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Biomedical subjects

C Franke

Publications and source records attributed to C Franke.

At least 109 records · Page 6Linked to original sources
PubMed1991

Altered Na+ channel activity and reduced Cl- conductance cause hyperexcitability in recessive generalized myotonia (Becker).

Intact muscle fibers or resealed fiber segments from 7 patients with recessive generalized myotonia were studied in vitro. All fibers had normal resting membrane potentials and normal resting [Ca2+]i several hours after removal. Contractions were characterized by slowed relaxation which was due to electrical after-activity. Often spontaneous depolarizations were recorded intracellularly. In all fibers, the steady state voltage-current relationship was abnormal, due to a reduced Cl- conductance. However, this conductance ranged from 0% to 66% of the total membrane conductance, whereas, in normal muscle, it was 80%. Theoretically, myotonic after-discharges would not appear until the Cl- conductance is below 20%. Thus, the membrane hyperexcitability must be due to another defect, at least in the preparations in which the Cl- conductance was only slightly reduced. In all patches from all patients investigated with the patch clamp technique, we observed reopenings of the Na+ channels throughout depolarizing pulses (such behavior was absent in normal muscle). If a patch was polarized to potentials less negative than the resting potential, the duration of the reopenings increased. We conclude that a combination of reduced Cl- conductance and the reopenings of Na+ channels underlie the electrical after-activity in recessive generalized myotonia.

Chlorides↗
PubMed1991

Altered gating and conductance of Na+ channels in hyperkalemic periodic paralysis.

Electrophysiological studies on muscle fibres from patients with hyperkalemic periodic paralysis with myotonia have shown that the episodes of weakness are caused by a sustained depolarization of the sarcolemma to potentials between -40 and -60 mV. In muscle fibre segments from three such patients this sustained depolarization was caused by noninactivating Na+ channels with reduced single-channel conductance blocked by TTX and procainamide. As the chloride conductance was normal, myotonia may be best explained with the abnormal reopenings of the Na+ channels. The recently described genetic linkage between hyperkalemic periodic paralysis with myotonia and the gene coding for the TTX-sensitive Na+ channel suggests an altered primary structure of this channel causing its abnormal function.

Electric Conductivity↗
PubMed1991

Steep concentration dependence and fast desensitization of nicotinic channel currents elicited by acetylcholine pulses, studied in adult vertebrate muscle.

Skeletal muscles of adult mice and frogs were dissociated enzymatically and prepared for patch-clamping within less than 6 h. Outside-out patches were superfused with repetitive pulses of acetylcholine (ACh) with switching times of about 0.2 ms. Peak responses were reached within 1 ms. In mouse muscle the average channel conductance was 65 pS and the average open time 1 ms (20 degrees C). Between 1 and 10 microM ACh, the peak responses increased proportional to the second to third power of the ACh concentration, and less steeply between 10 and 1000 microM ACh. The apparent Km of the dose-response curve was about 100 microM. After the peak, channel opening probability declined with time constants decreasing from about 1 s with 1 microM ACh to 15-50 ms with 1 mM ACh. After 100 ms desensitization the channel opening had decreased to less than 1/300 peak value. The rate of desensitization increased with rising temperature, with Q10 values of 1.7-2.5 between 10 and 30 degrees C. The desensitization characteristics of channels from frog muscle were similar to that from mice. With pulses of 100 microM ACh the channels opened with a probability of 0.55, the open probability declining with a time constant of about 60 ms and dropping to less than 0.001 after 300 ms. The results support the view that three binding steps of ACh are necessary for opening of the channel. Desensitization in the presence of high ACh concentrations is slower than the decay of synaptic currents.

Acetylcholine↗
PubMed1991

Altered sodium channel behaviour causes myotonia in dominantly inherited myotonia congenita.

The cause of increased excitability in autosomal dominant myotonia congenita (MyC) was studied in resealed greater than 3-cm long segments of muscle fibres from eight patients. Three hours after biopsy only about 50% of the fibre segments had regained a normal resting potential. This differs from our experiences with normal muscle or other disorders of myotonia (e.g. recessive generalized myotonia) where nearly all cut fibres reseal and repolarize during this time. When the depolarized MyC fibre segments were placed in a solution containing 1 microM tetrodotoxin (TTX) they repolarized to -80 to -90 mV. In fibre segments with normal resting potential, in the absence of TTX, spontaneous myotonic runs were recorded intracellularly, occasionally with double spikes. For only one of the eight patients, the Cl- conductance was reduced (50% of the total membrane conductance vs the usual 75%), for the rest of the patients the steady-state current-voltage relationship was normal. Sodium currents through single membrane channels were recorded with a patch clamp. For every patient re-openings of the Na+ channels were observed throughout 10-ms depolarizing pulses. These are very uncommon in normal muscle. At potentials positive to the resting potential, the duration of the re-openings increased, but the current amplitude was the same. It is concluded that in myotonia congenita re-openings of Na+ channels are the major cause of hyperexcitability and that Cl- conductance is normal. If it is reduced in rare cases, it may potentiate the myotonia.

Action Potentials↗
PubMed1991

Kinetic constants of the acetylcholine (ACh) receptor reaction deduced from the rise in open probability after steps in ACh concentration.

Outside-out patches of enzymatically dissociated adult and denervated mouse muscle fibers were superfused repetitively by pulses of acetylcholine (ACh) containing solution. Up to 300 channels opened simultaneously 300 microseconds after the beginning of a 1,000 microM ACh pulse corresponding to a peak current i of almost -1 nA. Single responses to ACh were averaged and the concentration dependence of i and of the rise time tr from 0.1 i to 0.9 i was measured. In adult receptors, i increased proportional to the second to third power of ACh concentration, whereas in embryonic-type receptors it was proportional to the first to the second power. tr increased from approximately 0.3 ms at 1,000 microM ACh to a plateau value of approximately 5 ms for adult and of approximately 10 ms for embryoniclike receptors at concentrations less than 10 microM ACh. The concentration dependence of i and tr was simulated using the standard model of ACh binding with different combinations of rate constants and two and three binding sites for ACh. The calculated curves were compared to the measurements and a set of well fitting rate constants was determined for adult and embryoniclike receptors. Three binding sites for ACh were necessary to fit the dose response for i for adult receptors. A method for deriving rate constants in a model of ACh-receptor interaction is described that avoids analysis of open-closed kinetics of single channels, which in rapid systems, as the ones studied here, are at the limit of the frequency response of the current measurement.

Acetylcholine↗
PubMed1991

Glutamate-activated channels in adult rat ventral spinal cord cells.

1. Currents in response to rapid application of glutamate and its agonists were studied in cells dissociated from the ventral spinal cord of adult rats. 2. Glutamate activated an inward current that desensitized in less than 15 ms. 3. Responses to quisqualate and to DL-alpha-amino-3-hydroxy-5-methyl-isoxeazolepropionic acid (AMPA) also desensitized with time constants ranging from 7 to 18 ms in whole cell configuration and from 3.4 to 4.3 ms in outside-out configuration. Desensitization rate was independent of membrane potential. Single-channel conductance was 12 pS. 4. Currents in response to N-methyl-D-aspartate activation also desensitized; the time constants ranged from 15 to 50 ms. Single-channel conductance was 23 pS. 5. Kainate responses did not desensitize appreciably. Single-channel conductance was 17 pS. 6. These data obtained from adult cells are similar to values reported for cultured embryonic and neonatal neurons, indicating minimal postnatal changes in these aspects of glutamate receptors.

2-Amino-5-phosphonovalerate↗
PubMed1991

Desensitization and resensitization rates of glutamate-activated channels may regulate motoneuron excitability.

1. Single-channel properties of desensitizing glutamate-activated channels were analyzed in outside-out patch-clamp recordings from a motoneuron-enriched cell fraction from embryonic chick. A piezo-driven device was used to achieve fast solution exchange at the electrode tip, resulting in maximum activation within 2 ms. 2. Quisqualate/AMPA receptors, with a 13-pS conductance, desensitized rapidly; the desensitization rate depended on agonist concentration but not on membrane potential. When quisqualate was applied slowly, the quisqualate-activated channels desensitized without prior channel opening, indicating desensitization from the closed state. After a 10-ms refractory period, resensitization of all channels required up to 300 ms; resensitization rate did not depend on the duration of the preceding quisqualate application. 3. At agonist concentrations less than or equal to 1 mM, kainate receptors, with a 20-pS conductance, did not desensitize. At kainate concentrations greater than or equal to 1 mM, though, kainate receptors desensitized to a low steady-state conductance within approximately 200 ms. Resensitization of all channels required as long as 3 s, which could render kainate receptors inexcitable during high-frequency activation. 4. Desensitization rates of whole-cell currents were similar to those observed in outside-out mode. Glutamate- and quisqualate-activated responses were similar, suggesting that the rapidly desensitizing quisqualate-sensitive receptor type may dominate the kinetics of whole-cell excitatory postsynaptic currents (EPSCs) in this preparation. 5. It may be concluded that the efficacy of glutamate-mediated synaptic transmission is modulated by differences in the rates of desensitization and resensitization.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1991

Ionic currents of cultured olfactory receptor neurons from antennae of male Manduca sexta.

Whole-cell and single-channel voltage-clamp techniques were used to identify and characterize the ionic currents of insect olfactory receptor neurons (ORNs) in vitro. The cells were isolated from the antennae of male Manduca sexta pupae at stages 3-5 of adult development and maintained in primary cell culture. After 2-3 weeks in vitro, the presumptive ORNs had resting potentials of -62 +/- 12 mV (n = 18) and expressed at least 1 type of Na+ channel and at least 3 types of K+ channels. Na+ currents, recorded in the whole-cell mode, were reversibly blocked by 0.1 microM tetrodotoxin. The predominant type of K+ channel observed was a voltage-activated K+ channel (gamma = 30 pS) with characteristics similar to those of the delayed rectifier. The activity of the 30-pS K+ channel could be inhibited by the application of nucleotides to the cytoplasmic face of inside-out patches of membrane. The nucleotides had relative potencies as follows: ATP greater than cGMP greater than cAMP, with an inhibition constant for ATP of Ki = 0.18 mM. Raising the intracellular Ca2+ concentration from 0.1 to 5 microM induced the opening of a Ca2(+)-activated K+ channel (gamma = 66 pS at 0 mV) that had a low voltage sensitivity. A third, transient type of K+ channel (gamma = 12-18 pS) could be activated by depolarizing voltage steps from very negative resting potentials. Properties of this channel were similar to those of the "A-channel." These results support the conclusion that M. sexta ORNs differentiate in vitro and provide the basis for studying primary mechanisms of olfactory transduction.

Animals↗
PubMed1990

Two different types of potassium channels in human skeletal muscle activated by potassium channel openers.

The inside-out patch clamp technique was used to record the effects of K+ channel openers (EMD 52692, RP 49356 and Cromakalim) on single channel currents in membrane blebs of human skeletal muscle. Two types of K+ channels were activated by these drugs: an ATP-sensitive K+ channel which was inhibited by 3 mM ATP and 5 microM Glibenclamide and an ATP insensitive K+ channel. The open probability of both types was strongly increased by K+ channel openers. Glibenclamide antagonized the action of the K+ channel openers.

Adenosine Triphosphate↗
PubMed1990

Glutamatergic channels in locust muscle show a wide time range of desensitization and resensitization characteristics.

Outside-out patches of membrane were excised from locust muscle. 10 mM L-glutamate applied to such patches in short pulses elicited rapidly a peak of channel opening, followed by desensitization with time constants between 1 ms and 20 ms for different channel subtypes. Slowly rising glutamate concentrations could produce almost complete desensitization without any channel openings. Desensitization thus proceeds from a closed channel state, prior to channel opening. Resensitization, i.e. recovery from desensitization, was tested by applying pairs of glutamate pulses, varying the pulse interval. Resensitization has a biphasic time course. While in rapidly desensitizing channels more than 50% of resensitization was achieved within a few milliseconds, in slowly desensitizing channels resensitization took seconds.

Animals↗
PubMed1990

Schwartz-Jampel syndrome: II. Na+ channel defect causes myotonia.

Skeletal muscle fibers from a patient with Schwartz-Jampel syndrome were studied in vitro. The fibers had normal resting membrane potentials, but their resting [Ca2+]i was elevated. The resting potentials were unstable and spontaneous depolarizations caused twitching in all fibers. Stimulated contractions were characterized by markedly slowed relaxation which was due to electrical after-activity. Neither curare (0.7 microM), tocainide (50 microM), nor phenytoin (80 microM) had an effect on the myotonic activity. In contrast, procainamide (200 microM) suppressed the hyperexcitability without affecting the twitch amplitude. The steady-state current-voltage relation was normal in 5 fibers, but altered in 3 others. These latter fibers had an increased specific membrane resistance owing to a decreased Cl- conductance. The Na+ channels were investigated in the cell-attached patch clamp mode. In all patches on either type of fiber, depolarizing pulses elicited delayed, synchronized openings of Na+ channels. These abnormal openings occurred even after the surface membrane repolarized. We hypothesize that these altered membrane conductances are responsible for the hyperexcitability and the associated slowed relaxation.

Child, Preschool↗
PubMed1990

Characteristics of single Na+ channels of adult human skeletal muscle.

The patch-clamp technique was used to study Na+ channels of human skeletal muscle. Preparations were from biopsies of quadriceps muscle from adults who were not suffering from neuromuscular diseases. Activity of Na+ channels was recorded from inside-out patches when the membrane potential was stepped from a holding potential of -110 mV to potential above a threshold of about -65 mV. Single channel activity increased within minutes after hyperpolarizing the patch due to recovery from ultra-slow inactivation. Up to ten Na+ channels were active in individual patches. Macroscopic currents were reconstructed by averaging single channel currents. The time-to-peak current declined from 1.6 ms at -60 mV to 0.5 ms at + 10 mV. The currents decayed mono-exponentially with time constants between 12.1 ms at -60 mV and 0.4 ms at + 10 mV (21 C). The conductance of single Na+ channels was 1.65 pS and the mean open time was voltage-dependent. At -50 mV, the mean open time was 0.4 ms, while positive to -10 mV it increased to values above 1 ms. In the threshold potential range, the number of openings per depolarizing pulse was larger than the number of channels under the patch-clamp pipette, indicating reopening of Na+ channels at this potential. Openings could be observed only rarely 10 ms after onset of depolarization and the macroscopic current produced by late openings was less than 0.1% of the peak current. Human skeletal muscle is thus suitable for investigation with the patch-clamp technique and the determination of properties of Na+ channels with this technique could be the basis for an assessment of possible defects of these channels in diseased muscle.

Biopsy↗
PubMed1990

Rapid activation, desensitization, and resensitization of synaptic channels of crayfish muscle after glutamate pulses.

Completely desensitizing excitatory channels were activated in outside-out patches of crayfish muscle membrane by applying glutamate pulses with switching times of approximately 0.2 ms for concentration changes. Channels were almost completely activated with 10 mM glutamate. Maximum activation was reached within 0.4 ms with greater than or equal to 1 mM glutamate. Channel open probability decayed with a time constant of desensitization of 2 ms with 10 mM glutamate and more rapidly at lower glutamate concentrations. The rate of beginnings of bursts (average number of beginnings of bursts per time bin) decayed even faster but approximately in proportion to the glutamate concentration. The dose-response curve for the channel open probability and for the rate of bursts had a maximum double-logarithmic slope of 5.1 and 4.2, respectively. Channels desensitized completely without opening at very low or slowly rising glutamate concentrations. Desensitization thus originates from a closed channel state. Resensitization was tested by pairs of completely desensitizing glutamate pulses. Sensitivity to the second pulse returned rapidly at pulse intervals between 1 and 2 ms and was almost complete with an interval of 3 ms. Schemes of channel activation by up to five glutamate binding steps, with desensitization by glutamate binding from closed states, are discussed. At high agonist concentrations bursts are predominantly terminated by desensitization. Quantal currents are generated by pulses of greater than 1 mM glutamate, and their decay is determined by the duration of presence of glutamate and possibly by desensitization.

Animals↗
PubMed1990

Characteristics of Na+ channels and Cl- conductance in resealed muscle fibre segments from patients with myotonic dystrophy.

1. Electrical and contractile properties of resealed fibre segments were investigated by a variety of in vitro techniques. The preparations were removed from skeletal muscles of normal subjects and of eight patients with myotonic dystrophy. 2. Several hours after removal, fibre segments from normal subjects and those patients in whom myotonia was the primary symptom had resting membrane potentials of approximately -80 mV. In contrast, fibre segments obtained from patients in whom muscle dystrophy was more expressed were depolarized (-60 to -70 mV). 3. Contractions induced in fibre segments of myotonic muscle which had normal potentials were characterized by slowed relaxation which was due to electrical after-activity. 4. After single stimuli, long-lasting (3-100) runs of action potentials were recorded intracellularly from the myotonic muscle. In some of these fibre segments complex repetitive discharges were observed: multiple sites of locally gated currents were identified. 5. The three-electrode voltage clamp was used to determine the total membrane conductance, gm, and the ion component conductances. All fibres of a particular patient had similar conductances. However, the Cl- conductance varied from patient to patient from normal (74% of gm) to low values (30% of gm). The K+ conductance was normal in all fibres of all patients. 6. The patch-clamp technique was used to record currents through single Na+ channels of the sarcolemma. After treatment of the fibre segments with collagenase gigaohm seals were routinely obtained. The rate of success was greater when using the cell-attached mode than the inside-out mode. 7. Sodium channel currents were elicited by depolarizing voltage steps which produced an initial burst of Na+ channel openings. Up to ten channels were activated simultaneously when the patch was depolarized to potentials more positive than -30 mV. The Na+ channels re-opened very rarely in controls. The macroscopic sodium current, INa, was reconstructed by averaging depolarizing pulses. The time constant of rapid decay of INa reflecting macroscopic inactivation, the onset of INa and the amplitude of INa were voltage dependent. The mean amplitude of the current produced by re-openings was on average only 0.11 +/- 0.04% of the amplitude of the peak current. 8. Late openings of the Na+ channels were frequent in patches on the myotonic fibre segments. The amplitude of the current produced by re-openings was as high as about 0.75 +/- 0.11% of the amplitude of the peak current.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1990

[Anamnesis, inhalation histamine test and 2 stress tests in the diagnosis of exercise-induced asthma].

UNLABELLED: 32 patients (15 men, 17 women) of between 15 and 28 years of age were examined in a prospective study using a staged histamine test and a running test. A special anamnesis was prepared. Following a methodic preliminary examination two methods of the exercise test were compared with one another in 16 patients based on running freely and on the endless tape ergometer-in the form of a two-stage test with measurement of the heart rate. An anamnestic pointer was seen in each of 7 patients with pronounced bronchial hyperreactivity; in moderate hyperreactivity only 1 out of 9 patients had an anamnestic pointer. In 29 of 43 patients, anamnesis and the result of the exercise test agreed. Of 9 patients with exercise induced asthma, 3 had moderate hyperreactivity and 6 a very pronounced one. Free run and two-stage test on the endless tape yield the same result in respect of exercise induced asthma in all 16 patients. CONCLUSIONS: 1. Anamnesis has a good predictive value in respect to exercise induced asthma. 2. This is confirmed: no exercise induced asthma without bronchial hyperreactivity, but the former is not necessarily always a sequence of the latter. 3. It is possible to standardise the running test on the endless tape with definite load and estimation of the efficiency; however, the simple running test in which the patient adjusts the load in accordance with his subjective feeling, is equally valid for arriving at a clinical diagnosis of exercise induced asthma.

Adolescent↗
PubMed1989

Similarities between the effects of lindane (gamma-HCH) and picrotoxin on ligand-gated chloride channels in crayfish muscle membrane.

Effects of lindane (gamma-HCH) and picrotoxin have been tested on excised outside-out patches of crayfish stomach muscle membrane. The pulsed application of 10(-3) M gamma-aminobutyric acid (GABA) elicited an inward CL- current of several 100 pA, which could be reversibly reduced by the simultaneous application of lindane or picrotoxin. The rise time and desensitization time constant of the GABA response did not seem to be affected. The apparent KI was 5 x 10(-6) M for lindane and 15 x 10(-6) M for picrotoxin. Also Cl- currents induced by glutamate or acetylcholine could be reversibly reduced by lindane or picrotoxin. The evaluation of single Cl- channel openings which were elicited by the simultaneous application of glutamate and lindane showed no effect of lindane on the unitary conductance and on the mean open time of these channels. A mechanism for the antagonistic action of lindane and picrotoxin is discussed.

Animals↗