Structure and functional characterization of neuronal alpha 1E calcium channel subtypes.

Williams, Mark E.; Marubio, Lisa M.; Deal, C.; Hans, Michael; Brust, Paul F.; Philipson, Louis H.; Miller, Richard J.; Johnson, Edwin C.; Harpold, Michael M.; Ellis, Steven B.

doi:10.1016/s0021-9258(17)31796-9

Cited by 256 publications

(30 citation statements)

References 36 publications

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“…Ni 2+ regulation of I A was dose-dependent and the apparent IC 50 (∼44.1 μM) approximated values reported for Cav2.3 (27.4-66.0 μM) ( SFig. 4A ) (77). Ni 2+ did not affect voltage dependence of inactivation for I A , suggesting a distinct mechanism in hippocampus relative to cerebellum ( SFig.…”

Section: Resultssupporting

confidence: 62%

R-type voltage-gated Ca²⁺channels mediate A-type K⁺current regulation of synaptic input in hippocampal dendrites

Murphy

Gutzmann

Lin

et al. 2020

Preprint

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15The transient voltage-gated K + current (IA) mediated by Kv4.2 in CA1 hippocampal pyramidal 16 neurons regulates dendritic excitability, synaptic plasticity, and learning. Here we report that Ca 2+ 17 entry mediated by the voltage-gated Ca 2+ channel subunit Cav2.3 regulates Kv4.2 function in CA1 18 pyramidal neurons through Ca 2+ binding auxiliary subunits known as K + channel interacting 19 proteins (KChIPs). We characterized an interaction between Cav2.3 and Kv4.2 using 20 immunofluorescence colocalization, coimmunoprecipitation, electron microscopy, FRAP, and 21 FRET. We found that Ca 2+ -entry via Cav2.3 increases Kv4.2-mediated whole-cell current due in 22 part to an increase in Kv4.2 surface expression. In hippocampal neurons, pharmacological block 23 of Cav2.3 reduced whole-cell IA. We also found reduced IA in Cav2.3 knockout mouse neurons 24 with a loss of the dendritic IA gradient. Furthermore, the Cav2.3-Kv4.2 complex was found to 25 regulate the size of synaptic currents and spine Ca 2+ transients. These results reveal an 26 intermolecular Cav2.3-Kv4.2 complex impacting synaptic integration in CA1 hippocampal 27 neurons. 28 29 30 31 32 33 34 93 and Kv4.2 that regulates Kv4.2 function by a novel mechanism. 94 95 96 97 98 99 100 101 4 RESULTS 102 103 Cav2.3 and Kv4.2 form an ion channel complex in hippocampal neurons 104The characteristic kinetics of neuronal IA requires expression of KChIP and DPP 105 accessory subunits that had been identified and described nearly two decades ago (27, 28). In 106 an effort to identify novel Kv4.2 protein interactions and modifying enzymes in hippocampal 107 neurons we used tandem affinity purification (TAP) and protein identification with mass 108 spectrometry (MS) (58). As expected, Kv4.2 pulled down Kv4.1, Kv4.2, Kv4.3, and previously 109 described auxiliary subunits (DPPs and KChIPs) confirming the specificity of the assay (Figure 1110 -Figure supplement 1A). In addition to known binding partners, we also identified peptides 111 representing a significant proportion of the Cav2.3 amino acid sequence (Figure 1 -Figure 112 supplement 1B,C). Cav2.3 was the only voltage-gated Ca 2+ channel identified in this screen.113 To confirm the presence of a hippocampal Cav2.3 and Kv4.2 molecular complex we 114 carried out several immunological measurements using hippocampal tissue. Broad neuropil 115 colocalization of endogenous Cav2.3 and Kv4.2 immunofluorescent signal was consistent with 116 enrichment in the dendrite layers of the hippocampus as has been previously reported for each 117 channel separately (Figure 1A) (51, 59). Both Cav2.3 and Kv4.2 are also known to be expressed 118 in dendrites and spines. To evaluate Cav2.3-Kv4.2 colocalization, we transfected cultured 119 hippocampal neurons with Cav2.3-GFP (i), Kv4.2-myc (ii), and mCherry (iii) plasmids (Figure 1B). 120In addition to immunofluorescent colocalization in dendrites, both channels were enriched in 121 spines when compared to the free cytosolic mCherry fluorescent protein (Figure 1C,D). 122Immunofluorescent...

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Section: Resultssupporting

confidence: 62%

R-type voltage-gated Ca²⁺channels mediate A-type K⁺current regulation of synaptic input in hippocampal dendrites

Murphy

Gutzmann

Lin

et al. 2020

Preprint

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“…Transcripts of the ␣1E voltage-gated Ca 2ϩ channels are detected in rat, rabbit, mouse, and human brain and in other organs from different species (Williams et al 1994;Marubio et al 1996;Vajna et al 1998). In mammalian brain, more than four different isoforms can be predicted on the basis of cloned (Niidome et al 1992;Soong et al 1993;Schneider et al 1994;Williams et al 1994) and PCR-amplified ␣1E cDNA fragments (Marubio et al 1996;Pereverzev et al 1998;Vajna et al 1998;Kamphuis and Hendriksen 1998). In cerebellum, three different longer ␣1E isoforms are identified by RT-PCR, which have been described as ␣1Ed, ␣1Ee, and ␣1Ef.…”

Section: Discussionmentioning

confidence: 99%

“…In human, rat, and rabbit cerebellum, the expression of ␣1E has been investigated in the past by in situ hybridization (Soong et al 1993;Wakamori et al 1994;Williams et al 1994;Volsen et al 1995) by antibodies raised against a fragment of the II-III loop (aa 958-1098 of human ␣1E; see Volsen et al 1995) or by peptide-directed antibodies derived from two highly divergent regions in the intracellular II-III loop of rat ␣1E (Yokoyama et al 1995). Recently, ␣1E was found by an antisense riboprobe in the cell bodies of cerebellar Purkinje cells and in the granule cell layer (Volsen et al 1995).…”

Section: Expression Of ␣1e Isoforms In the Cerebellummentioning

confidence: 99%

Immunohistochemical Detection of α1E Voltage-gated Ca²⁺ Channel Isoforms in Cerebellum, INS-1 Cells, and Neuroendocrine Cells of the Digestive System

Grabsch

Pereverzev²,

Weiergräber³

et al. 1999

J Histochem Cytochem.

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SUMMARY Polyclonal antibodies were raised against a common and a specific epitope present only in longer ␣ 1E isoforms of voltage-gated Ca 2 ϩ channels, yielding an "anti-Ecom" and an "anti-E-spec" serum, respectively. The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected HEK-293 cells or membrane proteins derived from them. Cells from the insulinoma cell line INS-1, tissue sections from cerebellum, and representative regions of gastrointestinal tract were stained immunocytochemically. INS-1 cells expressed an ␣ 1E splice variant with a longer carboxy terminus, the so-called ␣ 1Ee isoform. Similarily, in rat cerebellum, which was used as a reference system, the anti-E-spec serum stained somata and dendrites of Purkinje cells. Only faint staining was seen throughout the cerebellar granule cell layer. After prolonged incubation times, neurons of the molecular layer were stained by anti-E-com, suggesting that a shorter ␣ 1E isoform is expressed at a lower protein density. In human gastrointestinal tract, endocrine cells of the antral mucosa (stomach), small and large intestine, and islets of Langerhans were stained by the anti-E-spec serum. In addition, staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae. We conclude that the longer ␣ 1Ee isoform is expressed in neuroendocrine cells of the digestive system and that, in pancreas, ␣ 1Ee expression is restricted to the neuroendocrine part, the islets of Langerhans. ␣ 1E therefore appears to be a common voltagegated Ca 2 ϩ channel linked to neuroendocrine and related systems of the body.

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“…The primary structure of the cardiac ␣1Ee isoform shows major differences from the cloned neuronal ␣1Ed (Schneider et al 1994) in the II/III loop and from the carboxy terminus of cloned ␣1E-3 (Williams et al 1994). Within the II/III loop, insert 1 probably represents a duplication of an adjacent arginine-rich sequence: a stretch of 19 amino acids is lacking in ␣1Ee and starts at position aa 749 with RERRRR-HHMS, resembling the adjacent sequence at position aa 768 of human ␣1E, RERRRRHHMS.…”

Section: Transcripts Of Voltage-gated Ca 2ϩ Channels In Rat Heartmentioning

confidence: 99%

Immunodetection of α1E Voltage-gated Ca²⁺ Channel in Chromogranin-positive Muscle Cells of Rat Heart, and in Distal Tubules of Human Kidney

Weiergräber

Pereverzev

Vajna

et al. 2000

J Histochem Cytochem.

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S U M M A R YThe calcium channel ␣ 1E subunit was originally cloned from mammalian brain. A new splice variant was recently identified in rat islets of Langerhans and in human kidney by the polymerase chain reaction. The same isoform of ␣ 1E was detected in rat and guinea pig heart by amplifying indicative cDNA fragments and by immunostaining using peptide-specific antibodies. The apparent molecular size of cardiac ␣ 1E was determined by SDS-PAGE and immunoblotting (218 Ϯ 6 kD; n ϭ 3). Compared to ␣ 1E from stably transfected HEK-293 cells, this is smaller by 28 kD. The distribution of ␣ 1E in cardiac muscle cells of the conducting system and in the cardiomyoblast cell line H9c2 was compared to the distribution of chromogranin, a marker of neuroendocrine cells, and to the distribution of atrial natriuretic peptide (ANP). In serial sections from atrial and ventricular regions of rat heart, co-localization of ␣ 1E with ANP was detected in atrium and with chromogranin A/B in Purkinje fibers of the conducting system in both rat atrium and ventricle. The kidney is another organ in which natriuretic peptide hormones are secreted. The detection of ␣ 1E in the distal tubules of human kidney, where urodilatin is stored and secreted, led to the conclusion that the expression of ␣ 1E in rat heart and human kidney is linked to regions with endocrine functions and therefore is involved in the Ca 2 ϩ -dependent secretion of peptide hormones such as ANP and urodilatin.

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Structure and functional characterization of neuronal alpha 1E calcium channel subtypes.

Cited by 256 publications

References 36 publications

R-type voltage-gated Ca²⁺channels mediate A-type K⁺current regulation of synaptic input in hippocampal dendrites

R-type voltage-gated Ca²⁺channels mediate A-type K⁺current regulation of synaptic input in hippocampal dendrites

Immunohistochemical Detection of α1E Voltage-gated Ca²⁺ Channel Isoforms in Cerebellum, INS-1 Cells, and Neuroendocrine Cells of the Digestive System

Immunodetection of α1E Voltage-gated Ca²⁺ Channel in Chromogranin-positive Muscle Cells of Rat Heart, and in Distal Tubules of Human Kidney

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Structure and functional characterization of neuronal alpha 1E calcium channel subtypes.

Cited by 256 publications

References 36 publications

R-type voltage-gated Ca2+channels mediate A-type K+current regulation of synaptic input in hippocampal dendrites

R-type voltage-gated Ca2+channels mediate A-type K+current regulation of synaptic input in hippocampal dendrites

Immunohistochemical Detection of α1E Voltage-gated Ca2+ Channel Isoforms in Cerebellum, INS-1 Cells, and Neuroendocrine Cells of the Digestive System

Immunodetection of α1E Voltage-gated Ca2+ Channel in Chromogranin-positive Muscle Cells of Rat Heart, and in Distal Tubules of Human Kidney

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R-type voltage-gated Ca²⁺channels mediate A-type K⁺current regulation of synaptic input in hippocampal dendrites

R-type voltage-gated Ca²⁺channels mediate A-type K⁺current regulation of synaptic input in hippocampal dendrites

Immunohistochemical Detection of α1E Voltage-gated Ca²⁺ Channel Isoforms in Cerebellum, INS-1 Cells, and Neuroendocrine Cells of the Digestive System

Immunodetection of α1E Voltage-gated Ca²⁺ Channel in Chromogranin-positive Muscle Cells of Rat Heart, and in Distal Tubules of Human Kidney