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I M Cooke

Publications and source records attributed to I M Cooke.

At least 37 records · Page 2Linked to original sources
PubMed1990

Characterization of Ca current underlying burst formation in lobster cardiac ganglion motorneurons.

1. The anterior motorneurons of the cardiac ganglion of Homarus americanus were ligated less than 300 microns from the soma. This removes impulse-generating membrane and sites of synaptic input while preserving the ability of the soma to generate the burst-forming potentials termed "driver potentials" regenerative, slow (250-ms duration) depolarizations (to -20 mV) in response to brief, depolarizing stimuli. At stimulus intervals corresponding to rates of bursting observed in spontaneously active, intact ganglia (0.3-1.2/s), driver potential amplitude increases with increasing stimulus interval. 2. A two-electrode voltage clamp was used to characterize inward current observable from the ligated neurons in tetrodotoxin (TTX)-tetraethylammonium (TEA)-containing salines. The amplitude of inward current shows a hyperbolic relation to [Ca]o that is well fitted by a form of the Michaelis-Menten equation. Inward current is maintained but not augmented when Ca2+ is replaced by Ba2+ or Sr2+. It is concluded that the inward current, to be referred to as ICa, is mediated by voltage-dependent Ca channels. 3. Contamination of ICa by early outward current (IA) was evaluated by addition of 4-aminopyridine (4-AP, 4 mM). In the presence of 4-AP, the net inward current is increased and the potential at which maximum ICa occurs is shifted 10 mV more positive. 4. Subtraction of outward currents recorded in Mn2(+)-containing saline from overall currents in the absence of Mn2+ provided another means to separate inward from outward current. I-V curves from such "Mn-subtracted" records show ICa approaches a saturating value for steps to -5 mV and more depolarized. The time to peak ICa is voltage dependent. The largest inward currents (up to 240 nA) and minimal time to peak (4 ms) are observed for steps from holding potentials of -50 to -60 mV. 5. Decline of ICa during depolarized steps observed in Mn-subtracted records represents inactivation rather than development of competing outward current. Inactivation is slow and incomplete; the rate and fractional amount of inactivation are not directly voltage dependent. Nonsubtracted responses to 500-ms depolarizations to potentials evoking little outward current show that an initial rapid decline of ICa (tau approximately 40 ms) is followed at approximately 80 ms by a slower phase of decline (tau approximately 180 ms). With repetitive clamps, the early phase proved labile.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Studies on the crustacean cardiac ganglion.

1. An overview of studies on the decapod crustacean cardiac ganglion is given emphasizing contributions to questions of general interest in cellular neurophysiology. 2. John Welsh, in 1951, introduced this 9-celled, semi-autonomous ganglion as a preparation offering physiologists unique experimental possibilities. 3. It exhibits remarkable reliability and stability in rhythmic pattern generation. The neurons show endogenous burst-forming capability mediated by "driver potentials". 4. These regenerative, Ca-mediated potentials are restricted to the soma, while impulse-generating membrane is segregated to the distal axon. 5. Thus, voltage-clamp analysis of the ionic currents underlying the burst-forming potentials is possible by isolating the soma with a ligature. 6. The isolated ganglion is spontaneously active, but the normal mechanism of pacemaking remains to be clarified, including the possible contribution of stretch-sensitive dendrites. 7. The activity of the ganglion is subject to modulation by neurohumors. These include the transmitter at intraganglionic synapses, transmitters of the pair of inhibitory and the two pairs of acceleratory fibers, and neurohormones released from the pericardial organs. The transmitters are not established. 8. Effects on the ganglion of substances isolated from the pericardial organs have been described. 9. These include 5-hydroxytryptamine, dopamine, octopamine, and two peptides. 10. One of these, proctolin, produces a long-lasting sequence of effects. 11. The work continues to raise new questions for which the ganglion offers excellent research material.

Animals↗
PubMed1987

Structure and localization of synaptic complexes in the cardiac ganglion of a portunid crab.

The cardiac ganglion of Portunus sanguinolentus exhibits spontaneous rhythmic activity when isolated. The ganglion contains five large and four small intrinsic neurons and is innervated by three pairs of fibres originating in the thoracic ganglia. We have identified the processes of the large neurons in electron micrographs by injecting these cells with two electron-dense markers, horseradish peroxidase (HRP) and Procion Rubine (PR). In addition we have studied the processes of the four smaller neurons by light microscopy serial reconstructions and by electron microscopy of selected regions. Both markers were found only in neuronal processes and not in glial cells nor in the extracellular space, except close to the soma of the injected cell. We found contacts between the small secondary (collateral) processes of the large cells but not between their somata or their primary processes (axons and dendrites). Two specialized structures present at the contacts between the collateral processes were small membrane close appositions, possibly the site of electrotonic junctions, and chemical synapses. Contacts between processes marked by HRP and those marked by PR were common, as were contacts between processes marked by either HRP or PR and those of the other intrinsic neurons. Adjacent processes stained by PR could contain PR deposits of different densities, but it is unclear whether this finding was due to intercellular diffusion of the dye or to its diffusing at different rates into branches of the same process. Identified processes of all the intrinsic neurons contained the same type of vesicles, which were different from those found in processes of the extrinsic fibres. Chemical synapses were present at contacts between processes of the extrinsic and intrinsic neurons, as well as at contacts between processes of the intrinsic neurons. The axons of three small cells made a series of contacts at which extensive arrays of membrane close appositions, but not chemical synapses, were found. These three axons also formed contacts, either directly or through their collateral branches, with processes of the large cells, at which both membrane close appositions and chemical synapses were present. The axon of the fourth small cell could not be followed in our series.

Animals↗
PubMed1987

Somatostatin and altered medium osmotic pressure elicit rapid changes in prolactin release from the rostral pars distalis of the tilapia, Oreochromis mossambicus, in vitro.

Prolactin (PRL) cells in the rostral pars distalis of the tilapia Oreochromis mossambicus respond to somatostatin (SRIF) and reduced medium osmotic pressure within 10-20 min of exposure during perifusion incubation. Pieces of rostral pars distalis tissue were removed from freshwater-adapted tilapia and were preincubated in [3H]leucine in static culture (355 m phi smolal) for 48 hr. Following preincubation, they were placed in the perifusion apparatus and baseline release was established for 3 hr in hyperosmotic medium (355 m phi smolal). Exposure to hyposmotic medium (280 m phi smolal) resulted in a rapid and steep rise in the release of [3H]PRL, which remained elevated for more than 2 hr. When SRIF was added simultaneously with hyposmotic medium, the rise in PRL release normally initiated by reduced osmotic pressure was prevented. Somatostatin also quickly reduced release that had been previously elevated by exposure to hyposmotic medium. The time course of these changes suggests that SRIF and altered osmotic pressure act on PRL secretion in at least partial independence of effects which they may have on PRL synthesis in the tilapia pituitary.

Animals↗
PubMed1987

Comparison of electrical responses of terminals, axons, and somata of a peptidergic neurosecretory system.

Spontaneous and evoked electrical activity was recorded intracellularly from somata, axons, and terminal dilatations of an isolated peptidergic neurosecretory system, the X-organ-sinus gland, of the crabs Cardisoma carnifex and Podophthalmus vigil in order to compare their electrical characteristics. Spontaneous impulse activity was present in most penetrations and included irregular and pacemaker-like firing, as well as patterned activity (bursting). Extracellular recording showed that spontaneous impulses and bursting originate in a proximal region of the axon tract. Somata vary from being electrically nonresponsive to having overshooting impulses with a relatively slow rate of rise. Overshooting impulses were consistently recorded from axons and terminals. Regional differences include (1) a longer action potential duration in terminals, (2) ability of axons and terminals but not somata to sustain repetitive firing, (3) presence of depolarizing afterpotentials in axons but of hyperpolarizing afterpotentials in somata and terminals, and (4) occurrence of impulse broadening during repetitive firing in some terminals but not in axons or somata. Somata and terminals sustained reduced and slowed, but regenerative impulses in nominally Na-free saline and showed alterations of waveform in nominally Ca-free salines, while axons showed no regenerative responses in Na-free saline and no change of impulse form in Ca-free saline. Terminal responses in the presence of tetraethylammonium chloride (TEA) (50 mM) or Ba (50 mM) exhibited long depolarized plateaus, while impulses of somata were much less prolonged. Bursts often took the form of impulses superimposed on a depolarized plateau. Bursts could be evoked by single stimuli applied to the axon tract but not by current passed intracellularly. After addition of TTX, axon tract stimulation evoked plateaus without superimposed impulses. Terminals exhibit specialization of their electrical responses by comparison to axons and somata in having long-duration action potentials attributable to participation of Ca, capability of sustained firing, impulse broadening, and channels supporting sustained inward currents, all of which might enhance the admission of Ca for initiation of peptide secretion.

Action Potentials↗
PubMed1986

Currents under voltage clamp of burst-forming neurons of the cardiac ganglion of the lobster (Homarus americanus).

Crustacean cardiac ganglion neuronal somata, although incapable of generating action potentials, produce regenerative, slow (greater than 200 ms) depolarizing potentials reaching -20 mV (from -50 mV) in response to depolarizing stimuli. These potentials initiate a burst of action potentials in the axon and are thus termed driver potentials. The somata of the anterior-most neurons (cells 1 or 2) were isolated by ligaturing for study of their membrane currents with a two-electrode voltage clamp. Inward current is attributed to Ca2+ by reason of dependence of driver potential amplitude on [Ca2+]0, independence of [Na+]0, resistance to tetrodotoxin, and inhibition by Cd (0.2 mM) and Mn (4 mM). Ca-mediated current (ICa) is present at -40 mV. It is optimally activated by a holding potential (Vh) of -50 to -60 mV and by clamps (command potential, Vc) to -10 mV. Time to peak (10-30 ms) and amplitude are strongly voltage dependent. Maximum tail-current amplitudes observed at -70 to -85 mV are ca. 100 nA. Inward tail peaks may not be resolved by our clamp (settling time, 2 ms). Tails relax with a time constant (tau) of approximately equal to 12 ms (at -70 to -85 mV). ICa exhibits inactivation in double pulse regimes. Recovery has a tau of approximately equal to 0.7 s. Tail current analyses indicate an exponential decline (tau approximately equal to 23 ms at -20 mV) toward a maintained amplitude of inward current tails. Analysis of outward currents indicates the presence of three conductance mechanisms having voltage dependences, time courses, and pharmacology similar to those of early outward current (IA), delayed outward current (IK), and outward current (IC) of molluscan neurons. Analysis of tail currents indicates a reversal potential for each of these near -75 mV, indicating that they are K currents. Early outward current, IA, shows a peak at 5 ms followed by rapid decline. Response to a second clamp given within 0.4 s is reduced; recovery is exponential, with a tau of approximately equal to 200 ms (at Vh = -50 mV). The amplitude of IA tested at 0 mV shows activation or deactivation by subthreshold shifts of Vh. The extent and rate of these changes shows voltage dependence (tau approximately equal to 100-500 ms for subthreshold prepulses). At the normal cell resting potential of -50 mV the amplitude of IA is 25% of that tested from -80 mV.(ABSTRACT TRUNCATED AT 400 WORDS)

4-Aminopyridine↗
PubMed1986

The role of calcium in prolactin release from the pituitary of a teleost fish in vitro.

The release of PRL from the pituitary of a teleost fish, the tilapia (Oreochromis mossambicus), has been previously shown to be dependent on calcium. However, the source(s) and specific action(s) of calcium in the secretory process have not been identified. Also undefined are the mechanisms by which regulators of PRL cell function may alter calcium distribution. In the present investigation, the elevation of medium K+ concentration during static incubations to a depolarizing concentration (56 mM) produced no change in cumulative PRL release over control levels during the 18-20 h of incubation. During perifusion incubation, exposure to high K+ concentrations briefly stimulated (less than or equal to 10 min) and then depressed PRL release. In contrast, reduced medium osmotic pressure elicited a rapid elevation in PRL release that was sustained for 2 h or more. D600, a calcium entry blocker, at 10(-5) M diminished the K+-induced pulses of PRL release. The same concentration, however, did not alter the release of PRL evoked by reduced osmotic pressure. In contrast, CoCl2, which blocks a range of calcium-mediated processes in addition to calcium influx, suppressed PRL release during perifusion and static incubations in hyposmotic medium. These findings suggest that while PRL secretion from the tilapia pituitary is calcium dependent, calcium entry through voltage-regulated plasmalemma channels may not be a prerequisite to the actions of reduced osmotic pressure.

Animals↗
PubMed1985

Electrophysiological characterization of peptidergic neurosecretory terminals.

Electrical activity recorded intracellularly from peptidergic neurosecretory terminal dilatations in the sinus gland of crabs (principally Cardisoma guanhumi and C. carnifex) is described. Recordings were made from the neurohaemal organ in situ on the neural tissue of the isolated eyestalk and from isolated sinus gland-sinus gland nerve preparations. Verification that electrodes penetrated terminals was obtained by dye marking. Resting potentials ranged between -30 and -80mV. Overshooting action potentials of long duration (5-20 ms at 1/2 amplitude) relative to those of non-secretory axons (less than 2ms) were recorded in approximately 70% of stable penetrations. Action potentials occurred spontaneously at slow (less than 0.2s-1) rates in 75% of penetrations. Sequential intra- and extracellular recordings with the same microelectrode, on the same terminal, indicate impulse generation by the terminal itself. Extracellular stimulation of the axon tract evokes an all-or-none action potential at distinct threshold and latency. At rates of stimulation exceeding 5s-1, discrete fluctuations in the form of responses occur. Similar waveforms occur spontaneously and can be evoked by passing current through the electrode. They are interpreted to be electrotonically recorded activity of other parts of a complex axonal terminal arborization. Some, but not all, terminals exhibit impulse broadening (up to three-fold at 1/2 amplitude) during repetitive firing exceeding 1s-1. The same terminals show reduced impulse duration with hyperpolarization and broadened impulses with imposed depolarization. The changes are due to altered repolarization rates. Terminals sustain steady impulse firing at rates (up to 5s-1) linearly related to the imposed depolarizing current. Regenerative potentials, though of reduced rate of rise and amplitude, can be evoked by depolarizing current passed through the electrode during perfusion with salines having 1/2 normal [Na+], or containing tetrodotoxin (10(-6)moll-1). However, these block axonal conduction. Nominally Ca-free saline causes increased spontaneity and depolarization of about 5 mV in half the preparations examined, but reaching 20 mV in the others, with resultant inactivation of regenerative activity. Impulses in low-Ca saline show alterations of the falling phase, it being faster initially and then slower than normal. Thus, while the action potentials of neurosecretory axons are Na dependent, the terminals support regenerative impulses mediated by both Na and Ca.

Action Potentials↗
PubMed1984

Ionic dependence of secretory and electrical activity evoked by elevated K+ in a peptidergic neurosecretory system.

Secretion of the octapeptide erythrophore- (red pigment-) concentrating hormone (ECH, RPCH) and extracellularly monitored electrical activity were followed simultaneously from individual, isolated sinus glands (neurohaemal organs), of the crab Cardisoma carnifex. Following introduction of saline having elevated [K], 100-196 mmol l-1 (5-11 X normal), secretion (bioassayed from 1-min fractions during continuous perfusion) increases from barely detectable (less than 1 fmol min-1) to a peak, average 31 fmol min-1, within 5 min, and immediately subsides. Additional responses are obtainable following a period, greater than 30 min, of normal saline perfusion. Secretory responses to K are Ca-dependent. If Ca is restored (in high K) following perfusion in 0-Ca, high K, only a small secretory response is observed. Addition of Mn (10 mmol l-1, normal Ca) reduces secretion to one-tenth. Increased net uptake of 45Ca of 2.5- to 6-fold is observed in individual sinus glands exposed to 10 X K compared to paired, unstimulated organs. The pattern and Ca-dependence of secretory responses to K are unaffected, but the amount of secretion is augmented in Na-deficient or TTX-containing salines. Intracellular recording confirms that brief (10-40 s) bouts of intense firing recorded extracellularly upon commencing a high K perfusion include repetitive firing by terminals, superimposed on rapid depolarization. Firing ceases as the membrane potential reaches a depolarized value (-18 to -15 mV for [K] 100-176 mmol l-1), which is then maintained until restoration of normal saline, when slow repolarization ensues. In 0-Ca, spontaneous impulse firing is increased, resting potential depolarized by 5 to 15 mV, but the bout of impulse firing and the maintained depolarization in response to K are similar. Thus, mechanisms of secretion of a crustacean peptide neurohormone appear closely similar to those of other systems characterized: responsiveness to elevated K, dependence on Ca, depolarization-, but not secretion-dependent inactivation, and lack of dependence on Na inward current. Intracellular recording here permits direct observation of electrical responses of terminals.

Animals↗
PubMed1979

Isolation and characterization of slow, depolarizing responses of cardiac ganglion neurons in the crab, Portunus sanguinolentus.

1. Tetrodotoxin-resistant, active responses to depolarization of the large cardiac ganglion cells were studied in semi-isolated preparations from the crab, Portunus sanguinolentus. Impulse activity was monitored with extracellular electrodes, simultaneous recordings from two or three large cells were made with intracellular electrodes, and current was passed via a bridge or second intracellular electrode. Preparations were continuously perfused with saline containing 3 x 10(-7) M tetrodotoxin (TTX). 2. About 20 min after introduction of TTX, small-cell impulses and resultant EPSPs in large cells cease, while rhythmic, spontaneous bursting of large cells continues. A pacemaker depolarization between bursts and slow depolarizations underlying the impulse bursts are prominent at this time. Shortly after, spontaneous burst rate slows, and at ca. 25 min, the ganglion becomes electrically quiescent. 3. In the quiescent, TTX-perfused ganglion, injection of depolarizing current into any one of the large cells results in active responses. At current strengths of sufficient intensity and duration (e.g., 20 nA, 20 ms; 5 nA, 500 ms) to depolarize a large cell by ca. 10 mV from resting potential (-53 mV, avg), the graded responses become regenerative and of constant form, provided the stimulation rate is less thna 0.15/s. Such responses have been termed "driver potentials." At more rapid rates, thresholds are increased and responses reduced. 4. Driver potentials of anterior large cells reach peak amplitudes of ca. 20 mV (to -32 mV), have maximum rates of rise of 0.45 V/s and of fall of 0.2 V/s, and a duration of ca. 250 ms. They are followed by hyperpolarizing afterpotentials, a rapidly decaying one (1 s) to -58 mV, followed by a slowly decaying one (7.5 s), -55 mV. Responses of posterior large cells are smaller (16 mV) and slower; the site of active response may be at a distance from the soma. 5. The ability of elicit near-synchronous responses and the identity of amplitude and form of responses among anterior cells and of posterior cells, regardless of which cell receives depolarizing current, indicates that all cells undergo active responses and are stimulated by electrotonic spread of depolarization. 6. The responses involve a conductance increase since memses during a driver potential are much reduced. 7. Depolarization by steady current increases the absolute threshold, decreases the maximum depolarization of the peak, and slows rates of rise and fall. Hyperpolarization increases rates of rise and fall; the absolute value reached by the peak depolarization is unchanged. Hyperpolarization reduces the amplitude of the rapid after-potential relative to the displaced resting potential. 8. Hyperpolarizing current pulses imposed during the rise and peak of driver-potential responses are followed by redevelopment of a complete response. Sufficiently strong hyperpolarization can terminate a response. The current strength needed to terminate a response decreases the later during the response the pulse is given...

Action Potentials↗
PubMed1979

Spontaneous electrical activity and interaction of large and small cells in cardiac ganglion of the crab, Portunus sanguinolentus.

1. Semi-isolated preparations of the nine-celled cardiac ganglion of the crab, Portunus sanguinolentus, were studied electrophysiologically, using simultaneous recording from extracellular and two or three intracellular electrodes. Nine penetrations of small cells were achieved. 2. Three large (80 x 120 micron) cells lie near the anterior end of the 5-mm main trunk; two large and four small (less than 50 micron) cells at the posterior end. Large-cell axons pass along the main trunk and then exit to innervate cardiac muscle; small-cell axons do not leave the ganglion. 3. The semi-isolated ganglion produces spontaneous electrical activity organized into regularly patterned, rhythmic bursts of large- and small-cell impulses recurring at rates of 0.3-0.6/s and lasting 500-800 ms. Small impulse activity commences and ends each burst. Small cells fire trains during the burst, but impulses are not synchronized among them. Large-cell trains are synchronous, are at about one-half the frequency, and have fewer impulses than small-cell trains. 4. Intracellular recordings from small cells show a slow, pacemaker depolarization from a maximum membrane potential of -54 mV leading with only a slight inflection at ca. -50 mV to a depolarized plateau at ca. -40 mV; nonovershooting impulses are superimposed on this but cease before it repolarizes. Impulses, therefore, arise at a site distant from the soma and do not invade it. Deflections suggesting synaptic potentials are not seen. 5. Intracellular recordings from large cells show complex depolarizations corresponding to extracellularly recorded bursts. These represent excitatory postsynaptic potentials (EPSPs) corresponding with individual small-cell impulses, attenuated, non-overshooting spikes, and an underlying slow depolarization; usually no pacemaker depolarization is apparent between bursts. Chemically mediated transmission is probable for the EPSPs because they show delay, increase in amplitude with hyperpolarization, sometimes show facilitation, and are reduced in saline having one-third Ca, 3 x Mg. 6. EPSPs, impulses, and the slow depolarization occur synchronously among the large cells. Potentials recorded from posterior cells are attenuated and slower than those of the anterior cells. This is interpreted to reflect sites of occurrence more distant from the soma in the posterior than in the anterior cells. Impulses do not invade the somata. 7. Intracellular recordings from large-cell axons 4 mm from the soma show overshooting action potentials arising sharply from a base line. EPSPs are absent or highly attenuated and there is little underlying depolarization (less than 2 mV). 8. Current passing with electrodes intracellular to two cells has established directly that all large cells are electrotonically coupled and that an anterior cell and a small cell are coupled. Changes of burst rate during current passing into any large cell indicate that all large cells and small cells are electrotonically coupled. 9...

Animals↗
PubMed1975

Neurohormonal alteration of integrative properties of the cardiac ganglion of the lobster Homarus americanus.

The spontaneous burst discharges of isolated lobster (Homarus americanus) cardiac ganglia were recorded with a spaced array of electrodes. Small regions (less than 1 mm) of the ganglion were exposed to the cardioexcitor neurohormone in extracts of pericardial organs (XPO) or to 10(-5) M 5-hydroxytryptamine (5HT). All axons were excited (increased mean firing frequency, f) by both substances, but only by applications in the region between the soma (but excluding it) and proximal site of impulse initiation. Units not so exposed changed their f relatively little despite f increases of as much as threefold in exposed units and changes in burst rate and overall length. Regularity and grouping of all impulse activity into bursts was never disturbed. 5HT increases burst rate at any point of application. The increases are larger if small cells are affected than if only large cells are exposed. Burst length decreases except when the pacemaker is affected. In contrast, XPO affects neither burst rate or length unless small cells are affected. Length is increased if non-pacemaker small cells are affected; both rate and length increase if the pacemaker is affected. The pacemaker usually exhibits an f of intermediate value. Rate changes are not simply related to its f. A small cell can "burst" in the absence of impulses from any other cells. XPO may enhance endogenous "driver potentials," while 5HT may excite by depolarizing at limited sites.

Action Potentials↗