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Biomedical subjects

D Bertrand

Publications and source records attributed to D Bertrand.

At least 163 records · Page 9Linked to original sources

A fast, high precision and inexpensive analog to digital board for PC-AT or compatible.

We describe a simple and inexpensive circuit for analog data acquisition with high speed and high resolution, for use in an IBM PC-AT or compatible. The circuit is suitable for a wide range of applications in electrophysiology. Features of the circuit include multichannel reading, programmable sampling rate, and direct memory access controlled dual buffering for continuous recording. A program written in the C programming language for control of data acquisition is also given.

Computers↗

Single-channel current simulation and recording using a photodiode as current generator.

A device which can generate rectangular currents in the picoampere range is described. The current generator is a photodiode connected to the head stage of a single-channel recording amplifier. The photodiode is activated by a light-emitting diode controlled by a computer or any other current source. The device can transmit signals corresponding to simulated single-channel behaviour. Since the kinetic parameters of the simulation are known, the user can test the data acquisition and analysis system under conditions similar to those prevailing during recording from a biological membrane. This current generator can also be used for the tuning of patch-clamp amplifiers; rectangular currents generated by the photodiode allow the frequency response of the amplifier to be properly adjusted.

Electric Stimulation↗

Expression of substance P and of a Ca2+-activated Cl- current in quail sensory trigeminal neurons.

A chloride current activated by an increase in intracellular calcium concentration is not present in all neurons of the trigeminal ganglion. It is not known whether the trigeminal neurons expressing calcium-activated chloride current belong to a defined class of neurons or whether they could belong to any class of sensory neurons. An answer to this question would be of importance because the physiological role of calcium-activated chloride current in neurons has not yet been completely established, nonetheless it is clear that this current, when activated, would act to modulate neuronal excitability. The goal of this study was to determine whether there was a difference in the expression of calcium-activated chloride current between neurons with and without substance P. The rationale was that the use of this morphological marker, which is present in a substantial fraction of embryonic trigeminal neurons, may give a first estimate of a possible inhomogeneity in the expression of calcium-activated chloride current among different classes of sensory neurons. The study was done on freshly dissociated neurons in order to minimize the influence of the culture conditions on the expression of the current or of substance P. By recording from large samples of neurons in cultures either enriched or depleted in substance P-containing neurons, we found that neurons with substance P expressed calcium-activated chloride current three times less frequently than neurons without substance P. This observation was confirmed by performing the immunocytochemical labelling for substance P immediately after the electrophysiological assessment of the presence or absence of calcium-activated chloride current. This result indicates that calcium-activated chloride current may not be randomly distributed in neurons of a sensory ganglion. It raises the possibility that neurons belonging to certain sensory modalities may need calcium-activated chloride current for their physiological functioning.

Animals↗

Characterization of dissociated monolayer cultures of human spinal cord.

Embryonic human spinal cord cells have been grown in dissociated monolayer cultures for 1 to 7 weeks. Using cell type specific markers, it was possible to show that the cultures contain neurons, astrocytes and fibroblasts. Electrical membrane properties were studied with patch electrodes using the whole cell recording technique. Neurons had short duration action potentials that could be blocked by tetrodotoxin. The membrane currents in these neurons were studied in voltage clamp experiments. Three types of voltage-dependent currents were observed: a sodium current; a potassium current made up of two components, IA and IK; and a calcium current. Both cholinergic and GABAergic neurons are present in the cultures. There is more choline acetyltransferase activity in cultures prepared from the anterior as compared to the posterior part of the spinal cord, suggesting that the cultures contain motoneurons. This tissue culture preparation was developed for the study of amyotrophic lateral sclerosis; we have been unable to detect the presence of any toxic agent from the serum of these patients on the cultured cells. Experiments are in progress to purify the motoneurons using Percoll gradients.

Acetylcholine↗

Electrophysiology of a chick neuronal nicotinic acetylcholine receptor expressed in Xenopus oocytes after cDNA injection.

Brain nicotinic acetylcholine receptors (nAChRs) are made up of protein subunits that differ from those constituting muscle nAChRs. To characterize the physiological properties of one class of avian brain nicotinic receptor, we injected the nuclei of Xenopus oocytes with full-length cDNAs for the ligand binding (alpha 4) and structural (n alpha) subunits. Injected oocytes had large ACh-induced currents in the microampere range that were insensitive to alpha-bungarotoxin, as expected for neuronal nAChRs. We found that these brain nAChRs incorporate at least two alpha 4 subunits and that their functional properties differ from muscle nAChRs in at least two respects: the elementary conductance is considerably smaller (20 pS), and channels in outside out patches stop functioning within a few minutes.

Animals↗

Membrane currents of rat satellite cells attached to intact skeletal muscle fibers.

Muscle satellite cells play an important role in the postnatal growth of skeletal muscle and in the regeneration of damaged muscle during adult life. Little is known about the physiological properties of satellite cells in their dormant state as they lie adjacent to the intact muscle fibers, underneath the basement membrane. Our recent experiments, using patch clamp techniques, indicate that no tight electrical coupling is present between satellite cells and the muscle fiber dissociated from rat flexor digitorum brevis. Satellite cells possess sodium channels with low sensitivity to tetrodotoxin and at a much lower density than muscle. In addition, satellite cells are insensitive to acetylcholine (ACh) for at least 24 hr after having been removed from the animal, even when detached from their muscle fiber. However, we could measure ACh-evoked currents from satellite cells 48-72 hr in culture, indicating that ACh sensitivity develops with time.

Acetylcholine↗

[Propofol versus propanidid for the conduction of suspension laryngoscopy].

Forty patients who where to undergo suspension laryngoscopy were randomly assigned to two groups, the first receiving 1 microgram . kg-1 fentanyl and a bolus of 2.5 mg . kg-1 propofol followed by 5 to 10 mg . kg-1 . h-1 propofol infusion, and the second 1 microgram . kg-1 fentanyl and 0.2 mg . kg-1 flunitrazepam with 8 mg . kg-1 propanidid in a bolus followed by 40 to 50 mg . kg-1 propanidid infusion. The following parameters were studied: length of apnoea, quality of anaesthesia, the time between stopping giving the anaesthetic and the moment when the patient opens the eyes, gives his name and date of birth, the heart rate, the systolic, diastolic and mean blood pressures, blood gases, before induction, during suspension and at stopping the infusion. Anaesthetic quality was the same for both protocols, and the variations of the haemodynamic parameters were very similar for both groups. Apnoea lasted twice as long with propofol as with the flunitrazepam-propanidid association (p less than 0.001), whereas recovery was twice as quick (p less than 0.001). This seemed to confirm that propofol is better indicated for this type of surgery than the previously used flunitrazepam-propanidid association.

Adult↗

[Effects of isoflurane on bleeding in microsurgery of the middle ear].

40 patients undergoing middle ear microsurgery were anaesthetized with a narconeuroleptanalgesia (droperidol, phenoperidine, thiopentone) associated with a randomly selected halogenated volatile anaesthetic, either halothane or isoflurane at a concentration of 1 MAC. Clinical and haemodynamic tolerance was perfect. Respiratory and haemodynamic parameters were not significantly different in either series at the different moments that were common to all the operations. There was less bleeding in the isoflurane group, and it was stable (p less than 0.01). Isoflurane would seem to have the double advantage of maintaining anaesthesia and producing a bloodless surgical field, so avoiding the use of added hypotensive agents.

Adult↗

Calcium-activated chloride current in cultured sensory and parasympathetic quail neurones.

1. Sensory (trigeminal and dorsal root) and autonomic (ciliary) ganglia from embryonic quail were dissociated and the neurones were grown in tissue culture. 2. Intracellular recordings were made in voltage clamp using patch electrodes and the whole-cell recording technique. In order to investigate a calcium-activated chloride current, the sodium and potassium currents were blocked. 3. Depolarizing voltage steps from a holding potential of -100 mV to a test potential of +20 mV triggered an early inward and a delayed outward current. The latter persisted as a long-lasting inward tail current when the membrane was depolarized to -100 mV. 4. These currents were all blocked by extracellular cobalt suggesting that they were calcium dependent. During a test depolarization to +20 mV, in the presence of intracellular EGTA (20 mM), the inward current persisted but the outward current was suppressed. EGTA (20 mM) also suppressed the long-lasting inward tail current at -100 mV. This suggested the presence of a calcium-activated current. 5. The reversal potential of the calcium-activated current was near the equilibrium potential for chloride ions and was shifted as predicted by the Nernst equation when the extracellular chloride concentration was changed. 6. The calcium-activated current was partially blocked by adding 4-acetamido-4'-isothiocyanatostilbene-disulphonic acid (SITS) at a concentration of 1 mM to the external superfusion medium. This effect of a compound known to interfere with chloride channels together with the results of point (5) suggested the existence of a calcium-activated chloride current (ICl(Ca)). 7. ICl(Ca) could be activated by transient and sustained components of the calcium current present in the cultured neurones. 8. ICl(Ca) was present in 80% of the sensory neurones but only in 10% of the parasympathetic neurones.

Animals↗

Calcitonin and vasopressin affect epithelial properties in a renal cell line.

We have used a stable clonal variant (D + Sc), isolated from the LLC-PK1 pig kidney-derived cell line and selected for its extensive capacity to form domes, in order to study the hormonal modulation of epithelial permeability in culture. Calcitonin, vasopressin, and other agents that raise intracellular adenosine 3',5'-cyclic monophosphate levels caused a rapid and dramatic decrease in the size and number of domes. This effect was independent of RNA and protein synthesis, and thus appeared unrelated to the production of urokinase, a proteinase synthesized by the cells in response to these agents. Calcitonin caused a decrease in transepithelial electrical resistance, suggesting that the effect of the hormone on domes was due to an increase in the permeability of a paracellular pathway. Thus, in addition to the wellknown effects of vasopressin on collecting duct permeability, part of the in vivo effect(s) of calcitonin and vasopressin on the renal tubule might also involve alterations of epithelial permeability related to those described here.

Animals↗

DATAC: a multipurpose biological data analysis program based on a mathematical interpreter.

The use of a mathematical command interpreter combined with the structural facility of the C-language allowed us to design a data treatment program having considerable flexibility and being able to handle any types of data (electrophysiological, biochemical and theoretical data). Ensembles of data are treated by the interpreter as if they were simple variables so that an elaborate computation can be performed on the spot by simply writing the appropriate equation on the terminal. These facilities combined with the ability of editing macrocommands at run time provide the user with data treatment possibilities that extend far beyond the possibilities actually implemented in the program. The originality of this program is that the user can easily implement the commands he most often needs, writing them in a language that most scientists will know, algebra.

Biometry↗

Evoked mechanical responses of isolated cochlear outer hair cells.

Intracellular current administration evokes rapid, graded, and bidirectional mechanical responses of isolated outer hair cells from the mammalian inner ear. The cells become shorter in response to depolarizing and longer in response to hyperpolarizing currents in the synaptic end of the cell. The cells respond with either an increase or decrease in length to transcellular alternating current stimulation. The direction of the movement with transcellular stimuli appears to be frequency dependent. Iontophoretic application of acetylcholine to the synaptic end of the cell decreases its length. The microarchitecture of the organ of Corti permits length changes of outer hair cells in a manner that could significantly influence the mechanics of the cochlear partition and thereby contribute to the exquisite sensitivity of mammalian hearing.

Acetylcholine↗

Voltage-dependent potassium currents in developing neurones from quail mesencephalic neural crest.

Neurones in explants cultured from quail mesencephalic neural crest were studied at different stages of their development using the voltage-clamp technique. A voltage-dependent outward current activated by membrane depolarization was identified as a potassium current by the sensitivity of its reversal potential to extracellular potassium. The voltage-dependent potassium current is made up of two components which differ in their sensitivity to 4-aminopyridine (4-AP) and tetraethylammonium (TEA). The component most sensitive to 4-AP has fast activation kinetics and inactivates quickly at sustained depolarized voltages. By analogy with a current described in other preparations, this current was called IA. The component most sensitive to TEA has slower activation kinetics and inactivates more slowly at sustained depolarized voltages. This current was called IK. IA and IK were already present in neurones cultured for 24 h. The ratio between the peak of IK and that of IA increased significantly between 24 h and 4 days in culture. This means that the two components of the voltage-dependent potassium current follow a different time course during development.

4-Aminopyridine↗

Systematic changes in perceptual reactance induced by physical fitness training.

The effect of life-change events on perceptual augmentation-reduction was studied in 72 subjects (40 men, 32 women). In three experiments the kinesthetic figural aftereffect was measured prior to and after either a physical fitness program, a course in yoga, or training in Transcendental Meditation. Each program lasted a minimum of 8 wk. Subjects completing fitness training, all of whom were initially classified as augmenters, became reducers by the end of their program. Subjects enrolled in yoga and meditation courses remained relatively stable in their perceptual tendencies.

Adolescent↗

Human spinal cord neurons in dissociated monolayer cultures: morphological, biochemical, and electrophysiological properties.

The preparation of dissociated monolayer cultures from embryonic human spinal cord is described. Optimal survival was achieved with embryonic tissue between the eighth and ninth week. The neurons survive for as long as 7 weeks in culture and they grow in a standard tissue culture medium which contains 13% decomplemented human serum. The neurons have been identified by indirect immunofluorescence techniques using antibodies to tetanus toxin and neurofilament protein. Our biochemical studies demonstrate the presence of cholinergic and GABAergic neurons. Cholinergic neurons develop in culture and are more numerous in the cultures prepared from the anterior part of the spinal cord as compared to those from the posterior part. Therefore, it is possible that a large part of the cholinergic neurons derive from the motoneuron pool. Electrical membrane properties were studied with patch electrodes using the whole cell recording technique. Neurons had short duration action potentials that could be blocked by tetrodotoxin (TTX). Voltage clamp experiments combined with the use of pharmacological blocking agents revealed the presence of several voltage- and time-dependent currents: a sodium current sensitive to TTX, a potassium current made up of two components, sensitive to tetraethylammonium and 4-aminopyridine, and a calcium current sensitive to cobalt. From a biochemical and electrophysiological point of view the properties of human spinal cord neurons in culture closely resemble the properties of spinal cord neurons from other species.

Acetylcholine↗