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Biomedical subjects

M V Bennett

Publications and source records attributed to M V Bennett.

At least 109 records · Page 6Linked to original sources
PubMed1986

Changes in gain of the vestibulo-ocular reflex induced by combined visual and vestibular stimulation in goldfish.

Adaptive changes in the vestibulo-ocular reflex (VOR) of goldfish were produced in a few hours by sinusoidally rotating restrained fish in the horizontal plane inside a vertically striped drum. The drum could also be sinusoidally rotated so that the gain of the VOR (the ratio of eye to head angular velocity) would have to increase to two or decrease to zero in order to maintain a stable retinal image. During 'training' towards two VOR gain measured at the stimulation frequency of 0.125 Hz increased rapidly over 6 h of stimulation to about 1.5 from an initial gain of 0.7. Half of that change occurred in the first 30 min. During training towards zero VOR gain measured at the stimulation frequency decreased to 0.15. About one-third of that change occurred in the first 30 min. Testing at different sinusoidal frequencies after 6 h stimulation showed that increases in VOR gain were generated across a 6-octave range; however, reductions in gain were produced over a narrow frequency range close to the training frequency. Gain reductions occurred more rapidly on a second day of stimulation. In a paradigm simulating reversing prisms, partial reversal of the VOR was observed in some fish. However, these fish also demonstrated spontaneous slow sinusoidal eye movements that may have represented a different means of adjusting eye movements to stabilize the retinal image. Goldfish provide a useful preparation for the study of adaptive gain changes in vertebrate oculomotor systems.

Adaptation, Physiological↗
PubMed1986

Changes in gain of the vestibulo-ocular reflex induced by sinusoidal visual stimulation in goldfish.

The effects of sustained sinusoidal visual stimulation on the vestibulo-ocular reflex (VOR) and the optokinetic reflex (OKR) were investigated. Goldfish were held stationary inside a striped drum rotating sinusoidally about the vertical axis for 3 h. The VOR gain, the ratio of eye to head rotational velocities, was measured in the dark with passive sinusoidal rotation of the fish and showed modest increases that were greatest at the stimulation frequency. Furthermore, the fish generated spontaneous sinusoidal eye movements at approximately the stimulation frequency, and these movements summated with the response to other frequencies of vestibular stimulation in the dark. It is hypothesized that the pathways of OK and VO stimuli converge and that the animal increases gain in a common part when it attempts to stabilize the visual image by increasing its response to the OK signal. Thus increases in gain of both OKR and VOR are produced.

Animals↗
PubMed1986

The giant fiber and pectoral fin adductor motoneuron system in the hatchetfish.

In the medulla of the hatchetfish each Mauthner fiber forms chemical synapses on a number of large myelinated axons termed giant fibers. The giant fibers form rectifying electrotonic synapses on pectoral fin adductor motoneurons, and in this fish bilateral pectoral fin adduction is an important component of the Mauthner fiber-mediated escape reflex. The branching patterns of giant fibers were determined by intracellular injection of Lucifer yellow. Dye coupling to the motoneuron somata was not observed, although a low level of transfer might have been obscured by autofluorescence. Individual giant fibers terminate primarily on pectoral fin motoneurons contralateral to their cell bodies, but may also send a branch back across the midline to ipsilateral motoneurons. The rostral process of each giant fiber ends on neurons presumably associated with cranial musculature. The number and geometry of the pectoral fin motoneurons were determined using Golgi and Nissl staining and serial reconstruction methods.

Animals↗
PubMed1986

Sensitivity of gap junctional conductance to H ions in amphibian embryonic cells is independent of voltage sensitivity.

In vertebrate embryos gap junctional conductance (gj) is reduced by transjunctional voltage (Vj) and by cytoplasmic acidification; in each case sensitivity is comparable to those of other channels gated by voltage and ligand-receptor binding. We show here that the mechanisms by which Vj and intracellular pH (pHi) gate gj are apparently independent. Partial reduction of gj by lowering pHi neither attenuates nor enhances further reduction by Vj. Certain drugs irreversibly (glutaraldehyde, 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) or reversibly (retinoic acid) abolish dependence of gj on pHi without appreciably affecting kinetic properties of voltage dependence or the shape of the steady-state Vj-gj relation. These findings suggest that the mechanisms by which pHi and Vj act on the gap junction are at least partially distinct and presumably involve separate regions of the junctional macromolecules.

Animals↗
PubMed1986

Isolated liver gap junctions: gating of transjunctional currents is similar to that in intact pairs of rat hepatocytes.

We have shown previously that conductance of rat liver gap junctions is blocked by an affinity-purified polyclonal antibody generated against rat liver junctional membranes, is not affected by moderate transjunctional or transmembrane potentials, and is reversibly decreased by cytoplasmic acidification and perfusion with octanol. We have now recorded currents from isolated liver gap junctions using patch electrodes dipped through a layer of mixed lipids whose concentrations match those of isolated liver appositional membranes. These currents are blocked by the same polyclonal antibody, are insensitive to moderate voltages imposed across the pipette tip, and are reversibly blocked by similar concentrations of H ions and octanol as are junctions in situ. The currents are likely to be gap junctional in origin; their block by low pH and other agents indicates that the gating mechanisms are intrinsic to the gap junctions themselves and presumably result from conformational change in the channel-forming protein.

Animals↗
PubMed1986

cAMP increases junctional conductance and stimulates phosphorylation of the 27-kDa principal gap junction polypeptide.

Membrane-permeant cAMP derivatives (dibutyryl- and 8-bromo-cAMP) increase gap-junctional conductance within minutes when applied to voltage-clamped pairs of rat hepatocytes. Glucagon also increases junctional conductances, but the response has a more rapid onset and is more rapidly reversible. The glucagon effect can be prevented by intracellular injection of the protein inhibitor of the cAMP-dependent protein kinase (Walsh inhibitor), indicating that the catalytic subunit of cAMP-dependent protein kinase is directly involved. The 27-kDa major gap junction polypeptide is phosphorylated when liver cells dissociated into small groups are incubated with 32P. Addition of 8-bromo-cAMP to cells increases the incorporation of 32P into the 27-kDa junctional protein. Serine is the amino acid residue that is phosphorylated. When isolated liver gap junctions are incubated in the presence of catalytic subunit of the cAMP-dependent protein kinase, the 27-kDa gap junction polypeptide is phosphorylated with low stoichiometry on serine. The rapid increases in gap junctional conductance caused by agents that elevate cAMP and phosphorylation of the gap junction protein by cAMP-dependent protein kinase suggest that cAMP-dependent phosphorylation of the gap junction channel modulates the conductance of liver gap junctions.

Animals↗
PubMed1985

Time course of miniature postsynaptic potentials at the Mauthner fiber giant synapse of the hatchetfish.

The hatchetfish Mauthner fiber is presynaptic to 8-14 large myelinated axons in the medulla; the large ('giant') synapses formed by these fibers appear to be nicotinic cholinergic. Miniature postsynaptic potentials (mPSPs) were recorded from single identified synapses. The mPSPs were averaged to more accurately determine their shape; the rise time was approximately 70 microseconds, and the fall usually was biphasic with time constants of decay for the two phases of 280 and 800 microseconds. In 25% of the records analyzed a third, slow tail of decay was seen which had an average decay constant of 4.2 ms. The biphasic decay of mPSPs largely accounts for the similar shape of the postsynaptic current following a presynaptic impulse, which is described in the accompanying paper.

Animals↗
PubMed1985

Postsynaptic currents at the Mauthner fiber giant synapse of the hatchetfish.

Postsynaptic currents (PSCs) at the giant synapse between Mauthner and giant fibers of the hatchetfish Gasteropelecus were studied under voltage clamp. This axo-axonic synapse lies in the central nervous system beneath the floor of the 4th ventricle where electrodes can be closely positioned both pre- and postsynaptically. Transmission is nicotonic cholinergic. The PSCs produced by Mauthner fiber impulses rise rapidly to a peak and decay in two phases; an early more rapid phase is followed by a late slower phase. The slope conductance of the peak amplitude of the PSCs declines at more inside positive potentials. The late phase of decay is exponential and voltage dependent, becoming faster for PSCs evoked at more inside positive potentials. At potentials positive to about -40 mV the late phase merges with the early phase. The decay rate constant of the slowest phase is exponentially related to voltage for potentials negative to about -10 mV, but becomes less voltage dependent for more positive potentials. The peak current is independent of whether it is evoked during inward or outward active currents of the electrically excitable membrane, and two phase decays are observed in PSCs of reduced quantal content. Thus, changes in slope conductance and two phase decays are not due to series resistance or interactions between quanta. PSCs can be modeled by a 3 state reaction scheme in which closed channels open when they bind transmitter and then can pass to a second closed state with receptor still bound such that they must return through the open state before losing their transmitter and returning to the resting, closed state.

Animals↗
PubMed1985

Cell junctions in early embryos of squid (Loligo pealei).

Squid embryos examined by freeze-fracture and thin-section electron microscopy exhibit identifiable gap junctions during mid-cleavage stages (stages 7-8), and junctional complexes composed of adherent appositions, elaborate septate junctions and gap junctions at slightly later stages (stages 12-13). During germinal layer establishment (stages 12-13) cytoplasmic bridges frequently link the embryonic cells. The presence of gap junctions in cleavage-stage embryos provides the morphological substrate for a demonstrated pathway of direct cell-cell communication that is modifiable by experimental treatments and may be physiologically regulatable. The existence of septate junctions and adherent contacts at later stages suggests that some functional specialization, perhaps the establishment of a strongly joined framework of cells at the surface of the embryo, accompanies the formation of germinal layers.

Animals↗
PubMed1985

Ultrastructure of the rectifying electrotonic synapses between giant fibres and pectoral fin adductor motor neurons in the hatchetfish.

Synapses formed by giant fibres on pectoral fin adductor motor neurons were identified by horseradish peroxidase (HRP) injection. The synapses were distributed in clusters on the somata and proximal dendrites of the motor neurons. All of the labelled synapses contained synaptic vesicles and often had clearly defined active zones characteristic of chemical synapses. Some synapses also showed gap junctions with the motor neuron soma, often directly adjacent to an active zone. The gap junctions were asymmetrical, with a thick layer of electron dense material on the postsynaptic side. Previous electrophysiological data indicate that giant fibre inputs to motor neurons are purely electrotonic and that these electrical synapses rectify.

Action Potentials↗
PubMed1985

Reduction of gap junctional conductance by microinjection of antibodies against the 27-kDa liver gap junction polypeptide.

Antibody raised against isolated rat liver gap junctions was microinjected into coupled cells in culture to assess its influence on gap junctional conductance. A rapid inhibition of fluorescent dye transfer and electrical coupling was produced in pairs of freshly dissociated adult rat hepatocytes and myocardial cells as well as in pairs of superior cervical ganglion neurons from neonatal rats cultured under conditions in which electrotonic synapses form. The antibodies have been shown by indirect immunofluorescence to bind to punctate regions of the plasma membrane in liver. By immunoreplica analysis of rat liver homogenates, plasma membranes, and isolated gap junctions resolved on NaDodSO4/polyacrylamide gels, binding was shown to be specific for the 27-kDa major polypeptide of gap junctions. This and similar antibodies should provide a tool for further investigation of the role of cell-cell communication mediated by gap junctions and indicate that immunologically similar polypeptides comprise gap junctions in adult mammalian cells derived from all three germ layers.

Animals↗
PubMed1985

Some electrical and pharmacological properties of gap junctions between adult ventricular myocytes.

Ventricular myocytes were isolated from adult rat hearts using the technique of Wittenberg and Robinson (Cell Tissue Res. 216: 231-251, 1981). These cells exhibited morphology, input resistance, time constant, and excitability expected for cells in intact cardiac tissue. Pairs of these cells were electronically coupled, and junctional conductance was unaffected by transjunctional potential or hyperpolarization of both cells. Brief exposure of cell pairs to medium equilibrated with 100% CO2 or containing 0.1 mM octanol quickly and reversibly decreased junctional conductance. We conclude that gap junctions between pairs of ventricular myocytes possess physiological properties like those of junctions in many other tissues. This preparation will be useful in evaluating drug action on junctional communication in heart.

Animals↗
PubMed1985

Regulation of gap junctional conductance.

Gap junctional conductance is regulated by the number of channels between coupled cells (the balance between formation and loss of these channels) and by the fraction of these channels that are open (gating mechanisms). A variety of treatments are known to affect junction formation. Adenosine 3',5'-cyclic monophosphate (cAMP) is involved in some cases, and protein synthesis may be required but precursor molecules can also exist. Junction removal occurs both by dispersion of particles and by internalization of junctional membrane. Factors promoting removal are not well understood. A variety of gating mechanisms exist. Coupling may be controlled by changes in conductance of nonjunctional membranes. Several kinds of voltage dependence of junctional conductance are known, but rat ventricular junctions at least are electrically linear. Cytoplasmic acidification decreases conductance of most gap junctions. Sensitivity in rat ventricular myocytes allows modulation of coupling by moderate changes near normal internal pH. Increasing intracellular Ca also decreases junctional conductance, but in the better studied cases sensitivity is much lower to Ca than H. A few data support low sensitivity to Ca in cardiac cells, but quantitative studies are lacking. Higher alcohols such as octanol block junctional conductance in a wide range of tissues including rat ventricular myocytes. An antibody to liver gap junctions blocks junctions between rat ventricular myocytes. Cross reactivity indicates at least partial homology between many gap junctions. Although differences among gap junctions are known, a general physiology is being developed, which may have considerable relevance to normal cardiac function and also to conduction disorders of that tissue.

Animals↗
PubMed1984

pH dependence of transmission at electrotonic synapses of the crayfish septate axon.

Gap junctions between segments of the crayfish septate axon mediate electrotonic transmission of impulses propagating along the length of the nerve cord. We simultaneously measured intracellular pH (pHi) and gap junctional conductance (gj) while axons were exposed to saline equilibrated with CO2, weak acids, and the weak base ammonium chloride. Normal pHi is about 7.1. When pHi is elevated, gj is unaffected. When pHi is reduced, gj declines with an apparent pK of about 6.7 and a Hill coefficient of about 2.7. We also measured effects of pHi on non-junctional conductance (gnj) and on the coupling coefficient, k. Over the pHi range 6.2-8, gnj increases approximately linearly with pHi. Since k is a function of gj and gnj, it reached a maximum at about pHi 7.1, decreasing at higher and lower pHi. The pHi dependence of gj in crayfish septate axon is less steep and has a lower apparent pK than the gj-pHi relation in two vertebrate embryos previously examined. This finding illustrates a difference in gating among analogous and possibly homologous membrane channels.

Animals↗
PubMed1984

The fine structure of identified electrotonic synapses following increased coupling resistance.

Gap junctions exist in the septa between the segments of the lateral giant axons in the ventral nerve cord of the crayfish Procambarus. A large increase in the resistance (uncoupling) of these gap junctions was brought about by mechanical injury to the axonal segments. Both thin sections and freeze-fracture preparations were used to monitor the morphological changes which occurred up to 45 min after injury. There was no apparent change in the organization (a loose polygonal array) of the intramembrane particles which make up the junctional complex up to 45 min after injury. In some instances, however, the intramembrane particles appeared to have moved away from the junctional area. Other junctional regions were internalized and appeared similar to what have been called annular gap junctions. Also at this time (20-25 min after injury), a dense cytoplasmic plug formed in uninjured axon near the junctional region. It is concluded that the gap junctions that exhibit a loose polygonal organization of the intramembrane particles may be either in a state of low resistance (coupled) or a state of high resistance (uncoupled).

Animals↗
PubMed1984

Developmental uncoupling between blastoderm and yolk cell in the embryo of the teleost Fundulus.

During cleavage and blastula stages of embryos of the teleost Fundulus heteroclitus all of the cells are both electotonically coupled and dye coupled to one another, as determined by microelectrode impalements and spread of Lucifer Yellow. At about the time that gastrulation begins we observed a specific loss of junctional coupling between the yolk cell and cells of the blastoderm. Passage of Lucifer Yellow between the yolk cell and blastoderm was reduced at stage 12 (late blastula), and not detected at stage 13 and thereafter, although cells of the blastoderm remain dye coupled to one another through gastrula stages. Also, junctional electrical coupling between the yolk cell and blastoderm became substantially reduced at stage 13 and thereafter. The loss of coupling at this specific cell apposition and time and the large size of the yolk cell may prove useful in analyzing the underlying cellular mechanisms.

Animals↗
PubMed1984

Gating of gap junction channels.

Gap junctional conductance ( gj ) in various species is gated by voltage and intracellular pH (pHi). In amphibian embryos, gj is reduced to half by a 14 mV transjunctional voltage ( Vj ), a change that in fish embryo requires approximately 28 mV. Crayfish septate axon and pairs of dissociated rat myocytes show no voltage dependence of gj over a range of Vj greater than +/- 50 mV. In fish and amphibian blastomeres , gj is steeply decreased by decrease in pHi (n, Hill coefficient: 4.5) and the apparent pKH (7.3) is in the physiological range. In crayfish septate axon the pKH is lower (6.7) and the curve is less steep (n = 2.7). Rises in cytoplasmic Ca can also decrease gj but much higher concentrations are required (greater than 0.1 mM in fish blastomeres). Voltage and pH gates on gap junctions in amphibian embryos appear independent. In squid blastomeres pH gates exhibit some sensitivity to potential, both transjunctional and between inside and outside. A pharmacology of gap junctions is being developed: certain agents block gj directly (aldehydes, alcohols, NEM in crayfish); others block by decreasing pHi (esters that are hydrolyzed by intrinsic esterases, NEM in vertebrates, and, as in the experiments demonstrating the effect of pHi, weak acids). Certain agents block pH sensitivity without affecting voltage dependence (retinoic acid, glutaraldehyde, EEDQ), further indicating separateness of pH and voltage gates. These studies demonstrate a dynamics of gap junctional conductance and variability in gating in a series of possibly homologous membrane channels.

Amphibians↗