Bilateral Section Research Articles

SPINAL IMPLANTS OF OLFACTORY ENSHEATHING CELLS PROMOTE AXON REGENERATION AND BLADDER ACTIVITY AFTER BILATERAL LUMBOSACRAL DORSAL RHIZOTOMY IN THE ADULT RAT

We performed spinal implantation of olfactory ensheathing cells to demonstrate dorsal root afferent regeneration as well as bladder activity restoration after lumbosacral L6 to S2 rhizotomy. Spinal segments receiving bladder innervation, usually L6, S1 and S2, were identified by bipolar stimulation of the ventral roots. Bilateral section of the identified dorsal roots L6 to S2 was performed in 18 male Wistar rats. Immediately after rhizotomy olfactory ensheathing cells or vehicle was unilaterally injected in the vicinity of the sacral parasympathetic nucleus in 9 rats each using a glass micropipette and air pulse system. The severed roots were reattached to the cord with fibrin glue and the animals recovered under antibiotic prophylaxis. Anatomical regeneration of bladder wall primary afferents was demonstrated by the presence of labeled wheat germ agglutinin-horseradish peroxidase fibers in the dorsal horn and sacral parasympathetic nucleus in 8 of 9 cases of olfactory ensheathing cell implantation but not in the 9 controls injected with vehicle. One week after surgery all rats had an atonic bladder on cystometrography. At 6 weeks 8 of the 9 olfactory ensheathing cell implanted rats had recovered bladder activity. No recovery was observed in controls, in which vehicle was injected instead of olfactory ensheathing cells. Regenerated primary afferent fibers from the bladder project to the sacral parasympathetic nucleus, where they presumably form synapses mediating the recovery of bladder activity. Thus, olfactory ensheathing cell implants in the adult rat promote sensory axon regeneration, target reinnervation and bladder activity restoration.

Reflex respiratory response to changes in upper airway pressure in the anaesthetized rat.

1. We examined the upper airway (UA) motor response to upper airway negative pressure (UANP) in the rat. We hypothesized that this response is mediated by superior laryngeal nerve (SLN) afferents and is not confined to airway dilator muscles but also involves an increase in motor drive to tongue retractor and pharyngeal constrictor muscles, reflecting a role for these muscles in stabilizing the UA. 2. Experiments were performed in 49 chloralose-anaesthetized, tracheostomized rats. Subatmospheric pressure in the range 0 to -30 cmH(2)O was applied to the isolated UA. Motor activity was recorded in separate experiments from the main trunk of the hypoglossal nerve (XII, n = 8), the pharyngeal branch of the glossopharyngeal nerve (n = 8), the medial and lateral branches of the XII (n = 8) and the pharyngeal branch of the vagus (n = 8). Afferent activity was recorded from the whole SLN in six experiments. 3. All UA motor outflows exhibited phasic inspiratory activity and this was significantly (P < 0.05) increased by UANP. Tonic end-expiratory activity increased significantly in response to pressures more negative than -20 cmH(2)O. Bilateral section of the SLN also increased (P < 0.05) motor activity and abolished the responses to UANP. Electrical stimulation of the SLN inhibited inspiratory XII activity. SLN afferents were tonically active and inhibited by UANP. 4. We conclude that UANP causes a reflex increase in motor drive to pharyngeal dilator, tongue retractor and pharyngeal constrictor muscles via afferent fibres in the SLN. Tonic activity in SLN afferent fibres at zero transmural pressure exerts a marked inhibitory effect on UA motor outflow.

Effects of vagal and splanchnic section on food intake, weight, serum leptin and hypothalamic neuropeptide Y in rat

Truncal vagotomy can cause reduced food intake and weight loss in humans and laboratory animals. In order to investigate some of the factors that might contribute to this effect, we studied changes in ingestive behaviour, whole body and organ weights, serum leptin and hypothalamic neuropeptide Y in rats with bilateral vagal section, bilateral splanchnic nerve section and combined vagotomy plus splanchnectomy. Pyloromyotomy was combined with vagotomy to lessen effects of vagotomy on gastric emptying. Animals with vagotomy or vagotomy plus splanchnectomy lost weight and decreased their daily food intake relative to animals with splanchnectomy alone, rats with bilateral sham exposure of one or both nerve, or rats with pyloromyotomy alone. Serum leptin and white fat mass, 4 weeks after vagotomy, were about 20% of the values in the sham-operated animals at this time. No effect for splanchnic nerve section alone was observed. Pyloromyotomy caused no reduction in weight or fat mass, but reduced serum leptin. Following vagotomy with or without splanchnic nerve section, neuropeptide Y was elevated in the arcuate nucleus relative to values for the other four groups. Changes in neuropeptide Y were inversely correlated with levels of serum leptin. It is concluded that the effect of vagotomy could be due to the loss of a feeding signal carried by vagal afferent neurons, or to changed humoral signals, for example, increased production of a satiety hormone. However, it cannot be attributed to signals that reduce feeding (for example, gastric distension) reaching the central nervous system via the splanchnic nerves. The changes were sufficient to cause weight loss even though serum leptin was decreased, a change that would be expected to increase food intake.

Whisker Deafferentation and Rodent Whisking Patterns: Behavioral Evidence for a Central Pattern Generator

Even in the absence of explicit stimulation, rats emit patterns of rhythmic whisking movements. Because of their stereotyped nature and their persistence after sensory denervation and cortical ablation, whisking movements have been assumed to reflect the output of a central pattern generator (CPG). However, identification of a movement pattern as the product of a CPG requires evidence that its generation, patterning, and coordination are independent of sensory input. To provide such evidence, we used optoelectronic instrumentation to obtain high-resolution records of the movement trajectories of individual whiskers in rats whose heads were fixed to isolate their exploratory whisking from exafferent inputs. Unconditioned whisking patterns were quantitatively characterized by a biometric analysis of the kinematics, rhythmicity, and coordination of bilaterally homologous vibrissa movements. Unilateral and bilateral sectioning of the infraorbital nerve, which innervates the whiskers, was then performed to block reafferent inputs generated by the animal's own whisking movements. Unilateral sectioning of the nerve has no effect on whisking kinematics but is followed by a significant but relatively transient bilateral increase in whisking frequency. However, bilateral deafferentation, when performed in a single-stage procedure, does not disrupt the generation, patterning, or bilateral coordination of whisking patterns in the rat. These findings provide strong behavioral evidence for a whisking CPG and are discussed in relation to its possible location and properties.

Delayed administration of NGF reverses nerve injury induced central alterations of primary afferents.

We have examined whether delayed exogenous NGF administered to an axotomised peripheral nerve reverses the increased transganglionic choleragenoid (CTB) labelling in lamina II. Two, four, eight or 18 weeks after bilateral sciatic nerve section, NGF was applied unilaterally for an additional 2-week period to the transected nerve stump. The transganglionic choleragenoid labelling and substance P (SP) expression were determined and compared to the contralateral axotomised side in the spinal cord dorsal horn. Delayed NGF administration reversed the transganglionic choleragenoid labelling in lamina II when administered 2 or 18 weeks after the sciatic nerve lesion, but not at 4 or 8 weeks. There was also a clear recovery of SP on the axotomised, NGF-treated side 2 or 18 weeks after the sciatic nerve lesion, but not at the intermediate survival times. At the longer survival time, however, there was a recovery of SP regardless of NGF treatment. These results suggest that there is a critical window as to when NGF administration can be effective in reversing axotomy-induced changes in the spinal cord.

Spinal proerectile effect of oxytocin in anesthetized rats.

The spinal cord contains the neural network that controls penile erection. This network is activated by information from peripheral and supraspinal origin. We tested the hypothesis that oxytocin (OT), released at the lumbosacral spinal cord level by descending projections from the paraventricular nucleus, regulated penile erection. In anesthetized male rats, blood pressure and intracavernous pressure (ICP) were monitored. Intrathecal (it) injection of cumulative doses of OT and the selective OT agonist [Thr(4),Gly(7)]OT at the lumbosacral level elicited ICP rises whose number, amplitude, and area were dose dependent. Thirty nanograms of OT and one-hundred nanograms of the agonist displayed the greatest proerectile effects. Single injections of OT also elicited ICP rises. Preliminary injection of a specific OT-receptor antagonist, hexamethonium, or bilateral pelvic nerve section impaired the effects of OT injected it. NaCl and vasopressin injected it at the lumbosacral level and OT injected it at the thoracolumbar level or intravenously had no effect on ICP. The results demonstrate that OT, acting at the lumbosacral spinal cord, elicits ICP rises in anesthetized rats. They suggest that OT, released on physiological activation of the PVN in a sexually relevant context, is a potent activator of spinal proerectile neurons.

Imiquimod-elicited emesis is mediated by the area postrema, but not by direct neuronal activation

The immunomodulator, imiquimod, has been shown to have antiviral and antitumor properties in animal models. It also has been reported to alter cytokine levels in both animals and humans. However, because imiquimod appeared to be emetic, studies were conducted to determine the degree of sensitivity, and the pathways involved. Subcutaneous administration of ≥10 mg/kg imiquimod to ferrets elicited emesis with latencies as short as 2′; 12 and 15 mg/kg were optimal doses. Emetic responsiveness was eliminated by complete ablation of the area postrema, but was unaffected by bilateral supradiaphragmatic section of the vagus nerve. This indicates that the emesis is produced by an activation of the chemoreceptor trigger zone B the area postrema. Ferret brain stem slices (450 μm) were preincubated in oxygenated Krebs-Ringer and then mounted in a submerged slice recording chamber. Extracellular recordings of spontaneous and ionophoretically evoked activity of area postrema neurons were obtained for up to 8 h, while the effect of bath-applied imiquimod was determined. Under control conditions, neurons showed a low frequency spontaneous discharge. Introduction of imiquimod (concentration range, 1 × 10−7 to 5 × 10−8M) had no effect on neuronal firing. With ionophoresis of glutamate from an independent micropipette, a brief excitatory response was obtained. We conclude that imiquimod does not directly excite area postrema neurons. It is likely that imiquimod causes synthesis and release of some unknown emetic substance(s), very possibly cytokines.

Vaginocervical stimulation-induced release of classical neurotransmitters and nitric oxide in the nucleus of the solitary tract varies as a function of the oestrus cycle

The effects of vaginocervical stimulation (VCS) on glutamate (GLU), aspartate (ASP), γ-aminobutyric acid (GABA), noradrenaline (NA), arginine (ARG) and nitric oxide (NO) (citrulline) release in the nucleus of the solitary tract (nTS) were measured in anaesthetised female rats as a function of the oestrus cycle. During pro-oestrus/oestrus (P/E), but not during met-oestrus/di-oestrus (M/D), VCS significantly increased concentrations of NA, ASP, GLU, NO (citrulline) and GABA, but not ARG. Basal NA concentrations were also increased in P/E. These effects were prevented by bilateral section of either the vagus nerve or pelvic and hypogastric nerves. Vagotomy also significantly decreased basal NO concentrations in M/D and P/E while pelvic and hypogastric nerve section significantly increased GABA concentrations. Our results therefore confirm that the nTS is a relay structure for the visceral afferents sending information from the uterus into the central nervous system. The ability of VCS to trigger classical transmitter release and NO in the female is influenced by the stage of the oestrous cycle and is routed both via the vagus and pelvic/hypogastric nerves.

Crossed unilateral lesions of the medial forebrain bundle and either inferior temporal or frontal cortex impair object–reward association learning in Rhesus monkeys

In an accompanying paper we showed that combined transection of the fornix, amygdala and temporal stem in monkeys produced dense amnesia, including an impairment in visual object–reward association learning. We proposed that this combined surgical section had its effect by isolating temporal cortex from the ascending projections of the basal forebrain and midbrain structures. To test this hypothesis, in the present experiment we disconnected the inferior temporal cortex from these basal forebrain and midbrain structures, while sparing cortical white matter, by crossed unilateral lesions of the medial forebrain bundle in one hemisphere and inferior temporal cortex in the opposite hemisphere. The aim of the medial forebrain bundle lesion was to section axons of cells, both those that project to the cortex via the medial forebrain bundle, and those which control the activity of these same structures. A single unilateral lesion alone had no effect on the ability to learn and remember visual object–reward associations, but the crossed unilateral lesions produced an impairment in this task which was equal in severity to the impairment seen earlier after bilateral section of the fornix, amygdala and temporal stem. The impairment was not an effect of interrupting fibres to the cortex from the ventromedial hypothalamus, or of unilateral sensory neglect. This supports the hypothesis that these midbrain and basal forebrain afferents to the inferior temporal cortex are important for new visual learning. Furthermore, an impairment of equal severity was demonstrated in a separate group of animals that received crossed unilateral lesions of the medial forebrain bundle in one hemisphere and of the frontal cortex in the opposite hemisphere. We propose that the frontal cortex acts to modulate basal forebrain activity which in turn reinforces object representations in the inferior temporal cortex during learning.

Dexrazoxane-induced reduction in mortality in mice subjected to severe forebrain ischemia

In this study we determined whether dexrazoxane reduces mortality in mice subjected to bilateral sequential common carotid artery sectioning. Under pentobarbital anesthesia, the left common carotid artery was ligated and sectioned. In one group of experiments, 32 days later mice were injected i.p. with dexrazoxane (16, 64, 256 mg/kg) 30 min before being reanesthetized to ligate and section the right common carotid artery. In the second group of animals dexrazoxane was given, at the same doses, 15 min after ligating and sectioning the right common carotid artery. Dexrazoxane significantly decreased the cumulated mortality rate compared with controls. When given 30 min before the second surgery dexrazoxane was active only at doses of 256 mg/kg. In contrast, when given 15 min after the second ischemic insult the protective effect of dexrazoxane was already evident at 16 mg/kg, increasing dose-dependently with an almost complete protection at 256 mg/kg during the first 24 h of observation; some degree of protection persisted up to day 4. One possible explanation for this striking difference in efficacy is that dexrazoxane passes through a compromised but not through an intact blood–brain barrier. We conclude that dexrazoxane has important neuroprotective properties against brain ischemia and that its clinical trial in stroke should be considered. Drug Dev. Res. 51:149–152, 2000. © 2001 Wiley-Liss, Inc.

Water stimulation of the posterior oral cavity induces inhibition of gastric motility.

The response of gastric motility to the administration of water and saline in the larynx and epiglottis was investigated in urethan-chloralose anesthetized rats. Administration of water inhibited motility of the distal stomach, but 0.15 M NaCl did not induce the inhibitory response. Bilateral sectioning of the superior laryngeal nerve (SLN) abolished the inhibitory response induced by water. Bilateral cervical vagotomies abolished the inhibitory responses, although spinal transection did not affect the inhibitory response. These inhibitory responses have been observed in immobilized animals. The degree of inhibition by water and hypotonic saline was negatively correlated with the sodium concentration. In contrast, the degree of inhibition to hypertonic saline was positively correlated with the sodium concentration. The proximal stomach also showed a reduction in intragastric pressure in response to the administration of water. These findings suggest that water-responsive afferent neurons in the SLN suppress gastric motility via the vagal efferent nerve.

Effects of unilateral or bilateral superior ovarian nerve section in infantile rats on follicular growth.

We report the effects that sectioning the superior ovarian nerve of infantile female rats has on their follicular development at different ages before puberty. Compared with the control group, sham-operated animals showed a significant decrease in the number of measured follicles in right and left ovaries, although no difference in the follicular atresia ratio was observed. Animals with a sectioned left superior ovarian nerve (SON), killed 12 days after surgery had a significant increase in the number of follicles in the ovaries. Most of the follicles were atretic. Sectioning the right SON induced contrasting effects in the ovaries of animals killed 4 and 16 days after surgery. Rats with a denervated (right) ovary showed a decrease in the number of follicles and a greater number of atretic follicles compared with the control group, whereas the innervated (left) ovary showed an increase in measured follicles compared with the control group. Bilateral sectioning had no apparent effect on the total number of follicles measured, although an increased number of atretic follicles in both ovaries was observed. Animals with a unilateral section of the SON, killed 8 and 12 days after surgery, showed a decrease in serum concentrations of estradiol. In turn, animals killed 16 days after surgery showed a significant increase in estradiol and a decrease in the progesterone serum concentration. These results suggest that sympathetic innervation of the ovary via the SON has a stimulatory role in the regulation and differentiation of follicular growth.

Functional anatomy of the vagal innervation of the cervical trachea of the dog.

The canine cervical trachea has been used for numerous studies regarding the neural control of tracheal smooth muscle. The purpose of the present study was to determine whether there is lateral dominance by either the left or right vagal innervation of the canine cervical trachea. In anesthetized dogs, pressure in the cuff of the endotracheal tube was used as an index of smooth muscle tone in the trachea. After establishment of tracheal tone, as indicated by increased cuff pressure, either the right or left vagus nerve was sectioned followed by section of the contralateral vagus. Sectioning the right vagus first resulted in total loss of tone in the cervical trachea, whereas sectioning the left vagus first produced either a partial or no decrease in tracheal tone. After bilateral section of the vagi, cuff pressure was recorded during electrical stimulation of the rostral end of the right or left vagus. At the maximum current strength used, stimulation of the left vagus produced tracheal constriction that averaged 28.5% of the response to stimulation of the right vagus (9.0 +/- 1.8 and 31.6 +/- 2.5 mmHg, respectively). In conclusion, the musculature of cervical trachea in the dog appears to be predominantly controlled by vagal efferents in the right vagus nerve.

Pelvic floor structures and sacral innervation influence rectal compliance

Structural anatomy and the surrounding pelvic floor musculature influence the compliance of the rectum. Additionally, afferent inputs and spinal reflexes from sacral spinal roots SI-S3 modulate rectal compliance. The individual contribution of these mechanisms has not been clearly established. Aims. The current study sought to evaluate the influence of pelvic floor muscles and sacral innervation, both afferent and efferent, on rectal compliance in the cat. Methods. Following surgical preparation, animals were decerebrated and ventilated. After a lumbo-sacral laminectomy, the sacral roots were exposed. All measures were obtained after stabilization of respiration and arterial pressure. Pressure-volume curves were determined during slow ramp distention (0 to 50 mmHg) in animals with intact pelvic floor (n= 10) and following isolation of the rectum from the pelvic floor musculature (n= 10). To assess the influence of spinal outflow and afferent inputs, rectal compliance was measured in the isolated rectum following transection of the dorsal and ventral sacral roots sequentially from SI to S3 and from dorsal to ventral (n=4). Results. Isolation of the rectum from the pelvic floor musculature resulted in a significant interaction effect (F(9,189)=6.12, p<O.OOI) due to a rightward shift of the pressure-volume curve in the lower range of distention (0-25 mmHg) without affecting compliance at higher pressure. Bilateral section of dorsal root SI further shifted the pressure-volume curve to the right (F(l,3)= 13.69, p=O.03). Subsequent sections to ventral root S3 failed to significantly influence rectal compliance. Section of ventral root S3 resulted in an additional rightward shift (F(l,3)=41.35, p=0.008) of the compliance curve. Conclusion. The rightward shift of the compliance curve in the lower distention range after isolation of the rectum from its pelvic floor anchors may have resulted from an alteration in adaptive relaxation of the longitudinal muscle. Section of sacral spinal afferents in the dorsal root S I and efferent inputs from ventral root S3 further shifted the compliance curve to the right. These observations suggest that afferent inputs and spinal reflexes may exert inhibitory influences on the rectum during distention, thereby initiating adaptive responses to rectal filling (Supported by DFG).

Baroreceptor control of heart rate in the awake toad: peripheral autonomic effectors and arterial baroreceptor areas

Systemic injection of sodium nitroprusside (30 μg/kg, i.v.) in the awake Bufo paracnemis toad induced a fall in arterial blood pressure and tachycardia. This tachycardia, but not the hypotension, was significantly reduced in toads with bilateral electrolytic lesion of the caudal and commissural regions of the solitary tract nucleus and in animals with transection of the spinal cord, 2 mm below the obex. This indicates that the tachycardia is reflex, depends on the integrity of the solitary tract nucleus and is due to descending spinal autonomic activation. Pretreatment with propranolol (4 mg/kg, i.v.) significantly reduced the tachycardia but did not block it completely, showing the importance of beta-adrenoceptors in its genesis. The reflex increase in heart rate induced by nitroprusside was not statistically significant in animals with bilateral section of the laryngeal nerve, whose baroreceptor fibers originate from the pulmocutaneous artery or in animals in which the bilateral section of the laryngeal nerve was performed together with section of the glossopharyngeal nerves, which incorporate fibers originating from the carotid labyrinth. The reduction of the reflex tachycardia was significant in toads with aortic arch denervation alone or combined with section of the laryngeal nerves or in animals with complete denervation of the three baroreceptors areas. These results suggest that the region of the aortic arch, when submitted to unloading, is the most important baroreceptor zone for cardiac compensation in toads.

Bilateral sequential common carotid artery sectioning in mice as a new model for testing neuroprotective drugs

The consequences of cerebral global ischemia induced by right common carotid artery (CCA) sectioning in mice previously subjected to a chronic deficiency of brain blood supply by sectioning of the contralateral artery, as well as the neuroprotector efficacy of 3 5-hydroxytryptamine_1A receptor agonists, were investigated in this study. Left CCA sectioning produced a remarkable increase in the diameter of arteries constituting the cerebral arterial circle; the effect was maximal at 32 days. No neurological deficit or significant brain damage were found in these animals. In mice with previous left CCA sectioning, sectioning of the contralateral artery gave rise to a wide range of motor-behavioral abnormalities, characteristic brain damage, and high mortality rate. Neurological abnormalities ranged from alterations in locomotor activity to lack of coordination, hemiparesis, circling, immobility, and unresponsiveness. Seventeen percent of the animals died within 15 minutes after the acute ischemic event, and the number of deaths progressively increased to 63 at 24 hours and 92 at day 8. In mice killed 72 hours after right artery sectioning, the incidence of infarction was 100, and was more pronounced in the right hemisphere. The 3 5-hydroxytryptamine_1A receptor agonists, tested reduced the severity of neurological alterations and decreased the cumulative mortality rate of the mice. The activity of ±8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide and indorenate was dose-dependent, whereas the protective action of buspirone was more pronounced with lower and higher doses than with intermediate ones. Maximal neuroprotection was obtained with ±8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide. This new model of global ischemia may be relevant to stroke in humans. It incorporates the idea of provoking an acute ischemic insult in animals subjected to chronic hypoperfusion and uses neurological and mortality endpoints to determine the effects of brain ischemia and the degree of neuroprotection. The present findings warrant continued efforts to refine our model to better reflect human stroke and to determine its usefulness in predicting the effectiveness of a particular treatment strategy.

Involvement of sympathetic activation and brown adipose tissue in calcitonin gene-related peptide-induced heat production in the rat

We previously reported that microinjection of calcitonin gene-related peptide (CGRP; 1.6–8.0 pmol, 0.2–1.0 μl) into the ventromedial hypothalamus (VMH) increased oxygen consumption ( VO 2), heart rate (HR), colonic temperature ( T co), and temperature of interscapular brown adipose tissue ( T IBAT). In the present study, we investigated whether the autonomic nervous system is involved in the CGRP-induced heat production in urethane-anesthetized rats. Intraperitoneal administration of the ganglion blocker hexamethonium (20 mg/kg) or the β-adrenergic antagonist propranolol (5 mg/kg) suppressed the CGRP-induced increases in VO 2, HR, T co, and T IBAT. Pretreatment with the α-adrenergic antagonist phentolamine (5 mg/kg) partly attenuated the heat production response but did not affect the tachycardiac response. Bilateral sectioning of the nerves supplying the IBAT attenuated the CGRP-induced increase in T IBAT but not significantly that in VO 2 or T co. In rats with adrenal demedullation, the effects of CGRP were similar to those in intact rats. These results suggest that the CGRP-induced heat production is mediated by the sympathetic nervous system and, at least in part, by the BAT through the α- and β-adrenoceptors.

Effect of tracheosyringeal denervation on call in greenfinch (Carduelis sinica)

After sections of left or right tracheosyringeal nerve (NXIIts), greenfinches may repeat everyday calls, with no effect on temporal properties. It is suggested that either side of syrinx may produce sound alone and ipsilateral innervation of NXIIts for the syringeal muscles. After section of left NXIIts, the bird produces the vocal pattern of partial tone increase, and effects on the sound intensity and sentence length average 1.4 and 2.8 times those after section of right NXIIts, suggesting that the innervation of NXIIts has left side dominance. After bilateral section of NXIIts, the call rhythm in company with expiratory motions is 98-146 times/min, on an average, and lose all sentence types and syllable structure of normal call. But the call spectra produced by tympaniform membrane vibrations without innervation still reserve frequency components similar to the tonic frequency and harmonics of normal calls.

Stereological assessment of the glial reaction to chronic deafferentation of the cochlear nuclei in the macaque monkey (Macaca fascicularis)

Neurectomy of the auditory nerve produces a massive deafferentation of the cochlear nuclei (CN) in the brainstem. Degenerating primary afferents are removed in the acute phase, and this is followed by a synaptic reorganization in the CN. As part of an ongoing study on the effect and applicability of auditory brain implants in the CN of Macaca fascicularis monkeys, we studied the chronic response of astrocytes in the CN to bilateral deafferentation of the VIIIth cranial nerve. Four control and five deafferentated animals were employed. The treated animals had a bilateral extradural section of the VIIIth cranial nerve and a survival of 3 months. Animals were euthanized and perfused, and the brainstem was serially sectioned. The astrocyte population of the CN was studied by glial fibrillary acidic protein (GFAP) immunohistochemistry and quantified by unbiased stereological methods. The total length of astrocyte processes, L(proc), was estimated as the product of nuclear volume V(nuc), which was estimated by the Cavalieri method, times the ratio L(V)(proc, nuc) of process length to nuclear volume. Mean nuclear volume was significantly lower in deafferented animals, whereas the mean ratio L(V)(proc, nuc) was higher (albeit no statistical significance was reached). However, the mean total astrocytic process length was virtually the same in both groups. The absence of a length increase in the glial processes indicates a decrease of the astrocytic reaction after the acute phase. No glial scar is present in the CN of the monkey after long-term deafferentation, so the usefulness of auditory brain implants to stimulate CN neurons directly as a means to overcome deafness resulting from direct damage to the VIIIth cranial nerve (i.e., acoustic neuromas) is plausible.

Fast and slow recovery phases of goldfish behavior after transection of the optic nerve revealed by a computer image processing system

As the goldfish is a common experimental animal for vision research, including psychophysical behavior, it is very important to quantitatively score fish behavior. We have previously developed a computer image processing system which can acquire the positional coordinates of goldfish moving freely in an aquarium and determine turning directions (go straight, right or left turn). In the present study, an algorithm to determine tilting angles of moving goldfish was constructed. We also made histograms for quantifying the interaction between pairs of goldfish (two-point distance). By using these histograms, we estimated the time-course of behavioral regeneration after optic nerve transection in goldfish. Control goldfish showed an equal percentage of right or left turns and maintained an upright position in a dorsoventral axis. When the optic nerve of a goldfish was unilaterally sectioned, the goldfish showed predominant turning and slight tilting toward the intact eye. The abnormal turning and tilting behaviors lasted for 10–14 days and then gradually decreased, returning to control behaviors by one month after the unilateral transection. When the optic nerve of a single goldfish was bilaterally sectioned, it did not show any preferential turning and tilting behavior, which is similar to what was observed in control goldfish. However, the trace maps showed that, after bilateral sectioning, fish preferred to cross the center of the tank, which was unlike control fish. In control pairs, one goldfish chased the other with a fixed small range of two-point distances. However, in pairs of goldfish with bilateral transection of the optic nerve, the blind goldfish behaved independently of each other, with a long two-point distance. The long two-point distance of the blind goldfish lasted for at least two months and then slowly returned to control two-point distance by four months after bilateral transection. Such fast and slow recovery in goldfish behaviors evoked after unilateral and bilateral transection of the optic nerve is discussed with respect to reconnection of regenerating optic nerves in the fish central nervous system. This computer image processing system is a useful tool with which we can quickly and easily quantify fish behavior.