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J T Buchanan

Publications and source records attributed to J T Buchanan.

51 records · Page 3Linked to original sources
PubMed1982

Activities of identified interneurons, motoneurons, and muscle fibers during fictive swimming in the lamprey and effects of reticulospinal and dorsal cell stimulation.

1. Application of D-glutamate to the isolated spinal cord of the lamprey produces phasic activity in ventral roots, which is similar to that of the muscles of the intact swimming animal (5,18). Therefore, the isolated spinal cord may be used as a convenient model for the investigation of the generation of locomotor rhythms in a vertebrate. 2. Almost all slow muscle fibers exhibited excitatory junctional potentials (EJPs) during swimming activity. The number of EJPs per cycle increased with the intensity of ventral root (VR) bursting. Few twitch fibers were active, and these fired action potentials only during high intensities of VR bursts. 3. As was found by Russell and Wallén (25), myotomal motoneurons had oscillating membrane potentials during fictive swimming which, on the average, reached a peak depolarization in the middle of the VR burst (phi = 0.21 +/- 0.05; phi = 0 is defined as the onset of the VR burst, and the duration of the cycle is set equal to 1). Membrane potential oscillations in fin motoneurons were antiphasic to those of nearby myotomal motoneurons (peak depolarization phi = 0.68 +/- 0.05). 4. Lateral interneurons had oscillating membrane potentials in synchrony with those of myotomal motoneurons (peak depolarization phi = 0.21 +/- 0.10). Interneurons with axons projecting contralaterally and caudally (CC interneurons) had oscillating membrane potentials that peaked significantly earlier in the cycle (peak depolarization phi = 0.06 +/- 0.12). 5. Edge cells were only weakly modulated during fictive swimming. Their peak depolarizations occurred near the end of the VR burst (phi = 0.33 +/- 0.10). Most giant interneurons were not phasically modulated during fictive swimming. 6. Repetitive intracellular stimulation of Müller cells during fictive swimming generally evoked an increased burst intensity in ipsilateral VRs and a decreased burst intensity in contralateral VRs. The cells M3, B1, and B2 also produced increases or decreases in the frequency of VR bursts. Repetitive intracellular stimulation of sensory dorsal cells could also change the intensities and timing of VR bursts. 7. This study is an initial survey of lamprey spinal interneurons that participate in swimming activity. Lateral interneurons and CC interneurons are active during fictive swimming and probably help coordinate the undulations of the body, but their roles in pattern generation are not known. The central pattern generator is subject to modification by descending and sensory inputs.

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PubMed1982

Identification of interneurons with contralateral, caudal axons in the lamprey spinal cord: synaptic interactions and morphology.

1. As part of a continuing investigation of the organization of the spinal cord of the lamprey, propriospinal interneurons with axons projecting contralaterally and caudally (CC interneurons) were surveyed with intracellular recordings. 2. CC interneurons were identified by recording their axon spikes extracellularly in the spinal cord during intracellular stimulation of the cell body. The axon projections of Cc interneurons were confirmed after intracellular injection and development of horseradish peroxidase. 3. Intracellular stimulation of CC interneurons produced synaptic potentials in myotomal motoneurons, lateral interneurons and other CC interneurons that lay caudally on the opposite side of the spinal cord. Most CC interneurons were inhibitory, but some were excitatory. 4. CC interneurons were divided into three classes on the basis of reticulospinal Müller cell inputs. CC1 interneurons were excited by the ipsilateral Müller cell B1 and the contralateral Mauthner cell. CC1 interneurons were inhibitory. They were excited polysynaptically by ipsilateral sensory dorsal cells and were inhibited by contralateral dorsal cells. They were distinguished morphologically by having no rostral axon branch and no contralateral dendrites. CC1 interneurons were phasically active during fictive swimming with their peak depolarizations preceding those of myotomal motoneurons by about 0.15 cycle. 5. CC2 interneurons were also inhibitory, but they were distinguished from CC1 interneurons by their excitation from the ipsilateral Müller cells B2-4 nd by their thin rostral and thicker caudal axonal branches on the contralateral side of the spinal cord. 6. CC3 interneurons were excitatory, and they were inhibited by the ipsilateral Müller cell I1. CC3 interneurons could have contralateral dendrites and bifurcating axons, and they had lower average axonal conduction velocities than CC1 and CC2 interneurons. 7. Inhibitory CC interneurons may be important for motor coordination in the lamprey. Movements of the lamprey body during reflexes and swimming consist of contraction and relaxation of myotomal muscles on opposite sides of the body. By being coactive with ipsilateral myotomal motoneurons, inhibitory CC interneurons could contribute to the inhibition of contralateral motoneurons during these movements.

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PubMedDate not supplied

Calcium currents and GABAB receptors in the dorsal sensory cells of the lamprey spinal cord.

Patch-clamp studies were performed on the isolated dorsal sensory cells of the spinal cords of three species of lamprey, Ichthyomyzon unicuspis, Petromyzon marinus, and Lampetra fluviatilis, to measure changes in the amplitudes of calcium current induced by GABA and its specific antagonists and agonists. The experiments showed that GABA (4 mM) reduced the peak amplitude of the calcium current by 28.5 +/- 4.9%, with subsequent recovery to 96.2 +/- 9.2% of control (n = 45). The GABAB agonist baclofen had similar effects. The GABAA agonists glycine and taurine had no effect on the Ca2+ current. The inhibitory effect of GABA was blocked by 2-hydroxysaclofen (a GABAB antagonist), but persisted in the presence of bicuculline (a GABAA antagonist). These results are evidence that the membranes of dorsal sensory cells contain GABAB receptors, which significantly increases our understanding of the mechanisms of presynaptic inhibition in the spinal cords of the cyclostomata.

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PubMedDate not supplied

The effects of baclofen on calcium channel currents in dorsal sensory cells of the spinal cord in the lamprey.

Dorsal sensory cells isolated from the spinal cord of the lamprey species Ichthyomyzon unicuspis and Lampetra fluviatilis were used for whole-cell patch-clamp studies of the effects of baclofen on calcium channel currents, evoked in conditions in which Na+, K+ currents were blocked, by depolarizing membranes from constant holding potentials of -100 or -80 mV to +30 mV. Ba ions were used as carriers of currents through calcium channels. These studies demonstrated that baclofen (0.5 mM) decreased the peak amplitude of the Ba2+ current by an average of 22.5 +/- 4.2% (n = 12) in dorsal sensory cells of the lamprey Ichthyomyzon unicuspis and by 28.4 +/- 3.3% in the dorsal sensory cells of Lampetra fluviatilis (n = 25). The conductivity of dorsal sensory cell membranes in the presence of baclofen (and GABA) did not change. The blocking action of baclofen persisted in the presence of bicuculline (100 microM) and was lifted by addition of delta-aminovaleric acid and 2-hydroxysaclofen to the perfusing solution. These results are interpreted as evidence for the presence of GABAB receptors in dorsal sensory cell membranes. The data were compared with published results, and the question of the functional significance of GABAB receptors in the dorsal sensory cells (primary afferent cells) of cyclostomata is discussed.

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PubMedDate not supplied

Improved attachment and spreading in primary cell cultures of the eastern oyster, Crassostrea virginica.

At present, establishment of a cell line from bivalve molluscs has been unsuccessful, and in vitro work is limited to primary cell cultures. We sought to improve attachment and spreading of cells of the eastern oyster, Crassostrea virginica, to aid primary cultures and to assist development of a bivalve cell line. Our objectives were to examine the effects of substrate on ventricle cell viability, attachment, and spreading by testing of collagen I, collagen IV, fibronectin, laminin, poly-D-lysine, and two types of uncoated tissue culture plates (Falcon and Corning). Experiments were conducted by incubating cells with the various substrates for 24 h and 5 d. An assay with a tetrazolium compound (MTS) was used to estimate cell numbers based on metabolic activity. Although differences in MTS assay values for substrate effect on cell viability were detected at 24 h and at 5 d (P > 0.0001), these were attributed to variations in metabolic activity due to different levels of attachment and spreading among treatments. Differences among treatments were detected in attachment and spreading at 24 h and 5 d (for all, P > 0.0001). At 24 h, poly-D-lysine induced the highest levels of attachment and spreading; no other factor performed better than the uncoated Falcon substrate, and collagen I performed most poorly. At 5 d, poly-D-lysine and the uncoated Corning substrate induced significantly higher levels of attachment and spreading than did the uncoated Falcons substrate, and collagen I performed most poorly. From these results, poly-D-lysine best promoted cell attachment and spreading. Fibronectin (at 24 h) and laminin (at 5 d) warrant further study. Along with improvements in medium composition, future work should involve screening of other attachment factors and combinations of factors, including those of bivalve origin.

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PubMedDate not supplied

The effects of serotonin on functionally diverse isolated lamprey spinal cord neurons.

The experiments reported here showed that application of serotonin (5-hydroxytryptamine, 5-HT) (100 microM) did not induce any significant current through the membranes of any of the spinal neurons studied (n = 62). At the same time, the membranes of most motoneurons and interneurons (15 of 18) underwent slight depolarization (2-6 mV) in the presence of 5-HT, which was not accompanied by any change in the input resistance of the cells. Depolarization to 10-20 mV occurred in some cells (3 of 18) of these functional groups, this being accompanied by 20-60% decreases in input resistance. The same concentration of 5-HT induced transient low-amplitude depolarization of most sensory spinal neurons (dorsal sensory cells), this changing smoothly to long-term hyperpolarization by 2-7 mV. The input resistance of the cell membranes in these cases showed no significant change (n = 8). Data were obtained which provided a better understanding of the mechanism by which 5-HT modulates the activity of spinal neurons. Thus, 5-HT facilitates chemoreceptive currents induced by application of NMDA to motoneurons and interneurons, while the NMDA responses of dorsal sensory cells were decreased by 5-HT. 5-HT affected the post-spike afterresponses of neurons. 5-HT significantly decreased the amplitude of afterhyperpolarization arising at the end of the descending phase of action potentials in motoneurons and interneurons and increased the amplitude of afterdepolarization in these types of cells. In sensory spinal neurons, 5-HT had no great effect on post-spike afterresponses. The results obtained here support the suggestion that 5-HT significantly modulates the activity of spinal neurons of different functional types. 5-HT facilitates excitation induced by subthreshold depolarization in motoneurons and some interneurons, facilitating the generation of rhythmic discharges by decreasing afterhyperpolarization. In sensory cells, 5-HT enhances inhibition due to hyperpolarization, suppressing NMDA currents. The differences in the effects of 5-HT on functionally diverse neurons are presumed to be associated with the combination of different types of 5-HT receptors on the membranes of these types of spinal neurons.

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PubMedDate not supplied

Serotonin modulates oscillations of the membrane potential in isolated spinal neurons from lampreys.

Studies were performed on spinal neurons from lampreys isolated by an enzymatic/mechanical method using pronase. The effects of 100 microM serotonin (5-HT) on membrane potential oscillations induced by a variety of excitatory amino acids were studied. 5-HT was found to depolarize branched cells (presumptive motoneurons and interneurons) by 2-6 mV without inducing membrane potential oscillations. However, when oscillations were already present because of an excitatory amino acid, 5-HT changed the parameters of these oscillations, increasing the amplitudes of all types of oscillations, increasing the frequency of irregular oscillations, and increasing the duration of the depolarization plateaus accompanied by action potentials. Serotonin modulation of the effects of excitatory amino acids and the electrical activity of cells in the neural locomotor network facilitates motor activity and leads to increases in the contraction of truncal muscles and more intense movements by the animal. The possible mechanisms of receptor coactivation are discussed, along with increases in action potential frequency and changes in the parameters of the locomotor rhythm.

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PubMedDate not supplied

[Potential-gated currents in isolated spinal cord neurons of the river lamprey Lampetra fluviatilis].

Potential-gated currents in the membranes of enzymatically isolated neurons from the lamprey spinal cord were investigated using the whole cell variant of the patch-clamp technique. As it was revealed the main currents underlying action potential (AP) in the dorsal sensory cells as well as in branched cells are inward Na(+)- and outward K(+)-currents. Sodium current has the duration 4-9 ms, it was rapidly activated and inactivated and blocked by TTX. Its activation and inactivation processes can be fit to modified Bolzmann equations. Outward potassium current can be subdivided into at least two components: 1) early transient rapidly inactivated component which was sensitive to 4-AP and 2) later noninactivated more longer one which was sensitive to TEA. Their function as in the other neurons may be to repolarize membranes after AP generation and to provide the interval between APs during rhythmic activity. Ca2(+)-current was investigated using Ba2+ as a divalent carrier. Judging on two different values of the threshold there are two types of the Ca2(+)-channels in the membranes of the lamprey spinal cells. Ba2(+)-current depend not only on the voltage but on the intracellular enzyme systems particularly on the G-proteins, ATP and CP-CPK. Under normal conditions Ca2(+)-current activated during AP is small but it increase up to 5-10 nA when the other voltage-gated currents are blocked. We conclude that lamprey spinal neurons retain after isolation the electrical properties of neurons in the spinal cord. The obtained results have testified that the kinetic and pharmacological properties of the basic currents in the cells under study are close to those in the other vertebrate neurons and these cells are suitable for investigation of neurotransmitter, neuromodulator and pharmacological agents action on voltage- and agonist-gated membrane currents in vertebrate neurons.

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PubMedDate not supplied

[Calcium current and GABA(B) receptors in dorsal sensory cells of the lamprey spinal cord].

GABA and GABAB receptor agonists were shown to reduce the peak calcium current amplitude with its subsequent recovery, whereas glycine and taurine, the GABAA receptor agonists, did not modify the current. The findings suggest that the GABAB receptors mediate a presynaptic inhibition by suppression of the Calcium currents in the cyclostome spinal cord.

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PubMedDate not supplied

[Effect of baclofen on calcium channel currents in dorsal sensory cells of the lamprey spinal cord].

delta-Baclofen reduced the peak amplitude of the barium current in the DSCs from Lunicuspis and L.fluviatilis. The membrane conductance was not altered either by baclofen or GABA. The reducing effect could be abolished with delta-aminovaleric acid or 2(OH)saclofen. The data obtained suggest presence of the GABAB receptors in the DSC membranes. Functional role of the GABAB receptors in primary afferent cells of cyclostomes, is discussed.

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