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K Appenteng

Publications and source records attributed to K Appenteng.

At least 19 recordsLinked to original sources

Monosynaptic EPSPs elicited by single interneurones and spindle afferents in trigeminal motoneurones of anaesthetized rats.

1. Our aim has been to quantify the monosynaptic connections of trigeminal interneurones and spindle afferents onto jaw-elevator motoneurones as a step towards identifying common features in organization of monosynaptic inputs onto motoneurones. We have used the intracellular variant of the spike-triggered averaging method to examine the connections of single identified trigeminal interneurones and jaw-elevator muscle spindle afferents onto single jaw-elevator motoneurones. The interneurones examined lay in the region immediately caudal to the trigeminal motor nucleus. The experiments were performed on rats anaesthetized with pentobarbitone, paralysed and artificially ventilated. 2. Ten EPSPs and eight IPSPs were obtained from examining the connections of seventeen interneurones to thirty-six motoneurones, suggesting a functional connectivity of 50% for individual interneurones onto elevator motoneurones. Fourteen EPSPs were obtained from examining the connections of thirteen spindle afferents onto twenty-seven motoneurones, giving a functional connectivity of 52% for individual spindle afferents onto elevator motoneurones. The amplitudes of the EPSPs elicited by interneurones ranged from 7-48 microV (mean = 17, S.D. = 12.5, n = 10) and from 7 to 289 microV (mean = 64, S.D. = 76.0, n = 14) for the spindle-mediated EPSPs; the difference in the two means was not significant (P = 0.07). 3. However, the amplitude of averaged responses obtained by signal averaging methods are dependent on the assumption that the postsynaptic response occurs following every impulse in the presynaptic neurone. We therefore estimated the percentage of sweeps which contained EPSPs triggered by the presynaptic neurone under study. In essence the method used consisted of visual inspection of the individual sweeps comprising an average in order to assess the occurrence of EPSPs within six separate time windows, each of duration +/- 0.3 ms. Five windows were placed at randomly selected times on average and were used to provide an estimate of the frequency of occurrence of randomly triggered EPSPs. The sixth window was centred on the start of the averaged EPSP and the frequency of occurrence of randomly triggered EPSPs was subtracted from the frequency of occurrence of EPSPs in this window to produce an estimate of the incidence of EPSPs triggered by the presynaptic neurone under study. 4. Values of the incidence of occurrence of EPSPs triggered by the presynaptic neurones ranged from 4.3 to 92% for the fifteen averaged EPSPs which could be analysed in this manner (two elicited by interneurones and thirteen by spindle afferents).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Quantitative analysis and postsynaptic targets of GABA-immunoreactive boutons within the rat trigeminal motor nucleus.

We have used the post-embedding immunogold labelling method using antibodies to gamma-aminobutyric acid (GABA) to obtain quantitative data on the distribution, frequency, postsynaptic targets and ultrastructural characteristics of GABA-immunoreactive (GABA-IR) boutons in the trigeminal motor nucleus of rats. We have also combined this method with horseradish peroxidase tracing to obtain specific evidence for termination of some GABA-IR boutons onto identified jaw-elevator motoneurones. Twenty-eight percent of all synapses in the motor nucleus involved GABA-IR boutons. Seventy-three percent of the GABA-IR boutons formed axo-dendritic synapses, 13% axo-somatic synapses and 14% axo-axonic synapses. Ninety-three percent of GABA-IR boutons formed symmetrical synapses. Overall, 58% of all boutons contained only flattened vesicles, while 26% contained round vesicles and 16% a mixture of vesicle types. Measurements of bouton cross sectional area, apposition length, and active zone length were obtained from serial reconstructions of 15 GABA-IR boutons and 30 unlabelled boutons. In each case mean values for GABA-IR boutons were significantly smaller than those for nonlabelled boutons.

Animals

Light and electron microscopical localisation of 5-HT-immunoreactive boutons in the rat trigeminal motor nucleus.

We have used pre-embedding EM immunohistochemical methods to obtain quantitative data on the frequency and post-synaptic targets of 5-hydroxytryptamine-immunoreactive (5-HT-IR) boutons within the rat V motor nucleus. Thirteen percent (69/531) of all synaptic contacts in the motor nucleus involved 5-HT-IR boutons. Seventy-four percent of 5-HT-IR boutons made axo-dendritic contacts, 20% axo-somatic contacts, and 6% axo-axonic contacts. We conclude that a significant fraction of boutons in the motor nucleus are 5-HT-IR and most contribute to postsynaptic rather than presynaptic effects on trigeminal motoneurones.

Animals

The morphology and electrical geometry of rat jaw-elevator motoneurones.

1. The aim of this work was to quantify both the morphology and electrical geometry of the dendritic trees of jaw-elevator motoneurones. To do this we have made intracellular recordings from identified motoneurones in anaesthetized rats, determined their membrane properties and then filled them with horseradish peroxidase by ionophoretic ejection. Four neurones were subsequently fully reconstructed and the lengths and diameters of all the dendritic segments measured. 2. The mean soma diameter was 25 microns and values of mean dendritic length for individual cells ranged from 514 to 773 microns. Dendrites branched on average 9.1 times to produce 10.2 end-terminations. Dendritic segments could be represented as constant diameter cylinders between branch points. Values of dendritic surface area ranged from 1.08 to 2.52 x 10(5) microns 2 and values of dendritic to total surface area from 98 to 99%. 3. At branch points the ratio of the summed diameters of the daughter dendrites to the 3/2 power against the parent dendrite to the 3/2 power was exactly 1.0. Therefore the individual branch points could be collapsed into a single cylinder. Furthermore for an individual dendrite the diameter of this cylinder remained constant with increasing electrical distance from the soma. Thus individual dendrites can be represented electrically as cylinders of constant diameter. 4. However dendrites of a given neurone terminated at different electrical distances from the soma. The equivalent-cylinder diameter of the combined dendritic tree remained constant over the proximal half and then showed a pronounced reduction over the distal half. The reduction in equivalent diameter could be ascribed to the termination of dendrites at differing electrical distances from the soma. Therefore the complete dendritic tree of these motoneurones is best represented as a cylinder over the proximal half of their electrical length but as a cone over the distal half.

Animals

Contrasting effects of urethane and pentobarbitone anaesthesia on the electrical properties of rat jaw-elevator motoneurones.

Our main finding is that elevator motoneurones do not show sustained firing to intracellular injections of depolarising current pulses in rats anaesthetised with urethane. In contrast, virtually all elevator motoneurones show sustained firing in pentobarbitone-anaesthetised rats. The differences in firing are not associated with significant differences in membrane potential, spike amplitudes, AHP amplitude or duration, input resistance, time constant or rheobase (P greater than 0.06 in all cases). However, there are clear differences in the extent of sag seen under the two anaesthetics and so we tentatively suggest that the anaesthetics may differ in their effects on the inward rectifier.

Anesthesia

Monosynaptic connexions of single V interneurones to the contralateral V motor nucleus in anaesthetised rats.

We have used the extracellular spike triggered averaging method to obtain evidence for a monosynaptic connexion of single V (trigeminal) interneurones, located in the region immediately caudal to the V motor nucleus, onto neurones within the contralateral V motor nucleus. The extracellular fields recorded in the contralateral nucleus are of smaller amplitude than those detected within the ipsilateral nucleus and the implications of this are discussed.

Action Potentials

The membrane properties and firing characteristics of rat jaw-elevator motoneurones.

1. We have determined the membrane and firing properties of fifty-six jaw-elevator motoneurones in rats that were anaesthetized with pentobarbitone, paralysed and artificially ventilated. 2. Forty-two neurones were identified as masseter motoneurones and fourteen as masseter synergist motoneurones. The membrane potentials for the sample ranged from -60 to -86 (mean = -68; S.D. = 7.3; n = 56), and spike amplitudes from 50 to 95 mV. The duration of the after-hyperpolarization following antidromic spikes in masseter motoneurones ranged from 15 to 50 ms (mean = 30; S.D. = 12.8) and their amplitudes from 1.0 to 4.5 mV (mean = 2.7; S.D. = 2.2; n = 42). 3. The mean input resistance for the total sample was 2.3 M omega (S.D. = 0.9; n = 56), membrane time constant 3.9 ms (S.D. = 0.9; n = 48) and rheobase 4.2 nA (S.D. = 2.6; n = 56). The distribution of these parameters was independent of membrane potential. We found no significant interrelationships between the membrane properties and one interpretation of this is that our sample may be drawn from a homogenous population of motoneurones. We also suggest that elevator motoneurones may have a lower Rm (specific membrane resistivity) value than cat hindlimb motoneurones because they have a similar range of input resistance values but only half the total surface area. 4. Forty-six out of forty-nine neurones fired repetitively to a depolarizing current pulse at a mean threshold of 1.6 x rheobase. Current-frequency plots were constructed for thirteen neurones and all but one showed a primary and secondary range in the firing of the first interspike interval. The mean slope in the primary range was 31 impulses s-1 nA-1 and 77 impulses s-1 nA-1 for the secondary range. The mean minimal firing frequency for steady firing was 26 impulses s-1 and, in response to an increase of stimulation, the rate increased monotonically with a slope of 11 impulses s-1 nA-1. 5. The dynamic sensitivity of twelve neurones was assessed from their response to ramp waveforms of current of constant amplitude but varying frequencies (0.2-2 Hz). Firing initially increased along a steep slope up to a frequency of between 40 and 60 impulses s-1 and then increased along a much shallower slope. Both the threshold for eliciting firing and the firing at the transition point of the two slopes remained constant with changes in ramp frequency.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials

The morphology of the axons and axon collaterals of rat jaw-elevator motoneurones.

We have made intracellular injections of horseradish peroxidase into the somata of jaw-elevator motoneurones and subsequently reconstructed the axonal morphology of 4 cells. In each case the axons gave off collaterals which were essentially restricted to the ventral portion of the V motor nucleus. This observation provides the first evidence that these motoneurones may exert recurrent synaptic effects.

Action Potentials

The monosynaptic excitatory connections of single trigeminal interneurones to the V motor nucleus of the rat.

1. We have used the extracellular spike-triggered averaging method to identify a population of trigeminal interneurones that make monosynaptic connections within the V motor nucleus. The experiments were performed on rats anaesthetized with pentobarbitone (60 mg/kg I.V.; supplementary doses given as necessary to maintain a deep level of anaesthesia). 2. A tungsten microelectrode (tip exposure of some 200 microns) was inserted into the masseter motoneurone pool to allow recording of extracellular activity. A glass electrode filled with DL-homocysteic acid was used to make simultaneous extracellular recordings of the firing of single neurones in the region immediately caudal to the motor nucleus. 3. Fifty-eight out of 166 interneurones tested gave unitary extracellular fields in the motor nucleus. The responses consisted of a terminal spike (presynaptic spike) followed by a negative field of duration approximately 3 ms and amplitude 0.4-10.8 microV. The mean latency between the positive peak of the terminal spike and the onset of the field (synaptic delay) was 0.43 ms (S.D. = 0.10 ms), and the mean latency from the onset of the interneurone spike to the positive peak of the presynaptic spike was 0.35 ms (S.D. = 0.22 ms). Thus the interneurones project directly to the motor nucleus where they then make monosynaptic connections. 4. The negative extracellular fields were taken to reflect an excitatory synaptic input onto neurones within the motor nucleus. The fields were of maximum amplitude at the point of maximum masseter motoneurone antidromic field, suggesting that the connection may be onto elevator motoneurones. 5. The location of the interneurone appeared to the main factor governing the likelihood of obtaining a field. Interneurones located more than 400 microns from the caudal border of the motor nucleus rarely produced fields whereas most of those located within this distance gave fields. This pattern of distribution is strikingly similar to that of trigeminal interneurones labelled by the transneuronal transport of wheatgerm agglutinin-horseradish peroxidase after an intramuscular injection of the tracer into the masseter muscle. We suggest that this provides electrophysiological support for the suggestion that the tracer does indeed label interneurones by means of retrograde transsynaptic transport.

Animals

Visualisation of CGRP and ChAT-like immunoreactivity in identified trigeminal neurones by combined peroxidase and alkaline phosphatase enzymatic reactions.

We report here a method that allows simultaneous visualisation of two antigens within single neurones. In essence this involves the combined use of horseradish peroxidase and alkaline phosphatase reactions to visualise two markers. Using this method we show that ChAT-and CGRP-like immunoreactivity can be co-localised within single neurones of the V to VII motor nuclei. In the case of the V motor nucleus, we show that each marker can be localised in motoneurones labelled with horseradish peroxidase.

Abducens Nerve

Transneuronal transport of wheat germ agglutinin-conjugated horseradish peroxidase into trigeminal interneurones of the rat.

Intramuscular injections of either horseradish peroxidase (HRP) or wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) were made into the masseter muscle of rats. Both tracers labeled primary sensory neurones in the V mesencephalic nucleus, motoneurones in the V motor nucleus, and some motoneurones in the facial motor nucleus. WGA-HRP labeled additional neurones in the V main sensory nucleus and the rostral pole of the V nucleus oralis. These were classed as interneurones because they lay in areas outside those known to contain either first-order afferent or motoneurone somata. We argue that these were labeled by retrograde transport of tracer because they lay close to the V motor nucleus, and from some of them processes could be followed into the region of the V motor nucleus.

Animals

'Fusimotor set': new evidence for alpha-independent control of gamma-motoneurones during movement in the awake cat.

The discharge activity of single muscle spindle receptors was recorded in freely moving cats. Large changes in responsiveness to length variations was observed in different types of movement. In separate stimulations on anaesthetised cats, the activity of the gamma-fusimotor neurones responsible for these changes was reconstructed. The results suggested that fusimotor action on a given spindle afferent during the movements studied was not rigidly alpha-linked, but 'set' by the CNS to steady levels, and that it could switch from largely static (gamma s) to largely dynamic (gamma d) according to the motor tasks performed.

Animals

Morphological and electrophysiological determination of the projections of jaw-elevator muscle spindle afferents in rats.

The fluorescent compound Lucifer Yellow was injected into the somata of nine identified jaw-elevator muscle spindle afferents, located in the V mesencephalic nucleus. Reconstructions of the central course of their axons were subsequently made from serial, transverse, sections to identify sites of projection. Three sites of termination were identified on the basis of collaterals that ended in varicosities and/or boutons. All afferents projected to the V nucleus oralis and, all but one, also to the V motor nucleus. Two out of nine afferents had terminations in the supra-trigeminal nucleus, though a further four appeared to send collaterals to this area. The relative density of projection, judged by the number of collaterals supplied to each area, decreased in the order: V nucleus oralis, V motor nucleus and supra-trigeminal nucleus. The central course of the afferent axons was such that impulses from the periphery would arrive first at the V motor nucleus, then the V nucleus oralis, the supra-trigeminal nucleus, and finally the afferent somata in the V mesencephalic nucleus. In animals in which the masseter nerve was exposed in-continuity for electrical stimulation, electrophysiological recordings were made in the three areas described above to identify units that received a monosynaptic input from spindles in the masseter muscle. Criteria were formulated on the basis of the pattern of responses on stimulation of the masseter nerve, and the morphology of labelled neurones, for differentiating between afferents, interneurones, and motoneurones. In the V motor nucleus, monosynaptic excitatory post-synaptic potentials (e.p.s.p.s) were obtained in both synergist and masseter motoneurones. These were assumed to arise from a masseter muscle spindle input as the thresholds for exciting such afferents and eliciting e.p.s.p.s were similar. Some interneurones, chiefly in the V nucleus oralis, were activated at thresholds close to that of muscle spindle afferents and could also fire in response to muscle stretch. As their latencies (measured extracellularly) were similar to that of e.p.s.p.s in motoneurones, they were assumed to receive a monosynaptic muscle spindle input. However, most interneurones were activated at longer latencies (up to 7 ms) and some also fired to muscle stretch. Arguments are advanced, based on the long rise time of e.p.s.p.s recorded in some, that the majority of these may also be candidates for monosynaptic activation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Tendon organ firing during active muscle lengthening in awake, normally behaving cats.

Recordings were obtained of the discharge of single tendon organ (Ib) and muscle spindle (Ia) afferents of the ankle extensor muscles during movement in normal cats. During very slow, smooth increases and decreases in muscle force, Ib afferents showed from one to five stepwise changes in firing rate, attributable to the recruitment of motor units inserting into the receptor capsule. These 'recruitment steps' in Ib firing rate became smoothed and tended to merge during faster variations in muscle force, and were rarely discernible in normal movements such as slow stepping. Rapid imposed stretches resulted in Ib firing patterns which fitted well a dynamic function of whole muscle force. Comparisons were made between the responses of Ib and Ia afferents during rapid, imposed muscle stretch. The segmentation of discharge typical of Ia afferents was not present in Ib afferents, despite segmentation of the e.m.g. of the receptor-bearing muscles. This would imply that Ib afferents exert a rapidly fluctuating reflex action against a relatively steady background of Ib input. Ankle extensor Ib firing during stepping was characterized by feeble firing during the swing phase and substantial, smoothly modulated firing during the stance phase. Taken together with previous chronic recordings, the data support the view that the ensemble of Ib afferents from a muscle signals a dynamic, non-linear function of whole muscle force over a wide range of normal movement.

Action Potentials

Effect of fusimotor stimulation on ia discharge during shortening of cat soleus muscle at different speeds.

1. In barbiturate-anaesthetized cats, the L7 and S1 dorsal and ventral roots were dissected to isolate functionally single afferents identified as primary endings of soleus muscle spindles, and motor filaments which exerted a fusimotor action on the afferents with limited action on extrafusal muscle. Up to seven filaments, with an action on a given primary ending, could be isolated and each was classified as exerting either a predominantly dynamic or static action.2. Combined stimulation of these filaments, at rates up to 200 impulses/s could maintain afferent firing during muscle shortenings at speeds up to 200 mm/s.3. Fusimotor stimulation could also maintain afferent firing at a target frequency of 100 impulses/s during muscle shortenings up to 200 mm/s. The timing, in relation to the onset of shortening, and the rates of fusimotor stimulation were found to be critical in achieving the target frequency.4. Sinusoidal modulation of the frequency of fusimotor stimulation was used to study the conditions required to achieve constant afferent firing in the face of imposed sinusoidal length changes.5. For given depths of modulation, the phase advance of fusimotor stimulation needed to produce minimum modulation of afferent firing (best compensation) increased with increasing frequency of the sinusoids. The compensation deteriorated with an increase in the frequency of the sinusoids and a change in the mean muscle lengths, although in some cases it could be restored by adjustments to the depth of modulation of fusimotor rate. This suggests that for movements of varying speeds and amplitudes, settings which are appropriate for shortening at a given velocity and mean muscle length, do not apply if either of these two variables are altered.6. These findings demonstrate that the fusimotor system is potentially capable of eliciting constant afferent firing as envisaged in the ;servo-assistance' hypothesis (Matthews, 1964, 1972; Stein, 1974). This, and the fact that constant afferent firing is not seen during normal unobstructed shortenings at velocities greater than 0.2 resting length/s (Prochazka, 1981), are used to argue that it is by choice rather than necessity that ;servo-assistance' (as defined above) is not employed during normal movements. However, servo-assistance of a different form (involving modulated spindle afferent feed-back from both agonists and antagonists) remains a viable alternative.

Animals

Proprioceptive input from the jaw muscles and its influence on lapping, chewing, and posture.

Experiments are described which define criteria for identifying fusimotor axons discharging in filaments of the masseter nerve in lightly anaesthetized cats. During reflex movements of the jaw two patterns of discharge were observed in different fusimotor fibres. One type, called "sustained," fired at a fairly constant increased rate. The other, called "modulated," fired at high frequencies during the extrafusal muscle contraction. Evidence from spindle primary and secondary recordings in similar experiments strongly suggests that the "sustained" type were dynamic fusimotor fibres and the "modulated" type were static fusimotor fibres. New spindle recordings in normal unanaesthetized cats indicate that the modulated pattern of static fusimotor discharge also occurs in these conditions. Its effect is to reduce the tendency for spindle afferents to become silent during muscle shortening. A proposal is made that the static fusimotor discharge in cyclic movements may represent a temporal "template" for the intended movement as directed by the central pattern generator.

Animals

Fusimotor activity in masseter nerve of the cat during reflex jaw movements.

1. Unit recordings have been made from the central ends of filaments of the masseter nerve in lightly anaesthetized cats. Evidence is presented to show that fusimotor activity may be distinguished from alpha motor activity. 2. During reflex cyclic movements induced by intra-oral stimulation, two distinct patterns of fusimotor firing emerged. One type of unit increased firing at the beginning and sustained this with little modulation throughout the movements. The other type was strongly modulated approximately in parallel with the alpha motor activity. 3. By comparison with records of jaw elevator spindle afferents under similar conditions, it was deduced that the sustained type of action was due to dynamic fusimotor neurones while the modulated type was due to static fusimotor neurones. 4. The patterns of fusimotor activity seen in these rhythmic movements under light anaesthesia agree well with the patterns deduced from spindle recordings in the conscious cat during mastication. 5. The results emphasize the importance of looking beyond a simple hypothesis of 'alpha-gamma co-activation' to explain fusimotor function. It is proposed that tonic dynamic fusimotor activity is set at the beginning of a movement to determine the incremental sensitivity of primary endings to stretch. The static fusimotor fibres are activated principally during shortening to help keep both primary and secondary endings active.

Animals