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C Lohr

Publications and source records attributed to C Lohr.

13 recordsLinked to original sources
PubMed2002

Calcium influx into dendrites of the leech Retzius neuron evoked by 5-hydroxytryptamine.

5-Hydroxytryptamine (5-HT) is a ubiquitous neurotransmitter and neuromodulator that affects neural circuits and behaviours in vertebrates and invertebrates. In the present study, we have investigated 5-HT-induced Ca(2+) transients in subcellular compartments of Retzius neurons in the leech central nervous system using confocal laser scanning microscopy, and studied the effect of 5-HT on the electrical coupling between the Retzius neurons. Bath application of 5-HT (50mM) induced a Ca(2+) transient in axon, dendrites and cell body of the Retzius neuron. This Ca(2+) transient was significantly faster and larger in dendrites than in axon and cell body, and was half-maximal at a 5-HT concentration of 5-12mM. The Ca(2+) transient was suppressed in the absence of extracellular Ca(2+) and by methysergide (100mM), a non-specific antagonist of metabotropic 5-HT receptors, and was strongly reduced by bath application of the Ca(2+) channel blocker Co(2+) (2mM). Injection of the non-hydrolysable GTP analogue GTPgammaS increased and prolonged the dendritic 5-HT-induced Ca(2+) transient. The non-selective protein kinase inhibitor H7 (100mM) and the adenylate cyclase inhibitor SQ22536 (500 mM) did not affect the Ca(2+) transient, and the membrane-permeable cAMP analogue dibutyryl-cAMP (500 mM) did not mimic the effect of 5-HT application. 5-HT reduced the apparent electrical coupling between the two Retzius neurons, whereas suppression of the Ca(2+) influx by removal of external Ca(2+) improved the transmission of action potentials at the electrical synapses which are located between the dendrites of the adjacent Retzius neurons. The results indicate that 5-HT induces a Ca(2+) influx through calcium channels located primarily in the dendrites, and presumably activated by a G protein-coupled 5-HT receptor. The dendritic Ca(2+) increase appears to modulate the excitability of, and the synchronization between, the two Retzius neurons.

Animals↗
PubMed2001

Activity-dependent accumulation of Ca2+ in axon and dendrites of the leech Leydig neuron.

We have investigated Ca2+ changes evoked by single action potentials (APs) in axon and dendrites of leech Leydig neurons. Dendritic Ca2+ transients induced by an AP were twice as large as in the axon, and Ca2+ recovery was significantly faster in the dendrites as compared to the axon. The AP-induced Ca2+ transients were blocked by Co2+ and suppressed in Ca2+-free saline, indicating Ca2+ influx through voltage-activated channels. During a train of APs, Ca2+ accumulated significantly more in the axon than in the dendrites. Suppression of the Ca2+ influx changed the shape of the action potential and increased the firing frequency. The results suggest a functional role of Ca2+ influx and Ca2+ accumulation during electrical activity in different neuronal subcompartments.

Action Potentials↗
PubMed2001

Olfactory receptor axons influence the development of glial potassium currents in the antennal lobe of the moth Manduca sexta.

In the olfactory (antennal) lobe of the moth Manduca sexta, olfactory receptor axons strongly influence the distribution and morphology of glial cells. In the present study, we asked whether the development of the electrophysiological properties of the glial cells is influenced by the receptor axons. Whole-cell currents were measured in antennal lobe glial cells in acute brain slices prepared from animals at different stages of metamorphic development (stages 3, 6, and 12). Outward currents were induced by depolarizing voltage steps from a holding potential of -70 mV. At all developmental stages investigated, the outward currents were partly blocked by bath application of the potassium channel blocker 4-aminopyridine (4AP, 10 mM) or by including tetraethylammonium (TEA, 30 mM) in the pipette solution. The relative contribution of the 4AP-sensitive current to the outward current increased from 18% at stages 3 and 6 to 42% at stage 12, while the TEA-sensitive current increased from 18% at stage 3 to 81% at stage 6, and then declined again to 40% at stage 12. In contrast, in the absence of receptor axons, these changes in the contribution of the TEA- and 4AP-sensitive currents to the total outward current did not occur; rather, the current profile remained in the most immature state (stage 3). The results suggest that olfactory receptor axons are essential for development of the mature pattern of glial potassium currents.

4-Aminopyridine↗
PubMed2001

Calcium transients in subcompartments of the leech Retzius neuron as induced by single action potentials.

Regional Ca(2+) influx into neurons plays an essential role for fast signal processing, yet it is little understood. We have investigated intracellular Ca(2+) transients induced by a single action potential (AP) in Retzius neurons in situ of isolated ganglia of the leech Hirudo medicinalis using confocal laser scanning microscopy in the cell body, in different axonal branches, and in dendrites. In the cell body, a single AP induced a Ca(2+) transient in submembrane regions, while in central regions no fluorescence change was detected. Burst activity evoked a much larger Ca(2+) influx, which elicited Ca(2+) signals in central somatic regions, including the cell nucleus. A single AP induced a Ca(2+) transient in distal branches of the axon and in dendrites that was significantly larger than in the proximal axon and in the cell body (p <.05), and the recovery of the Ca(2+) transient was significantly faster in axonal branches than in dendrites (p <.01). The AP-induced Ca(2+) transient was inhibited by Co(2+) (2 mM). The P/Q-type Ca(2+) channel blocker omega-agatoxin TK (500 nM) and the L-type Ca(2+) channel blocker nifedipine (20 microM) had no effect on the Ca(2+) transient, whereas the L-type Ca(2+) channel blocker methoxyverapamil (D600, 0.5-1 mM) irreversibly reduced the Ca(2+) transient by 37% in axons and by 42% in dendrites. Depletion of intracellular Ca(2+) stores following inhibition of endoplasmic Ca(2+)-ATPases by cyclopiazonic acid (10 microM) decreased the AP-induced Ca(2+) transient in the dendrites by 21% (p <.01), but not in axons, and increased the Ca(2+) recovery time constant (tau) in the axonal branches by 129% (p <.01), but not in dendrites. The results indicate that an AP evokes a voltage-gated Ca(2+) influx into all subcompartments of the Retzius neuron, where it produces a Ca(2+) signal of different size and/or kinetics. This may contribute to the modulation of electrical excitation and propagation of APs, and to different modes of synaptic and nonsynaptic processes.

Action Potentials↗
PubMed2001

Bursting activity in leech Retzius neurons induced by low external chloride.

We have investigated the bursting activity of Retzius neurons in the central nervous system of the leech Hirudo medicinalis as induced in Cl(-)-free saline by measuring membrane potential, membrane current and the intracellular calcium concentration ([Ca2+]i), using fura-2 or Oregon-Green488-Bapta-1. The Retzius neurons changed their low tonic firing to rhythmical bursting activity when the extracellular Cl- concentration ([Cl-]o) was lowered to 1 mM or less. In Cl(-)-free saline (Cl- exchanged by gluconate), bursting was accompanied by a rise in intracellular Ca2+ in both cell body and axon, which oscillated in synchrony with the bursts. The Ca2+ transients depended on the amplitude and duration of the depolarization underlying the burst, and were presumably due to Ca2+ influx through voltage-dependent Ca2+ channels. In Ca(2+)-free, EGTA-buffered saline or in the presence of Ca2+ channel blockers verapamil (1 mM) or diltiazem (500 microM) the depolarizations underlying the bursts in Cl(-)-free saline were enhanced in amplitude and duration. Bursting was not affected by depleting the intracellular Ca2+ stores with cyclopiazonic acid. The depolarization in Cl(-)- and Ca(2+)-free saline did not evoke intracellular Ca2+ changes. The burst-underlying membrane depolarization induced by Cl- removal was found to be due to a Na(+)-dependent persistent inward current and could be inhibited by saxitoxin (25-50 microM). The results suggest that a persistent Na+ current is generated in Cl(-)-free saline and induces the depolarization underlying rhythmic activity, and that presumably Ca(2+)-induced K+ currents modulate the bursting behaviour.

Animals↗
PubMed1999

Dendritic calcium transients in the leech giant glial cell in situ.

Glial cells have been shown to respond to neuronal activity with changes in the membrane potential and the intracellular Ca2+ concentration. In order to get closer to glial structures associated with neuronal synapses, we have now looked at Ca2+ signalling in the glial processes ("glial dendrites") in response to neurotransmitters and neuronal activity. Single giant glial cells in situ of isolated ganglia of the leech Hirudo medicinalis were filled iontophoretically with the Ca(2+)-sensitive dyes Oregon green 488 BAPTA-1 or Fluo-3. Relative Ca(2+)-dependent fluorescence changes in response to bath and focal application of the ionotropic glutamate receptor agonist kainate (50 microM) and of 5-hydroxytryptamine (5-HT, 100 microM) were recorded in glial dendrites, using confocal laser scanning microscopy. The amplitudes of the [Ca2+]i transients in the dendritic processes were 2-4 times larger than those recorded in the cell body. Electrical stimulation of a nerve root (20 Hz for 15 s) elicited [Ca2+]i transients in glial dendrites (n = 32) that were reduced by the ionotropic glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; n = 14). The results demonstrate that neuronal activity can evoke [Ca2+]i transients not only in glial cell bodies but also in glial dendrites, where these transients display regional variation. This may reflect local release of neurotransmitters like glutamate and 5-hydroxytryptamine and/or regional differences in the density of glial receptors.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1999

Leech giant glial cell: functional role in a simple nervous system.

The giant glial cell in the central nervous system of the leech Hirudo medicinalis has been the subject of a series of studies trying to link its physiological properties with its role in neuron-glia interactions. Isolated ventral cord ganglia of this annelid offer several advantages for these studies. First, single giant glial cells can easily be identified and are quite accessible to electrophysiological and microfluorometric studies. Second, only two giant macroglial cells are located in the neuropil of each ganglion, rendering them well suited for studying neuron-glia interactions. Third, many neurons can be identified and are well known with respect to their physiology and their roles in controlling simple behaviors in the leech. This review briefly outlines the major recent findings gained by studying this preparation and its contributions to our knowledge of the functional role of glia in nervous systems. Emphasis is directed to glial responses during neuronal activity and to the analysis of intracellular Ca(2+) and H(+) transients mediated by neurotransmitter receptors and ion-driven carriers. Among its numerous properties, the leech giant glial cell prominently expresses a large K(+) conductance, voltage-dependent Ca(2+) channels, ionotropic non-NMDA glutamate receptors, and an electrogenic, reversible Na(+)-HCO(3)(-) cotransporter.

Animals↗
PubMed1997

Intracellular Ca2+ release mediated by metabotropic glutamate receptor activation in the leech giant glial cell.

We have investigated the effects of glutamate and glutamate receptor ligands on the intracellular free Ca2+ concentration ([Ca2+]i) and the membrane potential (Em) of single, identified neuropile glial cells in the central nervous system of the leech Hirudo medicinalis. Exposed glial cells of isolated ganglia were filled iontophoretically with the Ca2+ indicator dye Fura-2. Application of glutamate (200-500 mumoll-1) caused biphasic membrane potential shifts and increases in [Ca2+]i, which were only partly reduced by either removing extracellular Ca2+ or blocking ionotropic glutamate receptors with 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 50-100 mumol l-1. Metabotropic glutamate receptor (mGluR) ligands had the following rank of potency in inducing a rise in [Ca2+]i: quisqualate (QQ, 200 mumol l-1) > glutamate (200 mumol l-1) > L(+)2-amino-3-phosphonopropionic acid (L-AP3, 200 mumol l-1 > trans-1-aminocyclopentane-1,3-dicarboxylic acid (t-ACPD, 400 mumol l-1). The mGluR-selective antagonist (RS)-alpha-methyl-4-carboxyphenylglycine [(RS)-MCPG, 1 mmol l-1] significantly reduced glutamate-evoked increases in [Ca2+]i by 20%. Incubation of the ganglia with the endoplasmic ATPase inhibitor cyclopiazonic acid (CPA, 10 mumol l-1) caused a significant (53%) reduction of glutamate-induced [Ca2+]i transients, while incubation with lithium ions (2 mmol l-1) resulted in a 46% reduction. The effects of depleting the Ca2+ stores with CPA and of CNQX were additive. We conclude that glutamate-induced [Ca2+]i transients were mediated by activation of both Ca(2+)-permeable ionotropic non-NMDA receptors and of metabotropic glutamate receptors leading to Ca2+ release from intracellular Ca2+ stores.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1996

Extracellular Ca2+ changes during transmitter application in the leech central nervous system.

Changes in extracellular Ca2+ concentration ([Ca2+]e) evoked by transmitters and transmitter agonists, respectively, and by elevation of bath K+ concentration were recorded in isolated segmental ganglia of the leech Hirudo medicinalis using Ca(2+)-selective microelectrodes. A 1-min bath application of kainate (10 microM), glutamate (1 mM), aspartate (1 mM), or carbachol (200 microM) decreased [Ca2+]e by up to 1 mM, whereas the inhibitory transmitters gamma-amino butyric acid (GABA, 100 microM) and serotonin (5-HT, 100 microM) did not change [Ca2+]e. The amplitude of the kainate-induced changes in [Ca2+]e increased with repetitive applications, and changes were blocked by 6-cyano-7-dinitroquinozaline-2,3-dione (CNQX). Elevation of bath K+ concentration from 4 to 40 mM led to a Ni(2+)-sensitive decrease in [Ca2+]e by 0.9 mM. Our results suggest that excitatory transmission in the leech central nervous system might be accompanied by substantial decreases in [Ca2+]e.

Animals↗
PubMed1995

Activity-induced Ca2+ transients in nerve and glial cells in the leech CNS.

We have measured activity-induced Ca2+ transients in Retzius neurones, neuropile glial cells, and extracellular spaces of isolated ganglia of the leech Hirudo medicinalis using the fluorescent dye fura-2 and Ca(2+)-sensitive microelectrodes. Neuronal activity, induced by electrical side nerve stimulation (20 Hz/1 min), elicited transient rises of intracellular Ca2+ in both neurones and glial cells, which amounted to 24 +/- nM (n = 15) and 17 +/- 14 nM (n = 7), respectively. The extracellular Ca2+ declined by 160 +/- 73 microM (n = 6) during stimulation. Intra- and extracellular Ca2+ transients were reduced by the glutamate/kainate receptor blocker CNQX (6-cyano-7-dinitroquinozaline-2,3-dione; 50 microM). Our results show that neuronal activity evokes Ca2+ signals not only in neurones, but also in glial cells and suggest that these Ca2+ transients are partly mediated via activation of glutamate/kainate receptors.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1990

Self-catalyzed irreversible inactivation of rat hepatic aryl sulfotransferase IV by N-hydroxy-2-acetylaminofluorene.

Rat hepatic aryl sulfotransferase IV catalyzes the sulfonation of the hepatocarcinogen, N-hydroxy-2-acetylaminofluorene. The resulting reactive N-O-sulfate ester is believed to be the ultimate carcinogenic species responsible for the induction of hepatic neoplasia. Previous studies have shown that dietary administration of either 2-acetylaminofluorene or N-hydroxy-2-acetylaminofluorene to rats is accompanied by a rapid decline in hepatic aryl sulfotransferase activity in vivo. In the present study, preincubation of purified rat hepatic aryl sulfotransferase IV with N-hydroxy-2-acetylaminofluorene resulted in rapid, time-dependent enzyme inactivation. This in vitro inactivation was not reversed by dialysis or gel filtration. Inclusion of excess nucleophile, methionine, resulted in considerable but not complete protection from inactivation. The inactivation was PAPS dependent and blocked by the sulfotransferase inhibitor, pentachlorophenol. The above observations and the apparent pseudo first-order kinetics observed suggest that the inactivation was in part mechanism based. Mechanism-based inactivation of the aryl sulfotransferases has not been previously reported. Furthermore, the results of the present study indicate that the previously reported in vivo decline in rat hepatic aryl sulfotransferase activity may be attributable in part to enzyme inactivation by its own reactive product.

2-Acetylaminofluorene↗
PubMedDate not supplied

Calcium signalling in glial cells.

Calcium signals are the universal way of glial responses to the various types of stimulation. Glial cells express numerous receptors and ion channels linked to the generation of complex cytoplasmic calcium responses. The glial calcium signals are able to propagate within glial cells and to create a spreading intercellular Ca2+ wave which allow information exchange within the glial networks. These propagating Ca2+ waves are primarily mediated by intracellular excitable media formed by intracellular calcium storage organelles. The glial calcium signals could be evoked by neuronal activity and vice versa they may initiate electrical and Ca2+ responses in adjacent neurones. Thus glial calcium signals could integrate glial and neuronal compartments being therefore involved in the information processing in the brain.

Animals↗