Biomedical subjects

R L Milton

Publications and source records attributed to R L Milton.

6 recordsLinked to original sources

PubMed1992

Fast and slow twitch skeletal muscle fibres differ in their distribution of Na channels near the endplate.

Sodium channel distributions were measured in fast and slow twitch rodent skeletal muscle fibres using the loose patch voltage clamp technique. Large differences were found between these fibre types with respect to Na channel density in the perijunctional region. Fast twitch fibres exhibited a large increase in Na channel density near the endplate, while slow twitch fibres did not.

Animals↗

PubMed1990

How do patch clamp seals form? A lipid bleb model.

Individual ion channels are electrically isolated and studied in living cells with the tight patch voltage clamp method. Channels are identified, categorized, and sometimes named on the basis of the biophysical properties obtained with this method. Although it is usually presumed that these recordings are from native, undisturbed membrane, the physical basis of this technique is not well established. Observations that lipid blebs readily form when suction is applied to patch clamp electrodes suggest that many single channel recordings are from ion channels in these blebs.

Animals↗

PubMed1990

Na current in membrane blebs: implications for channel mobility and patch clamp recording.

When suction was applied to loose patch clamp pipettes while recording from enzymatically dissociated muscle fibers, large membrane blebs formed within the pipettes. We initiated a study of these suction-induced blebs because ion channels in the blebs would complicate or possibly invalidate loose patch voltage clamp measurements of membrane current density. The low lateral mobility (Stühmer and Almers, 1982) and steep gradients of Na channels at the end-plate and tendon (Caldwell et al., 1986) imply tight binding of Na channels to cytoskeletal elements and led us to expect few, if any, Na channels in the blebs. Bleb formation produced an increase in membrane capacitance, as expected from the increase in membrane area. Bleb formation also increased the Na current, indicating that the blebs contained Na channels. Assuming that the increased capacitance and Na current were due to lipid and Na channels moving from membrane outside the pipette, ejection of the bleb from the pipette was expected to bring the capacitance and Na current back to their original values. Capacitance did return to its original value, but Na current was lower than expected. The decrease in Na current is explained by Na channels moving from the patch membrane into the bleb. Normalization of bleb and patch Na current to their respective capacitances revealed that bleb membrane had a Na channel density almost 50% that of normal surface membrane. Thus, bleb membrane is neither devoid of proteins nor truly representative of the normal surface membrane from which it arose. It is enriched in membrane lipids and is relatively protein poor. Two conclusions can be drawn.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Sodium channel distribution in normal and denervated rodent and snake skeletal muscle.

1. Sodium channel current density was measured using the loose-patch voltage clamp technique. Innervated rat, mouse and snake muscle had the highest density of Na+ channels in the end-plate region. These high Na+ channel densities were maintained in denervated muscle. 2. Perijunctional membrane had a Na+ current density 5- to 10-fold greater than the density several hundred micrometres from the end-plate. In all muscles this concentration of channels near the end-plate persisted following denervation. 3. At the tendon Na+ current density fell to low values (approximately 1 mA/cm2). The decrease in density began about 300-500 microns from the tendon. This pattern was found in all snake twitch fibres and fast-twitch (EDL) rat and mouse muscle fibres. This reduction in channel density near the tendon was not affected by denervation. 4. Sodium channels in all regions of innervated rat and snake muscle fibres were highly sensitive to tetrodotoxin (TTX). Sodium channels in snake muscle remained sensitive to TTX after denervation. Sodium channels that are relatively resistant to TTX appeared in rat muscle after denervation. TTX-resistant channels were even more concentrated near the end-plate than were TTX-sensitive channels in innervated muscle. At the tendon TTX-resistant Na+ channel density decreased. 5. We conclude that although the nerve presumably directs the localization of Na+ channels during development, the ability to maintain this distribution and to control the distribution of newly appearing channels persists long after the nerve has been removed.

Action Potentials↗

PubMed1987

Suppression of charge movement in frog skeletal muscle by D600.

Charge movements in intact frog twitch fibres were studied using a three-microelectrode voltage-clamp technique. When high potassium solution was applied transiently to the muscle fibres at low temperature in the presence of D600, the fibres became paralysed and, concomitantly, charge movement disappeared. The amount of charge suppressed by the paralysis treatment was about 70-100% of that in control experiments. This paralysing action of D600 is not shared by its derivative D890. The requirement of conditioning potassium contracture is, most likely, related to prolonged membrane depolarization, as voltage-clamped depolarization to 0 mV lasting tens of seconds also suppressed charge movement. When paralysed fibres were warmed, the main charge component (Q beta) was reprimed. By contrast, the hump charge component (Q gamma) was only reprimed in some of the fibres. Other than by warming, as paralysed fibre could be revived by stimulating it with large suprathreshold pulses but not by voltage-clamped hyperpolarization to -160 mV for tens of seconds. The paralysing action of D600 described here appears to be unrelated to its ability in blocking Ca2+ channels.

Animals↗

PubMed1985

Electrical properties of the myotendon region of frog twitch muscle fibers measured in the frequency domain.

The electrical properties of the end of a muscle fiber were determined using three microelectrodes, one passing sinusoidal current, the other two recording the resulting voltages. An electrical model was constructed from the morphology of the fiber, including the resistance of the extracellular space between cells; the parameters of this model were determined by fitting the model to the observed voltage responses. Our results, analyzed directly or by curve fits, show that the end of muscle fibers contains a large capacitance resulting from the extensive membrane folds at the myotendon junction. Analysis and simulations show that the extra capacitance at the myotendon junction has substantial effects on measurements of linear properties, in particular on estimates of the capacitance of the membranes. There is little qualitative effect on classical measurements of nonlinear charge movement (provided they were made with one set of electrode locations) if the linear components have been subtracted. Quantitative estimates of nonlinear charge movement and ionic currents are significantly affected, however, because these estimates are customarily normalized with respect to the linear capacitance.

Animals↗