Effects of the calcium antagonist nifedipine on the afferent impulse activity of isolated cat muscle spindles

The impulse activity of muscle spindles isolated from the cat tenuissimus muscle was investigated under varying concentrations of the L-type calcium channel blocker nifedipine. At a concentration of 25 microM nifedipine impulse activity was clearly diminished in both primary and secondary endings. However, low concentrations of the drug (5-10 microM) exerted unexpected excitatory effects. The dynamic properties of primary endings in particular were augmented; those of secondary endings were also increased, although only slightly. A detailed analysis of the afferent discharge patterns obtained under ramp-and-hold stretches yielded the following effects of 10 microM nifedipine. (1) The initial burst at the beginning of the ramp phase of a stretch was increased in primary endings; (2) the peak dynamic discharge frequency at the end of the ramp phase was considerably increased in most primary endings; (3) the sensitivity of the peak dynamic discharge value to varying amplitudes and velocities of stretch was significantly enhanced in primary endings, and also increased, although only slightly, in secondary endings; (4) the rise in the discharge frequency during the ramp phase of a stretch was augmented in both types of ending, the effect being again stronger in primary endings; (5) the fast adaptive decay of the impulse frequency following the ramp phase of a ramp-and-hold stretch was significantly increased in primary endings, but remained unaffected in secondary endings. The enhanced dynamic properties of primary endings were also observed under small sinusoidal stretch stimuli (10 microm, 40 Hz), where nifedipine induced a significant shift in the position of the 1:1 driven action potentials toward smaller phase values. In view of an increase in tension in the isolated muscle spindle and an increased initial burst in primary endings in the presence of nifedipine, it is suggested that the drug facilitates the attachment of cross-bridges in the poles of the intrafusal muscle fibers. The increase in the dynamic properties of primary endings points to the possibility that the drug preferentially affects the nuclear bag(1) fiber. The inhibitory effect on the afferent discharge rate at high doses of the drug is interpreted as the consequence of a calcium channel block in the membranes of the sensory endings. The membrane potential of sensory endings appears to be highly dependent on sustained Ca(2+) conductance.     

Effects of changes in pH on the afferent impulse activity of isolated cat muscle spindles

Muscle spindle activity has been shown to decrease in the sustained contracting muscle. The effect has been assumed to result from a declining fusimotor drive. Since accumulation of metabolites including H(+), lactate and CO(2) might also affect the receptor in the fatiguing muscle, the impulse activity of muscle spindles isolated from the cat tenuissimus muscle was characterized under varying degrees of extracellular pH, thus excluding any effect on fusimotor activity, blood supply and extrafusal muscle fibers. The isolated receptor was exposed to bathing fluids of pH 6.4, 7.4 and 8.4, and afferent discharge activity was recorded from the spindle nerve. Both primary and secondary endings responded similarly to changes in pH. Resting discharge frequency usually decreased with decreasing pH and increased with increasing pH. A sudden break-off in activity was observed with about 40% of primary endings and about 30% of secondary endings at pH 6.4. Experiments with slow stretch stimulation indicated that this effect was caused by a rising threshold of firing at the encoder site of the endings. With brief ramp-and-hold stretches, we tested the effects of changes in pH on the dynamic and static sensitivity of primary and secondary endings. When pH was reduced from 7.4 to 6.4, the initial burst activity at the beginning of the ramp phase increased in primary and secondary endings and the dynamic response increased in secondary endings, demonstrating that the dynamic properties of muscle spindle endings were usually augmented in the acidic milieu. The static properties rose as well because the static index of both types of ending increased significantly. By contrast, dynamic and static properties of both primary and secondary endings decreased significantly, when pH was increased from 7.4 to 8.4. The amplitude of tension that was measured during the passive stretch stimuli very slightly decreased in the acidic solution and very slightly increased in the alkaline solution. The decrease in the resting discharge activity at low pH supports those previous observations, which demonstrate a reduced peripheral input from muscle spindle afferents to the spinal motor nuclei during fatigue in the isometric contracting muscle. The present finding indicates that an attenuated afferent discharge is not only caused by a decreasing central activation of gamma-motorneurons, but may additionally be supported by a direct effect of protons on the muscle receptor itself. The accompanying augmentation of stretch sensitivity is suggested to correspond to the well-known increase in physiological tremor during exhaustive exercise.     

The effect of hypertonic solutions on the electrical activity of isolated frog muscle spindles]

Electrical activity of an isolated frog muscle spindle was investigated in hypertonic solutions obtained by adding 400 mM of sucrose, glucose or glycerine to the Ringer solution. In sucrose and glucose hypermedia the frequency of background impulse activity first increased and then decreased up to zero; receptor potentials and evoked impulse activity simultaneously decreased and disappeared. In the glycerine hypermedia impulse activity after some increase returned to normal and then a second rise of frequency to a constant supernormal level was observed. After returning to the Ringer solution from sucrose or glucose the background and evoked impulse activity progressively returned to normal. But after glycerine hypermedia a significant growth in the background impulse activity and a small change in frequency of evoked impulse activity were observed. The observed changes of electrical activity of the muscle spindle in hypermedia are discussed in terms of deformation of sensory endings and intrafusal muscles fibres leading to depolarization of the receptor membrane.     

Reversal of the static component of spindle potential by imposed depolarizing current in the frog muscle spindle

The static component of the spindle potential provoked during stretch of isolated muscle spindles of the frog was reversed during the application of depolarizing currents ranging from 0.2 to 5 nA in normal Ringer solution and also in Na+-free Ringer solution. In the same range of current intensities, spontaneous rhythmic hyperpolarizations due to [Ca2+]i-activated GK, an attenuation of membrane impedance, and an anomalous decrease in amplitude of the afferent spikes were observed. All 4 phenomena were abolished by K+ channel blockers (10 mM CsCl, 1-2 mM 4-aminopyridine (4-AP), or 20 mM tetraethylammonium chloride (TEA], Ca2+ channel blockers (5-10 mM CoCl2, MnCl2, 1-2 mM CdCl2 or 0.5 mM verapamil) or 0.1 mM quinine. The amplitude of the static component of the spindle potential was markedly increased at threshold concentration of the K+ channel blockers (5 mM CsCl, 0.1-0.5 mM 4-AP or 5-10 mM TEA), but the component disappeared at that of the Ca2+ channel blockers. The rhythmic hyperpolarizations are associated with the spindle potential, except for its dynamic component, which often triggers a hyperpolarizing deflection. We suggest that both the static component of the spindle potential and rhythmic hyperpolarizations are due to GK(Ca) in the intracapsular axon, either along the terminal or at the branching nodes, or both; and that the receptor potential contributes to, but is not the same as, the spindle potential.     

Vibration sensitivity of human muscle spindles and Golgi tendon organs

The responses of the various muscle receptors to vibration are more complicated than a naïve categorization into stretch (muscle spindle primary ending), length (muscle spindle secondary endings), and tension (Golgi tendon organs) receptors. To emphasize the similarity of responses to small length changes, we recorded from 58 individual muscle afferents subserving receptors in the ankle or toe dorsiflexors of awake human subjects (32 primary endings, 20 secondary endings, and six Golgi tendon organs). Transverse sinusoidal vibration was applied to the distal tendon of the receptor‐bearing muscle, while subjects either remained completely relaxed or maintained a weak isometric contraction of the appropriate muscle. In relaxed muscle, few units responded in a 1:1 manner to vibration, and there was no evidence of a preferred frequency of activation. In active muscle the response profiles of all three receptor types overlapped, with no significant difference in threshold between receptor types. These results emphasize that when intramuscular tension increases during a voluntary contraction, Golgi tendon organs and muscle spindle secondary endings, not just muscle spindle primary endings, can effectively encode small imposed length changes. Muscle Nerve, 2007     

The pathology of the human muscle spindle: effect of denervation

The effect of denervation on the morphology and innervation of human muscle spindles has been studied in mixed sensorimotor neuropathies and in sensory and motor denervation in 25 cases examined at necropsy.In sensorimotor denervation the changes found consisted of degeneration and atrophy of the intrafusal muscle fibres, which were often clumped together in the centre of the dilated spindle capsules, degeneration of the equatorial aggregations of sarcolemmal nuclei and, in longstanding denervation, of collagenous thickening of the spindle capsule and deposition of collagen around the intrafusal muscle fibres. There was a reduction in number of sensory and motor nerve fibres in the affected spindles and changes were found in both the sensory and the motor nerve endings.In sensory denervation there was atrophy of the equatorial regions of the intrafusal muscle fibres, with degeneration and loss of the equatorial nuclei and simplification and degeneration of the primary and secondary sensory endings. Excess intraluminal collagen was sometimes found in the equatorial regions. The polar parts of the spindle capsules and of the intrafusal muscle fibres, and the fusimotor innervation, were normal. In tabes dorsalis attempts at regeneration of the sensory endings were sometimes recognisable and there was swelling and attenuation of the terminal part of the primary and secondary sensory axons.In motor denervation degenerative changes were found in both gamma and alpha-collateral fusimotor nerve fibres and motor endings, and in some spindles no motor nerve fibres could be identified. The polar regions of the intrafusal muscle fibres were thin and atrophic, and, as in sensorimotor denervation, degeneration of nuclear chain fibres was more prominent than, and preceded, degeneration of nuclear bag fibres. This could not easily be related to the differing patterns of innervation of these two types of muscle fibre, but seemed to be due to structural differences in the fibres themselves. The equatorial regions of these intrafusal fibres and their sensory innervation were normal.     

Morphological and functional demonstration of rat dura mater mast cell-neuron interactions in vitro and in vivo

The effects of changes in temperature on primary and secondary endings of isolated cat muscle spindles were investigated under ramp-and-hold stretches and different degrees of pre-stretch. Temperature-induced alterations of the discharge frequency were compared over a temperature range of 25–35°C. Both primary and secondary endings responded to warming with increasing discharge frequencies when the spindle was pre-stretched by 5–10% of its in situ length. The following differences between the temperature effects on primary and secondary endings were observed: (1) The temperature coefficients (Q₁₀) obtained from the discharge frequencies during the dynamic and static phase of a stretch were similar for endings of the same type, but they were larger in primary endings (range of Q₁₀: 2.3–3.3; mean: 2.9) than in secondary endings (range of Q₁₀: 1.6–2.2; mean: 2.0); (2) With primary endings, but not with secondary endings, the temperature sensitivity (imp s⁻¹ °C⁻¹) was larger during the dynamic phase than during the static phase of a stretch; (3) In primary endings, the fast and slow adaptive components occurring in the discharge frequency during the static phase of a stretch clearly increased with warming while in secondary endings, the slow decay was less affected, and the fast decay showed no change; (4) In relaxed spindles, the excitatory effect of warming was overlaid by a strong inhibitory effect as soon as the temperature exceeded about 30°C, resulting in an abrupt cessation of the background activity in most secondary endings, but not usually in primary endings. In general, warming induced an enhanced stretch sensitivity in both types of ending, and additionally an inhibitory effect that is obvious only in secondary endings of relaxed spindles. The different effects of temperature on the discharge frequency of primary and secondary afferents are assumed to be caused by different properties of their sensory membranes.     

Effect of intrafusal muscle mechanics on mammalian muscle spindle sensitivity

Sensitivity differences between primary and secondary endings of mammalian muscle spindles under various conditions of stretch and fusimotor activation may be due to differences in their respective mechanoelectric transducers or to mechanical properties of the intrafusal muscle supporting those endings. This study of isolated cat muscle spindles examines the strain in individual intrafusal muscle fibers resulting from stretch and fusimotor stimulation. The degree of local stretch occurring at the sensory endings under these conditions was measured. The results support the hypothesis that the sensitivities of primary and secondary endings are quite similar. They are directly related to the local stretch of the underlying muscle which may be altered by changes in muscle stress and stiffness.     

Effects of chlorobutanol on primary and secondary endings of isolated cat muscle spindles

The effects of the preservative chlorobutanol on primary and secondary endings of muscle spindles isolated from the tenuissimus muscle of the cat were investigated in this study. Chlorobutanol was applied to the bathing solution in final concentrations of between 10 and 100 microg/ml. It induced a reversible and dose dependent decrease in the discharge frequency of both types of ending without any visible length change in the sensory region of the receptor. The initial activity, the peak dynamic discharge, the maximum static discharge value and the final static discharge value were evaluated from an ending's discharge pattern obtained during ramp-and-hold stretches. These four basic discharge frequencies decreased in parallel with increasing concentrations of chlorobutanol. Their sensitivities to chlorobutanol were similar (mean values: -0.11 to -0.29 imp/s per microg/ml chlorobutanol) and were independent of the amplitude of stretch. The dynamic response and the static response of both primary and secondary endings remained unchanged, indicating that the sensitivity of the spindle to stretch was not influenced by chlorobutanol. Chlorobutanol also reduced the discharge activity of the muscle spindle afferents during sinusoidal stretches. The amplitude of the receptor potential (AC component) remained unchanged under chlorobutanol. With the available recording technique it was not possible to measure slow shifts of the membrane potential. However, a hyperpolarization of the ending's membrane might explain why the afferent discharge frequency is reduced by chlorobutanol. The calcium dynamics of the spindle do not appear to be altered by CB, as the effect exerted on the afferent discharge by a change in the extracellular calcium concentration and a blockage of calcium channels was different from the CB effect. As the inhibitory effect of CB was reduced by ouabain, it is possible that CB activates the electrogenic Na/K pump or affects a mechanism that is closely related to the activity of the pump. The properties of the axonal membrane appear not to be altered, as chlorobutanol did not change the shape of action potentials.     

A novel stretch receptor in the jaw of the rat

We report here an unusual type of stretch receptor found on each side of the rat jaw. This receptor has unique morphological featurs: it is quite long (24–28 mm), lies in connective tissue in between masticatooy muscle, and extends between the medial pterygoid muscle-tendon on the maxilla and the masseter-tendon on the mandible through a zigzag course, forming a Greek capital letter sigma when viewed from the side. The receptor is neither in parallel nor in series with any masticatory muscles and receives multiple innervation. The receptor increases its length when the jaw closes and shortens when the jaw opens. Electron microscopy revealed axial structures composed of a central cellular core surrounded by tightly packed collagen bundles which are separated from the capsule by a wide capsular space. Most of the sensory endings are found among axial collagen bundles, some in between core cells. The core cells have many finger-like processes on their surface, being coupled by desmosomes. The origin and nature of these cells are unclear. The wide capsular space is filled with Alcian blue positive substrate, probably acid glycosaminoglycans. The structures of outer and inner capsules are similar to those of muscle spindles, the former being composed of three to ten layers of thin flattened cells. The response of the receptor was examined with in vivo as well as in vitro preparations. In in vivo experiments, impulse discharges from this receptor increased with the increase in jaw closing. When the jaw was fully opened the impulse discharge from this receptor disappeared. It is clear from these experiments that this receptor is a slowly adapting stretch receptor, being sensitive to jaw position and movement. With in vitro preparations, a linear relation was obtained between the frequency of impulse discharge measured during the static phase of stretch and the amplitude of stretch. Beyond 20% of stretch the response tended to saturate. The dynamic component of discharge was less marked, and comparable to the response of Golgi tendon organs or secondary endings of muscle spindles. The conduction velocity of innervating axons was 23–31 m/s at 35°C.     

Calcium and calcium-activated potassium currents of motor nerve endings in the frog

Evoked electrical responses of nerve endings were recorded in experiments on frog cutaneous-pectoris muscle under visual control. During superfusion by Ca2+-free solution with tubocurarine a late inward current was found in the evoked response of nerve endings recorded by CaCl2 filled electrode. With addition of the external solution of 4-aminopyridine the late current was changed into an outward one. This outward current depended on Ca2+ concentration in the electrode, decreased after local increase of K+ concentration and was abolished by Co2+ ions. Local ionophoretic application of tetraethyl-ammonium eliminated the outward current and revealed a strong and prolonged inward current. Similar currents were recorded by microelectrodes filled with SrCl2, BaCl2 and MgCl2. It is concluded that the late inward current is carried through potential-dependent calcium channels, while the late outward one--through Ca2+-activated K+ channels. The ratio of calcium and Ca2+-activated potassium currents in motor nerve endings and their role in the transmitter release are discussed.     

Cobalt-dependent potentiation of net inward current density in Helix aspersa neurons.

A low concentration of transition metal ions Co2+ and Ni2+ increases the inward current density in neurons from the land snail Helix aspersa. The currents were measured using a single electrode voltage-clamp/internal perfusion method under conditions in which the external Na+ was replaced by Tris+, the predominant external current carrying cation was Ca2+, and the internal perfusate contained 120 mM Cs+/0 K+; 30 mM tetraethylammonium (TEA) was added externally to block K+ current. In the presence of Co2+ (3 mM) or Ni2+ (0.5 mM) inward Ca2+ currents were stimulated normally by voltage-dependent activation of Ca2+ channels. There was a 5-10% decrease in the rate of rise of the inward current. The principal effect of Co2+ and Ni2+ in increasing the current density seems to be a decrease in the rate at which the inward currents decline during a depolarizing voltage pulse. The results may be due to a decrease in a voltage-dependent or Ca(2+)-dependent outward current and/or an inhibition of Ca2+ channel inactivation. Outward current under these conditions (zero internal K+) was significant and most likely due to Cs+ efflux through the voltage-activated or Ca(2+)-activated nonspecific cation channels. Co2+ is an extremely effective blocker of this outward current. These results are not an artifact of internal perfusion or the special ionic conditions. Intracellular recording of unperfused neurons in normal Helix Ringer's solution showed that the Ca(2+)-dependent action potential duration was increased significantly by low concentrations of Co2+. This result is consistant with the Co(2+)-dependent increase in inward (depolarizing) current seen in voltage-clamp experiments.     

Participation of the stretch reflex in human physiological tremor.

Microneurographic studies were made of normal human muscle spindle afferent activity to evaluate the role played by these endings, and by the segmental stretch reflex, in various types of physiological tremor. Primary endings are sufficiently sensitive to respond to the minute tremulous movements normally seen in contracting muscles. Human muscle spindle endings appear to possess the same non-linear sensitivity to small stretches as has been reported for cat muscle spindles. The findings also support the notion that the segmental stretch reflex plays an important role in enhanced physiological tremor, in so far as it tends to synchronize the motor outflow in such a way as to produce rhythmical contractions properly timed to reinforce the mechanical resonant properties of muscle spindle endings, the stretch reflex also serves to damp the larger oscillations of finger or hand which result from a brisk tap or muscle twitch. There is no evidence for alpha-gamma co-activation in these enhanced physiological tremors.     

Location and completeness of reinnervation by two types of neurons at a single target: The feline muscle spindle

Muscle spindles from the tenuissimus muscle of the cat were examined microscopically to assess the precision and completeness of reinnervation of intrafusal muscle fibers by efferent and afferent neurons. Positions of motor and sensory nerve terminals were charted relative to the cross-sectional area enclosed by the outer capsule of the spindle. Profiles of nerve endings were measured for normally innervated and reinnervated spindles. The tenuissimus was deprivedof innervation by freezing its nerve, sometimes in conjunction with either spinal ganglion removal or ventral rhizotomy. Sensory and motor terminals occupied separate locales along the length of normal muscle spindles. Nerve terminals of efferent and afferent neurons were located in appropriate positions along the length of spindles when axons of both types of neurons regrew-together and when either category of axon regenerated alone. Precise reinnervation of muscle spindles occurred in spite of a diminished diameter of intrafusal fibers. Repopulation of the spindle with motor endings was less complete than that by sensory endings, based on the proportion and size of the regenerated terminals. We conclude that under optimal conditions for axonal regrowth, efferent and afferent neurons reinnervate their respective regions along intrafusal muscle fibers but motor lags sensory reinnervation within the spindle. The mechanism by which positional specificity happens during reinnervation of intrafusal fibers requires neither an interaction between terminals of the two types of neurons nor target cells of normal bulk. © 1993 Wiley-Liss, Inc.     

Electrical 'tuning' in muscle spindle receptors

A variety of mechanical and electrical mechanisms have been shown to tune sensory receptors to selected stimuli. This study demonstrates the presence of electrical tuning in the mammalian muscle spindle. Apamin and tetraethyl ammonium ions (TEA), blockers of Ca2+-activated K+ channels, and ZnCl2, a Ca2+ channel blocker, were shown to change the dynamic behavior of the muscle spindle. We conclude that a Ca2+-activated K+ channel in the sensory nerve of the spindle provides negative feedback which alters its dynamic behavior tending to compensate for muscle dynamics. This mechanism is similar to that found in sensory hair cells in the cochlea and vestibule and may represent a general strategy for tuning in sensory receptors.     

Activation of κ-opioid receptors inhibits depolarisation-evoked exocytosis but not the rise in intracellular Ca in secretory nerve terminals of the neurohypophysis

Nerve endings of the magnocellular neurohypophysial neurones possess kappa-opioid receptors. Using a preparation of isolated terminals from the neurohypophysis we studied kappa-opioid effects on secretion of oxytocin and vasopressin and on intracellular Ca2+ concentration ([Ca2+]i) measured fluorimetrically or using digital video imaging with Fura-2. The dihydropyridine Ca(2+)-channel antagonist nicardipine reduced [Ca2+]i responses to K(+)-depolarisation (30-40 mM K+) by 55-75% and inhibited evoked secretion of oxytocin and vasopressin to a similar extent. The selective kappa-receptor agonist D-Pro10 Dynorphin A 1-11 (DPDYN) substantially inhibited K+ evoked secretion of oxytocin by 40-90% and secretion of arginine vasopressin (AVP) by 20-50%. DPDYN caused only a 10% reduction in the average total population [Ca2+]i response to K+ depolarisation. No sub-population of inhibitory responses was observed when samples of individual terminal [Ca2+]i responses were examined with imaging. Although kappa-receptors are coupled to Ca(2+)-channels at neuronal somata our data suggest that alternative effector mechanisms operate in these secretory nerve endings.     

Fast cholinergic efferent inhibition in guinea pig outer hair cells

Hair cells of inner ear are suggested to be inhibited by the activation of the alpha9-containing nicotinic acetylcholine (ACh) receptors (alpha9-containing nAChRs). Several studies have suggested that the native nicotinic-like ACh receptors (nAChRs) in hair cells display a significant permeability of Ca(2+) ions and unusual pharmacological properties. The activation of native nAChRs will initiate the hyperpolarization of hair cells by activation of the small conductance, Ca(2+)-activated K(+) channels (SK). In this work, the properties of the ACh-sensitive potassium current (IK(ACh)) in outer hair cells (OHCs) of guinea pigs were investigated by employing whole-cell patch-clamp. Followed by perfusion of ACh, OHCs displayed a rapid desensitized current with an N-shaped current-voltage curve (I-V) and a reversal potential of - 66 +/- 7 mV. The IK(ACh) was still present during perfusion of either iberiotoxin (IBTX, 200 nM) or TEA (5 mM) but was potently inhibited by apamin (1 muM), TEA (30 mM). The IK(ACh) demonstrated a strong sensitivity to alpha-bungarotoxin (alpha-BgTx), bicuculline and strychnine. These results suggested that OHCs display the well-known SK current, which might be gated by the alpha9-containing nAChRs. Two important changes were present after lowering the Ca(2+) concentration in the external conditions from 2 mM to 0.2 mM: one was a flattened N-shape I-V relationship with a maximum shifted toward hyperpolarized potentials from -20 approximately -30 mV approximately -40 to -50 mV, the other was a significant reduction in the agonist maximal response (percentage of maximal response 10.5 +/- 5.4). These results indicated that native nAChRs are both permeable to and modulated by extracellular Ca(2+) ions. Taken together, this work provides direct evidences that SK channels in OHCs of guinea pigs are gated by alpha9-containing nAChRs, which play an important role in the fast cholinergic efferent inhibition. This fast inhibition is both potently dependent on the permeability of Ca(2+) ions through the native nAChRs and modulated by Ca(2+) ions.     

Effects of anions on calcium component in sensory nerve terminal of frog muscle spindles

An increase in the Ca2+ component following individual sodium spike in spontaneous discharges was observed in the isolated sensory terminal of the frog muscle spindle perfused with isotonic sodium solution with a group of anions, of which the size in the aqueous solution ranged from 0.74 to 1.32 times that of hydrated sodium ion. The effects of these anions was counteracted with divalent cations. It is hypothesized that anions similar in size to hydrated sodium ions may form an anion-cation complex at the interaction site of the Na+ carrier, whereby the Na+ may enter the Ca2+ channel. This may be inactivated by the divalent cations.     

Swelling-activated calcium signalling in cultured mouse primary sensory neurons

The effects of hypo-osmotic membrane stretch on intracellular calcium concentration ([Ca(2+)](i)), cell volume and cellular excitability were investigated in cultured mouse primary sensory trigeminal neurons. Hypotonic solutions (15--45%) led to rapid cell swelling in all neurons. Swelling was accompanied by dose-dependent elevations in [Ca(2+)](i) in a large fraction of neurons. Responses could be classified into three categories. (i) In 57% of the neurons [Ca(2+)](i) responses had a slow rise time and were generally of small amplitude. (ii) In 21% of the neurons, responses had a faster rise and were larger in amplitude. (iii) The remaining cells (22%) did not show [Ca(2+)](i) responses to hypo-osmotic stretch. Slow and fast [Ca(2+)](i) changes were observed in trigeminal neurons of different sizes with variable responses to capsaicin (0.5 microM). The swelling-induced [Ca(2+)](i) responses were not abolished after depletion of intracellular Ca2+ stores with cyclopiazonic acid or preincubation in thapsigargin, but were suppressed in the absence of external Ca(2+). They were strongly attenuated by extracellular nickel and gadolinium. Hypotonic stimulation led to a decrease in input resistance and to membrane potential depolarization. Under voltage-clamp, the [Ca(2+)](i) elevation produced by hypotonic stimulation was accompanied by the development of an inward current and a conductance increase. The time course and amplitude of the [Ca(2+)](i) response to hypo-osmotic stimulation showed a close correlation with electrophysiological properties of the neurons. Fast [Ca(2+)](i) responses were characteristic of trigeminal neurons with short duration action potentials and marked inward rectification. These findings suggest that hypo-osmotic stimulation activates several Ca(2+)-influx pathways, including Gd(3+)-sensitive stretch-activated ion channels, in a large fraction of trigeminal ganglion neurons. Opening of voltage-gated Ca(2+) channels also contributes to the response. The pattern and rate of Ca(2+) influx may be correlated with functional subtypes of sensory neurons.     

Effects of Ruthenium ions on the sensory terminal discharges of the frog muscle spindle

The presence of a mixed Na+-Ca2+ spike along the sensory terminal of the frog muscle spindle was verified. When the terminal was perfused with Ringer's solution containing 0.1-0.5 mM ruthenium red (RuR), the amplitude and duration of the spike were increased, occurring as a prolonged or a long-lasting depolarization of up to 20-30 s duration following individual afferent spikes evoked spontaneously or antidromically by electrical stimulation. In an isotonic TEA solution, the amplitude and duration of the afferent spikes were increased; however, no prolonged depolarization occurred. Adding 0.2 mM RuR to the TEA solution produced the prolonged and long-lasting depolarization. All responses disappeared in the presence of 3 microM TTX or Na+-free Ringer's solution. An impedance decrease along the terminal was observed during the prolonged or long-lasting depolarization. The prolonged depolarization was blocked by the addition of Ca2+-blockers; the afferent spikes remained. In preparations preincubated with 0.1 mM RuR, increasing CaCl2 in Ringer's solution from 0.2 mM, resulted in shortening of the duration of individual spikes with prolonged depolarization and in increase in the maximum rate of rise (MRR) of the spikes. Preincubation with higher concentrations of RuR produced higher sensitivities in the modifications of the duration and MRR to the change in [Ca2+]O. The responses were retained by adding RuR or RuCl3 to Ca2+-free Ringer's solution containing 0.1-5 mM EGTA, although all responses disappeared in Ca2+-free EGTA Ringer's solution. It is concluded that the RuR-induced prolonged response is produced by an influx of Na+.