The Journal of neuroscience : the official journal of the Society for Neuroscience, Jan 15, 1999
Low voltage-activated Ca2+ channels play important roles in pacing neuronal firing and producing ... more Low voltage-activated Ca2+ channels play important roles in pacing neuronal firing and producing network oscillations, such as those that occur during sleep and epilepsy. Here we describe the cloning and expression of the third member of the T-type family, alpha1I or CavT.3, from rat brain. Northern analysis indicated that it is predominantly expressed in brain. Expression of the cloned channel in either Xenopus oocytes or stably transfected human embryonic kidney-293 cells revealed novel gating properties. We compared these electrophysiological properties to those of the cloned T-type channels alpha1G and alpha1H and to the high voltage-activated channels formed by alpha1Ebeta3. The alpha1I channels opened after small depolarizations of the membrane similar to alpha1G and alpha1H but at more depolarized potentials. The kinetics of activation and inactivation were dramatically slower, which allows the channel to act as a Ca2+ injector. In oocytes, the kinetics were even slower, sugg...
Pancreatic islets have a central role in blood glucose homeostasis. In addition to insulin-produc... more Pancreatic islets have a central role in blood glucose homeostasis. In addition to insulin-producing beta-cells and glucagon-secreting alpha-cells, the islets contain somatostatin-releasing delta-cells. Somatostatin is a powerful inhibitor of insulin and glucagon secretion. It is normally secreted in response to glucose and there is evidence suggesting its release becomes perturbed in diabetes. Little is known about the control of somatostatin release. Closure of ATP-regulated K(+)-channels (K(ATP)-channels) and a depolarization-evoked increase in cytoplasmic free Ca(2+) concentration ([Ca(2+)](i)) have been proposed to be essential. Here, we report that somatostatin release evoked by high glucose (>or=10 mM) is unaffected by the K(ATP)-channel activator diazoxide and proceeds normally in K(ATP)-channel-deficient islets. Glucose-induced somatostatin secretion is instead primarily dependent on Ca(2+)-induced Ca(2+)-release (CICR). This constitutes a novel mechanism for K(ATP)-channel-independent metabolic control of pancreatic hormone secretion.
Pharmacological and electrophysiological studies have established that there are multiple types o... more Pharmacological and electrophysiological studies have established that there are multiple types of voltage-gated Ca2+ channels. Molecular biology has uncovered an even greater number of channel molecules. Thus, the molecular diversity of Ca2+ channels has its basis in the expression of many alpha 1 and beta genes, and also in the splice variants produced from these genes. This ability to mix and match subunits provides the cell with yet another mechanism to control the influx of calcium. Future studies will describe new subunits, the subunit composition of each type of channel, and the cloning of new Ca2+ channel types.
Graefe's Archive for Clinical and Experimental Ophthalmology, 2005
Voltage-dependent Ca(2+) channels trigger and control important cellular processes like neurotran... more Voltage-dependent Ca(2+) channels trigger and control important cellular processes like neurotransmitter release and secretion, long-term potentiation, and gene expression in excitable cells. During retinal signal perception and processing, presynaptic Ca(2+) channels facilitate neurotransmitter release in photoreceptors and bipolar neurons, at nonspiking synapses which generate graded potentials. The nature of voltage-gated Ca(2+) channels involved in retinal signal transduction is investigated in the present report by recording the electroretinogram (ERG) from the isolated and perfused bovine retina. Transcripts of the E/R- and T-type Ca(2+) channels are detected by RT-PCR. Using the Ca(2+) channel antagonists (+/-)-isradipine, NiCl(2), mibefradil, and SNX-482 results in either stimulatory or inhibitory effects on the ERG b-wave amplitude. On the transcript level, mRNA is detected for the E/R-type and a T-type voltage-gated Ca(2+) channel containing Ca(v)2.3 and Ca(v)3.1 as ion-conducting subunits, respectively. Blocking of the E/R-type Ca(2+) channels by NiCl(2) (10 microM) and SNX-482 (30 nM) contributes to the stimulatory effect, whereas antagonism of T-type as well as L-type Ca(2+) channels meditates the inhibitory action on the b-wave amplitude. Thus, a novel function for E/R-type voltage-gated Ca(2+) channels is probably associated with the visual signal transduction in the mammalian retina.
Relatively little has been published on the pharmacology of R-type and T-type Ca(2+) channels. He... more Relatively little has been published on the pharmacology of R-type and T-type Ca(2+) channels. Here, whole-cell Ca(2+) channel currents were recorded from human embryonic kidney 293 cell-lines transfected with either alpha1E subunits (R-type currents) or alpha1G or alpha1I subunits (T-type currents). R-type currents were inhibited by sipatrigine and the related compound 202W92 (R-(-)-2,4-diamino-6-(fluromethyl)-5-(2,3,5-trichlorophenyl)pyrimidine) with IC(50) 10 and 56 microM, respectively. A therapeutic concentration of lamotrigine (10 microM) inhibited R-type currents (30%) but was without effect on alpha1I-mediated T-type currents. Lamotrigine was also a weak inhibitor of T-type currents mediated by alpha1G subunits (<10% inhibition by 100 microM).
Glucagon release upon hypoglycemia is an important homeostatic mechanism utilized by vertebrates ... more Glucagon release upon hypoglycemia is an important homeostatic mechanism utilized by vertebrates to restore blood glucose to normal. Glucagon secretion itself is triggered by Ca2+ influx through voltage-gated ion channels, and the gene inactivation of R-type Ca2+ channels, with Ca(v)2.3 as the ion conducting subunit, has been shown to disturb glucose homeostasis. To understand how glucagon release may be affected in Ca(v)2.3-deficient mice, carbachol, insulin and glucose induced glucagon response was investigated. While the rise of insulin and glucose induced by carbachol is normal, mutant mice show an impaired glucagon-response. Further, the effect of insulin injection on glucagon levels was altered by the loss of the Ca(v)2.3 subunit. Ca(v)2.3-deficient mice are characterized by an impaired glucose suppression of glucagon release. This was most obvious at the level of isolated islets suggesting that Ca(v)2.3 containing R-type voltage-gated Ca2+ channels are involved in the glucose-mediated signalling to glucagon release in mice.
Pathologic ECG events are known to accompany seizures and to persist in several chronic epilepsy ... more Pathologic ECG events are known to accompany seizures and to persist in several chronic epilepsy syndromes. The contribution of antiepileptic drugs (AEDs) to these events and the implications in the etiology of sudden-unexpected death in epilepsy (SUDEP) continue to be a matter of debate. We therefore investigated cardiac parameters during kainic-acid (KA) induced experimental epilepsy and antiepileptic treatment with lamotrigine (LTG). Epilepsy was induced in seven C57Bl/6 mice by injections of KA (20 mg/kg) on days 1 and 5, which produced severe acute seizures and spontaneous seizures 10 days later. Treatment with LTG (30 mg/kg) was initiated on day 11 and repeated on day 12. Continuous ECGs and ECoGs were collected telemetrically from freely moving mice. Mice displayed pre-ictal but not ictal tachycardia. The squared coefficient of variation (SCV) of R-R intervals was significantly elevated 30s before and during seizures compared to control conditions. LTG produced a significant reversible increase in SCV and LF/HF ratio during slow-wave sleep (SWS), potentially indicative of sympatho-vagal imbalance during this state of vigilance, in which epileptic patients are known to be particularly vulnerable to SUDEP. The KA model used in this study permits the investigation of cardiac phenomena during epilepsy, as it features many effects found in human epileptic patients. Increased LF/HF, a known risk factor for cardiac disease, which is often found in epileptic patients, was observed as a side-effect of LTG treatment during SWS, suggesting that LTG may promote imbalance of the autonomous nervous system in epileptic mice.
Multiple types of voltage-activated Ca2+ channels (T, L, N, P, Q, R type) coexist in excitable ce... more Multiple types of voltage-activated Ca2+ channels (T, L, N, P, Q, R type) coexist in excitable cells and participate in synaptic differentiation, secretion, transmitter release, and neuronal plasticity. Ca2+ ions entering cells trigger these events through their interaction with the ion channel itself or through Ca2+ binding to target proteins initiating signalling cascades at cytosolic loops of the ion conducting subunit (Cava1). These loops interact with target proteins in a Ca2+-dependent or independent manner. In Cav2.3-containing channels the cytosolic linker between domains II and III confers a novel Ca2+ sensitivity to E-type Ca2+ channels including phorbol ester sensitive signalling via protein kinase C (PKC) in Cav2.3 transfected HEK-293 cells. To understand Ca2+ and phorbol ester mediated activation of Cav2.3 Ca2+ channels, protein interaction partners of the II-III loop were identified. FLAG-tagged II-III - loop of human Cav2.3 was over-expressed in HEK 293 cells, and the molecular chaperone hsp70, which is known to interact with PKC, was identified as a novel functional interaction partner. Immunopurified II-III loop-protein of neuronal and endocrine Cav2.3 splice variants stimulate autophosphorylation of PKCa, leading to the suggestion that hsp70--binding to the II-III loop--may act as an adaptor for Ca2+ dependent targeting of PKC to E-type Ca2+ channels.
The Journal of neuroscience : the official journal of the Society for Neuroscience, Jan 15, 1999
Low voltage-activated Ca2+ channels play important roles in pacing neuronal firing and producing ... more Low voltage-activated Ca2+ channels play important roles in pacing neuronal firing and producing network oscillations, such as those that occur during sleep and epilepsy. Here we describe the cloning and expression of the third member of the T-type family, alpha1I or CavT.3, from rat brain. Northern analysis indicated that it is predominantly expressed in brain. Expression of the cloned channel in either Xenopus oocytes or stably transfected human embryonic kidney-293 cells revealed novel gating properties. We compared these electrophysiological properties to those of the cloned T-type channels alpha1G and alpha1H and to the high voltage-activated channels formed by alpha1Ebeta3. The alpha1I channels opened after small depolarizations of the membrane similar to alpha1G and alpha1H but at more depolarized potentials. The kinetics of activation and inactivation were dramatically slower, which allows the channel to act as a Ca2+ injector. In oocytes, the kinetics were even slower, sugg...
Pancreatic islets have a central role in blood glucose homeostasis. In addition to insulin-produc... more Pancreatic islets have a central role in blood glucose homeostasis. In addition to insulin-producing beta-cells and glucagon-secreting alpha-cells, the islets contain somatostatin-releasing delta-cells. Somatostatin is a powerful inhibitor of insulin and glucagon secretion. It is normally secreted in response to glucose and there is evidence suggesting its release becomes perturbed in diabetes. Little is known about the control of somatostatin release. Closure of ATP-regulated K(+)-channels (K(ATP)-channels) and a depolarization-evoked increase in cytoplasmic free Ca(2+) concentration ([Ca(2+)](i)) have been proposed to be essential. Here, we report that somatostatin release evoked by high glucose (>or=10 mM) is unaffected by the K(ATP)-channel activator diazoxide and proceeds normally in K(ATP)-channel-deficient islets. Glucose-induced somatostatin secretion is instead primarily dependent on Ca(2+)-induced Ca(2+)-release (CICR). This constitutes a novel mechanism for K(ATP)-channel-independent metabolic control of pancreatic hormone secretion.
Pharmacological and electrophysiological studies have established that there are multiple types o... more Pharmacological and electrophysiological studies have established that there are multiple types of voltage-gated Ca2+ channels. Molecular biology has uncovered an even greater number of channel molecules. Thus, the molecular diversity of Ca2+ channels has its basis in the expression of many alpha 1 and beta genes, and also in the splice variants produced from these genes. This ability to mix and match subunits provides the cell with yet another mechanism to control the influx of calcium. Future studies will describe new subunits, the subunit composition of each type of channel, and the cloning of new Ca2+ channel types.
Graefe's Archive for Clinical and Experimental Ophthalmology, 2005
Voltage-dependent Ca(2+) channels trigger and control important cellular processes like neurotran... more Voltage-dependent Ca(2+) channels trigger and control important cellular processes like neurotransmitter release and secretion, long-term potentiation, and gene expression in excitable cells. During retinal signal perception and processing, presynaptic Ca(2+) channels facilitate neurotransmitter release in photoreceptors and bipolar neurons, at nonspiking synapses which generate graded potentials. The nature of voltage-gated Ca(2+) channels involved in retinal signal transduction is investigated in the present report by recording the electroretinogram (ERG) from the isolated and perfused bovine retina. Transcripts of the E/R- and T-type Ca(2+) channels are detected by RT-PCR. Using the Ca(2+) channel antagonists (+/-)-isradipine, NiCl(2), mibefradil, and SNX-482 results in either stimulatory or inhibitory effects on the ERG b-wave amplitude. On the transcript level, mRNA is detected for the E/R-type and a T-type voltage-gated Ca(2+) channel containing Ca(v)2.3 and Ca(v)3.1 as ion-conducting subunits, respectively. Blocking of the E/R-type Ca(2+) channels by NiCl(2) (10 microM) and SNX-482 (30 nM) contributes to the stimulatory effect, whereas antagonism of T-type as well as L-type Ca(2+) channels meditates the inhibitory action on the b-wave amplitude. Thus, a novel function for E/R-type voltage-gated Ca(2+) channels is probably associated with the visual signal transduction in the mammalian retina.
Relatively little has been published on the pharmacology of R-type and T-type Ca(2+) channels. He... more Relatively little has been published on the pharmacology of R-type and T-type Ca(2+) channels. Here, whole-cell Ca(2+) channel currents were recorded from human embryonic kidney 293 cell-lines transfected with either alpha1E subunits (R-type currents) or alpha1G or alpha1I subunits (T-type currents). R-type currents were inhibited by sipatrigine and the related compound 202W92 (R-(-)-2,4-diamino-6-(fluromethyl)-5-(2,3,5-trichlorophenyl)pyrimidine) with IC(50) 10 and 56 microM, respectively. A therapeutic concentration of lamotrigine (10 microM) inhibited R-type currents (30%) but was without effect on alpha1I-mediated T-type currents. Lamotrigine was also a weak inhibitor of T-type currents mediated by alpha1G subunits (<10% inhibition by 100 microM).
Glucagon release upon hypoglycemia is an important homeostatic mechanism utilized by vertebrates ... more Glucagon release upon hypoglycemia is an important homeostatic mechanism utilized by vertebrates to restore blood glucose to normal. Glucagon secretion itself is triggered by Ca2+ influx through voltage-gated ion channels, and the gene inactivation of R-type Ca2+ channels, with Ca(v)2.3 as the ion conducting subunit, has been shown to disturb glucose homeostasis. To understand how glucagon release may be affected in Ca(v)2.3-deficient mice, carbachol, insulin and glucose induced glucagon response was investigated. While the rise of insulin and glucose induced by carbachol is normal, mutant mice show an impaired glucagon-response. Further, the effect of insulin injection on glucagon levels was altered by the loss of the Ca(v)2.3 subunit. Ca(v)2.3-deficient mice are characterized by an impaired glucose suppression of glucagon release. This was most obvious at the level of isolated islets suggesting that Ca(v)2.3 containing R-type voltage-gated Ca2+ channels are involved in the glucose-mediated signalling to glucagon release in mice.
Pathologic ECG events are known to accompany seizures and to persist in several chronic epilepsy ... more Pathologic ECG events are known to accompany seizures and to persist in several chronic epilepsy syndromes. The contribution of antiepileptic drugs (AEDs) to these events and the implications in the etiology of sudden-unexpected death in epilepsy (SUDEP) continue to be a matter of debate. We therefore investigated cardiac parameters during kainic-acid (KA) induced experimental epilepsy and antiepileptic treatment with lamotrigine (LTG). Epilepsy was induced in seven C57Bl/6 mice by injections of KA (20 mg/kg) on days 1 and 5, which produced severe acute seizures and spontaneous seizures 10 days later. Treatment with LTG (30 mg/kg) was initiated on day 11 and repeated on day 12. Continuous ECGs and ECoGs were collected telemetrically from freely moving mice. Mice displayed pre-ictal but not ictal tachycardia. The squared coefficient of variation (SCV) of R-R intervals was significantly elevated 30s before and during seizures compared to control conditions. LTG produced a significant reversible increase in SCV and LF/HF ratio during slow-wave sleep (SWS), potentially indicative of sympatho-vagal imbalance during this state of vigilance, in which epileptic patients are known to be particularly vulnerable to SUDEP. The KA model used in this study permits the investigation of cardiac phenomena during epilepsy, as it features many effects found in human epileptic patients. Increased LF/HF, a known risk factor for cardiac disease, which is often found in epileptic patients, was observed as a side-effect of LTG treatment during SWS, suggesting that LTG may promote imbalance of the autonomous nervous system in epileptic mice.
Multiple types of voltage-activated Ca2+ channels (T, L, N, P, Q, R type) coexist in excitable ce... more Multiple types of voltage-activated Ca2+ channels (T, L, N, P, Q, R type) coexist in excitable cells and participate in synaptic differentiation, secretion, transmitter release, and neuronal plasticity. Ca2+ ions entering cells trigger these events through their interaction with the ion channel itself or through Ca2+ binding to target proteins initiating signalling cascades at cytosolic loops of the ion conducting subunit (Cava1). These loops interact with target proteins in a Ca2+-dependent or independent manner. In Cav2.3-containing channels the cytosolic linker between domains II and III confers a novel Ca2+ sensitivity to E-type Ca2+ channels including phorbol ester sensitive signalling via protein kinase C (PKC) in Cav2.3 transfected HEK-293 cells. To understand Ca2+ and phorbol ester mediated activation of Cav2.3 Ca2+ channels, protein interaction partners of the II-III loop were identified. FLAG-tagged II-III - loop of human Cav2.3 was over-expressed in HEK 293 cells, and the molecular chaperone hsp70, which is known to interact with PKC, was identified as a novel functional interaction partner. Immunopurified II-III loop-protein of neuronal and endocrine Cav2.3 splice variants stimulate autophosphorylation of PKCa, leading to the suggestion that hsp70--binding to the II-III loop--may act as an adaptor for Ca2+ dependent targeting of PKC to E-type Ca2+ channels.
Uploads
Papers by Toni Schneider