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Biomedical subjects

I Parker

Publications and source records attributed to I Parker.

At least 127 records · Page 7Linked to original sources
PubMed1981

Calcium transients in normal and denervated slow muscle fibres of the frog.

1. Intracellular changes in free Ca2+ concentration were recorded from slow muscle fibres in the pyriformis muscle of Rana temporaria, using the dye arsenazo III. Fibres were voltage clamped, and arsenazo signals were recorded in response to depolarizing pulses. 2. The size of the arsenazo response to depolarizing pulses of 100 msec duration was a sigmoid function of membrane potential over the range -45 to 0 mV, and remained constant with further depolarizations up to +100 mV. 3. The peak size of the arsenazo signal to supramaximal depolarizations increased with increasing pulse length. The initial rising phase during a pulse was much slower than in twitch fibres, and this phase was followed by an even slower rise. Following short pulses the decay of the response was exponential, with a time constant of about 1.4 sec, while after long pulses the decline became much slower. 4. Decreasing free Ca2+ concentration in the bathing medium to very low levels, using EGTA , did not affect the responses to short (100 msec) depolarizations. 5. Slow fibres bathed in Ringer's solution containing 12 mM-Ca2+ showed a well maintained arsenazo response to supramaximal depolarizations lasting over 1 min. Reduction of external Ca2+ to 1.8 and (nominally) 0 mM caused the response to become progressively more transient. 6. After denervation, slow fibres developed action potentials, but non of the parameters of the arsenazo response was significantly changed. During the early phase of reinnervation by a mixed nerve, when fast conduction axons begin to innervate slow fibres, the ability to give a maintained response during long depolarizations was reduced. 7. It is concluded that intracellular Ca2+ transients in slow muscle fibres are probably generated by a similar mechanism as in twitch fibres and entry of external Ca2+ is not an appreciable factor. The slow time course of the transients may be important in determining the time courses of tension development and relaxation.

Animals↗
PubMed1981

Single glutamate-activated channels recorded from locust muscle fibres with perfused patch-clamp electrodes.

1. Glutamate-activated single channels have been examined with conventional and internally perfused patch-clamp electrodes applied to the extrajunctional membrane of locust muscle fibres which were usually treated with concanavalin A to reduce desensitization. Channels opened by glutamate and other agonists have been compared.2. Recording patches were selected where there appeared to be only one active channel under the pipette. The conductance for single glutamate-activated channels was 150 pS and was not markedly dependent on clamp potential. The lifetimes of the channels were usually exponentially distributed with a mean of tau(glutamate) = 2.3 +/- 0.12 msec, T = 23 degrees C, V(m) = -60 mV.3. Channels opened by fluoroglutamate had a mean lifetime of tau(fluoroglutamate) = 1.4 +/- 0.1 msec; channels opened by quisqualate had a mean lifetime of tau(quisqualate) = 6.4 +/- 1.0 msec. The conductances of channels opened by fluoroglutamate, quisqualate and glutamate were not significantly different.4. The behaviour of individual receptor-channel complexes has been examined at various concentrations of glutamate. Drug solutions were applied through an internal perfusion pipette which allowed exchange of the solution in the patch-electrode tip within 10 sec. The distribution of channel closed times could be fitted with a single exponential. Channel lifetime was not markedly dependent on glutamate concentration (30-600 mum) whereas the channel closed time decreased with increasing glutamate concentration.5. The reciprocal of channel closed time vs. glutamate concentration had a slope value of 1.85 on logarithmic co-ordinates. The approximately second power dependence of net forward reaction rate on glutamate concentration suggests that at least two glutamate molecules activate a single receptor-channel complex.6. The apparent dissociation constant for the glutamate-receptor complex is large, being about 300-500 muM. If the receptors have an equally low affinity for neurally released transmitter, then only a small amount of the transmitter packet is expected to bind to receptors. Quisqualate and glutamate have similar receptor affinities whereas receptor affinity for fluoroglutamate is smaller.

Animals↗
PubMed1980

Procollagen mRNA metabolism during the fibroblast cell cycle and its synthesis in transformed cells.

Procollagen mRNA was isolated from mouse embryos and used for the synthesis of a highly labelled cDNA probe complementary to collagen mRNA. This probe was used for the investigation of procollagen mRNA metabolism during the cell cycle of 3T6 mouse embryo fibroblasts in culture. Titration hybridization experiments revealed that procollagen mRNA was present throughout the cell cycle following stumulation of confluent monolayers. Procollagen mRNA levels of sparse cultures appeared similar to those of unstimulated monolayers. The fluctuating levels of collagen synthesis during the cell cycle can be ascribed to changes in the amount of collagen mRNA present. In mouse sarcoma virus transformed 3T3 cells only 20--30% of the amount of procollagen mRNA in 3T3 cells is present indicating that the decline in collagen synthesis is due to mRNA availability.

Animals↗
PubMed1980

The synthesis and isolation of DNA complementary to histone mRNA from mouse embryos.

A 9S poly(A)-RNA preparation isolated from mouse embryos was shown to stimulate the synthesis of histones in an ascites cell free extract. This RNA preparation was used for the synthesis of a highly labelled cDNA probe complementary to histone mRNA. Hybridisation of this cDNA probe to rRNA showed that 52% of the cDNA consisted of sequences complementary to rRNA. The histone mRNA specific cDNA was purified by hybridising the impure cDNA to rRNA followed by removal of the single-stranded histone cDNA by hydroxylapatite chromatography.

Animals↗
PubMed1980

Histone mRNA metabolism during the mouse fibroblast cell cycle.

The synthesis of histone mRNA in the nucleus and the eventual appearance of histone mRNA in the cytoplasm of 3T6 Swiss mouse embryo fibroblasts was investigated with a highly labelled cDNA probe complementary to histone mRNA. Very low levels of histone mRNA were found in G1 phase nuclei, with even lower levels in the cytoplasm. At the onset of DNA synthesis, there is a rapid increase in histone mRNA synthesis, with a concomitant increase of histone mRNA in the cytoplasm followed by its decline in both nucleus and cytoplasm on entering the G2 phase of the cell cycle. The decline in the nucleus is much more rapid than in the cytoplasm.

Animals↗
PubMed1980

Transmitter induced calcium entry across the post-synaptic membrane at frog end-plates measured using arsenazo III.

1. The Ca2+ influx occurring across the post-synaptic membrane during transmitter action was studied at the frog neuromuscular junction, using the Ca sensitive dye arsenazo III to monitor the resulting changes in free myoplasmic Ca2+ concentration. 2. Calibration experiments showed a linear relationship between the amount of Ca2+ injected by ionophoresis into a muscle fibre, and the peak size of the arsenazo light absorbance record. 3. Ionophoretic application of acetylcholine (ACh) to voltage clamped end-plates gave rise to an arsenazo signal. The size of this response varied with the Ca2+ concentration in the bathing solution. 4. The arsenazo light response increased in size steeply, and non-linearly, with hyperpolarization of the end-plate membrane, even when the end-plate current increased approximately linearly with hyperpolarization. The voltage dependence of the light response could be fitted well by an exponential with a voltage constant of 28 mV. Changes in Ca2+ concentration of the bathing medium had little effect on this relationship. 5. At end-plates bathed in isotonic CaCl2 solution the voltage dependence of both the arsenazo light response, and the end-plate current showed a closely similar, non-linear relationship. 6. Addition of 12 mM-Co2+ to a bathing solution initially containing 12 mM-Ca2+ substantially reduced the size of the arsenazo light response, and the voltage dependence of this response became more linear. 7. Arsenazo light responses were also recorded in response to transmitter release evoked by nerve stimulation. The size of the nerve evoked light response showed a non-linear voltage dependence, whilst the end-plate current was a linear function of membrane potential.

Acetylcholine↗
PubMed1980

Effects of strontium ions on end-plate channel properties.

1. Changes in end-plate channel properties resulting from substitution of Sr2+ for Ca2+ in the Ringer solution have been analysed at the voltage clamped frog end-plate, by recording m.e.p.c.s and ACh induced noise variance. 2. In 2 mM-Sr2+--Ringer the peak size of m.e.p.c.s showed a very small increase, and the time constant of the decay phase (tau m.e.p.c.), at any given voltage, was increased by a factor of about two compared to control Ringer. The voltage dependence of tau m.e.p.c. was the same in both solutions. 3. Addition of increasing amounts of CaCl2 to 2 mM-Sr2+--Ringer produced a progressive shortening of tau m.e.p.c., with no change in voltage dependence. 4. Estimates of single channel properties from noise analysis showed that the elementary conductance appeared to be slightly increased in 2 mM-Sr2+--Ringer, whilst the mean channel life-time was prolonged by a factor of about two. These changes in single channel properties are sufficient to account for the observed changes in m.e.p.c.s. 5. Following inhibition of cholinesterase activity by neostigmine, similar effects on m.e.p.c.s and single channel properties were still observed on changing to 2 mM-Sr2+--Ringer. The shapes of m.e.p.c.s in Sr2+ + neostigmine Ringer were often altered, and showed flat 'plateaus'. 6. The observed effects of Sr2+--Ringer on channel life-time cannot be explained on the basis of changes in surface charge density on the membrane, and suggest that divalent cations have an additional, and more direct, influence on receptor channel properties.

Acetylcholine↗
PubMed1979

Globin gene expression in MSV-transformed fibroblasts.

The activation of globin gene expression on viral transformation of 3T3 cells was investigated. Globin mRNA was determined using a radioactive complementary DNA probe. No difference was found between 3T3 and transformed 3T3 cells. There does not therefore appear to be a random activation of extensive regions of the cellular genome.

Animals↗