Eupatilin improves cilia defects in human CEP290 ciliopathy models.

The photoreceptor outer segment is a highly specialized primary cilium essential for phototransduction and vision. Biallelic pathogenic variants in the cilia-associated gene CEP290 cause non-syndromic Leber congenital amaurosis 10 (LCA10) and syndromic diseases, where the retina is also affected. While RNA antisense oligonucleotides and gene editing are potential treatment options for the common deep intronic variant c.2991+1655A>G in CEP290 , there is a need for variant-independent approaches that could be applied to a broader spectrum of ciliopathies. Here, we generated several distinct human models of CEP290 -related retinal disease and investigated the effects of the flavonoid eupatilin as a potential treatment. Eupatilin improved cilium formation and length in CEP290 LCA10 patient-derived fibroblasts, in gene-edited CEP290 knockout (CEP290 KO) RPE1 cells, and in both CEP290 LCA10 and CEP290 KO iPSCs-derived retinal organoids. Furthermore, eupatilin reduced rhodopsin retention in the outer nuclear layer of CEP290 LCA10 retinal organoids. Eupatilin altered gene transcription in retinal organoids, by modulating the expression of rhodopsin, and by targeting cilia and synaptic plasticity pathways. This work sheds light into the mechanism of action of eupatilin, and supports its potential as a variant-independent approach for CEP290 -associated ciliopathies.

Targeting MYC in multiple myeloma.

Multiple myeloma (MM) is a plasma cell tumor marked by clonal evolution and preceded by a premalignant stage, which progresses via molecular pathway deregulation, including MYC activation. This activation relates to translocation or gain of the MYC locus and deregulation of upstream pathways such as IRF4, DIS3/LIN28B/let-7, or MAPK. Precision medicine is an approach to predict more accurately which treatment strategies for a particular disease will work in which groups of patients, in contrast to a "one-size-fits-all" approach. The knowledge of mechanisms responsible for MYC deregulation in MM enables identification of vulnerabilities and therapeutic targets in MYC-driven tumors. MYC can be targeted directly or indirectly, by interacting with several of its functions in cancer. Several such therapeutic strategies are evaluated in clinical trials in MM. In this review, we describe the mechanism of MYC activation in MM, the role of MYC in cancer progression, and the therapeutic options to targeting MYC.

The route to pathologies in chronic inflammatory diseases characterized by T helper type 2 immune cells.

T helper type 2 (Th2)-characterized inflammatory responses are highly dynamic processes initiated by epithelial cell damage resulting in remodelling of the tissue architecture to prevent further harm caused by a dysfunctional epithelial barrier or migrating parasites. This process is a temporal and spatial response which requires communication between immobile cells such as epithelial, endothelial, fibroblast and muscle cells and the highly mobile cells of the innate and adaptive immunity. It is further characterized by a high cellular plasticity that enables the cells to adapt to a specific inflammatory milieu. Incipiently, this milieu is shaped by cytokines released from epithelial cells, which stimulate Th2, innate lymphoid and invariant natural killer (NK) T cells to secrete Th2 cytokines and to activate dendritic cells which results in the further differentiation of Th2 cells. This milieu promotes wound-healing processes which are beneficial in parasitic infections or toxin exposure but account for increasingly dysfunctional vital organs, such as the lung in the case of asthma and the colon in ulcerative colitis. A better understanding of the dynamics underlying relapses and remissions might lead ultimately to improved therapeutics for chronic inflammatory diseases adapted to individual needs and to different phases of the inflammation.

Radiation therapy education for rural and remote GPs.

INTRODUCTION: Radiation therapy (RT) plays an important role in the treatment of many malignancies, either as a primary treatment or adjuvant modality. The referral base for many New South Wales (NSW) radiation oncology departments extends across rural and remote NSW regions. There are limited resources and support available to GPs in these rural areas to assist them in caring for patients considering or about to undergo RT, and those who have completed RT treatment and have returned to their rural residence. The project described aimed to develop an electronic learning (e-learning) information resource for: (1) both health professionals and patients in the NSW rural sector on general RT information; and (2) GPs on the specifics of radiation induced skin reactions. In order to produce a comprehensive information package and resource for rural GPs and their patients, a needs assessment was conducted on a sample GP population. METHODS: The needs assessment was conducted via distribution of a survey to 1700 rural GPs throughout NSW. The survey was developed using patient and clinician input; and SPSS software (SPSS Inc; Chicago, IL, USA) was used for data analysis. The collected data were analysed descriptively to quantify the GPs' responses. RESULTS: A response rate of 22% was achieved. Of the respondents, 93.9% had previously cared for a patient undergoing radiation therapy, and 76.7% felt they had insufficient information to support their patient through this process. In total, 96.1% of the GPs indicated the need for information about acute and chronic radiation induced skin reactions. The need for educational material to be available in both hard copy and electronically was identified. CONCLUSION: The results indicate that most GPs have cared for an RT patient and few felt they had sufficient information. There was genuine interest from the respondents in obtaining an information and resource package that would assist them in the care of these patients.

Terpenoids from Picris echoides.

A new monoterpenoid glycoside, named (-)-cis-chrysanthenol-beta-D-galactopyranoside (1), was isolated from the air-dried parts of Picris echoides along with the known compounds (-)-cis-chrysanthenol and 1alpha-hydroxyeudesma-2,4(15),11(13)-trien-5alpha,7alphaH-12-oic acid.

Differential distribution of interneurons in the neural networks that control walking in the mudpuppy (Necturus maculatus) spinal cord.

Locomotor behavior is believed to be produced by interneuronal networks that are intrinsically organized to generate the underlying complex spatiotemporal patterns. In order to study the temporal correlation between the firing of individual interneurons and the pattern of locomotion, we utilized the spinal cord-forelimb preparation from the mudpuppy, in which electrophysiological recordings of neuronal activity were achieved during walking-like movement of the forelimb induced by bath application of N-methyl- D-aspartate (NMDA). Intra- and extracellular recordings were made in the C2 and C3 segments of the spinal cord. These segments contain independent flexor and extensor centers for the forelimb movement about the elbow joint during walking. Among the 289 cells recorded in the intermediate gray matter (an area between the ventral and dorsal horns) of the C2 and C3 segments, approximately 40% of the cells fired rhythmically during "walking." The firing rates were 6.4+/-0.4 impulses/s (mean +/- SE). These rhythmically active cells were classified into four types based on their phase of activity during a normalized step cycle. About half the rhythmic cells fired in phase with either the flexor (F) or extensor (E) motoneurons. The rest fired in the transitions between the two phases (F-->E and E-->F). Longitudinal distributions of the four types of interneurons along the spinal cord were in agreement with observations that revealed distinct but overlapping flexor and extensor centers for walking. Some cells triggered short-latency responses in the elbow flexor or extensor muscles and may be last-order interneurons. These observations suggest that there is a differential distribution of phase-specific interneurons in the central pattern generator of the mudpuppy spinal cord for walking.

Intrafascicular electrodes for stimulation and recording from mudpuppy spinal roots.

This paper presents a technique for stimulating and recording from multiple intact spinal roots in the in vitro mudpuppy (Necturus maculatus) spinal cord-forearm preparation using fine wire electrodes, a modified intrafascicular electrode. We found that multiple spinal roots of the preparation could be implanted with these modified electrodes for independent stimulation or recording of the roots without inducing mechanical vibrations, disrupting conduction, or obscuring the view of or access to the spinal cord. Recording and stimulation performance using these electrodes was compared with results obtained using conventional hook electrodes. We found that intrafascicular electrodes were more efficient than hook electrodes for stimulating nerve fibers, being able to produce equivalent levels of activation using stimulation levels that were an order of magnitude smaller. Compound action potential signals recorded from electrodes implanted in the spinal roots were found to be larger than those from hook electrodes placed around the corresponding spinal nerve, showing that intrafascicular electrodes are more efficient at recording activity in the nerve. Moreover, it was possible to record evoked activity from cutaneous mechanoreceptors, even though the signal to noise ratio was low. Rough estimates of the conduction velocities for the fastest components in the compound action potentials were calculated and found to be around 17.5 m/s for both dorsal and ventral roots.

Effects of inhibitory neurotransmitters on the mudpuppy (Necturus maculatus) locomotor pattern in vitro.

Effects of inhibitory neurotransmitters on the locomotor rhythm and pattern generation were investigated using an in vitro preparation isolated from the mudpuppy (Necturus maculatus). The preparation consisted of the first five segments of the spinal cord and the right forelimb attached by the brachial nerves. During N-methyl-d-aspartate (NMDA)-induced locomotion, the rhythmic motor output (EMG) was recorded unilaterally from elbow flexor and extensor muscles. While neither glycine nor gamma-aminobutyric acid (GABA)-related substances induced locomotion in the absence of NMDA, they modulated NMDA-induced locomotion. Bath application of glycine and GABA suppressed the rhythmic motor pattern induced by NMDA. Addition of glycine receptor antagonist strychnine or GABA(A) receptor antagonist bicuculline disrupted the phase relationship between antagonistic motor pools during ongoing locomotion, thereby changing the normal alternating pattern into synchronous EMG bursts. Both the GABA(A) receptor agonist muscimol and GABA(B) receptor agonist baclofen mimicked the effects of GABA as they either slowed down or stopped locomotion. Nipecotic acid, a GABA uptake blocker, had a similar effect. This suggested that an endogenous release of GABA modulated the locomotor rhythm. The endogenous release was antagonized by the GABA(A) and GABA(B) receptor antagonists bicuculline and CGP-35348, respectively. Immunocytochemistry revealed that glycine and GABA-positive neurons and fibers were present in mudpuppy spinal cord. Although the GABAergic neurons were more numerous than glycinergic neurons, both cell types contributed processes directed towards the white matter and occasionally towards the ependymal lining of the central canal. Our results suggest that inhibitory neurotransmitters exert powerful actions upon the neuronal network governing forelimb locomotion in the mudpuppy. The effects we observed may be mediated by a network of segmentally distributed glycinergic and GABAergic spinal neurons.

Identification, localization, and modulation of neural networks for walking in the mudpuppy (Necturus maculatus) spinal cord.

We tested the hypothesis that the neural networks for walking in the mudpuppy can be divided into a flexor and an extensor center, each of which contains collections of interneurons localized in the vicinity of their motoneuron pools. Combining a battery of techniques, we identified and localized the elbow flexor center and its motoneuron pool in the C2 segment and the elbow extensor center and its motoneuron pool in the C3 segment. Rhythmic flexion or extension of the limb in isolation could be induced by continuous trains of current pulses of the C2 or C3 segments, respectively. Independent activation could also occur after application of glutamate receptor agonist NMDA. Part of segment C2 in isolation generated rhythmic elbow flexor bursts, whereas part of segment C3 in isolation generated rhythmic elbow extensor bursts. An isolated region spanning the C3 roots generated both flexor and extensor bursts. The step cycle was modulated in a phase-dependent manner by stimulation of the dorsal roots, the ventral roots, or either of the two centers. The effects of ventral root stimulation were removed by deafferentation to block reafferent input attributable to muscle contraction induced by the stimulation. We conclude that the neural networks for walking contain at least a flexor and an extensor generator that are localized in close apposition to the motoneuron pools, that the two centers can work independently despite the fact that there are reciprocal inhibitory interconnections between them, and that sensory input interacts with the spinal neural networks to reset the ongoing walking rhythm in a phase-dependent manner.

Serotonergic modulation of the mudpuppy (Necturus maculatus) locomotor pattern in vitro.

The aims of the present study were to: (1) study the role of serotonin (5-HT) in modulating the central pattern generator (CPG) underlying locomotion in the mudpuppy (Necturus maculatus); (2) investigate whether there is an intrinsic spinal serotonergic system. These aims were achieved by the use of pharmacological and immunocytochemical methods. To study modulation of the locomotor pattern and rhythm, we applied 5-HT, its uptake blocker zimelidine, and a variety of 5-HT receptor agonists and antagonists to an in vitro brainstem-spinal cord preparation isolated from the mudpuppy. The preparation consisted of the first five segments of the spinal cord and the right forelimb attached by the brachial plexus. The spinal CPG for locomotion was activated chemically by adding NMDA to the superfusing solution. During locomotion, bipolar electromyographic (EMG) recordings were made unilaterally from flexor and extensor ulnae muscles. 5-HT on its own did not induce locomotion, but it did have a profound modulatory effect on NMDA-induced locomotion. 5-HT produced a dose-dependent increase in the overall cycle duration and enhanced the EMG burst duration. Use of zimelidine indicated that there is an endogenous release of 5-HT which modulated the locomotor rhythm. The endogenous release was antagonized by 5-HT1/5-HT2 receptor antagonist methiothepin. Immunocytochemical analysis, in which the entire spinal cord of the mudpuppy was used, revealed that there were more than one type of spinal serotonergic neuron. They were differentiated according to the cell diameter, shape, and arborization pattern of their processes. These neurons were located within the central gray matter ventrolateral to the central canal. Our results suggest that 5-HT plays an important role in modulating the locomotor CPG in the mudpuppy, by acting through a well-developed spinal serotonergic system. This is in contrast to what has been reported in higher vertebrates, where serotonergic innervation is derived from supraspinal structures.

Workload and use factor of medical linear accelerators in radiotherapy.

An important factor in the design of primary protective barriers is the use factor. The present study was aimed at obtaining historical data on the use factor of two dual modality linear accelerators in a radiotherapy department. Gantry angle, field size, and beam modifiers were recorded for all radiation qualities in use at two medical linear accelerators with 6 MV and 18 MV x-rays and multiple electron energies ranging from 4 MeV to 20 MeV. The data for one year of clinical use was extracted from a record and verifying system and an estimate of the physics workload on the machines was obtained by going through the quality assurance records and machine log books. Of the total dose of approximately 37,000 Gy delivered in one year at isocenter on each unit 80% was given as 6 MV x-rays. As can be expected, most x-ray beams were directed at the four cardinal gantry angles with the angular distribution for 6 MV and 18 MV x-rays being very similar. Electron fields were broadly distributed around the gantry pointing down position. Less than 25% of all clinical x-ray treatment fields extended beyond a field size of 200 cm2.

Fusimotor responses to fatiguing muscle contractions in non-denervated hindlimb of decerebrate cats.

Changes in discharge rate of 21 fusimotor neurons to medial gastrocnemius muscle during long-lasting fatiguing contractions of lateral gastrocnemius and soleus muscles were recorded in decerebrate cats with innervation of the same hindlimb preserved. Both the spontaneous activity and reflex responses of fusimotor neurons differed from those found previously in preparations with denervated hindlimb. Higher proportion of units fired at rest at rates above 20 impulses/s, the initial increase in discharge rate at the onset of muscle contraction was markedly prolonged, lasting in the majority of units throughout the muscle contraction, while the late increase in discharge rate developing with muscle fatigue was either absent or short-lasting. It is suggested that the increase in spontaneous firing rate occurs in dynamic fusimotor neurons being supported by afferent inflow from secondary muscle spindle endings from non-contracting muscles, the enhancement of the early responses to be primarily due to recurrent disinhibition and the differences in changes of early and late responses to reflect their partly different origin. The possibility is raised that the late reflex responses are lacking more often in static than in dynamic fusimotor neurons. From the functional point of view the differences in fusimotor reflex responses in innervated versus denervated hindlimb may indicate their susceptibility to modifications by changes in afferent inflow according, supposedly, to the current demands of motor control of the active and/or fatigued muscle.

The activity of interneurons during locomotion in the in vitro necturus spinal cord.

1. Less than two segments of the cervical spinal cord of the mudpuppy (Necturus maculatus) is sufficient to generate a locomotor rhythm with application of N-methyl-D-aspartic acid (NMDA). We have recorded intracellularly from rhythmically active interneurons in these segments and classified them according to their phase of firing within the step cycle and their afferent input. 2. Four classes of interneurons were found: flexor, flexor-->extensor, extensor, and extensor-->flexor. Interneurons that burst during the transition from flexion to extension or vice versa are referred to as "transitional" interneurons and represent the majority (68%) of rhythmically active interneurons studied in the mudpuppy spinal cord. 3. All flexor interneurons received only inhibitory input from cutaneous and dorsal root afferents, whereas the flexor-->extensor interneurons that responded received only excitatory input from dorsal root and cutaneous afferents. All extensor interneurons and all but one extensor-->flexor interneuron received no afferent input from the cutaneous or dorsal root afferents we stimulated. 4. Other interneurons have been classified as "tonic" cells. They fire continuously when the mudpuppy is walking and are silent when the mudpuppy is not walking. These interneurons receive no afferent input from the sources tested and may be responsible for turning locomotion on and off. 5. In conclusion, the presence of many transitional interneurons with specific patterns of afferent input may be required for the phasing of legged locomotion. We believe the in vitro preparation of the mudpuppy spinal cord and forelimb is an excellent model for studying the firing properties of interneurons during legged locomotion.

Membrane potential changes of skeletomotor neurons in response to random stretches of the triceps surae muscles in decerebrate cats.

The properties of membrane potential changes of skeletomotor neurons (S, FR, and FF) innervating triceps surae muscles during pseudorandom stretching of these muscles were studied in decerebrate cats. Peak amplitudes of pseudorandom muscle stretches ranged from 119 microns to 4.15 mm peak-to-peak. Sequences of ten identical stretching periods were applied for averaging. Shapes of membrane potential changes and probability density distribution of amplitudes of the input and output signals and power spectra suggest that the skeleto-motor neuron membrane has nonlinear properties. First- and second-order Wiener kernels were determined by applying the cross-correlation (Lee-Schetzen) method. The results suggest that the transfer function between muscle stretches and subthreshold membrane potentials is a Wiener-type cascade. This cascade is consistent with a linear, second-order, underdamped transfer function followed by a simple quadratic nonlinearity [linear (L) system followed by nonlinear (N) system, or LN cascade]. Including the nonlinear component calculated from the second-order Wiener kernel improved the model significantly over its linear counterpart, especially in S-type motoneurons. Qualitatively similar results were obtained with all types of motoneurons studied.

Spike discharges of skeletomotor neurons during random noise modulated transmembrane current stimulation and muscle stretch.

Spike discharges of skeletomotor neurons innervating triceps surae muscles elicited by white noise modulated transmembrane current stimulation and muscle stretch were studied in decerebrated cats. The white noise modulated current intensity ranged from 4.3 to 63.2 nA peak-to-peak, while muscle stretches ranged from 100 microns to 4.26 mm peak-to-peak. The neuronal responses were studied by averaging the muscle length records centered at the skeletomotor action potentials (peri-spike average, PSA) and by Wiener analysis. Skeletomotor spikes appeared after a sharp peak in PSA of the injected current, preceded by a longer-lasting smaller wavelet of either depolarizing or hyperpolarizing direction. The PSA amplitude was not related to the injected current amplitude nor showed any differences related to the motor unit type. The PSA amplitudes were virtually independent of the stretching amplitude sigma, after an initial increase with stretching amplitudes in the range of 15-40 microns (S.D.), or 100-270 microns peak-to-peak. Analyses of cross-spectra indicated a small or absent increase in gain with frequency in response to injected current, but about 20 dB/decade in the range 10-100 Hz in response to muscle stretch. The peaks of both Wiener kernels in response to current injection appear to decrease with the amplitude of injected current, but this decrease was not statistically significant. The narrow first-order kernels suggest that the transfer function between the current input and spike discharge is lowpass with a wide passband, i.e. there is very little change in dynamics. The values of the second-order kernels appear to be nonzero only along the main diagonal.(ABSTRACT TRUNCATED AT 250 WORDS)

Sensory nerve recording for closed-loop control to restore motor functions.

A method is developed for using neural recordings to control functional electrical stimulation (FES) to nerves and muscles. Experiments were done in chronic cats with a goal of designing a rule-based controller to generate rhythmic movements of the ankle joint during treadmill locomotion. Neural signals from the tibial and superficial peroneal nerves were recorded with cuff electrodes and processed simultaneously with muscular signals from ankle flexors and extensors in the cat's hind limb. Cuff electrodes are an effective method for long-term chronic recording in peripheral nerves without causing discomfort or damage to the nerve. For real-time operation we designed a low-noise amplifier with a blanking circuit to minimize stimulation artifacts. We used threshold detection to design a simple rule-based control and compared its output to the pattern determined using adaptive neural networks. Both the threshold detection and adaptive networks are robust enough to accommodate the variability in neural recordings. The adaptive logic network used for this study is effective in mapping transfer functions and therefore applicable for determination of gait invariants to be used for closed-loop control in an FES system. Simple rule-bases will probably be chosen for initial applications to human patients. However, more complex FES applications require more complex rule-bases and better mapping of continuous neural recordings and muscular activity. Adaptive neural networks have promise for these more complex applications.

Changes in discharge rate of cat hamstring fusimotor neurones during fatiguing contractions of triceps surae muscles.

Changes in the discharge rate of fusimotor neurones to hamstring muscles during long-lasting, fatiguing, contractions of the triceps surae muscles were studied in decerebrate cats. Discharges of fusimotor neurones were recorded from the nerve filaments. Muscle contractions were elicited by electrical stimulation of the muscle nerves applied until the muscle tension fell to about 30% of its initial value. Early and late changes could be recognized in fusimotor discharge rate. The early changes, at the onset of muscle contraction, occurred in 9 out of 22 neurones and varied in both sign and duration among the cells. The late change, encountered in 16 fusimotor neurones, was an increase in discharge rate developing towards the end of the muscle contraction and outlasting it. When the contracting triceps muscle was made ischaemic the late increase in discharge rate developed earlier, as did the muscle tension fall and started to subside after the arterial clamp was removed. After severing the muscle nerves their stimulation provoked either no changes or a slight sustained decrease in fusimotor discharge rate. It is supposed that the late increase in discharge rate of fusimotor neurones to hamstring muscles appears due to reflex excitation by discharges in group III and IV afferent fibres from the triceps muscle provoked and/or enhanced by metabolic products liberated during its fatiguing contraction. The possibility is raised that the excitation is elicited primarily by the discharges from chemosensitive afferent fibres. Its functional role in muscle fatigue is discussed.

Changes in discharge rate of fusimotor neurones provoked by fatiguing contractions of cat triceps surae muscles.

1. Changes in discharge rate of thirty-one fusimotor neurones to triceps surae muscles during long-lasting, fatiguing contractions of these muscles were studied in decerebrate cats. Discharges of fusimotor neurones were recorded from the nerve filaments. Muscle contractions were elicited by electrical stimulation of either the muscle nerves (twenty-one neurones) or the corresponding ventral roots (ten neurones), until the muscle tension fell to about 30% of its initial value. 2. Early and late changes could be recognized in fusimotor discharge rate during long-lasting muscle contraction. The early changes obviously not related to muscle fatigue, consisted of an initial increase at the onset of muscle contraction and a subsequent decrease to or below the resting discharge level. The late change in discharge rate, supposedly related to muscle fatigue, was an increase developing gradually towards the end of muscle contraction, ranging at its peak from 2 to 15 impulses/s (mean value 5.5 impulses/s, n = 31) and outlasting the contraction for 20-320 s. 3. When the contracting muscle was made ischaemic the late increase in fusimotor discharge rate started earlier and was maintained until the arterial clamp was removed. After severing the muscle nerves distal to the site of stimulation no changes, a slight sustained increase, or else a decrease in fusimotor discharge rate occurred during electrical stimulation of either muscle nerves or ventral roots. At its cessation the spontaneous firing rate was reassumed immediately. Stimulation of the distal stumps of the severed nerves elicited no changes in fusimotor discharge rate. 4. It is proposed that the late increase in fusimotor discharge rate may appear due to autogenetic excitation of fusimotor neurones by discharge from group III and IV muscle afferent fibres provoked and/or enhanced by metabolic products liberated in muscle tissue during the fatiguing contraction. The fusimotor firing was estimated to remain elevated to a level twice that of the spontaneous activity on average for approximately 120 s after the muscle contraction. Its functional role in muscle fatigue is discussed.

Time coupling of skeletomotor discharges in response to pseudo-random transsynaptic and transmembrane stimulation.

Firing pattern of skeletomotor neurones innervating triceps surae muscles in response to pseudo-random muscle stretching and white noise modulated transmembrane current stimulation was investigated in decerebrate cats. Pseudo-random muscle stretching (upper cut-off frequency 60 Hz, amplitude sigma (standard deviation) ranging from 18.5 microns to 40 microns) was applied to triceps surae muscles. Membrane potential changes and action potentials of skeletomotor neurones were recorded intracellularly. White noise modulated current was applied through the same (recording) microelectrode. Sequences of ten identical 5 s periods of either muscle stretching or transmembrane current stimulation were applied. Skeletomotor neurones belonging to slow motor units (rheobase less than 8.5 nA) generated action potentials in response to both pseudo-random muscle stretching and transmembrane current stimulation, while firing threshold of those belonging to fast motor units could not be reached by the muscle stretches applied. Peri-spike averaging of muscle length and injected current records showed that the action potentials appeared at the peak of either depolarizing current wave or muscle stretching both preceded by a change in opposite direction (the spikes coinciding with the peak in muscle length PSA being actually elicited by muscle spindle action potentials triggered at the moment of the peak stretching velocity). Time coupling of action potentials occurred during both muscle stretching and transmembrane stimulation, being more tight in the latter case as well as when larger amplitudes of the stimuli were applied.(ABSTRACT TRUNCATED AT 250 WORDS)