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PubMed1974

Differential reaction of fast and slow alpha-motoneurones to axotomy.

1. The properties of medial gastrocnemius (m.g., fast alpha) and soleus (sol., slow alpha) motoneurones of the cat were examined with intracellular electrodes 8-119 days after section of the muscle nerves.2. The axonal conduction velocity was significantly decreased in both m.g. and sol. motoneurones after chronic section of the muscle nerves.3. The amplitude of overshoot of action potentials was significantly increased in both m.g. and sol. motoneurones following section of the muscle nerves.4. No significant changes in the resting membrane potential or the input resistance were observed for sol. motoneurones, whereas m.g. motoneurones showed a slight decrease in the resting potential and a slight increase in the input resistance.5. The duration of after-hyperpolarization was significantly decreased in sol. motoneurones, whereas that in m.g. motoneurones remained virtually unchanged or increased slightly following section of the muscle nerves.6. The changes described above were not seen in the preparations examined 29-46 days after section of the lumbosacral dorsal roots, suggesting that alterations in the motoneurone properties observed after section of the muscle nerves resulted from axotomy of the motoneurones rather than from sensory deprivation.7. The differences in electrophysiological properties between m.g. and sol. motoneurones were less prominent in axotomized animals than in control, unoperated cats.8. It is concluded that fast (m.g.) and slow (sol.) alpha-motoneurones have qualitatively different properties. A possible ;dedifferentiation' of fast and slow alpha-motoneurones by axotomy is discussed.

Action Potentials↗
PubMed1975

Detachment of structurally intact nerve endings from chromatolytic neurones of rat superior cervical ganglion during the depression of synaptic transmission induced by post-ganglionic axotomy.

1. Electrophysiological studies showed that injury of post-ganglionic nerve fibres leads to severe and prolonged depression of synaptic transmission through the rat superior cervical ganglion, beginning within 24 h. This is in line with the results of previous studies in other species and upon other neurones. 2. electron microscopy after post-ganglionic axotomy revealed nerve endings of presynaptic type with all the specialized membrane-related features of a synaptic zone, but which were not apposed to any post-synaptic nervous element. These umusual profiles were interpreted as detached presynaptic nerve endings. In normal and control ganglia, such profiles formed at most 0-5% of all vesicle-containing profiles of presynaptic type; in ganglia with all major post-ganglionic branches cut the proportion rose to approximately 7%, between 3 and 7 d post-operatively. Over this period, the mean incidence of chromatolytic neurones was 74-6%. 3. Concomitantly, the incidence of synapses within the ganglion fell by about 75%, reaching its lowest levels between 3 and 7 d post-operatively. There was strikingly little evidence of persistence of post-synaptic membrane specializations ('membrane thickenings') following detachment of synapses. 4. At longer survival intervals the incidence of synapses gradually increased, and that of detached nerve endings gradually decreased; recovery was well advanced by 42 d. 5. The fall in the incidence of synapses was closely paralleled by a fall in the incidence of desmosome-like attachments in the ganglion; the incidence of such attachments was found to be correlated to a significant degree with that of synapses. 6. It is concluded that most or all of the synapses upon sympathetic neurones become physically dissociated during the chromatolytic reaction of these neurones to axotomy. The failure to persist of ultrastructurally specialized post-synaptic sites, and the loss of desmosomes (particularly marked for those involving purely post-ganglionic nervous elements) suggest that the post-ganglionic neurone is losing all its specializations for attachment. 7. Some evidence suggests that the satellite cells may effect the final separation between pre- and post-synaptic structures.

Action Potentials↗
PubMed1981

The effects of axotomy on the conduction of action potentials in peripheral sensory and motor nerve fibres.

Medial gastrocnemius and sural nerves in one hindlimb of the cat were transected and prevented from regenerating. After periods ranging from 29-273 days, compound action potentials were recorded from axotomised and contralateral control nerves. The amplitude and integrated area of action potentials decreased and conduction velocity slowed following axotomy. The area under compound action potentials generated by stimulating sensory fibres declined significantly faster than that generated by stimulating motor fibres. Analysis of changes in whole nerve conduction velocity distributions showed that the velocities of fast conducting sensory fibres decreased at the most rapid rate. The conduction velocities of motor fibres and slow sensory fibres declined at significantly slower rates. The loss of electrical activity in the largest sensory nerve fibres following axotomy, may play a role in determining the faster rate at which their action potentials deteriorate.

Action Potentials↗
PubMed1986

Altered excitability of goldfish mauthner cell following axotomy. I. Characterization and correlations with somatic and axonal morphological reactions.

Axonal transection 7-10 mm distal to the cell body of the goldfish Mauthner (M) cell induced alterations in its excitability; namely, the antidromic spike recorded in the soma was converted from a single-component axon-hillock response to a larger amplitude, two-component impulse. The mean spike amplitude of the axotomized cells was approximately 50% greater (59.6 +/- 15.1 mV, n = 94) than that in controls (39.4 +/- 6.3 mV, n = 73). The onset of the induced increase in spike amplitude occurs at approximately 20 days postaxotomy, and the transition to a reactive spike is complete by approximately 30-35 days. Eighty-three percent of the M-cells axotomized for more than 30 days were physiologically reactive as judged by their large spike amplitudes and/or the presence of an additional spike component. Concomitant with the enhanced spike amplitudes, there was a depression of excitability in the initial segment-axon hillock region of the axotomized cells. This depression was suggested by a decrease in the initial segment (IS) spike height (from 39.4 +/- 6.3 mV, n = 73, in controls to 27.5 +/- 5.6 mV, n = 13, in axotomized cells), a decrease in its maximum rate of rise (from 153.6 +/- 24 V/s, n = 15, to 112.5 +/- 30 V/s, n = 29), and frequent failure of antidromic invasion into the initial segment and axon hillock. These changes in excitability could not be attributed to alterations in passive membrane properties, since the mean resting potential (77.8 +/- 5.2 mV, n = 37, control; 76.9 +/- 7.8 mV, n = 87, axotomized) and input resistance (170 +/- 21.3 K omega, n = 13, control; 176 +/- 26.6 K omega, n = 21, axotomized) were not altered significantly by axotomy. Threshold voltage was also unaffected (13.4 +/- 3.2 mV, n = 11, control; 11.9 +/- 2.5 mV, n = 11, axotomized). Sequential recordings of spike amplitudes from the axon hillock, soma, and lateral dendrite suggest that the generator of the axotomy-induced component is localized to the normally passive soma and proximal dendrite. In addition, the presumed soma-dendritic In addition, the presumed soma-dendritic component contributes very little if anything to the action potentials recorded in the axon. The onset and occurrence of alterations in excitability and cell body morphology (chromatolysis and nuclear associated changes) were compared in different M-cell populations and in the same identified M-cells. The comparisons suggested that these two events tend to occur in parallel.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1997

Differential effects of NGF and BDNF on axotomy-induced changes in GABA(A)-receptor-mediated conductance and sodium currents in cutaneous afferent neurons.

The effects of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) on injury-induced changes in the electrophysiological properties of adult rat cutaneous afferent dorsal root ganglion (DRG) neurons were examined. Whole cell patch-clamp techniques were used to study gamma-aminobutyric acid-A (GABA(A))-receptor-mediated conductance, voltage-dependent sodium currents, and action potential waveform in cutaneous afferent neurons (35-60 microm diam) cultured from control and axotomized animals. Cutaneous afferent neurons were identified by retrograde labeling with hydroxy-stilbamidine (Fluoro-gold, a fluorescent retrograde axonal tracer); the sciatic nerve was transected 1 wk after Fluoro-gold injection and L4/L5 DRG neurons were cultured 2-3 wk after axotomy. NGF, BDNF, or Ringer (vehicle) solution was delivered in vivo directly to the transected sciatic nerve stump in axotomized rats via an osmotic pump. Recordings were obtained from neurons 5-24 h after culture. Axotomized neurons from rats treated with vehicle solution displayed a twofold increase in GABA-induced conductance and a prominent reduction in the proportion of neurons expressing action potentials that had inflections on the falling phase. The expression of kinetically slow tetrodotoxin (TTX)-resistant sodium current was markedly reduced and an increased expression of kinetically fast TTX-sensitive current was observed in neurons from vehicle-treated, axotomized rats. Treatment with NGF (0.25 microg/microl at 12 microl/day for 14 days) in axotomized animals resulted in an increase in the proportion of neurons expressing TTX-resistant, slow sodium currents and inflected action potentials, but had no effect on GABA-induced conductance. Treatment with BDNF (0.5 microg/microl at 12 microl/day for 14 days) attenuated the axotomy-induced increase in GABA(A)-receptor-mediated conductance while minimally affecting action potential waveform. The observed neurotrophin effects occurred independently of cell size changes. These findings indicate a differential regulation of GABA(A) receptor and sodium channel properties in axotomized rat cutaneous afferent neurons by specific neurotrophic factors.

Animals↗
PubMedDate not supplied

Rescue of motoneurons from axotomy-induced cell death by polymer encapsulated cells genetically engineered to release CNTF.

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) results from the progressive loss of motoneurons, leading to death in a few years. Ciliary neurotrophic factor (CNTF), which decreases naturally occurring and axotomy-induced cell death, may result in slowing of motoneuron loss and has been evaluated as a treatment for ALS. Effective administration of this protein to motoneurons may be hampered by the exceedingly short half-life of CNTF, and the inability to deliver effective concentration into the central nervous system after systemic administration in vivo. The constitutive release of CNTF from genetically engineered cells may represent a solution to this delivery problem. In this work, baby hamster kidney (BHK) cells stably tranfected with a chimeric plasmid construct containing the gene for human or mouse CNTF were encapsulated in polymer fibers, which prevents immune rejection and allow long-term survival of the transplanted cells. In vitro bioassays show that the encapsulated transfected cells release bioactive CNTF. In vivo, systemic delivery of human and mouse CNTF from encapsulated cells was observed to rescue 26 and 27% more facial motoneurons, respectively, as compared to capsules containing parent BHK cells 1 wk postaxotomy in neonatal rats. With local application of CNTF on the nerve stump and by systemic delivery through repeated subcutaneous injections, 15 and 13% more rescue effects were observed. These data illustrate the potential of using encapsulated genetically engineered cells to continuously release CNTF to slow down motoneuron degeneration following axotomy and suggest that encapsulated cell delivery of neurotrophic factors may provide a general method for effective administration of therapeutic proteins for the treatment of neurodegenerative diseases.

Amyotrophic Lateral Sclerosis↗
PubMed1982

The effects of axotomy upon the extrasynaptic acetylcholine sensitivity of an identified motoneurone in the cockroach Periplaneta americana.

The effects of axotomy on the sensitivity of the fast coxal depressor motoneurone (Df) of the cockroach (Periplaneta americana) to applied acetylcholine (ACh) and carbamylcholine (CCh) have been investigated. ACh and CCh applied to the soma membrane either by bath perfusion or by ionophoresis caused depolarization; repeated application of large doses of these agonists resulted in a relatively rapid desensitization and depression of the response. Axotomy performed 3-10 days before recordings were made caused an approximately threefold increase in the sensitivity to ACh but had no affect upon the sensitivity to CCh. The resting potential, input resistance and membrane time-constant remained within the normal range. In addition there was no change in the rise-times of the responses or of the ACh reversal potential, or in the apparent number of ACh molecules needed to combine with individual cholinoceptors to produce a response. The anticholinesterases physostigmine and neostigmine potentiated the ACh response at relatively low concentrations (10(-7) M-10(-6) M). This potentiation was significantly greater in the normal cells than in the axotomized cells. It is therefore concluded that the increase in ACh sensitivity of this motoneurone results at least partly from a fall in the activity of cholinesterase in the region of the applied drug.

Acetylcholine↗
PubMed1979

[Molecular forms of acetylcholinesterase in the chicken ciliary ganglion; changes after denervation, axotomy and double section].

Four main molecular forms of acetylcholinesterase (AChE), with sedimentation coefficients of 5, 7.5, 11.5 and 20 S, are found in Chicken ciliary ganglion. After denervation, the loss in 11.5 and 20 S forms occuring in 48 hrs coincides with the disappearance of presynaptic structures. In contrast, axotomy induces an early and durable increase in 7.5 S form. From these results, it is inferred that 11.5 and 20 S forms are predominant in presynaptic structures and 7.5 S form is mainly postsynaptic. In addition, the effects observed after simultaneous denervation and axotomy show a reciprocal control between pre- and postsynaptic elements. Finally, a trans-synaptic effect is exerted on 20 S AChE in controlateral ganglion after preganglionic sections.

Acetylcholinesterase↗
PubMed1975

Cholinesterase activity in the hypoglossal nucleus of the rat and the changes produced by axotomy: a light and electron microscopic study.

Within the hypoglossal nucleus large amounts of acetylcholinesterase (AChE) activity are present in all the neurons, whereas intracellular butyrylcholinesterase (BuChE) activity occurs only within a ventro-caudally situated cluster of cells. AChE activity within the neurons occurs mainly in the cisternae of the granular endoplasmic reticulum but there is some in the intermembranous space of the nuclear envelope and in the Golgi complexes. In the neuropil, reaction product is seen along some axonal and synaptic membranes. The distribution of BuChE in the ventro-caudal cells is identical with that of AChE except that BuChE activity is absent from the neuropil. The level of intraneuronal AChE activity falls rapidly during the first few days after axotomy. The fall is due partly to a dissolution and peripheral migration of the E.R. but also to a decrease in AChE content of the E.R. that remains. Return of staining begins in the 4th week and continues as the E.R. reassembles. Staining in the neuropil falls more slowly, but recovers less completely. The ventro-caudal group of cells shows the same kinds of change, but more dramatically. BuChE activity returns only erratically and never completely. The similarity in normal distribution, and in response to axotomy, of the two cholinesterases suggests that their functions are related.

Acetylcholinesterase↗
PubMed1992

Brain-derived neurotrophic factor administration protects basal forebrain cholinergic but not nigral dopaminergic neurons from degenerative changes after axotomy in the adult rat brain.

Cell culture studies with dissociated primary cultures from embryonic rat brain revealed that brain-derived neurotrophic factor (BDNF) promotes the developmental differentiation of both basal forebrain cholinergic and mesencephalic dopaminergic neurons. These studies suggested that, in the adult brain, BDNF may be able to protect cholinergic and dopaminergic neurons from degenerative changes induced by axotomy, similar to the known protective action of NGF in cholinergic neurons. Testing this hypothesis, we found that intraventricular administration of recombinant human BDNF (rhBDNF) to adult rats with transections of the fimbria significantly reduces axotomy-induced degenerative changes of the cholinergic cells in the basal forebrain. No such effect was seen on the dopaminergic neurons of the ventral mesencephalon after transection of their axons ascending in the medial forebrain bundle. Injected in equal amounts, rhBDNF and recombinant human NGF had quantitatively different effects on the cholinergic neurons. BDNF sustained only part of the population of cholinergic neurons affected by the lesion, whereas the entire population was protected by NGF treatment.

Animals↗
PubMed1991

[Effect of acupuncture on neuronal loss induced by axotomy in the rat hypoglossal nuclei].

It was reported previously that acupuncture promotes regeneration of rat's peripheral nerve. In order to study the effect of acupuncture on recovery of motoneuronal lesion and regeneration of central nervous system, we studied the effect of acupuncture on neuronal loss induced by axotomy in the rat hypoglossal nuclei. A total of 6 pairs of rats (acupunctured and controls in pairs) were used, (each pair of) the rats were paired of the same litter and sex. After the right hypoglossal nerves were severed, rats were reared for 14 days. In this period, we gave acupuncture therapy to the acupunctured rats but nothing to the controls. After perfusion, serial paraffin sections of brain stems of rats were cut and stained with cresyl violet. Serial sections per rat were used for counting the perikaryon of the hypoglossal nuclei. The reduced percentage of neuronal population followed axotomy was calculated. It was found that neuronal loss was less severe in acupunctured rats. Difference in the reduced percentage of neuronal population between acupunctured rats and controls was significant statistically. It was shown that acupuncture increased survival of lesioned neurons, and thus acupuncture may be helpful to the recovery of neuronal lesion. The detailed mechanism is going to be explored.

Animals↗
PubMed1989

Lack of in vitro spontaneous activity following axotomy of hamster sensory neurones.

Although peripheral axotomy of dorsal root ganglion cells in mice, rats and cats has been reported to generate spontaneous activity in sensory nerves, we did not find evidence for such activity in the hamster. In vitro, intracellular recording from L6-S1 dorsal root ganglion cells up to 6 weeks after axotomy did not reveal any evidence for either increased membrane excitability or spontaneous activity. Also, in the sciatic nerve-sectioned hamsters, there was a total absence of the self-mutilatory behaviour which has been reported in other rodents. These results support the hypothesis that species specific factors are important for the development of ongoing activity in sensory nerves following injury.

Animals↗
PubMed1989

Naturally occurring cell death and differentiation of developing spinal motoneurons following axotomy.

The purpose of this study was to examine the effects of axon transection on the development and differentiation of spinal motoneurons in the bullfrog (Rana catesbeiana) tadpole. The 3 ventral roots (VRs) that innervate the hindlimb were transected, and the animals were killed 6-7 weeks later (reinnervation took place within 3 weeks). At early stages of development, axotomy resulted in an increase in the number of spinal motoneurons on the operated side. By histological criteria, these motoneurons appeared more differentiated than those in normal tadpoles. Axotomy was effective in increasing motoneuron number only during the period of naturally occurring cell death. Similar effects were seen when the transected VRs were ligated to prevent regeneration. Hindlimb amputation without VR transection had no effect on motoneuron number or differentiation. Thus, target removal is neither a necessary nor a sufficient condition for hyperplasia of the lateral motor column. An extreme loss of spinal motoneurons was seen if the operated tadpole entered into metamorphic climax during the 6-7-week postoperative survival period. Motoneuron loss occurred although the injured motoneurons had reconnected to the hindlimb. In contrast, tadpoles allowed to survive up to 6 months showed no loss of motoneurons if they did not enter metamorphic climax. From these data, it appears axon transection in developing spinal motoneurons exerts its effects on motoneuron number and differentiation by altering the metabolic state of the motoneuron (axon reaction) rather than by depriving it of contact with its target.

Amputation, Surgical↗
PubMed1988

Ultrastructure of the Eimer's organs of the Japanese shrew mole, Urotrichus talpoides (Insectivora, Mammalia) and their changes following infraorbital axotomy.

The Eimer's organ and adjacent structures and their changes after infraorbital axotomy was examined with LM, TEM and SEM in the Japanese shrew mole, Urotrichus talpoides. Approximately 3,000 of Eimer's organ covers the hairless snout tip of this animal. It protrudes to the corium from epidermis, and consists of column-shaped core and the cylindrical peripheral structures. About a dozen of naked fibers ascend vertically in the core; one or a few of them runs amidst the core and the others along its circumferential part. Each fiber has shelf-like endings one in each of 5 to 7 succeeding cells in the upper part of the core. At the uppermost part of the core structure is an opaque degenerating cell. Neurites were considered to grow up with the epidermal cells to which their endings are attached. This pattern of nerve ending was clearly observed for the first time in this study. Usually 3 Merkel cells lie at the base of an Eimer's organ contacting with a soup-plate-shape nerve terminal. In the dermis, an encapsulated corpuscle enveloping a nerve endings is found underneath each Eimer's organ. After axotomy, ipsilateral nerve fibers and their endings totally disappeared from Eimer's organ in a week. Perforation in the domes, flattening of the dome surface, and degeneration of the encapsulated corpuscles appeared on the transected side. In addition to this, several features of degeneration also appeared on the contralateral side. It might be considered as an effect of disuse from the disorders of the other side.

Animals↗
PubMed1977

[Dynamics, following axotomy, of the A1 and A2 median protocerebral neurosecretory cells of the African locust. II. Ultrastructural study].

In the African locust after axotomy of the A1 and A2 median protocerebral neurosecretory cells (M-NSC), the decrease of the basophil neurosecretory products into the pericaryons does not result from an increase of the granulolysis but results from an inhibition of the production. The A1 and A2 neurosecretory products, stored anteriorly and posteriorly to the site of the axonal section, seem destroyed by: 1. a suspected entrance of lytic enzymes into the granule (individual granulolysis); 2. a vacuolar inclusion of small or big cytoplasmic areas rich in granules (partial or total collective granulolysis). The axons of the M-NSC, regenerated without reconnexion to the corpora cardiaca (CC), are not separated by glial elements. This glial deficiency could be at the origin of the inhibition of the neurosecretory dynamics observed in the A1 and A2 M-NSC after axotomy and regeneration without reconnexion to the CC.

Axons↗
PubMed1982

Blockade of retrograde axonal transport delays the onset of metabolic and morphologic changes induced by axotomy.

Axotomy-induced increase in 2-deoxyglucose (2-DG) uptake by motor nuclei and neuronal chromatolytic changes were studied after subepineural injection of colchicine into the motor nerve. Hypoglossal nuclei of either cats or rats were axotomized bilaterally, while one of the nerves was injected with colchicine or saline proximal to the site of nerve transection and the other was left intact or injected with saline. Colchicine abolished or decreased the uptake of 2-DG by axotomized nuclei and delayed the onset of chromatolysis. The decrease in 2-DG uptake was observed in rat hypoglossal nuclei between 24 and 48 hr but not 5 days after drug treatment. In turn, a delay in the onset of chromatolysis was observed in cat hypoglossal nuclei at 14 days but not 30 days after treatment. Saline did not prevent chromatolysis nor the increased uptake of 2-DG. Colchicine injected intraneurally in intact preparations did not result in chromatolysis or in increased 2-DG uptake. Following colchicine injection, the drug remained localized near the site of injection and blocked retrograde axonal transport of horseradish peroxidase in the hypoglossal nerve. These findings suggest that the onset of chromatolysis and of the increase in 2-DG uptake after axotomy are partly dependent upon retrograde axonal transport.

Afferent Pathways↗
PubMed1982

[Effects of distal axotomy on field potentials of the external oculomotor nucleus in the waking cat].

The electrophysiology of abducens nucleus antidromic field potentials from onset of peripheral axotomy up to reinnervation has been studied in the alert cat. Nerve section induced a reduction of 80% in the amplitude of the antidromic field potential. This reduction began from 2 to 5 days after axotomy and returned to control values from 20 to 30 days later, in parallel with recovery of oculomotor function. During the critical chromatolytic period, the latency of the antidromic potentials increased, while the amplitude of the antidromic field potential could be easily modified by adequate visual and/or vestibular stimulation. In contrast to control motoneurons, axotomized cells showed a noticeable variability in their activation latency, as well as a higher threshold for the antidromic invasion of the IS-SD compartments.

Abducens Nerve↗
PubMed1981

[Facilitating effects of axotomy of the glossopharyngeal nerve on the motor reflex response of the frog medulla oblongata].

In the previous papers we have reported that the innervation of the frog lingual mucosa exercises a presynaptic inhibition at the bulbar region in order to integrate the mechanoreceptive information coming from the tongue. The present investigation was undertaken to establish if also the complementary tactile innervation of fungiform papillae has a similar ability. First of all it has been assayed the evocation of PAD tests by papillary innervation; then the facilitating effects, induced by the glossopharyngeal axotomy, on the afferents mass discharges recorded from hypoglossal nerve have been examined. Electrical stimulation of mechanical neurons at the root of IX produced DRP and DRR both at glossopharyngeal and hypoglossal nerves. After 5 and 10 days from glossopharyngeal axotomy becomes evident a considerable enhancement of efferent responses in many diastaltic arches of bulb. These effects strongly suggest a very evident inhibitory activity at the bulbar projection of papillary innervation. From all these observations it follows that presynaptic inhibition is joint to all the lingual mechanoreceptive afferences.

Animals↗