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Biomedical subjects

G A Lnenicka

Publications and source records attributed to G A Lnenicka.

16 recordsLinked to original sources

Intrinsic differences in axonal growth from crayfish fast and slow motoneurons.

The motoneurons innervating the fast and slow flexor muscles in the abdomen of crayfish form morphologically distinct motor terminals. Axons of the fast flexor (FF) motoneurons, which innervate the large (fast) flexor muscle, produce extensive motor terminal arbors with many branches. Axons of the slow flexor (SF) motoneurons, which innervate the thin (slow) flexor muscle, produce small terminal arbors with many fewer branches. To determine whether intrinsic factors contribute to these differences in terminal arbors, we compared regenerating axonal arbors from these two populations of motoneurons. We used an explant of the crayfish nerve cord in which axons from the FF and SF motoneurons regenerate on a homogeneous substrate. We found that regardless of the substrate, FF motor axons produced arbors with a greater total length and a greater number and density of branches than SF motor axons. These differences in regenerated arbors persisted in defined medium and in the absence of impulse activity, indicating that they result from intrinsic, neuron-specific factors. The greater branching of the FF motor axons may be related to differences in growth cones: growth cones of FF axons were significantly larger with more filopodia than growth cones of SF axons.

Animals

Activity-dependent reduction in voltage-dependent calcium current in a crayfish motoneuron.

The effect of increased impulse activity upon voltage-dependent Ca2+ currents was studied in the cell body of a crayfish phasic motoneuron using two-electrode voltage-clamp technique. Increased electrical activity in this relatively inactive motoneuron produces a short-term and long-term reduction in the voltage-dependent Ca2+ current. Both forms of activity-dependent reduction in Ca2+ current are Ca2+ dependent. The short-term reduction in Ca2+ current appears to involve the Ca(2+)-dependent inactivation of Ca2+ channels, previously described in a variety of neurons. The long-term reduction in Ca2+ current is produced by prolonged Ca2+ influx and persists for days: in vivo stimulation of the phasic motor axon at 5 Hz for 1 hr results in a 30% reduction in Ca2+ current density, which persists for at least 3 d. Both the short-term and long-term reductions in Ca2+ current appear to result from changes in a single type of high-voltage-activated (HVA) Ca2+ channel. Inhibition of protein synthesis attenuates the long-term reduction in Ca2+ current and has no effect upon the short-term Ca2+ current reduction. During the long-term reduction in Ca2+ current, it appears that Ca2+ channels located distant to the site of Ca2+ influx are affected. The relationship of these results to a previously described Ca(2+)-dependent reduction in transmitter release is discussed.

Animals

Seasonal differences in motor terminals.

1. Motor terminals undergo growth- and age-related changes throughout the lifetime of the animal in both vertebrates and invertebrates. 2. Motor terminals also show seasonal differences in transmitter release and morphology in both vertebrates and invertebrates. 3. Seasonal differences in motor terminals are likely to result from seasonal changes in motor activity and hormonal levels.

Animals

Seasonal differences in the physiology and morphology of crayfish motor terminals.

The physiology and morphology of identified crayfish motor terminals were compared at different seasons. We examined initial excitatory postsynaptic potential (EPSP) amplitudes, synaptic fatigue, and the frequency of synaptic varicosities along the motor terminals of an identified phasic motoneuron in animals collected over a period of 5 years. The physiology and morphology of identified crayfish motor terminals are different for animals collected in summer and winter. In winter animals, phasic axon motor terminals in the claw closer muscle produce large EPSPs initially, but show dramatic synaptic fatigue during repetitive stimulation. In summer animals, these terminals produce smaller initial EPSPs, but are more fatigue resistant. Due to their greater fatigue resistance, synaptic terminals have a greater over-all capacity for transmitter release in summer animals than do those of winter animals. Morphologically, terminals in summer animals have more synaptic varicosities, this result supports earlier studies that have shown that fatigue-resistant motor terminals have more synaptic varicosities. Experiments in which the electrical activity of the motoneuron was experimentally altered suggest that these differences in motor terminals may be due to seasonal differences in activity.

Acclimatization

Activity-dependent development of synaptic varicosities at crayfish motor terminals.

Tonically and phasically active crayfish motor terminals have well-characterized differences in synaptic physiology. During repetitive activation, the tonic terminals show facilitation and no depression, while the phasic terminals show dramatic synaptic fatigue. It has been proposed that this greater capacity for transmitter release from tonic terminals may be largely due to the presence of large synaptic varicosities along tonic terminals that contain large mitochondria, synapses, and numerous synaptic vesicles. In addition, a recent study indicates that in vivo tonic stimulation of a phasic terminal in young animals increases the fatigue resistance of the neuromuscular synapses and increases the number of synaptic varicosities, as well as the size of mitochondria and synapses. Because these previous morphological studies of crustacean motor terminals were performed using transmission electron microscopy, only short lengths of terminal were observed, and changes in synaptic varicosity frequency and size could not be thoroughly examined. In order to examine the synaptic varicosities along the terminal, motor terminals were injected with HRP, and the morphology of contrast-enhanced light microscopic images was examined. We report here that tonic terminals have much larger and more numerous synaptic varicosities than phasic terminals. In vivo stimulation of a phasic motoneuron results in an increase in the frequency of synaptic varicosities. This represents an increase in the total number of varicosities because terminal length appears to remain constant. These synaptic varicosities appear to form along preexisting terminals and persist for days after the final in vivo stimulation. The results indicate that the greater transmitter-releasing capabilities of tonic terminals compared to phasic terminals are due to their larger and more frequent synaptic varicosities. In addition, the development of these synaptic varicosities is activity dependent.

Animals

The refinement of invertebrate synapses during development.

Evidence is provided that during invertebrate development synapses undergo a period of refinement during which there are changes in synaptic connectivity and specific synaptic properties. It appears that extrinsic cues such as competition and neural activity are involved in guiding these synaptic changes in invertebrates. Comparisons are made with findings in the vertebrate literature.

Animals

Impulse activity of a crayfish motoneuron regulated its neuromuscular synaptic properties.

1. Previous studies have demonstrated that initial transmitter release, fatigability, and the morphology of identified crayfish neuromuscular synapses adapt to long-term changes in motoneuron impulse activity. 2. Experiments were performed to determine whether these long-term, adaptive alterations in neuromuscular synaptic physiology are triggered by changes in neuromuscular synaptic activity, muscle activity, or neuronal impulse activity. The fast closer excitor of the crayfish claw, a phasic motoneuron, was studied. Either the central or the peripheral region of the motoneuron was selectively stimulated in vivo by blocking impulse activity midway along the motor axon with localized application of tetrodotoxin and stimulating either central or distal to the blocked region. 3. Neither muscle activity nor transmitter release from the neuromuscular synapses was required to trigger the changes in synaptic physiology. Stimulation central to the block induced changes in neuromuscular transmission that included a long-lasting decrease in initial transmitter release and increased fatique resistance. 4. Because peripheral stimulation also produced decreased initial transmitter release, it appears that increased impulse activity in either region of the motoneuron can produce the synaptic changes. These results along with earlier findings suggest that neuronal depolarization induces adaptive, long-term changes in synapses. 5. These results are discussed in relation to findings at vertebrate and invertebrate synapses.

Adaptation, Physiological

Early experience influences the development of bilateral asymmetry in a lobster motoneuron.

The development of functional asymmetry between a pair of homologous motoneurons of the claw closer muscles in lobsters, Homarus americanus, was studied. In juvenile lobsters, 3-5 years old, where the paired claws are highly specialized into a major (crusher) and minor (cutter) type, the fast closer excitor (FCE) motoneuron fired longer bursts of spikes in the crusher claw compared to those in its cutter counterpart. The intraburst impulse frequency was greater for the cutter FCE and its neuromuscular synapses showed greater facilitation at these high impulse frequencies compared to that of the crusher claw. However, such asymmetry in firing patterns and synaptic facilitation was absent in lobsters raised without a substrate and having paired cutter claws. In the earliest juvenile stage, synaptic facilitation was similar between the paired claws and then developed in either an asymmetric or symmetric manner depending on whether the lobsters experienced a substrate or not. In a substrate-free environment asymmetry could be produced by exercising one of the claws during development, implicating bilateral differences in the reflexive activity of the claws as a control mechanism.

Action Potentials

Long-term changes in neuromuscular synapses with altered sensory input to a crayfish motoneuron.

Prolonged changes in crayfish motoneuron electrical activity result in adaptations in neuromuscular synapses which are consistent with findings at other synapses. In this study we establish that this long-term adaptation (LTA) of crayfish neuromuscular synapses to increased activation of the motoneuron does not require the activation of any other neurons. Selectively increasing the impulse activity of the relatively inactive fast closer excitor motoneuron (FCE) over a period of 7 days results in a 41% reduction in initial amplitude of the excitatory postsynaptic potential (EPSP), and a 42% decrease in synaptic fatigue. These changes in EPSP properties have been previously shown to be due to decreased initial transmitter release and greater sustained release of transmitter during prolonged stimulation. Chronic stimulation of sensory receptors known to produce subthreshold synaptic potentials in the central processes of the FCE elicits LTA of its neuromuscular synapses. The initial EPSP is decreased by 21%, and the synaptic fatigue is reduced by 17%. These results lead to the hypothesis that the primary event leading to LTA of neuromuscular synapses is depolarization of the motoneuron.

Adaptation, Physiological

Neuronal experience modifies synaptic long-term facilitation.

In a crayfish phasic neuromuscular junction, we have demonstrated low-frequency depression (LFD), high-frequency depression (HFD), and long-term facilitation (LTF) in response to different regimens of stimulation. Chronic stimulation of the phasic axon supplying the closer muscle of the claw in Procambarus clarkii resulted in diminished expression of HFD and LTF. Conversely, when impulse production in the phasic motoneuron was reduced by claw immobilization, both HFD and LTF were enhanced. LFD was insensitive to these manipulations. These results provide further evidence for long-term adaptation of the phasic neuromuscular junction to ongoing levels of impulse activity and illustrate the importance of a neuron's past history for synaptic plasticity. The ability of the neuron to adjust its short-term plasticity in response to altered experience constitutes an adaptive response that could be of general significance.

Adaptation, Physiological

Morphological transformation of synaptic terminals of a phasic motoneuron by long-term tonic stimulation.

In vivo stimulation of a relatively "silent" phasic crayfish motoneuron changes the ultrastructure of its synaptic terminals to a more tonic phenotype. The closer muscle of the crayfish claw is supplied by only 2 excitatory motoneurons, one of which is phasic and the other tonic. The ultrastructures of conditioned phasic, unconditioned phasic, and tonic motor terminals were compared. The terminals of the tonic motor axon were larger in cross-sectional area, had larger mitochondria, greater synaptic contact area, and were more varicose than unconditioned phasic terminals. Following long-term tonic stimulation of the phasic axon, its terminals became more varicose, mitochondrial cross-sectional area more than doubled, and synapses and mitochondria came into closer proximity, although mean terminal cross-sectional area did not change. Thus, the conditioned phasic terminals became more similar to those of the tonic motor axon. These changes in ultrastructure correlate with, and may be causally linked to, previously reported changes in neuromuscular synaptic physiology produced by in vivo tonic stimulation of this motoneuron. We conclude that the ongoing level of impulse activity can affect the ultrastructural differentiation of synaptic terminals and synapses of the phasic motoneuron.

Animals

Long-term facilitation and long-term adaptation at synapses of a crayfish phasic motoneuron.

Stimulation of the phasic (fast) motor axon of the isolated crayfish claw preparation at relatively low frequency (0.1 Hz) leads to depression of the excitatory junction potential (EJP) recorded from single muscle fibers. When the same stimulation is delivered following depression of the EJP at a higher frequency (5 Hz), a potentiated EJP appears, which is more resistant to low frequency depression. The potentiation appears to be analogous to "long-term facilitation" observed after stimulation of a tonic motor axon in crayfish and crabs. Long-term facilitation can be detected in preparations made from claws of animals in which the phasic motoneuron was stimulated at 5 Hz for 2 h in situ. This effect lasts for at least one day after one conditioning trial. Long-term facilitation is observed after stimulation of decentralized axons in situ, indicating that the change is attributable to local changes in terminal regions of the axon, and does not require the cell body. When electrodes are implanted in situ and the phasic motoneuron stimulated at 5 Hz for 2 h each day, synaptic depression becomes less pronounced and initial EJP amplitude becomes smaller over a period of several days. The latter changes, which adapt the neuron to a more tonic activity pattern, usually require several days for completion. Adaptation of fatigability occurs more rapidly than adaptation of initial EJP amplitude, and once established, remains for many days without further superimposed activity. Long-term adaptation does not occur in decentralized axons. Long-term facilitation and long-term adaptation are different responses of the neuron to enhanced activity. The former can occur in isolated or decentralized axons and leads to enhancement of EJP amplitude for a period of several hours to at least one day after a single episode of conditioning. The latter requires more time to be established, and leads to reduction of initial EJP amplitude and to lessened fatigability which persists for many days.

Adaptation, Physiological

Age-dependent long-term adaptation of crayfish phasic motor axon synapses to altered activity.

Crustacean tonic and phasic motoneurons have neuromuscular synaptic properties corresponding with their functional requirements. Phasic axon synapses produce large excitatory postsynaptic potentials (EPSPs) which depress rapidly during repetitive activation. Tonic axon synapses generally produce smaller EPSPs which are more resistant to fatigue. To test whether nerve impulse activity of the motoneuron plays a role in the establishment of these synaptic properties, a phasic axon was tonically stimulated in vivo. The "fast" closer excitor of the crayfish claw, which normally fires few impulses, was stimulated for 2 hr/day at 5 Hz, through implanted electrodes. In young crayfish, this stimulation produced an 11-fold decrease in synaptic fatigue at the fast axon's neuromuscular synapses, as determined from measurements of EPSPs during 5 Hz stimulation of the fast axon for 30 min. In comparison with EPSPs of the contralateral control claw, the initial EPSP amplitude was 44% smaller and the final EPSP amplitude was 4.3 times larger for the chronically stimulated fast axon. These changes in EPSP amplitude are due to changes in transmitter release. This long-term adaptation of the fast axon to imposed tonic activity persists for at least 10 days after the effect has been established. The same chronic stimulation regimen produces significant, although less dramatic, results in adult crayfish. Compared to the contralateral control, the chronically stimulated fast axon showed no change in initial EPSP amplitude and only a 2-fold increase in the EPSP amplitude after 30 min of stimulation at 5 Hz. Thus, the decrease in synaptic fatigue was only 2- to 3-fold, much less than in young crayfish.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological

Changes in electrical properties and quantal current during growth of identified muscle fibres in the crayfish.

The muscle fibre electrical properties, miniature excitatory junctional current (m.e.j.c.) and miniature excitatory junctional potential (m.e.j.p.) were studied during growth of an identified crayfish muscle fibre from a diameter of 20 to 400 microns. The specific membrane resistance (Rm), and the specific internal resistance (Ri), of the muscle fibre were independent of fibre diameter (d) during growth. The current-voltage relation has a similar shape in large and small fibres, indicating that voltage dependence of Rm does not change during growth. The input resistance (R0) was approximately proportional to d-1.5, as predicted theoretically. The specific membrane capacitance (Cm) and the membrane time constant (Tm) increased linearly with fibre diameter, apparently as a result of the contribution of the tubular capacitance to Cm. The decrease in R0 and the increase in Tm should have resulted in a 90-fold decrease in m.e.j.p. amplitude during growth of the fibre from a diameter of 20 to 240 microns. However, m.e.j.p. amplitude was found to decrease only 21-fold. This discrepancy was shown to result from an increase in m.e.j.c. amplitude and duration during growth. There was 2.9-fold increase in m.e.j.c. amplitude and a 2.7-fold increase in m.e.j.c. duration over the range of muscle fibre growth studied. This increase in the m.e.j.c. apparently results from an increase in the magnitude and duration of the synaptic conductance change produced by a quantum of transmitter. Throughout the range of muscle fibre diameters studied, the muscle fibre effective input impedance for the m.e.j.c. was 17-19% of R0. This is due to the relatively large Cm and the short duration of the m.e.j.c.

Action Potentials

Transmitter release during normal and altered growth of identified muscle fibres in the crayfish.

During growth of identified crayfish muscle fibres from a diameter of 20 to 400 micron, the excitatory junctional potential (e.j.p.) amplitude was found to be independent of diameter. Thus, e.j.p. amplitude was maintained during growth in spite of a 21-fold decrease in miniature excitatory junctional potential (m.e.j.p.) amplitude previously reported (Lnenicka & Mellon, 1983). The maintenance of e.j.p. amplitude was found to be partially due to a 5-fold increase in quantal release at 'active sites' during growth. In order to determine whether the increase in transmitter release can be regulated by the rate of muscle fibre growth, the rate of growth was experimentally reduced. By decreasing the resting length of the muscle during growth, the rate of increase in the diameter was reduced by approximately 50% compared with the contralateral control muscle fibres. The input resistance and the m.e.j.p. were appropriately larger in the smaller-diameter experimental fibres. However, e.j.p. amplitude in the experimental fibres was not significantly different from that in the contralateral control fibres. This was apparently due to the significantly smaller quantal release at active sites on the experimental fibres compared with control fibres. Thus, experimental alteration of the rate of muscle fibre growth results in regulation of transmitter release, suggesting that the muscle fibre may control the increase in transmitter release seen during normal growth.

Action Potentials