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PubMed1984

Glucose utilization is unchanged in red nucleus after axotomy.

Separate series of adult rats were subjected to unilateral high cervical and low thoracic section of the rubrospinal tract and sacrificed 1-30 (cervical series) and 3-100 days (thoracic series) later. Local cerebral glucose utilization ([14C]2-DG method of Sokoloff et al.) was determined in the red nucleus and in the inferior colliculus, nucleus interpositus and sensorimotor cortex of both sides in operates and controls. Although severe atrophy of rubral neurons follows cervical tractotomy while reversible chromatolytic alterations occur after thoracic lesions, glucose utilization did not differ in the red nucleus of operated and control rats. However, glucose utilization increased slightly in the inferior colliculus of all operated animals, a finding of indeterminate significance. The failure of axotomized intrinsic neurons of red nucleus and their surround to show altered glucose utilization stands in sharp contrast to the marked increase which occurs in cranial nerve nuclei after axotomy of their contained extrinsic neurons. The data are held to constitute another indication that there is a fundamental difference in the metabolic responses of extrinsic and intrinsic mammalian neurons to axotomy and may support the contention that, in mammals, the axon reaction of intrinsic neurons is fundamentally different from that of extrinsic nerve cells. This difference may have significance for failure of axon regeneration in mammalian CNS.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

The effect of axotomy and denervation on calmodulin content in the superior cervical sympathetic ganglion of the rat.

Calmodulin (CaM) in the superior cervical sympathetic ganglion (SCG) of the rat and its changes after transection of the pre- or postganglionic nerves were studied biochemically and immunohistochemically. The concentration of CaM in the SCG was assayed using the extent of activation of CaM-dependent adenylate cyclase from bovine neural retina. In the SCG, CaM concentration was 4.5 micrograms/mg protein, a level similar to that in another peripheral ganglion, the nodose ganglion (4.9 micrograms/mg protein). Following denervation of the SCG, the total amount of ganglionic CaM did not change significantly within a week. On the other hand, CaM content per ganglion increased gradually 3 days after axotomy. Immunohistochemical examination revealed that CaM in unoperated SCG is present both in the neuronal fibers and in the cell soma of sympathetic neurons. But the intensity of the staining with CaM-antibody in the cytoplasmic soma varied among cells. The gross profile of the CaM immunostaining seemed to be unchanged one week after either denervation or axotomy, but the immunoelectron microscopic study showed heavy staining of the innercellular membranes of irregular shape in the axotomized SCG.

Adenylyl Cyclases↗
PubMed1989

Incorporation of plasma [14C]palmitate into the hypoglossal nucleus following unilateral axotomy of the hypoglossal nerve in adult rat, with and without regeneration.

Incorporation of plasma [14C]palmitate into the hypoglossal nucleus of the rat following transection, with and without regeneration of the hypoglossal nerve, was studied using quantitative autoradiography. The left hypoglossal nerve of 3-month-old, male Fischer-344 rats was transected using either: (1) an R-operation, which allowed nerve regeneration; or (2) a D-operation, in which regeneration was prevented. One to 84 days after axotomy, [14C]palmitate was injected intravenously and its rates of incorporation into stable structures of the left and right hypoglossal nuclei were measured at 4 h after injection. Following the R-operation, incorporation into the left hypoglossal nucleus was increased during and following axonal regeneration (up to 23% compared to control side), whereas incorporation was decreased 6-7% in the absence of regeneration, using the D-operation. The time courses of incorporation in both cases corresponded to histological changes, especially cell membrane changes following axotomy and suggest that [14C]palmitate incorporation reflects regenerative and degenerative neuronal changes associated with changes in lipid synthesis.

Animals↗
PubMed1990

Enzyme histochemical profiles of fish spinal motoneurons after cordotomy and axotomy of motor nerves.

Histochemical profiles were made of identified spinal motoneurons from normal adult zebrafish and from animals subjected to cordotomy or unilateral axotomy of the motor nerves. The lesions caused an increase of the myotomal area with oxidative muscle fibers. We studied the question: do changes in the myotomal muscle configuration concur with changes in the enzyme histochemical profiles of innervating motoneurons? Based on the location and size of cell somata, two categories of motoneurons are distinguished: large white (W) motoneurons that innervate the deep fast, glycolytic muscle fibers, and smaller red and intermediate (RI) motoneurons that innervate the superficial slow oxidative and intermediate muscle fibers. In normal animals, glucose-6-phosphate dehydrogenase activity is high in the large W motoneurons and relatively low in the small RI motoneurons. The reverse holds for succinate dehydrogenase activity is high in the large W motoneurons and relatively low in the small RI motoneurons. The reverse holds for succinate dehydrogenase activity. W and RI motoneurons show similar nicotinamide adenine dinucleotide diaphorase activity. Short- (2 weeks) and long- (8 weeks) term effects of lesions were studied. The results show that: (1) the 3 types of lesions lead to prolonged changes in the enzyme histochemical profiles of spinal motoneurons. The type of change depends on the type of lesion and on the type of motoneuron; (2) unilateral axotomy of the motor nerves affects the histochemical characteristics of spinal motoneurons and the myotomal muscle fiber type configuration on the ipsi- and contralateral side. The contralateral effects are conceived as adaptations to maintain a left-right symmetry in the motor output.

Animals↗
PubMed1990

Axotomy decreases conduction velocity of unmyelinated sensory fibres.

We have examined the effect of axotomy on the conduction velocity of unmyelinated sensory fibres in the vagus nerve of the rabbit. In an in vitro preparation of the nodose ganglion, conduction velocity was calculated for each cell from the latency of the intracellularly recorded action potential evoked by vagus nerve stimulation. The average velocity of sensory fibres conducting at less than 1.1 m/s, was 0.53 m/s in 287 control cells and 0.45 m/s in 269 neurones recorded 10 days after cutting the vagus nerve. Therefore, peripheral axotomy decreases conduction velocity in mammalian unmyelinated sensory axons.

Action Potentials↗
PubMed1993

Different stability of neurofilaments for trypsin treatment after axotomy in the dorsal motor nucleus of the vagal nerve and the hypoglossal nucleus.

In an attempt to obtain information about changes of neurofilaments in motor neurons after axotomy, we immuno-histochemically investigated the accumulated neurofilaments in the dorsal motor nucleus of the vagal nerve, which shows nerve cell loss and degenerative changes after axotomy, and in the hypoglossal nucleus, which shows regenerative changes. Affected neurons in the hypoglossal nucleus showed intensified immunoreactivities for neurofilament antibodies phosphorylated at the carboxy-terminal, and these reactions disappeared with trypsin treatment. Accumulated neurofilaments in the neuronal perikarya in the dorsal motor nucleus of the vagal nerve and axons in brain stem also showed intensified immunoreactivities for the same antibodies, and these reactions remained positive after trypsin treatment. Anti-ubiquitin antibody preferentially stained accumulated neurofilaments in the affected vagal neurons, while no reaction was found in the affected hypoglossal neurons. Phosphorylated neurofilaments in hypoglossal neurons are vulnerable to trypsin treatment probably because of the blocking of polymerization or the disassembly of neurofilaments due to amino-terminal phosphorylation. In vagal neurons, the deteriorated amino-terminal phosphorylation or hyperphosphorylation at the carboxy-terminal seems to cause the cross-linkage and polymerization of neurofilaments, and densely packed polymerized neurofilaments probably fail in axonal transport resulting in nerve cell degeneration and death in the dorsal motor nucleus.

Animals↗
PubMed1994

Axotomy-induced changes in cytochrome oxidase activity in the cat trochlear nucleus.

Following a unilateral section of the trochlear nerve, the effects of axotomy on cytochrome oxidase levels in the trochlear nucleus were studied. Cytochrome oxidase levels in the axotomized nucleus were significantly lower than in the control nucleus. The maximal decrease was observed at 2 weeks. Following partial restoration during weeks 3 and 4, cytochrome oxidase levels stabilized at levels only slightly below normal. Since a significant number of trochlear motoneurons die following axotomy, the restoration of cytochrome oxidase levels close to normal suggests that the surviving neurons may compensate for an increased load with a permanent increase in oxidative metabolism.

Animals↗
PubMed1994

Specific expression of type II protein kinase c after axotomy in the dorsal motor nucleus of the vagus nerve and the hypoglossal nucleus.

Protein kinase C (PKC) and growth-associated protein-43 (GAP-43) were investigated immunohistochemically in the dorsal motor nucleus of the vagus nerve and the hypoglossal nucleus after axotomy using monoclonal antibodies against type I, II and III PKC and GAP-43. In the control side of both nuclei, anti-type I and II PKC weakly stained neuronal cell bodies, while anti-type III PKC did not show any reaction with neurons. In the axotomized side of both nuclei, anti-type II PKC antibody intensely stained affected nerve cell bodies as well as plasma membrane. Some of the severed neurons showed intensified reactions for both anti-type II PKC and anti-GAP-43 antibodies in the serial sections. These findings suggest that axotomy increases the type II PKC of the severed neurons, and type II PKC seems to phosphorylate some protein, such as GAP-43, and plays some role in the retrograde neuronal reaction.

Animals↗
PubMed1995

Expression of pituitary adenylate cyclase-activating polypeptide in dorsal root ganglia following axotomy: time course and coexistence.

Pituitary adenylate cyclase-activating polypeptide (PACAP) has recently been demonstrated in sensory neurons. In the present study on rat 17.5% of all neurons, mainly of small size, contained PACAP in normal dorsal root ganglia (DRGs). Transection of the sciatic nerve induced a rapid and strong upregulation in PACAP peptide and mRNA levels which could be seen already after 15 h. After 3 days more than 51.5% of neurons of different sizes expressed PACAP. However, the intensity of PACAP-LI in the DRG neurons declined after 10 days. Thirty days after axotomy, 56.7% of the DRG neurons still expressed PACAP, but with a low intensity, in fact even lower than in normal controls. No VIP- or NPY-positive neurons were observed in normal or axotomized DRGs at 15 h. However a distinct increase in VIP and NPY levels were seen 3 days after the lesion, and their levels were considerably higher after 30 days. PACAP was often present in neurons expressing VIP, NPY and/or galanin. Thus, 3 days after injury, PACAP was present in 84.4%, 95.7%, and 76.8% of the VIP-, NPY-, and galanin-positive neurons, respectively. PACAP was also found in nerve fibers in control sciatic nerves. After nerve ligation, accumulation of PACAP was seen mainly proximal to the injury but also distally, suggesting both anterograde and retrograde transport of the peptide. Also a moderate increase (about 20%) in PACAP levels was found in the superficial spinal dorsal horn 3 days after nerve transection. Taken together, our results suggest that PACAP is involved in the response to nerve injury. The very high levels of expression in different populations of DRG neurons after axotomy, and its different time course as compared to galanin, NPY and VIP indicate that it may play a complementary and/or different role than these peptides in the adaptation to nerve injury, especially in its early phase.

Animals↗
PubMed1996

Effects of peripheral axotomy on neuropeptides and nitric oxide synthase in dorsal root ganglia and spinal cord of the guinea pig: an immunohistochemical study.

The effect of axotomy (3, 10 and 21 days) on the expression of some neuronal markers was analysed in dorsal root ganglia and spinal cord of guinea-pigs using immunohistochemistry. Three weeks following injury, substance P-like immunoreactivity (-LI) was slightly reduced in the DRGs of the ipsilateral side, whereas a marked increase in neuropeptide Y(NPY)-LI could be detected ipsilaterally and a smaller increase contralaterally. NPY-LI was mainly expressed in small, but also some medium-sized and large neuron profiles after axotomy. Galanin-LI showed a moderate bilateral increase. No significant changes could be observed in DRGs for calcitonin gene-related peptide (CGRP)-, vasoactive intestinal polypeptide-, peptide histidine isoleucine- or nitric oxide synthase-LIs. In the ventral horn CGRP-LI was slightly increased bilaterally in motoneurons, most pronounced on the injured side. Autotomy behaviour was seen in seven of the nine animals in the twenty-one day group. The present results demonstrate that also in guinea-pigs several peptides undergo distinct changes in their expression after peripheral nerve injury. However, in contrast to rats and monkeys, galanin-LI is only moderately increased in guinea-pigs. Neuropeptide Y showed a dramatic increase mainly in small neurons, in contrast to the upregulation in large neurons in the rat. Thus, distinct species differences exist with regard to the cellular response to nerve injury.

Animals↗
PubMed1996

Partial coexistence of neuropeptide Y and calbindin D28k in the trigeminal ganglion following peripheral axotomy of the inferior alveolar nerve in the rat.

Immunohistochemistry was applied to examine the correlation between neuropeptide Y (NPY) and the two calcium binding proteins (CaBPs) parvalbumin (PV) and calbindin D28k (CB) in the trigeminal ganglion following peripheral axotomy of the inferior alveolar nerve (IAN) in the rat. Five days following transection and application of FluoroGold (FG) to the cut end of the IAN, approximately 14.8% (80/539) and 18.6% (90/483) of FG-labeled IAN neurons in the trigeminal ganglion showed PV-like immunoreactivity (-LI) and CB-LI, respectively. The mean +/- S.D. area of FG-labeled PV-like immunoreactive (-IR) cells (FG/PV-IR cells) and FG/CB-IR cells were 835.9 +/- 303.1 mu m2 and 712.7 +/- 246.0 mu m2, respectively. FG/PV-IR cells were significantly larger than FG/CB-IR cells. Fourteen days following peripheral axotomy of the IAN, NPY-LI appeared in the medium- to large-sized cells. Double immunostaining revealed that approximately 3.3% (52/1569) of NPY-IR cells in the axotomized trigeminal ganglion displayed PV-LI, while approximately 26.7% (371/1392) of NPY-IR cells displayed CB-LI. The mean +/- S.D. cross-sectional areas of PV-IR and CB-IR trigeminal ganglion cells displaying NPY-LI were 819.5 +/- 265.6 mu m2 and 766.5 +/- 279.7 mu m2, respectively. There were no significant differences in the cross-sectional areas either between NPY/PV-IR cells and NPY/CB-IR cells, or between FG/PV-IR cells and NPY/PV-IR cells, or between FG/CB-IR cells and NPY/CB-IR cells. The present results indicate that injury-evoked medium- to large-sized NPY neurons were a different population from large-sized PV neurons, and NPY was partly co-localized with CB.

Animals↗
PubMed1996

Ca2+/calmodulin-dependent protein kinase type IV in dorsal root ganglion: colocalization with peptides, axonal transport and effect of axotomy.

Using the indirect immunofluorescence technique, the distribution of Ca2+/calmodulin-dependent protein kinase IV (CaM kinase IV) was studied in dorsal root ganglia (DRGs) and the sciatic nerve under normal circumstances and after axotomy and nerve ligation. CaM kinase IV-like immunoreactivity (-LI) was observed mainly in small DRG neurons but also in some large ones with the immunoreactivity mainly confined to the cell nuclei and with varying levels in the cytoplasm. CaM kinase IV-LI was present in around 1/4 of all CGRP-positive neurons and in the vast majority of the somatostatin-positive neurons. The enzyme levels decreased markedly after axotomy. The enzyme was also observed in axons in the sciatic nerve and accumulated both proximal and distal to a ligation. The present results suggest that CaM kinase is not of direct importance for upregulation of neuropeptides in DRG neurons after nerve injury. In addition to a nuclear function it may also play a role in the peripheral processes of DRG neurons.

Animals↗
PubMed1996

A continuous striatal infusion of 6-hydroxydopamine produces a terminal axotomy and delayed behavioral effects.

Rat models of Parkinson's disease typically employ a rapid nigral injection of 6-hydroxydopamine (6-OHDA) to produce a near-complete loss of nigrostriatal dopamine neurons, and thus, model end stage disease. The present report describes the use of a continuous, low dose infusion of 6-OHDA into the striatum which produces a terminal axotomy of nigrostriatal dopamine neurons and protracted behavioral response. A solution of 6-OHDA in 0.4% ascorbate, delivered at 37 degrees C from osmotic minipumps, was stable for 8 days as determined by its retained toxicity to a dopaminergic neuroblastoma cell line. The continuous infusion of 0.2 mu g 6-OHDA per h did not affect the striatal uptake of [3H]%GABA, [3H]choline, or [3H]glutamate but reduced [3H]dopamine uptake by 55% within 1.5 days after the start of the infusion. The striatal infusion of 6-OHDA produced a dose-dependent reduction of striatal dopamine and DOPAC levels but did not alter HVA, 5-HT, or 5-HIAA. An increase in amphetamine-induced ipsiversive rotations occurred within 1.5 days after the acute striatal injection of 20 mu g or 30 mu g of 6-OHDA but required 4 days to develop with the continuous 6-OHDA infusion. The topography of the lesion mapped by [3H]mazindol binding showed that, beginning by 1.5 days, a diffuse depletion of terminals encompassed much of the striatum in the 30 mu g acute injection group, whereas in the continuously infused rats, the lesion was apparent only by 4 days and was restricted to a smaller and more completely lesioned area. Unlike acutely lesioned animals, continuously infused rats revealed no obvious loss of dopamine neurons in the pars compacta by 5 weeks after 6-OHDA. The continuous striatal infusion of 6-OHDA can produce a topographically limited terminal axotomy of dopamine neurons and a protracted behavioral impairment.

Animals↗
PubMed1987

Proteins of fast axonal transport in regenerating rat sciatic sensory axons: a conditioning lesion does not amplify the characteristic response to axotomy.

The regeneration rate of axons following injury can be influenced by the past history of the neuron: if an axon received a "conditioning" lesion its regeneration rate from a subsequent "test" lesion is increased. Because regeneration is associated with the increased synthesis and fast axonal transport of growth-associated proteins of unknown functions, we investigated whether the increased regeneration rate (to 81%) seen in conditioned rat sciatic sensory axons following a test lesion was associated with an amplification of the increase in growth-associated protein expression. The test lesion did not produce any amplification of the increase in growth-associated protein expression resulting from the preceding conditioning lesion. We conclude that this aspect of the cell body reaction to axotomy is not involved in the conditioning lesion effect on regeneration rate, and, as a corollary, it appears that fast-transported proteins whose expression alters after axotomy are not involved in the regulation of the regeneration rate.

Animals↗
PubMed1990

The effects of age on enzyme activities in the rat facial nucleus following axotomy: acetylcholinesterase and cytochrome oxidase.

Advancing age affects the ability of motor neurons to regrow axons after the facial nerve is crushed. In rats, it requires 14 days after injury for 3-month-old animals to resume normal whisker activity, compared to at least 19 days in 15-month-old animals. The present study examines central enzymatic responses of facial motor neurons to axotomy. During the postoperative period from 1 day through 8 weeks, alternate frozen sections of brain stem are histochemically reacted to demonstrate activities of acetylcholinesterase (AChE) or cytochrome oxidase (COX) and the reactions are quantified using computerized image analyzing densitometry. AChE activity is evaluated separately in perikaryal cytoplasm and neuropil, while COX is assayed in the facial nucleus as a whole. Coincident with the initiation of axon outgrowth the activities of these enzymes decrease in the neurons. For AChE the decrease is greater in the older animals; for COX the decrease is equivalent in both age groups. With regard to the perikaryal AChE and the neuropil AChE, the recovery patterns are different in the two locations. In the perikarya AChE activity begins to recover after 4 days in both age groups; however, AChE activity in the neuropil remains decreased until after functional recovery of whisker activity, when it recovers rapidly in the 3-month-old animals, but more gradually in the 15-month-old animals. In both age groups, COX activity gradually decreases in response to axotomy. In the 3-month-old animals it recovers rapidly following return of whisker activity, while in the 15-month animals COX activity is maintained at the decreased level through 28 days post-crush, before it begins its gradual recovery. The study demonstrates that age differences are most apparent after the reestablishment of functional connections. This age-related deficiency may be related to deficiencies in retrogradely transported signals arising from the reinnervated target or in the older neuron's ability to respond to such signals.

Acetylcholinesterase↗
PubMed1989

Axotomy-like electrophysiological alterations in spinal motoneurons in beta,beta'-iminodipropionitrile neuropathy.

Motoneurons (MNs) exhibit characteristic electrophysiological alterations following axotomy which are concomitants of perikaryal remodeling induced by the axonal injury. beta,beta'-Iminodipropionitrile (IDPN) neurotoxicity, which produces proximal axonal swellings in the first internodes of motor fibers, was studied as a model of perikaryal electrophysiological properties in axonal pathologies without axonal degeneration. Similarities between parameters of MN excitability (delayed depolarizations, repetitive discharge, and the afterhyperpolarization potential, AHP) known to occur in axotomized MN and those in IDPN neuropathy were examined in type-identified spinal MN of cats during the evolution (7 to 35 days) of proximal axonal swellings. Delayed depolarization potentials were observed frequently in fast MN types throughout the neuropathy but only at 35 days in slow MN types. Similarly, repetitive firing occurred most prominently in fast MNs early in the neuropathy. Concomitantly, AHP duration decreased as early as 7 days in all MN types and was significantly shortened in types FF, FR, and S motoneurons. AHP peak amplitude and current declined continuously from 7 to 35 days of the neuropathy and were significantly (p less than 0.05) decreased at 35 days in types FF and S MNs. These results suggest that not only are fast MN types vulnerable early in IDPN neuropathy, but also that all MN types exhibit electrophysiological changes strikingly similar to those following mechanical axotomy. The possibility is raised that IDPN may initiate electrophysiological changes, analogous to perikaryal remodeling, by mechanism(s) unrelated to axonal degeneration.

Action Potentials↗
PubMed1993

Medullary afferent vagal axotomy disrupts NaCl-induced short-term taste aversion learning.

The effect of medullary afferent vagal axotomy on NaCl-induced short-term and long-term taste aversion learning (TAL) was examined to assess the relevance of the vagus nerve in drug-induced TAL. The results show that medullary afferent vagal axotomy disrupts NaCl-induced short-term (nondelayed) TAL, while having no effect on learning acquired with the same product in long-term (delayed) TAL protocols. Acquisition of learning in delayed discrimination tasks may be mediated by alternative mechanisms of nonvagal nature, e.g., the humoral system. The possibility that short-term and long-term TAL may be mediated by different neurobiological substrates is discussed.

Animals↗