PubMed Health⌕ Search

Biomedical subjects

D Swandulla

Publications and source records attributed to D Swandulla.

At least 19 recordsLinked to original sources
PubMed1999

Expression of early hippocampal CA1 LTP does not lead to changes in AMPA-EPSC kinetics or sensitivity to cyclothiazide.

We have analysed whether the expression of long-term potentiation (LTP) in rat hippocampal CA1 neurons involves a change in the kinetics of (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated excitatory postsynaptic currents (EPSCs) (AMPA-EPSCs) or their susceptibility to the AMPA receptor modulator cyclothiazide. AMPA-EPSCs in the CA1 region were evoked by alternate stimulation of two independent Schaffer collateral-commissural inputs of slices of adult rat hippocampus. In the current-clamp mode a strong tetanus (100 Hz, 1 s) applied to one input (input I) induced stable LTP of AMPA-EPSCs in this input, while the control input (input II) remained unaffected. For neither input were EPSC rise time and decay kinetics significantly changed. The application of cyclothiazide prolonged the rise time and the decay time constants of the AMPA-EPSCs in both control and potentiated inputs to the same extent (Input I-rise time: 198+/-8%, decay: 148+/-12%; input II-rise time: 212+/-14%, decay: 144+/-19%; n=8). Furthermore, when present during tetanization cyclothiazide did not occlude LTP, suggesting that cyclothiazide and tetanic stimulation enhance AMPA-EPSCs via independent mechanisms. Our findings argue against changes in (de-)activation or desensitization of AMPA receptors as the molecular basis for the expression of LTP.

Animals↗
PubMed1998

Synaptic modulation of oscillatory activity of hypothalamic neuronal networks in vitro.

1. Rhythmic bursts of action potentials in neurosecretory cells are a key factor in hypothalamic neurosecretion. Rhythmicity and synchronization may be accomplished by pacemaker cells synaptically driving follower cells or by a network oscillator. 2. In this review we describe a hypothalamic cell culture which may serve as a model for a hypothalamic network oscillator. An overview is given of neurochemical phenotypes, synaptic mechanisms and their development, properties of receptors for fast synaptic transmission, and membrane properties of cells in dissociated rat embryonic hypothalamic culture. 3. Rhythmic activity spreads in the cultured network through synapses that release glutamate, activating a heteromultimeric AMPA-type receptor containing a GluR2 subunit which is associated with a high-conductance channel for Na+ and K+. Rhythmic activity is controlled by synapses that release GABA to activate GABAA receptors. The presumed function of the two receptor types is facilitated by their respective location, GABAA receptors predominating near the soma and AMPA receptors being abundant in dendrites. 4. Network oscillators may be more reliable for the presumed function than single-cell oscillators. They are controlled through synaptic modulation, which may prove to represent a process important for the release of hormones.

Animals↗
PubMed1997

Fractional Ca2+ currents through capsaicin- and proton-activated ion channels in rat dorsal root ganglion neurones.

1. Capsaicin and protons cause excitation and sensitization of primary nociceptive afferents. In a subset of dorsal root ganglion (DRG) neurones, which probably represent nociceptive neurones, both capsaicin and protons induce slowly inactivating non-selective cation currents. Whole-cell as well as single channel currents activated by these two stimuli share many biophysical and physiological properties in these neurones. This has lead to the suggestion that protons and capsaicin might activate the same ion channels. 2. In this study we simultaneously measured fluorescence signals and whole-cell currents activated by capsaicin or protons in acutely isolated DRG neurones filled with a high concentration (1 mM) of the Ca2+ indicator dye fura-2. From these measurements the fractional contribution of Ca2+ (Pf; the portion of the whole-cell current carried by Ca2+) to capsaicin- and two types of proton-induced (fast and slowly inactivating) membrane currents was determined. 3. Capsaicin- and slowly inactivating proton-induced currents were accompanied by a change in fluorescence that was dependent on the presence of extracellular Ca2+. With 1.6 mM extracellular Ca2+ and at a holding potential of -80 mV Pf of capsaicin-induced currents (at pH 7.3) was 4.30 +/- 0.17% (mean +/- S.E.M.; no. of experiments, n = 16) and of slowly inactivating proton-induced currents (at pH 5.1) was 1.65 +/- 0.11% (n = 17). Pf of fast inactivating proton-induced currents was negligible. 4. Pf of capsaicin- and slowly inactivating proton-induced currents increased with increasing extracellular Ca2+ concentration (0.5-4.8 mM). 5. Pf of both current types decreased linearly with decreasing extracellular pH by about 0.7% per pH unit over the pH range investigated. When determined at the same extracellular pH Pf values were significantly different for the two current types at all pH values tested. 6. In summary, our results provide evidence that capsaicin and protons activate ion channels which are markedly permeable to Ca2+. The fractional contribution of Ca2+, however, was significantly different for capsaicin- and slowly inactivating proton-induced currents. This strongly suggests that the two stimuli activate different populations of ion channels and supports the possibility that Ca2+ influx through these channels may be important for Ca(2+)-dependent sensitization of primary nociceptive neurones.

Animals↗
PubMed1997

Modulation of excitatory synaptic transmission by nociceptin in superficial dorsal horn neurones of the neonatal rat spinal cord.

1. The modulatory actions of nociceptin/orphanin FQ on excitatory synaptic transmission were studied in superficial dorsal horn neurones in transverse slices from 7 to 14 day old rats. 2. Glutamatergic excitatory postsynaptic currents (e.p.s.cs) were recorded from the somata of the neurones in the whole-cell patch-clamp configuration. E.p.s.cs were evoked by extracellular electrical stimulation (100 microns, 3-10 V) of the ipsilateral dorsal root entry zone by use of a glass electrode. E.p.s.cs with constant short latency (< 2.3 ms) and with no failures upon stimulation were assumed to be monosynaptic. These e.p.s.cs occurred with an average latency of 1.72 +/- 0.098 ms and exhibited a fast decay with a time constant, tau, of 4.8 +/- 0.53 ms (n = 30). 3. Nociceptin reversibly reduced the amplitudes of e.p.s.cs in a concentration-dependent manner in 25 out of 27 cells tested. Average maximum inhibition was 51.6 +/- 5.7% (mean +/- s.e.mean; n = 9), at concentrations > 3 microM. EC30 was 485 +/- 47 nM and the Hill coefficient was 1.29 +/- 0.09. 4. Inhibition of synaptic transmission by nociceptin (10 microM) was insensitive to the non-specific opioid receptor antagonist naloxone (10 microM) indicating that nociceptin did not act via classical opioid receptors. 5. In order to determine the site of action of nociceptin spontaneous miniature e.p.s.cs (m-e.p.s.cs) were recorded. Nociceptin reduced the frequency of m-e.p.s.cs in 6 out of 7 cells but had no effect on their amplitude distribution or on their time course. These findings suggest a pre- rather than a postsynaptic modulatory site of action. This is in line with the finding that current responses elicited by extracellular application of L-glutamate (10 microM) were not affected by nociceptin (10 microM; n = 7). 6. No positive correlation was found between the degree of inhibition by nociceptin (10 microM) and by the mixed delta- and mu-receptor agonist methionine-enkephalin (10 microM). This suggests that both neuropeptides acted on different but perhaps overlapping populations of synaptic connections. 7. Our results indicate that nociceptin inhibits excitatory synaptic transmission in the superficial layers of the rat dorsal horn by acting on presynaptic, presumably ORL1 receptors. This may be an important mechanism for spinal sensory information processing including nociception.

Animals↗
PubMed1997

Synaptic connectivity in cultured hypothalamic neuronal networks.

We have developed a novel approach to analyze the synaptic connectivity of spontaneously active networks of hypothalamic neurons in culture. Synaptic connections were identified by recording simultaneously from pairs of neurons using the whole cell configuration of the patch-clamp technique and testing for evoked postsynaptic current responses to electrical stimulation of one of the neurons. Excitatory and inhibitory responses were distinguished on the basis of their voltage and time dependence. The distribution of latencies between presynaptic stimulation and postsynaptic response showed multiple peaks at regular intervals, suggesting that responses via both monosynaptic and polysynaptic paths were recorded. The probability that an excitatory event is transmitted to another excitatory neuron and results in an above-threshold stimulation was found to be only one in three to four. This low value indicates that in addition to evoked synaptic responses other sources of excitatory drive must contribute to the spontaneous activity observed in these networks. The various types of synaptic connections (excitatory and inhibitory, monosynaptic, and polysynaptic) were counted, and the observations analyzed using a probabilistic model of the network structure. This analysis provides estimates for the ratio of inhibitory to excitatory neurons in the network (1:1.5) and for the ratio of postsynaptic cells receiving input from a single GABAergic or glutamatergic neuron (3:1). The total number of inhibitory synaptic connections was twice that of excitatory connections. Cell pairs mutually connected by an excitatory and an inhibitory synapse occurred significantly more often than predicted by a random process. These results suggests that the formation of neuronal networks in vitro is controlled by cellular mechanisms that favor inhibitory connections in general and specifically enhance the formation of reciprocal connections between pairs of excitatory and inhibitory neurons. These mechanisms may contribute to network formation and function in vivo.

Animals↗
PubMed1997

A simple computer model to simulate ECG based on ionic channels.

The presented computer model reconstructs the ECG with 500 myocardium cells based on seven ionic channels. The low hardware-requirement and the exact demonstration of physiological and pathophysiological coherence make this simulation model very useful for medical scientific education.

Cell Hypoxia↗
PubMed1996

Calcium channel types contributing to excitatory and inhibitory synaptic transmission between individual hypothalamic neurons.

The contribution of L-, N-, P- and Q-type Ca2+ channels to excitatory and inhibitory synaptic transmission and to whole-cell Ba2+ currents through Ca2+ channels (Ba2+ currents) was investigated in rat hypothalamic neurons grown in dissociated cell culture. Excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) were evoked by stimulating individual neurons under whole-cell patch-clamp conditions. The different types of high-voltage-activated (HVA) Ca2+ channels were identified using nifedipine, omega-Conus geographus toxin VIA (omega-CTx GVIA), omega-Agelenopsis aperta toxin IVA (omega-Aga IVA), and omega-Conus magus toxin VIIC (omega-CTx MVIIC). N-, but not P- or Q-type Ca2+ channels contributed to excitatory as well as inhibitory synaptic transmission together with Ca2+ channels resistant to the aforementioned Ca2+ channel blockers (resistant Ca2+ channels). Reduction of postsynaptic current (PSC) amplitudes by N-type Ca2+ channel blockers was significantly stronger for IPSCs than for EPSCs. In most neurons whole-cell Ba2+ currents were carried by L-type Ca2+ channels and by at least two other Ca2+ channel types, one of which is probably of the Q-type and the others are resistant Ca2+ channels. These results indicate a different contribution of the various Ca2+ channel types to excitatory and inhibitory synaptic transmission and to whole-cell currents in these neurons and suggest different functional roles for the distinct Ca2+ channel types.

Animals↗
PubMed1996

Interactions of 2,3-benzodiazepines and cyclothiazide at AMPA receptors: patch clamp recordings in cultured neurones and area CA1 in hippocampal slices.

1. The 2,3-benzodiazepines GYKI 52466, GYKI 53405 and GYKI 53655 antagonized AMPA-induced currents in cultured superior colliculus neurones in a non use-dependent manner (steady state IC50s: GYKI 52466 9.8 +/- 0.6 microM; GYKI 53405 3.1 +/- 0.6 microM; GYKI 53655 0.8 +/- 0.1 microM). 2. Higher concentrations of all three antagonists slowed the onset kinetics and quickened the offset kinetics of AMPA-induced currents indicative of an allosteric interaction with the AMPA recognition site. 3. Cyclothiazide (3-300 microM) dramatically slowed desensitization of AMPA-induced currents and potentiated steady state currents (EC50 10.0 +/- 2.5 microM) to a much greater degree than peak currents. Both tau on and tau off were also increased by cyclothiazide in a concentration-dependent manner (EC50: tau on 42.1 +/- 4.5 microM; tau off 31.6 +/- 6.6 microM). 4. Cyclothiazide (10-100 microM) shifted the concentration-response curves of the 2,3-benzodiazepines to the right. For example, with 10 microM cyclothiazide the IC50s of GYKI 52466 and GYKI 53405 on steady-state AMPA-induced currents were 57.9 +/- 9.5 and 41.6 +/- 1.5 microM, respectively. 5. GYKI 53405 and GYKI 52466 concentration-dependently reversed the effects of cyclothiazide (100 microM) on offset kinetics (GYKI 53405 IC50 16.6 +/- 4.2 microM). However, the 2,3-benzodiazepines were unable to reintroduce desensitization in the presence of cyclothiazide and even concentration-dependently slowed the onset kinetics of AMPA responses further (GYKI 53405 EC50 8.0 +/- 2.8 microM). 6. GYKI 52466 decreased the peak amplitude of hippocampal area CA1 AMPA receptor-mediated excitatory postsynaptic currents (e.p.s.cs) (IC50 10.8 +/- 0.8 microM) with no apparent effect on response kinetics. Cyclothiazide prolonged the decay time constant of AMPA receptor-mediated e.p.s.cs (EC50 35.7 +/- 6.5 microM) with less pronounced effects in slowing e.p.s.c. onset kinetics and increasing e.p.s.c. amplitude. 7. Cyclothiazide (330 microM) shifted the concentration-response curve for the effects of GYKI 52466 on AMPA receptor-mediated e.p.s.c. peak amplitude to the right (GYKI 52466 IC50 26.9 +/- 9.4 microM). Likewise, GYKI 52466 (30-100 microM)) shifted the concentration-response curve for the effects of cyclothiazide on AMPA receptor-mediated e.p.s.c. decay time constants to the right. 8. In conclusion, cyclothiazide and the 2,3-benzodiazepines seem to bind to different sites on AMPA receptors but exert strong allosteric interactions with one another and with other domains such as the agonist recognition site. The interactions of GYKI 52466 and cyclothiazide on AMPA receptor-mediated e.p.s.cs in area CA1 of hippocampal slices provide evidence that the decay time constant of these synaptic events are not governed by desensitization.

Animals↗
PubMed1996

Ca2+ permeability of the sustained proton-induced cation current in adult rat dorsal root ganglion neurons.

1. Microfluorometric Ca2+ measurements using FURA-2 and whole cell patch-clamp recordings were performed to investigate the Ca2+ permeability of ion channels underlying the proton-induced sustained cation current in adult rat dorsal root ganglion neurons. 2. In a subpopulation of these neurons, extracellular application of acidic solutions (pH 5.1) elicited a sustained cation current and a concomitant reversible rise in the intracellular free Ca2+ concentration ([Ca2+]i), which depended on the presence of external Ca2+. Ruthenium red (10 microM) reduced both the current and the rise in [Ca2+]i to about the same extent. 3. In the presence of 2 mM external Ca2+, sustained proton-induced currents reversed sign at -4.6 +/- 1.2 (SE) mV, with external Na+ and internal Cs+ as the major charge carriers. Increasing the external Ca2+ concentration to 30 mM shifted the reversal potential (Erev) by 3.0 +/- 0.9 mV toward more positive values, suggesting a permeability ratio of Ca2+/Cs+ of 0.41. A similar value (0.35) could be obtained from Erev (-21 mV) under bi-ionic conditions with 100 mM external Ca2+ and 154 mM internal Cs+. 4. These results demonstrate that the proton-activated cation channels investigated here are moderately permeable to Ca2+. This may lead to pathophysiologically relevant increases in [Ca2+]i on prolonged exposure of the cells to an acidic environment in inflamed or ischemic tissue.

Animals↗
PubMed1995

Activation of Ca2+ signaling in neutrophils by the mast cell-released immunophilin FKBP12.

The immunophilins of the FK506-binding protein (FKBP) family are intracellular proteins that bind the immunosuppresants FK506 and rapamycin. In this study we show that HMC-1 mast cells sensitized with IgE release FKBP12 upon stimulation with anti-IgE. The release is rapid and not affected by actinomycin D or cycloheximide, suggesting that it is due to exocytosis from a storage compartment. FKBP12 from HMC-1 mast cells exhibits biological activity. When applied extracellularly to human neutrophils, it induces transient changes in the intracellular Ca2+ concentration ([Ca2+]i) due to Ca2+ release from intracellular stores. Inhibition of [Ca2+]i changes by ruthenium red and ryanodine indicates that ryanodine receptor/Ca2+ release channels are involved in FKBP12-induced Ca2+ signaling. Neutrophil activation by mast cell-derived FKBP12 is prevented by complexing FKBP12 with FK506 or rapamycin. These results demonstrate that extracellular FKBP12 functions as a cytokine in cell-to-cell communication. They further suggest a pathophysiological role for FKBP12 as a mediator in immediate or type I hypersensitivity and may have implications for novel therapeutic strategies in the treatment of allergic disorders with FK506 and rapamycin.

Amino Acid Sequence↗
PubMed1995

Synaptic feedback excitation has hypothalamic neural networks generate quasirhythmic burst activity.

1. Dissociated neurons from embryonic rat hypothalamus form synaptically coupled networks when cultured for several weeks. When synaptic inhibition is suppressed by the gamma-aminobutyric acid-A (GABAA) antagonist picrotoxin, neurons exhibit "rhythmic" burst activity that is blocked by the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). 2. We have examined whether this type of activity is generated by pacemaker cells driving follower cells synaptically or whether it is generated by a neuronal network through feedback excitation by Ca2+ imaging of Fura-2/AM-loaded neurons and single-cell recording. 3. IN the presence of the GABAA antagonist picrotoxin (20 microM), a large majority of neurons (> 85%) displayed quasirhythmic increases in intracellular concentration of Ca2+ ([Ca2+]i). Single-cell recording revealed that these increases in [Ca2+]i correspond to burst discharges in individual neurons. When excitatory synaptic transmission was blocked by CNQX (20 microM), none of the imaged neurons exhibited significant oscillations in [Ca2+]i. 4. Investigation of the intervals between bursts from single-cell recording revealed a random distribution over a range of 400% from the minimum interval. Poincaré maps of burst intervals, i.e., graphs of all burst intervals versus their preceding burst interval, revealed that a burst interval is unpredictable from its predecessor. 5. When synaptic coupling was attenuated by low concentrations of CNQX (50-500 nM), the mean burst interval was considerably increased without a change in the random character of burst activity. 4-Aminopyridine (1 mM), known to increase synaptic efficiency, reduced the mean burst interval.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Modulation of fast excitatory synaptic transmission by cyclothiazide and GYKI 52466 in the rat hippocampus.

The effects of cyclothiazide, a drug which potentiates AMPA receptor-mediated responses and GYKI 52466, a non-competitive AMPA receptor antagonist, were studied on fast glutamatergic transmission in rat hippocampal slices. Cyclothiazide prolonged the decay of AMPA receptor-mediated EPSCs (AMPA-EPSCs) in a concentration-dependent manner. GYKI 52466 reduced the peak amplitude of AMPA-EPSCs and blocked the induction of LTP. When GYKI 52466 was applied in the presence of cyclothiazide it still reduced the peak amplitude of AMPA-EPSCs but was not able to reverse the cyclothiazide induced prolongation of AMPA-EPSC duration. These data suggest that GYKI 52466 and cyclothiazide probably mediate their effects on the AMPA receptor via different binding sites.

2-Amino-5-phosphonovalerate↗
PubMed1994

Calcium-activated non-selective channels in the nervous system.

In the decade, since the first description of calcium-activated non-selective (CAN) channels in cardiac myocytes, pancreatic acini and neuroblastoma, this type of channel has been shown to have a ubiquitous distribution across a variety of tissues. Recently, their role in the function of cells of the nervous system has become better delineated. Because CAN channels pass depolarizing current, respond to cytoplasmic Ca2+ activity and do not inactivate, they are capable of producing maintained depolarization of neurons. This property endows upon CAN channels an important role in both physiological functions and pathological processes of the nervous system.

Animals↗
PubMed1994

Molecular pathways for presynaptic calcium signaling.

The results presented in this article describe two distinct, Ca-regulated molecular pathways in presynaptic terminals and implicate these two pathways in differentially mediating neurotransmitter secretion and PTP. Our current view of the Ca-dependent triggering of secretion and PTP is shown in Fig. 9. According to this scheme, differential activation of these two pathways is achieved by a combination of diffusion-based dilution of Ca that enters the terminal through voltage-gated Ca channels and by coupling these pathways to Ca receptors with different affinities for Ca ions. A simple way to achieve these conditions is to position these two receptors at different distances from the Ca channels, as shown in Fig. 2. Given that Ca ions are involved in activating many different presynaptic processes (Fig. 1), we propose that closer scrutiny of the molecular physiology of nerve terminals will reveal a wide variety of Ca-activated pathways responsible for producing these diverse processes.

Amino Acid Sequence↗
PubMed1993

Molecular properties of the glutamate receptor mediating synaptic excitation in rat hypothalamic neurons.

AMPA-type glutamate receptors (GluRs) mediate synaptic excitation in networks of cultured rat hypothalamic neurons [18, 25]. Under voltage clamp the agonists quisqualate and AMPA induce current responses which consist of a maintained and/or transient component depending on the concentrations applied. The current-voltage relationship for both components is linear. The biphasic response patterns are due to receptor desensitization which is fast and does not require intracellular second messengers for its activation. Several GluR-subtype-encoding transcripts were found in these neurons using polymerase chain reaction (PCR) methods. While mRNAs encoding the GluR2 and 3 flip forms are expressed early, mRNAs encoding the GluR1, 2 and 3 flop forms and the GluR4 flip form appear only in cultures older than 3 weeks. By comparison to recombinant receptors, the properties of the native receptor can be accommodated by a heteromeric receptor containing GluR2 as one of the subunits.

Animals↗
PubMed1993

Characterization of neurochemical phenotypes in cultured hypothalamic neurons with immunohistochemistry and in situ hybridization.

The expression of neurochemical phenotypes was studied in long-term cultures of dissociated embryonic neurons from rat hypothalamus. With time in culture, these neurons establish a complex dendritic and axonal network, as indicated by staining with antibodies against microtubulin-associated protein (MAP2) and neurofilaments (SMI32 and SMI33) as well as GABA and glutamate decarboxylase mRNA immunoreactivity. Neurons expressing neuropeptide Y (NPY) mRNA and NPY peptide and opioid-like peptides as well as vasopressin were observed. Further, weakly acetylcholinesterase- and NADPH diaphorase (nitric-oxide synthase)-labelled neurons were present. In conclusion, the neurochemical phenotypes reported for hypothalamic neurons in vivo can be observed in these cultures. This indicates that the culture conditions allow morphological and molecular differentiation. These findings support the view that long-term hypothalamic cultures provide a valuable model for studying mechanisms of neurosecretion in hypothalamic networks.

Acetylcholinesterase↗