3) channel. To determine the distribution of SK3 channels within the SON, we used immunocytochemistry in rats and in transgenic mice with a regulatory cassette on the SK3 gene, allowing regulated expression with dietary doxycycline (DOX). In rats and wild-type mice, SK3 immunostaining revealed an intense lacy network surrounding SON neurons, with weak staining in neuronal somata and dendrites. In untreated, conditional SK3 knockout mice, SK3 was overexpressed, but the pericellular pattern in the SON was similar to that of rats. DOX-treated transgenic mice exhibited no SK3 staining in the SON. Double staining for oxytocin or vasopressin neurons revealed weak co-localization with SK3 but strong staining surrounding each neuron type. Electron microscopy showed that SK3-like immunoreactivity was intense between neuronal somata and dendrites, in apparent glial processes, but weak in neurons. This was confirmed by using confocal microscopy and double staining for glial fibrillary acidic protein (GFAP) and SK3: many GFAP-positive processes in the SON, and in the ventral dendritic/glial lamina, were shown to contain SK3-like immunoreactivity. These studies suggest a prominent role of SK3 channels in astrocytes. Given the marked plasticity in glial/neuronal relationships, as well as studies suggesting that astrocytes in the central nervous system can generate prominent CA2+ transients to various stimuli, a CA2+-dependent K+ channel may help SON astrocytes with K+ buffering whenever astrocyte intracellular CA2+ is increased. © 2005 Wiley-Liss, Inc
Regional variation in angioplasty practice in the United States: A report from the Hirulog angioplasty study
Pla general de la font anomenada Homenatge
al poble, ubicada a la plaça Molina. A la part
superior de la font, a cada costat, hi ha un escut cisellat. Sota un d'aquests es troba la frase Gratitud al Ayuntamiento.Realitzada en pedra abans de 1874
A Conformation Change in the Extracellular Domain that Accompanies Desensitization of Acid-sensing Ion Channel (ASIC) 3
Acid-sensing ion channels (ASICs) are thought to trigger some forms of acid-induced pain and taste, and to contribute to stroke-induced neural damage. After activation by low extracellular pH, different ASICs undergo desensitization on time scales from 0.1 to 10 s. Consistent with a substantial conformation change, desensitization slows dramatically when temperature drops (Askwith, C.C., C.J. Benson, M.J. Welsh, and P.M. Snyder. 2001. PNAS. 98:6459–6463). The nature of this conformation change is unknown, but two studies showed that desensitization rate is altered by mutations on or near the first transmembrane domain (TM1) (Coric, T., P. Zhang, N. Todorovic, and C.M. Canessa. 2003. J. Biol. Chem. 278:45240–45247; Pfister, Y., I. Gautschi, A.-N. Takeda, M. van Bemmelen, S. Kellenberger, and L. Schild. 2006. J. Biol. Chem. 281:11787–11791). Here we show evidence of a specific conformation change associated with desensitization. When mutated from glutamate to cysteine, residue 79, which is some 20 amino acids extracellular to TM1, can be altered by cysteine-modifying reagents when the channel is closed, but not when it is desensitized; thus, desensitization appears to conceal the residue from the extracellular medium. D78 and E79 are a pair of adjacent acidic amino acids that are highly conserved in ASICs yet absent from epithelial Na+ channels, their acid-insensitive relatives. Despite large effects on desensitization by mutations at positions 78 and 79—including a shift to 10-fold lower proton concentration with the E79A mutant—there are not significant effects on activation
Transversely Driven Charge Density Waves and Striped Phases of High-T Superconductors: The Current Effect Transistor
We show that a normal (single particle) current density {\em
transverse} to the ordering wavevector of a charge density
wave (CDW) has dramatic effects both above and {\em below} the CDW depinning
transition. It exponentially (in ) enhances CDW correlations, and
exponentially suppresses the longitudinal depinning field. The intermediate
longitudinal I-V relation also changes, acquiring a {\em linear} regime. We
propose a novel ``current effect transistor'' whose CDW channel is turned on by
a transverse current. Our results also have important implications for the
recently proposed ``striped phase'' of the high-T superconductors.Comment: change of title and minor corrections, 4 RevTeX pgs, to appear in
Phys. Rev. Lett., 81, 3711 (1998
A Spectacular H Complex in Virgo: Evidence for a Collision Between M86 and NGC 4438 and Implications for Collisional ISM Heating of Ellipticals
Deep wide-field H+[NII] imaging around the Virgo cluster giant
elliptical galaxy M86 reveals a highly complex and disturbed ISM/ICM. The most
striking feature is a set of H filaments which clearly connect M86 with
the nearby disturbed spiral NGC 4438 (23=120 kpc projected away), providing
strong evidence for a previously unrecognized collision between them.
Spectroscopy of selected regions show a fairly smooth velocity gradient between
M86 and NGC 4438, consistent with the collision scenario. Such a collision
would impart significant energy into the ISM of M86, probably heating the gas
and acting to prevent the gas from cooling to form stars. We propose that cool
gas stripped from NGC 4438 during the collision and deposited in its wake is
heated by shocks, ram pressure drag, or thermal conduction, producing most of
the H filaments. Some H filaments are associated with the
well-known ridge of bright X-ray emission to the NW of the nucleus, suggesting
that the collision is responsible for peculiarities of M86 previously ascribed
to other effects. M86 is radio-quiet, thus AGN heating is unlikely to play a
significant role. The M86 system has implications for understanding the role of
gravitational interactions in the heating of the ISM in ellipticals, and how
collisions in clusters transform galaxies.Comment: 6 pages, 2 figures. For high-resolution images, see
http://www.astro.yale.edu/tal/research/index.htm
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