Granular and dysgranular retrosplenial cortices provide qualitatively different contributions to spatial working memory: evidence from immediate-early gene imaging in rats

The present study revealed striking task-dependent differences in immediate-early gene activity in the two main subregions (granular and dysgranular) of the retrosplenial cortex. In addition, there were activity differences along the rostro-caudal axis of both subregions. Two groups of rats were trained on a working memory task in a radial-arm maze, one group in the light, the other in the dark. Each working memory group had two sets of yoked controls. Working memory consistently increased retrosplenial immediate-early gene activity ( c-fos and zif268 ), although systematic differences occurred in the granular and dysgranular subregions. Both c-fos and zif268 expression increased in granular cortex irrespective of whether the spatial memory task was in the light or dark. In contrast, only in the light did spatial memory increase dysgranular cortex activation. Correlations based on the counts of Fos-positive cells helped to reinforce the particular association between the dysgranular retrosplenial cortex and radial-arm maze performance in the light. These results provide clear evidence for proposed functional differences between the major retrosplenial subregions: the granular cortex contributes to spatial learning and navigation based on both internal and external cues (light and dark), while dysgranular cortex is more selectively involved when distal visual cues control performance (light only).     

Biochemical mapping of noradrenergic nerves arising from the rat locus coeruleus

Norepinephrine (NE) concentration was measured in discrete brain regions of the rat following unilateral electrolytic lesions of the locus coeruleus, to determine the distribution of its noradrenergic neurons. Discrete brain nuclei and subdivisions were dissected from frozen sections, and norepinephrine was measured by a sensitive radio-isotopic assay.A significant reduction by 29–63% of control values in norepinephrine content was observed ipsilateral to the lesion in the following areas: all portions of the cerebral cortex examined (entorhinal, hippocampal, cingulate, parietal, and occipital areas), anterior half of the cerebellar cortex, hypothalamic periventricular and paraventricular nuclei, anterior ventral thalamic nucleus, ventral thalamic nucleus, and habenula. It appears that these regions receive unilateral innervation by axons from the locus coeruleus. In 3 regions (the medial geniculate body, inferior colliculus, and posterior half of the cerebellum), NE was reduced bilaterally in a pattern suggesting bilateral innervation from the locus coeruleus. Since no reduction in NE concentration occurred in the medial preoptic nucleus, nucleus interstitialis stria terminalis (ventralis), dorsomedial hypothalamic nucleus, or medial forebrain bundle, axons from noradrenergic neurons in the locus coeruleus do not appear to innervate these regions.The biochemical mapping of noradrenergic nerves from the locus coeruleus is discussed in relation to distribution studies based on the histofluorescence method.     

Retinal lesions induce fast intrinsic cortical plasticity in adult mouse visual system

Neuronal activity plays an important role in the development and structural–functional maintenance of the brain as well as in its life‐long plastic response to changes in sensory stimulation. We characterized the impact of unilateral 15° laser lesions in the temporal lower visual field of the retina, on visually driven neuronal activity in the afferent visual pathway of adult mice using in situ hybridization for the activity reporter gene zif268. In the first days post‐lesion, we detected a discrete zone of reduced zif268 expression in the contralateral hemisphere, spanning the border between the monocular segment of the primary visual cortex (V1) with extrastriate visual area V2M. We could not detect a clear lesion projection zone (LPZ) in areas lateral to V1 whereas medial to V2M, agranular and granular retrosplenial cortex showed decreased zif268 levels over their full extent. All affected areas displayed a return to normal zif268 levels, and this was faster in higher order visual areas than in V1. The lesion did, however, induce a permanent LPZ in the retinorecipient layers of the superior colliculus. We identified a retinotopy‐based intrinsic capacity of adult mouse visual cortex to recover from restricted vision loss, with recovery speed reflecting the areal cortical magnification factor. Our observations predict incomplete visual field representations for areas lateral to V1 vs. lack of retinotopic organization for areas medial to V2M. The validation of this mouse model paves the way for future interrogations of cortical region‐ and cell‐type‐specific contributions to functional recovery, up to microcircuit level.     

Modified coeruleo-cortical noradrenergic neurotransmission after serotonin depletion by PCPA: electrophysiological studies in the rat

To detect eventual modifications in the efficacy of the noradrenergic (NA) coeruleo-cortical system after serotonin (5-HT) depletion by parachlorophenylalanine (PCPA), three electrophysiological parameters were investigated in urethane-anesthetized rats which were treated for 2 days with daily injections of this inhibitor of 5-HT synthesis. 1) The spontaneous activity of locus coeruleus (LC) noradrenergic neurons showed a significant increase in PCPA-treated compared to control rats (4.3 vs. 2.6 Hz). 2) The sensitivity of NA autoreceptors was measured in the LC by the effect of intravenous administrations of clonidine or microiontophoretic applications of NA on spontaneous neuronal firing. In treated rats, clonidine and NA induced a lesser reduction of LC neuron firing than in the controls (27 vs. 75% decreases and 1,367 vs. 280 nC, respectively). 3) The responsiveness of cortical neurons to electrical stimulation of the LC was assessed by peristimulus time histograms in the dorsal fronto-parietal cortex. Following stimulation at 2 or 4 Hz, a majority of spontaneously firing cortical units was inhibited by electrical stimulation of the LC, but the percentage of such units was reduced and showed a decreased responsiveness after PCPA treatment. These findings suggest that following 5-HT depletion by PCPA, cortical NA neurotransmission is markedly reduced in its efficacy in spite of some increase in the spontaneous activity of coeruleo-cortical NA neurons.     

Enhancement of learning four weeks after stimulation of the nucleus locus coeruleus in the rat: Differential effects of dorsal noradrenergic bundle lesion and lesion of the locus coeruleus proper

In previous studies we showed that electrical stimulation of the nucleus locus coeruleus produced, four weeks later, a significant improvement in performance in acquisition of food-reinforced operant conditioning. In the two experiments reported here, we tested the role of the dorsal noradrenergic bundle and of the locus coeruleus proper in this long-term effect. Lesioning the dorsal noradrenergic bundle did not have a clear and consistent effect, whereas lesion of the nucleus coeruleus proper suppressed almost totally the beneficial effect of the stimulation. In the first experiment, the dorsal noradrenergic bundle was lesioned by local bilateral injection of 6-hydroxydopamine, 8 days before stimulation on the locus coeruleus. Four weeks after the stimulation, the rats were tested for acquisition of the operant task. Three control groups were used: not lesioned but stimulated, lesioned but not stimulated, and not lesioned/not stimulated. The locus coeruleus stimulation produced the same improvement of performance at the beginning of the acquisition, whether or not the dorsal noradrenergic bundle had been lesioned. However, a significant decrement of performance was observed in lesioned and stimulated rats during the last 40 min of the acquisition. In the second experiment, the locus coeruleus proper was destroyed by bilateral local injection of 6-hydroxydopamine and the locus coeruleus region was stimulated 15 days later. Three control groups were used, as in the first experiment. All the rats were tested 4 weeks later for acquisition of the operant task. The locus coeruleus lesion significantly attenuated the beneficial effect of the stimulation; however, the performance of the lesioned and stimulated rats was still significantly superior to that of the lesioned but not stimulated rats. These results suggest that the noradrenergic locus coeruleus system is involved in the long-term effect, but that the rostral projections passing through the dorsal bundle, in front of the lesion, are not critically involved in the observed effect.     

Dysregulation of gene induction in corticostriatal circuits after repeated methylphenidate treatment in adolescent rats: differential effects on zif 268 and homer 1a

Psychostimulants and other dopamine agonists produce molecular changes in neurons of cortico-basal ganglia-cortical circuits, and such neuronal changes are implicated in behavioural disorders. Methylphenidate, a psychostimulant that causes dopamine overflow (among other effects), alters gene regulation in neurons of the striatum. The present study compared the effects of acute and repeated methylphenidate treatment on cortical and striatal gene regulation in adolescent rats. Changes in the expression of the immediate-early genes zif 268 and homer 1a were mapped in 23 striatal sectors and 22 cortical areas that provide input to these striatal sectors, in order to determine whether specific corticostriatal circuits were affected by these treatments. Acute administration of methylphenidate (5 mg/kg, i.p.) produced modest zif 268 induction in cortical areas. These cortical zif 268 responses were correlated in magnitude with zif 268 induction in functionally related striatal sectors. In contrast, after repeated methylphenidate treatment (10 mg/kg, 7 days), cortical and striatal gene induction were dissociated. In these animals, the methylphenidate challenge (5 mg/kg) produced significantly greater gene induction (zif 268 and homer 1a) in the cortex. This enhanced response was widespread but regionally selective, as it occurred predominantly in premotor, motor and somatosensory cortical areas. At the same time, striatal gene induction was partly suppressed (zif 268) or unchanged (homer 1a). Thus, repeated methylphenidate treatment disrupted the normally coordinated gene activation patterns in cortical and striatal nodes of corticostriatal circuits. This drug-induced dissociation in cortical and striatal functioning was associated with enhanced levels of behavioural stereotypies, suggesting disrupted motor switching function.     

Induction of the learning and plasticity-associated gene Zif268 following exposure to a discrete cocaine-associated stimulus

We investigated whether the expression of the plasticity-associated gene, zif268, was associated with memories retrieved by exposure to a discrete stimulus that had been associated with cocaine, either self-administered or administered noncontingently. In the absence of drug, passive presentation of a cocaine-associated light stimulus induced changes in the expression of zif268 measured by in situ hybridization within a limbic cortical-ventral striatal circuit that was not only regionally selective but related to whether the rats had originally received response-contingent or noncontingent stimulus-drug pairings. In rats that had self-administered drug, the cocaine-conditioned stimulus (CS) increased zif268 expression in neurons of the ventral tegmental area, nucleus accumbens core and shell, and basal nucleus of the amygdala but not hippocampus, prelimbic area of the medial prefrontal cortex or amygdala central nucleus. The same CS that had been associated with cocaine administered noncontingently additionally increased zif268 mRNA levels in area Cg1 of the anterior cingulate cortex, ventral and lateral regions of the orbitofrontal cortex and lateral nucleus of the amygdala. Zif268 induction was related to the predictive relationship between the stimulus and cocaine as no changes were seen in cocaine-experienced rats that had received unpaired light and drug presentations during training. Thus, zif268 expression is increased by appetitively (drug) conditioned stimuli after Pavlovian learning. Zif268 may participate in the molecular mechanisms underlying the reconsolidation or re-encoding of Pavlovian stimulus-drug associations across a distributed limbic cortical-ventral striatal neural network and that may contribute to the basis of the enduring drug-seeking behaviour produced by environmental cues.     

Regulation of axonal ingrowth into area dentata as studied by sequential, double intraocular brain tissue transplantation

The anterior eye chamber was used as a model environment to study, in isolation, the interaction of embryonic area dentata transplants with transplants of one of three important sources of in situ innervation: entorhinal cortex, locus coeruleus or septal nuclei. None of these brain regions significantly affected the morphogenesis or in oculo growth of area dentata transplants. All three brain regions innervated the area dentata transplant. Entorhinal cortical transplants sent nerve fibers into a limited, and apparently specific, region of area dentata that was adjacent to the entorhinal transplant. This light innervation contrasts to the predominant innervation of area dentata by entorhinal cortex in situ. The fluorescent, noradrenergic neurons of locus coeruleus provided the area dentata transplant with an abundance of fine varicose nerve fibers. Given about 100 noradrenergic neurons in the locus coeruleus transplant and 4 to 6 months joint survival, the area dentata transplant was noradrenergically hyperinnervated. The cholinergic neurons of the septal nuclei transplant had a prolific ingrowth of acetylcholinesterase (AChE)-positive nerve fibers to the area dentata transplant. There appeared to be a mutual exclusion between the extrinsic AChE-positive fibers and the intrinsic Timm's-positive granule cell mossy fibers in the area dentata transplant. We conclude that isolated replicas of the coeruleo-, septo-, and entorhinal cortico-dentate pathways can be made through sequential intraocular double grafting. The nature of the in oculo connectivity between these replicates offers clues as to the mechanisms that might account for the regulation of nerve growth.     

Effects of lesioning noradrenergic neurones in the locus coeruleus on conditioned and unconditioned aversive behaviour in the rat

1. The brain noradrenergic system may have a role in anxiety disorder. This study has examined the effect of bilateral 6-hydroxydopamine lesions of the noradrenergic neurones in the locus coeruleus (LC) of male Lister hooded rats on behaviour produced by unconditioned and conditioned aversive stimuli. 2. The 6-hydroxydopamine (4 microg) lesions markedly reduced the noradrenaline content of the locus coeruleus hypothalamus, frontal cortex and the periaqueductal grey area without altering the levels of either dopamine or 5-hydroxytryptamine measured 14 days after administration. 3. Exposure to ultrasound (20 kHz at 98 dB for 60 sec), an unconditioned aversive stimulus, induced a defence response in the rats characterised by an increase in activity (running and jumping) followed by a period of inactivity (freezing). 4. Lesioning of the LC significantly attenuated the duration of freezing but was without effect on the active phase of the response. A similar reduction in freezing behaviour was seen with LC lesions when rats were exposed (3 hours after the acquisition) to the contextual cue of the conditioned emotion response paradigm. 5. These findings confirm that the locus coeruleus is involved in the regulation of fear-related behaviour in the rat both in an unconditioned and a conditioned model. Furthermore the results indicate that noradrenaline modifies defence behaviour rather than being the principle activating mechanism.     

Grafts of fetal locus coeruleus neurons in rat amygdala-piriform cortex suppress seizure development in hippocampal kindling

Hippocampal kindling was investigated in rats with a 6-hydroxydopamine-induced lesion of the forebrain catecholamine system after implantation of neural tissue from the fetal locus coeruleus region either bilaterally into the amygdala-piriform cortex (i.e., distant to the kindling site) or unilaterally into the hippocampus (close to the kindling site). Lesioned animals with either sham grafts or control grafts consisting of fetal striatal tissue showed a kindling rate much faster than that of normal controls. In contrast, in rats with bilateral locus coeruleus grafts in the amygdala-piriform cortex (implanted at three sites) the development of seizures was similar to that of controls and significantly slower than that in lesioned animals with sham grafts. All these animals had bilateral surviving grafts with a mean of 125 noradrenergic cells per implantation site. In the animals with locus coeruleus grafts in the stimulated hippocampus the kindling rate did not differ from that in the lesioned animals with control grafts. Most of these animals had large surviving grafts and showed a dense noradrenergic reinnervation of the implanted hippocampus. The present findings indicate that grafting of fetal pontine tissue (rich in noradrenergic neurons) to a site distant to the stimulation focus, but important for the generalization and spread of seizures, can retard the development of seizures in hippocampal kindling. Together with the data of our previous report this study also indicates that noradrenergic reinnervation of both hippocampi is important for the seizure-suppressant action in hippocampal kindling of locus coeruleus grafts implanted in the hippocampus.     

The involvement of noradrenaline in motor activity as shown by rotational behaviour after unilateral lesions of the locus coeruleus.

Unilateral electrolytic lesions in the region of the locus coeruleus in rats result in contraversive circling behaviour following the administration of systemic apomorphine or dexamphetamine. In a large group of such animals there was a very good correlation between histological damage to the locus coeruleus and the presence of such rotation. There was a progressive increase in the percentage of rats showing this drug-induced behaviour as ipsilateral cerebral cortical noradrenaline content decreased. Unilateral injections of 6-OHDA produce more selective damage to the noradrenergic locus coeruleus and result in a similar, although less marked, apomorphine- and dexamphetamine-induced rotation. Unlike the rats with electrolytic lesions of the locus coeruleus, those with 6-OHDA injections did not show an increase in ipsilateral striatal dopamine content. It seems likely that the motor asymmetry seen with lesions which involve the locus coeruleus is due to damage to its noradrenergic cells. A possible explanation of the differences between electrolytic and 6-OHDA induced lesion is that the 6-OHDA produces damage to the ventral noradrenergic bundle as well as the locus coeruleus and results in an increase in ipsilateral cerebral cortical 5-hydroxytryptamine.     

Unilateral striatal dopamine depletion: time-dependent effects on cortical function and behavioural correlates

Previously, we showed that unilateral blockade of D1 dopamine receptors in the striatum inhibits immediate-early gene expression bilaterally throughout large parts of the cortex, including sensory-evoked expression in the barrel cortex. To further investigate this dopamine regulation of cortical function, we examined the effects of dopamine depletion on cortical gene regulation and behavioural correlates. Two days after unilateral infusion of 6-hydroxydopamine into the midbrain, rats displayed a (to some degree) bilateral reduction in cortical zif 268 expression that was more pronounced on the lesioned side. This decrease was found across motor, somatosensory, insular and piriform, but not cingulate, cortex, similar to the effects of blockade of striatal D1 receptors. Furthermore, whisker stimulation-evoked c-fos and zif 268 expression in the barrel cortex ipsilateral to the lesion was also attenuated by acute dopamine depletion. These cortical deficits were accompanied by a breakdown of spontaneous behaviours in an open-field test. In contrast, 21 days after dopamine depletion, both basal and sensory-evoked gene expression in the cortex were near-normal. This cortical recovery was paralleled by recovery in locomotion and in sensory-guided behaviour (scanning) related to the hemisphere contralateral to the lesion, but not in scanning by the dopamine-depleted hemisphere. Our results suggest that striatal dopamine exerts a widespread facilitatory influence on cortical function that is necessary, but not sufficient, for normal behaviour. Moreover, the mechanisms mediating this cortical facilitation appear to be subject to substantial neuroplasticity after dopamine perturbation.     

Functional consequences of unilateral lesion of the locus coeruleus: A quantitative [C]2-deoxyglucose investigation

The functional consequences, as reflected in local rates of glucose utilization, of ablation of the locus coeruleus (the nucleus from which a major portion of the ascending noradrenergic fibres arise) have been examined in conscious rats with the quantitative autoradiographic [¹⁴C]2-deoxyglucose technique. Measurements of glucose utilization were made 72 h after histologically verified unilateral electrolytic lesions of the locus coeruleus. In the overwhelming majority of the 35 grey matter regions examined, the rate of glucose utilization was unaltered by lesions of the locus coeruleus, and in the limited number of CNS regions in which significant alterations were observed, the magnitude of the changes was invariably modest (less than 20% different from sham-operated control animals). Reductions in glucose use were observed in ipsilateral ventral (by 14%) and lateral thalamic nuclei (by 17%), and rates of glucose utilization in most regions of cerebral cortex were significantly lower (about 10%) in the ipsilateral hemisphere relative to the hemisphere contralateral to the lesion. In one region, the median raphe nucleus, glucose utilization was significantly elevated (by 19%) following lesions of the locus coeruleus. Attempts to accentuate the effects of locus coeruleus lesions by pharmacological manipulation of CNS adrenoreceptors by means of the systemic administration of phenoxybenzamine (30 mg/kg, 40 min prior to measurement of glucose use) in animals bearing unilateral locus coeruleus lesions were unsuccessful; the modest alterations in glucose utilization observed following locus coeruleus lesion alone were even less pronounced in lesioned animals receiving phenoxybenzamine. The alterations in local glucose utilization provoked by phenoxybenzamine were similar in sham-lesioned and locus coeruleus-lesioned animals. It would appear that the functional consequences, in terms of glucose utilization, are much less pronounced when a single neurotransmitter system (in the present studies, noradrenergic neurones) is lesioned than when a multiple neurotransmitter, functionally integrated pathway (such as the visual system) is disrupted.     

Electrophysiological identification of neurons in locus coeruleus

In 17 urethane-anesthetized rats 35 neurons, histologically verified as being situated in the locus coeruleus, were driven antidromically (latency, 44 msec) by electrical stimulation of the supracallosal bundle. Neurons of the locus coeruleus were also activated antidromically by stimulation of sites along the dorsal noradrenergic bundle in the midbrain (8-msec latency) and the hypothalamus (12-msec latency), and by stimulation of sites in the olfactory bulb (latency, 39 msec). Conduction velocity from these sites to the locus coeruleus was estimated to be 0.4 to 0.6 m/sec. Refractory periods of fibers in the dorsal noradrenergic bundle were determined at twice threshold and in the supracallosal bundle at intensities just above threshold; refractory periods ranging from 4 to 20 msec were observed. Because neurons both in and near the locus coeruleus were antidromically activated by stimulation of the dorsal noradrenergic bundle whereas stimulation of the supracallosal bundle antidromically activated only neurons in the locus coeruleus, stimulation of the dorsal noradrenergic bundle could not be used to identify locus coeruleus neurons. It is concluded that a subpopulation of neurons in the locus coeruleus can be identified by their slow, steady firing rate (2.6 per second) and long-latency antidromic response to stimulation of the supracallosal bundle. The electrophysiological properties of locus coeruleus neurons are considered in relation to neuroanatomical and functional studies of the locus coeruleus.     

Transhemispheric cortical reorganization in rat SmI and involvement of central noradrenergic system

Responses of single units in the hindpaw representational area of the left primary somatosensory cortex to electrical stimulation of both hindpaws and the right forepaw were recorded under urethane anaesthesia in three groups of adult male rats: a control group and two groups in which the right hindpaw representational area had been ablated 3–4 weeks previously, immediately after intraperitoneal injection of saline vehicle or DSP4, to destroy cortical noradrenergic terminals arising from the locus coeruleus. The lesion increased the overall number of neurones responding within 500 ms after the stimulation of the contralateral hindpaw (from 64 to 91%), and the proportion exhibiting short-latency response increased from 41 to 61%. Interestingly, the proportion of neurones with bilateral representation increased from 3 to 10% after the cortical lesioning. The changes were prevented by injection of DSP4 prior to lesioning and therefore depended on an intact central noradrenergic system. The increase in bilateral representation could not have been due to direct interhemispheric connections between corresponding representational areas because it occurred after lesioning of the homologous area in the contralateral hemisphere. The phenomenon was termed ‘transhemispheric reorganization’ and because it was somatotopically oriented (e.g. to either hindpaw); its function may be to ensure that when a sensory cortical area is damaged, its basic sensory functions are ‘taken over’ by the corresponding contralateral area.     

Changes in the incorporation of (14C) orotic acid into brain RNA of visually deprived rats following exposure to light

Newly-weaned rats that had been kept in the dark from birth were injected intraventricularly with [¹⁴C]orotic acid. Experimental rats were exposed to light for 2 hr and dark controls were returned to a dark environment for 2 hr. It was found that the relative specific activity of the RNA in the visual cortex of the former was significantly higher than that in the visual cortex of the latter. No difference was seen in the frontal cortex. The RNA content of the visual cortex of the rat exposed to light was significantly lower than that of the dark controls. This was not the case in the frontal cortex.Adult rats deprived of light for periods of 4–10 days also exhibited an increased RNA relative specific activity in the visual cortex after being exposed to light although the RNA content did not alter significantly. No changes were seen in the frontal cortex. Blind rats (of the “Campbell” strain) showed no changes in RNA metabolism in the visual cortex on being transferred from a dark to a light environment.     

Circadian covariation of norepinephrine and serotonin in the locus coeruleus and dorsal raphe nucleus in the rat

We report robust correlations between concentrations of the neurotransmitters norepinephrine and serotonin in the locus coeruleus and the dorsal raphe nucleus of the brainstem in rats analyzed at 7 different time points over 24 h. We found similar circadian rhythmicities for both monoamines with acrophases just before the onset of the dark period. The monoamine concentrations diminished and the significant intercorrelation between norepinephrine and serotonin in the locus coeruleus disappeared during the night suggestive of a noradrenergic stimulation of dorsal raphe serotonin during the day. Timing of experiments is crucially important in studies on brain monoaminergic indices and their interrelationships.     

Identification and localization of functional subdivisions in the visual cortex of the adult mouse

We investigated the anatomical characteristics of the mouse visual system through in situ hybridization for the neuronal activity marker zif268. Our main goal was to delineate the full extent of the cortical region processing visual information and additionally to identify the monocularly and binocularly driven subregions therein. We therefore analyzed the neocortex of monocularly and binocularly enucleated mice versus visually stimulated control mice. These visual manipulations revealed eye‐specific parcellations at the neocortical level. In binocularly enucleated mice we detected an unambiguous lateral border between visually driven and nonvisual cortex based on the clear deprivation‐induced reduction in zif268 expression in the first. However, medially a transition zone of intermediate intensity was found between primarily visual, that is V1 and multimodal retrosplenial cortex. Also in monocularly enucleated mice, the visual cortex contralateral to the deprived eye clearly displayed distinct regions of lower signal than the ipsilateral cortex. Yet interspersed between these regions of basal activity we could clearly identify a zone of high activity spanning the V1‐V2L border. A second zone of higher activity was noticeable near the medial border of visual cortex. Comparison with binocularly enucleated mice indicates the presence of both binocular input as well as nonvisual input in this medial cortical region and thus confirms the transitional nature of the recently described rostromedial areas. J. Comp. Neurol. 514:107–116, 2009. © 2009 Wiley‐Liss, Inc.     

Changes in projection from locus coeruleus to lateral geniculate nucleus following ablation of visual cortex in adult rats

The visual cortex of adult rats was unilaterally ablated. A histofluorescence study revealed an increase of noradrenergic terminals in the lateral geniculate nucleus (LGN) ipsilateral to the decortication, confirming the previous report. Corresponding to this, the frequency for encountering neurons in the locus coeruleus (LC) activated antidromically from the LGN was increased. We suggest that LC neurons whose axon terminals were damaged by ablation of the visual cortex formed new axons or axon collaterals (pruning effect) in the LGN, thus contributing, at least partly, to the increase of noradrenergic terminals therein.     

NGF and anti-NGF: evidence against effects on fiber growth in locus coeruleus from cultures of perinatal CNS tissues.

The present study examines whether the developing noradrenergic neurons of locus coeruleus depend on endogenous nerve growth factor (NGF) for nerve fiber production and if exogenous NGF stimulates fiber growth in this nucleus, using a collagen gel tissue culture technique. Lucus coeruleus from perinatal rat brain was used in three culture experiments: (1) lucus coeruleus, parietal cerebral cortex, and the superior cervical ganglion, prepared from newborn rats and cultured in different sectors of the same dishes; (2) locus coeruleus and parietal cerebral cortex from 17-day-old rat fetuses cultured in the same manner, and (3) locus coeruleus from 17-day-old rat fetuses co-cultured with spinal, sympathetic and ciliary ganglia from 8-day chick embryos. Experiments 1 and 2 were run with and without addition of NGF and anti-NGF, experiment 3 with and without anti-NGF. Total fiber production in all cultured tissues was evaluated daily by dark field and phase contrast microscopy during 4 days. Adrenergic nerve fiber production was then studied in the same locus coeruleus and superior cervical ganglia from the rats by Falck-Hillarp fluorescence histochemistry. Locus coeruleus and cortex cerebri from fetal rats produced dense fiber halos in culture. Locus coeruleus from newborn rats produced considerably less fibers, newborn cortex only few fibers. Superior cervical ganglia from the same newborn animals produced no or almost no fibers. Addition of NGF was not able to stimulate fiber growth in locus coeruleus nor in cortex cerebri as observed both in the living cultures and by fluorescence microscopy. Likewise, addition of anti-NGF did not affect fiber production in the CNS areas. The negative results with NGF on newborn locus coeruleus and cortex cerebri were in sharp contrast to the strong, highly significant fiber growth response demonstrated by the superior cervical ganglion from the same animals cultured in the same dishes. The third experiment tested whether locus coeruleus in tissue culture contained or produced nerve growth factor or any one of the three chick embryo ganglia. No response whatsoever in these three ganglia was observed. It is concluded that the developing locus coeruleus area does not contain or produce NGF, does not depend on NGF for fiber production, and is not stimulated by exogenous NGF.