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Biomedical subjects

R M Pitman

Publications and source records attributed to R M Pitman.

At least 37 records · Page 2Linked to original sources
PubMed1989

Post-embryonic development of rectifying electrical synapses in the crayfish: ultrastructure.

The post-embryonic development of the rectifying Giant Fibre-Motor Giant (GF-MoG) synapse and the Giant Fibre-Segmental Giant (GF-SG) synapse has been investigated using electron-microscopy. In adults, the MoG and SG neurons make contact with the GFs by sending narrow 'finger-like' processes through the glial and connective tissue sheath surrounding each GF. The junctional region is characterized by closely apposed membranes (approximately 4 nm separation) traversed by regularly spaced connections, and large (60-80 nm) spherical vesicles in the presynaptic fibre. In newly hatched crayfish junctional contact is made over extensive areas of flat membrane apposition, due to the absence of a thick connective sheath around the giant fibres. Initially the junctional region is dominated by contacts which are morphologically indistinguishable from chemical synapses, i.e. 1. The apposed membranes are separated by a cleft of approximately 20-30 nm (an order of magnitude larger than the cleft distance at electrotonic synapses). 2. There is pre- and post-synaptic thickening of the junctional membranes with a dense cytoplasmic material. 3. Small (25-40 nm) pleomorphic vesicles are found on the presynaptic side of the junction, commonly in association with a dense presynaptic bar. Regions of junctional contact displaying the adult electronic-type morphology first appear at approximately one week post-hatching. At this age they are limited in distribution and occupy a central position in the area of contact surrounded by a broad 'chemical-like' annulus. During subsequent development these sites with electrotonic-type morphology grow in relative size, so that the 'chemical-like' sites become compressed towards the edges of the regions of contact. The adult type of morphology, in which the 'chemical-like' regions are vestigial, is achieved approximately two months after hatching.

Animals↗
PubMed1989

Ionic currents in the soma of an identified cockroach motoneurone recorded under voltage-clamp.

1. Membrane currents have been recorded from the soma of a bifunctional basalar/coxal depressor motoneurone in the metathoracic ganglion of the cockroach (Periplaneta americana) using a two-electrode voltage-clamp technique. 2. This motoneurone cell body is normally inexcitable when studied under current-clamp. Appropriate depolarizing command steps evoke rapid transient outward currents and late outward currents. 3. Late outward currents are dominated by a Ca-dependent component that confers an N-shaped I-V relationship on the neurone. 4. The Ca-dependent outward current is suppressed by Cd2+ (1 mM), Mn2+ (5 mM) or verapamil (50 microM). 5. Externally applied tetraethylammonium ions (TEA+) (25 mM) block the Ca-dependent current, but also appear to suppress a component of the late outward current that is independent of Ca2+. 6. Aminopyridines cause only minor suppression of late outward currents, but shift the peak in the N-shaped I-V relationship to more negative potentials. 7. The reversal potential of tail currents recorded following pre-pulses to +50 mV were dependent upon the pre-pulse duration; increasing the duration from 10 to 50 msec caused a +17 mV shift in tail current reversal potential. 8. A five-fold increase in the K+ concentration of the solution bathing the preparation only produced small and inconsistent changes in the reversal potential of tail currents. 9. Five-fold reduction in external Cl- caused no change. 10. The dependence of tail current reversal potential upon pre-pulse duration and the limited effect of alterations in the composition of the bathing solution are discussed in the context of restricted ion movements near the external surface of the cell membrane.

Aminopyridines↗
PubMed1987

Conduction block silences parts of a chemical synapse in the leech central nervous system.

1. The pressure (P) sensory neurones innervating the ventral skin of the medicinal leech have receptive fields comprising a central region of skin innervated by two thicker axons and two neighbouring regions innervated by two thinner axons. Impulses originating in the thinner axons may fail to propagate through the central ganglion, apparently blocked at the branch point of large and small axons. 2. The P neurone excites the longitudinal (L) motoneurone, and blocked impulses originating in the anterior fine axon produce e.p.s.p.s that are less than one-half normal amplitude. Blocked impulses in the posterior fine axon are typically ineffective. 3. The branches of P and L neurones, marked with intracellularly injected horseradish peroxidase or with Lucifer Yellow, make synaptic contact at up to sixty-six sites within the neuropile. Of P neurone branches emerging from two fine axons, those from the posterior axon make fewer contacts, usually one or two at most, while branches from the anterior axon represent no more than half the total contacts. From cell to cell there is some variation in the total number of contacts, the distribution of branches, and the strength of transmission. 4. The locations of contacts measured morphologically correlate well with their distributions as predicted from reductions in e.p.s.p. amplitude during conduction block.

Animals↗
PubMed1984

The versatile synapse.

'Typically' chemical synaptic transmission takes place when an influx of calcium ions during a presynaptic nerve impulse triggers exocytosis of neurotransmitter substance from synaptic vesicles. The neurotransmitter diffuses across the synaptic cleft and occupies receptors embedded in the subsynaptic membrane. This interaction (directly or via a second messenger) operates characteristic ion channels and produces an increase in the postsynaptic membrane permeability to particular ions. Depending on the ionic species to which the postsynaptic membrane becomes more permeable, the physiological response will be an excitatory or an inhibitory postsynaptic potential. The action of neurotransmitters may be terminated either by enzymic inactivation or by cellular uptake mechanisms. Over the last decade it has become clear that a neurotransmitter substance may exert a number of different actions on a single postsynaptic neurone. These may involve opening or closure of either voltage-independent or voltage-dependent ion channels. It is also possible that in some instances transmitters may act on neuronal biochemical systems to modify the physiology of postsynaptic cells without directly altering their electrical characteristics. Analysis of the postsynaptic actions of neurotransmitter substances has become further complicated by the increasing body of evidence which indicates that more than one transmitter substance (one of which may be a peptide) can be released from a single presynaptic neurone. The significance of such dual transmitter systems has yet to be fully elucidated. The efficacy of transmission across many synapses may be modified by either presynaptic or postsynaptic mechanisms; both transmitter release and postsynaptic responsiveness may depend on the recent history of a single synapse, on synaptic inputs from other neurones or on circulating neuroactive substances.

Animals↗
PubMed1982

The effects of axotomy upon the extrasynaptic acetylcholine sensitivity of an identified motoneurone in the cockroach Periplaneta americana.

The effects of axotomy on the sensitivity of the fast coxal depressor motoneurone (Df) of the cockroach (Periplaneta americana) to applied acetylcholine (ACh) and carbamylcholine (CCh) have been investigated. ACh and CCh applied to the soma membrane either by bath perfusion or by ionophoresis caused depolarization; repeated application of large doses of these agonists resulted in a relatively rapid desensitization and depression of the response. Axotomy performed 3-10 days before recordings were made caused an approximately threefold increase in the sensitivity to ACh but had no affect upon the sensitivity to CCh. The resting potential, input resistance and membrane time-constant remained within the normal range. In addition there was no change in the rise-times of the responses or of the ACh reversal potential, or in the apparent number of ACh molecules needed to combine with individual cholinoceptors to produce a response. The anticholinesterases physostigmine and neostigmine potentiated the ACh response at relatively low concentrations (10(-7) M-10(-6) M). This potentiation was significantly greater in the normal cells than in the axotomized cells. It is therefore concluded that the increase in ACh sensitivity of this motoneurone results at least partly from a fall in the activity of cholinesterase in the region of the applied drug.

Acetylcholine↗
PubMed1979

Intracellular citrate or externaly applied tetraethylammonium ions produce calcium-dependent action potentials in an insect motoneurone cell body.

1. Electrophysiological observations have been made upon the cell body of an identified motoneurone of the cockroach, Periplaneta americana. Normal responses were compared with those observed after intracellular injection of citrate anions or when the preparation was bathed in solutions containing tetraethylammonium ions (TEA+).2. Normally when depolarized, the motoneurone soma gave a series of damped oscillations; the amplitude of these responses increased with increase in the applied current.3. After citrate ions had been injected into the neurone soma, all-or-none action potentials were evoked by depolarization; such responses appeared about 5-10 min after the onset of citrate injection. Injection of EGTA produced similar effects. Citrate and EGTA probably produce their effect through a reduction in the intracellular free calcium concentration.4. When preparations were bathed in saline solution containing 50 mM-TEA+, soma depolarization produced prolonged all-or-none action potentials (up to approximately 100 msec duration).5. The action potentials produced by citrate injection or externally applied TEA+ appeared to have a similar ionic mechanism; they were not depressed by sodium-free solutions or by tetrodotoxin (4 x 10(-6)M) but were reversibly blocked in saline solution containing 40 mM-manganous chloride.6. The overshoot amplitude of action potentials recorded after injection of citrate anions or in solutions containing TEA+ showed a 22.5 mV change for a ten-fold change in the external calcium concentration.7. Both intracellular citrate and external TEA+ caused a significant increase in the input resistance and membrane time constant of the motoneurone.8. It is concluded that action potentials generated under various experimental conditions in the soma of this insect motoneurone map have differing ionic mechanisms.

Action Potentials↗
PubMed1975

The ionic dependence of action potentials induced by colchicine in an insect motoneurone cell body.

1. The ionic requirements of the action potential recorded in the cell of an identified cockroach (Periplaneta americana) motoneurone following pre-treatment of the animal with colchicine have been studied. 2. Small cubes of gelatin containing 1% colchicine were implanted into one metathoracic leg near to the nerve trunk containing the axon of the identified motoneurone. 3. Electrophysiological experiments were performed 4--10 days after this treatment, when action potentials which frequently overshoot zero potential can be recorded from the cell body. Such action potentials are not normally seen in untreated animals. 4. Sodium-free solution reversibly abolished the action potential within 5 min. 5. Tetrodotoxin (10(-8)M) reversibly depressed the action potential. It was totally abolished by 10(-7)M tetrodotoxin, but this effect was not reversible. 6. Saline solution containing 40 mM manganous chloride either had no effect on the action potential amplitude, or caused a slight increase. It also caused prolongation of the falling phase and loss of the after-hyperpolarization. These effects were all reversible. 7. It is concluded that sodium carries a major proportion of the inward current of the action potential in this neurone. Some calcium probably enters also, and may, at least in part, be responsible for triggering the delayed rise in potassium conductance during the action potential.

Action Potentials↗
PubMed1972

Electrical responses of insect central neurons: augmentation by nerve section or colchicine.

Intracellular recording from the somata of central motor neurons in the cockroach Periplaneta americana normally shows little or no electrical response evoked by soma depolarization or by antidromic stimulation. Within 4 days after either cutting the axon or administration of colchicine, large action potentials can regularly be recorded from cell bodies of metathoracic motor neurons. Each experimental procedure evokes formation of a dense, perinuclear ribonucleic acid ring in the soma of neurons showing augmented electrical responses.

Action Potentials↗
PubMed1972

Branching of central neurons: intracellular cobalt injection for light and electron microscopy.

Cobalt chloride can be injected into an identified nerve cell body in an insect ganglion and reacted with ammonium sulfide to stain the soma and its branches with a black precipitate. The stained cell body and its branches throughout the neuropil are visible in both the light and electron microscope. In whole mount preparations, the resolution of neurites within the neuropil is of a quality that permits the comparison of branching patterns between cells and during various functional states.

Action Potentials↗