Synaptogenesis in the postnatal rat fascia dentata is influenced by 5-HT1a receptor activation.

Neonatal serotonin (5-HT) depletion or 5-HT1a receptor blockade results in a significant and permanent reduction in the number of dentate granule cell dendritic spines. The purpose of this study was to determine whether the loss of spines was accompanied by a reduction in molecular layer synaptic profile density. Rat pups were treated with parachloroamphetamine (PCA), 5,7-DHT or the 5-HT1a receptor antagonist NAN-190. The synaptic profile density (profiles/100 microm2) of the dentate molecular layer was estimated on P14, P21 and P60. Molecular layer synaptic profile density the was significantly reduced in each treatment group on P14 and P21. By P60, the NAN-190 and PCA groups had reached control values, but the 5,7-DHT group remained significantly lower than control. The most dramatic changes were observed among synapses terminating on dendritic spines. Numbers of profiles forming simple spine contacts were significantly reduced by all treatments at P14 and P21, but returned to normal by P60 in the PCA and NAN-190 groups. Simple spine synapses in the 5,7-DHT group remained significantly below control, but numbers of complex spine synapses were higher than either control or the other treatment groups at each age. These results indicate that the loss of dendritic spines observed following 5-HT depletion or 5-HT1a antagonist treatment is accompanied by a decrease in synaptic profile density. This effect appears to be a retardation of synaptogenesis since recovery occurs once 5-HT1a receptor activity resumes. Data from the 5,7-DHT group shows that complex spine synapse formation may represent an effort to attain some degree of functional compensation when synaptogenesis is slowed.

Dentate granule cell function after neonatal treatment with parachloroamphetamine or 5,7-dihydroxytryptamine.

In vitro, extracellular electrophysiological recording was used to test granule cell responses in P60 rats after neonatal PCA and 5, 7-DHT. Granule cell population EPSP and spike responses were in the normal range for both PCA and 5,7-DHT groups. However, the degree of paired pulse facilitation was reduced in both of these groups relative to control, reflecting a diminished synaptic drive. Synaptic potentiation in the 5,7-DHT group was not different from control, but was significantly reduced in slices from PCA-treated rats.

Serotonin regulates synaptic connections in the dentate molecular layer of adult rats via 5-HT1a receptors: evidence for a glial mechanism.

The present study sought to verify effects of 5-HT on synaptic density at the ultrastructural level, to determine whether the 5-HT1a receptor is important for the maintenance of synaptic connections and to obtain evidence implicating S100 beta in the apparent neurotrophic actions of 5-HT. Reduction of hippocampal 5-HT with para-chloroamphetamine (PCA) resulted in a significant decline in the synaptic density of the dentate molecular layer. Reduction of norepinephrine with DSP-4 produced a slight decrease in the number of molecular layer synapses, but this difference was not statistically different from control values. 5-HT1a antagonist treatment resulted in a decline in synaptic density comparable to that observed following PCA treatment. These observations suggest that 5-HT functions to maintain synaptic connections in the dentate molecular layer via a 5-HT1a mechanism. To determine whether the change in synaptic density was due to the action of 5-HT on neuronal receptors or astrocytic receptors, a monoclonal antibody against S100 beta was infused into the lateral ventricle for seven days. Controls received infusions of normal goat serum. Half of the rats from the anti-S100 beta and control groups also received daily injections of NAN-190. Anti-S100 beta infusion resulted in a significant (p < 0.01) decrease in synapses compared to serum controls. Concomitant NAN-190 administration did not enhance synapse loss in the anti-S100 beta group. The results of this study suggest that the maintenance of synaptic connections in the dentate molecular layer is influenced by S100 beta levels that are controlled by 5-HT stimulation of astrocytic 5-HT1a receptors.

Effects of neonatal serotonin depletion on the development of rat dentate granule cells.

The appearance of serotonergic (5-HT) neurons and projections early in central nervous system (CNS) development has resulted in the hypothesis that 5-HT is an important factor in neuronal differentiation and synaptogenesis. Studies of the effects of 5-HT on the development of molluscan and mammalian neurons in vitro support this hypothesis, but mammalian in vivo studies have produced equivocal results. The present study reinvestigated the role of 5-HT in CNS development using the dentate granule cell as a model. Dentate granule cells were chosen for this study of the effects of 5-HT depletion on neuronal development because they are generated in the early postnatal period. Thus, 5-HT depletion could be effected by the treatment of rat pups with either parachloroamphetamine (PCA) or 5,7-dihydroxytryptamine (5,7-DHT) thereby avoiding problems inherent in maternal treatment paradigms. The morphology of Neurobiotin-filled granule cells was studied on P14, P21, P60 and P120 (P0 = day of birth). The parameters measured were total dendritic length, number of dendritic segments and dendritic spine density (number of spines/50 microns dendritic length). Granule cells from vehicle-treated controls were similar to those previously reported in studies of normal granule cell development in all respects. In particular, the decrease in dendritic spine density from P14 to P120 observed in Golgi preparations was verified in our population of intracellularly filled granule cells. Transient depletion of 5-HT by neonatal PCA treatment resulted in a decrease dendritic length that was not statistically different from control values. However, dendritic spine density was reduced by about 27% at all ages studied. 5,7-DHT treatment produces a permanent, severe depletion of 5-HT. Spine densities in granule cells from 5,7-DHT-treated pups were also about 38% lower than controls. Total dendritic length in cells from 5,7-DHT-treated rats was reduced to a degree comparable to that observed in PCA-treated pups. The number of granule cell dendritic segments was also less than that observed in control and PCA-treated rats but this difference was not statistically significant. These observations suggest that reduction of 5-HT in the early postnatal period can result in changes in the morphology of dentate granule cells, particularly at the level of the synapse as reflected by the permanent reduction in synaptic spine density. The comparison of results from cases with permanent and transient reduction of 5-HT indicates that the developmental influence of 5-HT is most important during the first three postnatal weeks.

5-HT1a receptors mediate the neurotrophic effect of serotonin on developing dentate granule cells.

We have previously reported that neonatal (P3) serotonin (5-HT) depletion results in a significant decrease in the number of dendritic spines per 50 microns of dendritic length on dentate granule cells. This effect is specific and permanent. Neither total dendritic length nor the number of dendritic segments is affected by 5-HT depletion. The area dentata contains a dense 5-HT1a receptor population that is present in the at birth. Therefore, 5-HT1a receptors represented a likely candidate for the mediation of the effects of 5-HT on developing granule cells. The present study used the drugs buspirone and NAN-190, which have been shown to be an agonist and antagonist respectively at postsynaptic 5-HT1a receptors in vivo, to test the idea that neurotrophic actions of 5-HT result from 5-HT1a receptor stimulation. Following 5-HT depletion with PCA, pups received daily injections of buspirone (1.0 mg/kg) from P5 to P14. Granule cell morphology was then studied using intracellular filling with Neurobiotin on P14, P21 and P60. Buspirone treatment prevented the loss of dendritic spines previously shown to follow 5-HT depletion with PCA. No other morphological parameters were significantly changed by buspirone treatment. Naive pups received daily injections of NAN-190 from P3 to P14. One group received 1.0 mg/kg while a second group received 3.5 mg/kg. Both doses of NAN-190 resulted in dendritic spine loss comparable to that obtained with neonatal PCA treatment. This loss was permanent suggesting that the first two postnatal weeks may represent a critical period for the action of 5-HT on developing granule cells. Significant, dose-dependent changes in total dendritic length and number of dendritic segments reminiscent of the effects of norepinephrine depletion were also observed in NAN-190-treated rats. We suspect that this change is the result of the action NAN-190 at alpha receptors and is therefore distinct from the specific effect of 5-HT on the number of dendritic spines. The NAN-190 experiment also shows that the loss of dendritic spines is a function of decreased stimulation of 5-HT1a receptors and not the loss of 5-HT terminal membrane.

Neuronal dye coupling in the developing rat fascia dentata.

The present study describes dye coupling among neurons of the developing rat fascia dentata following impalement and intracellular filling with Neurobiotin. The number of neuronal impalements resulting in dye-coupled cells decreases from P14 to P120. The most rapid decline in dye coupling was observed between P14 and P21, the beginning of the most active period of synaptogenesis in the dentate molecular layer. Dye coupling between granule cells and axo-axonic interneurons (chandelier cells) accounts for about 10% of the dye-coupled neuronal population acquired in slices from P14 and P21 rats and declines to less than 5% by P60 and P120. Our data suggest that dye coupling is related reciprocally to the number of synapses formed on granule cells. Thus the relationship of dye coupling to synaptic density in the developing fascia dentata is similar to that reported in studies of the aging fascia dentata. Also the observation of axo-axonic interneurons coupled to granule cells at all ages suggests an interesting neuronal arrangement with the potential of limiting granule cell discharge to discrete neuronal assemblies in response to perforant path input.

Evidence for the elaboration of multiple axons by developing dentate granule cells.

A sample of 185 Neurobiotin-filled dentate granule cells has been collected from rats aged P14 to P120 in a study of the effects of serotonin (5-hydroxytryptamine, 5-HT) depletion on granule cell development. A small number (5.9%) of these neurons exhibit more than one axon. These neurons have morphologies consistent with that described for dentate granule cells. One axon typically arises from the soma while additional axons take origin from either dendrites or the soma. Both axons may be mossy fibers or one axon may be a mossy fiber and the second assume a morphology and distribution similar to that described for interneurons. These latter neurons therefore exhibit a mixed phenotype by having a granule cell morphology and an interneuronal axon type. These data suggest that some granule cells give rise to multiple mossy fibers or express a mixed axonal phenotype during maturation.

An endogenous dopaminergic neurotoxin: implication for Parkinson's disease.

Oxidation of dopamine by monoamine oxidase results in the endogenous metabolite 3,4-dihydroxyphenylacetaldehyde (DOPAL). The toxicity of DOPAL for dopaminergic neurons was investigated using rat neostriatal synaptosomes, PC-12 cells and cultures of fetal rat dissociated mesencephalon. The Na(+)-dependent uptake of [3H]DOPAL in synaptosomes was inhibited by mazindol. DOPAL selectively inhibited dopamine uptake but not [14C]GABA uptake, induced membrane damage and liberation of dopamine into the medium. Incubation of PC-12 cells with 6.5 microM of DOPAL for 24 h caused degeneration of the neuritic process, and the number of viable cells were reduced by 25% of control. There were practically no surviving cells after 24 h of incubation with 33 microM of DOPAL. After 8 h of treatment with 33 microM of DOPAL, dopamine and 3,4-dihydroxyphenylacetic acid content in the cells were reduced by 38% and 53% of control. DOPAL-induced cell damage released lactic acid dehydrogenase into the incubation media. This toxic effect of DOPAL was time- and concentration-dependent. In mesencephalic cultures, after exposure to 33 microM of DOPAL, the surviving TH+ cells showed rounded cell body, and fibre network was highly reduced. These results indicate DOPAL is a neurotoxin and may be involved in the degeneration of dopaminergic neurons.

The raccoon spinocervical and spinothalamic tracts: a horseradish peroxidase study.

The locations of the cells of origin of the spinocervical tract (SCT) and spinothalamic tract (STT) were examined in relation to the somatotopic and laminar organization of the cervical enlargement of the raccoon dorsal horn (DH). In different animals, either the lateral cervical nucleus or the lateral thalamus was injected with a 2% solution of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Following 24 h or 4 days, respectively, the animals were sacrificed and both injection and target sites (spinal cord segments C6-T2) were processed using the TMB method. All labelled cells were counted in every fifth 50 microns section. Following injection of WGA-HRP into the lateral cervical nucleus, all labelled SCT cells were located ipsilateral to the injection sites. Most (84%) were in laminae III and IV, laminae known from other studies to contain cells preferentially responsive to light tactile stimulation, with very few (3%) in lamina I. Nearly 50% of labelled cells were located in the medial 1/3 of the DH, the region of representation of the glabrous surfaces of the raccoon forepaw. The mean number of labelled SCT cells per section was 4.19. After tracer injections of the lateral thalamus, more than 75% of STT cells were located contralateral to the injection sites. Forty-three percent were located in lamina I and 24% were in lamina V, laminae whose cells have been shown to be responsive to more intense forms of stimulation, as well as to light touch. Only 22% were located in the medial 1/3 of the DH. The mean number of labelled STT cells per section was 0.83. The results suggest that the SCT may play a more critical role in relaying discriminative light tactile and nociceptive information from the glabrous surfaces of the forepaw, but that there may be a greater role for the STT in relaying nociceptive information from the forelimb as a whole.

Transient reduction in hippocampal serotonergic innervation after neonatal parachloroamphetamine treatment.

This study examined the effects of parachloroamphetamine on neonatal forebrain serotonergic (5-HT) innervation. Rat pups were treated with PCA on P3 and P4. Significant reductions in 5-HT content were observed in the hippocampal formation, frontal cortex and entorhinal cortex on P5 and P7. By P14, neocortical 5-HT had returned to normal levels while hippocampal 5-HT values remained less than control. Hippocampal 5-HT content reached normal range by P21. High affinity 5-HT uptake in hippocampal synaptosomal preparations was similarly reduced on P5 and P7 suggesting that 5-HT terminals were being lesioned by PCA. 5-HT uptake recovered significantly by P14 perhaps reflecting the extraordinary plasticity of the 5-HT projections in the neonate. However, in contrast to the complete restoration of hippocampal 5-HT content, 5-HT uptake values remained significantly less than control. No change in 5-HT content was observed in either the hypothalamus or midbrain raphe at any age studied. Thus, the rapid onset of effects, regional selectivity and transient reduction of 5-HT levels recommend the use of PCA in studies of the role of 5-HT in hippocampal development.

Hippocampal serotonin levels influence the expression of S100 beta detected by immunocytochemistry.

The response of astrocytes to pharmacological alterations of the serotonin (5-HT) content of the dorsal area dentata was studied by immunocytochemistry for S100 beta. Stereological analysis revealed changes in the density of astrocytic elements in conditions of 5-HT depletion and elevated 5-HT levels. There was a direct relationship between the expression of S100 beta and the level of 5-HT in the hippocampal formation. Thus when 5-HT was reduced by PCPA treatment, S100 beta immunoreactivity was also reduced. The converse of these observations was obtained in cases where the 5-HT levels were increased by fluoxetine administration. These immunocytochemical data are consistent with previous reports of the response of astrocytes to 5-HT in vitro.

Effects of para-chloroamphetamine upon the serotonergic innervation of the rat hippocampus.

The ability of hippocampal serotonergic (5-HT) axons to proliferate in response to damage by para-chloroamphetamine (PCA) was examined in this study. Synaptosomal uptake of 5-HT in the hippocampal formation was decreased to 40% of control 3 days after systemic administration of PCA. Six weeks after PCA, uptake values were 44% of control. Retrograde tracing combined with 5-HT immunocytochemistry showed a significant reduction (18% of control) in the number of 5-HT raphe neurons projecting to the hippocampus 3 days after PCA. The number of 5-HT neurons projecting to the hippocampal formation increased to 69% of control by 6 weeks. The dorsal raphe nucleus was not retrogradely labeled after PCA; the increase in labeled neurons was observed in the median raphe nucleus. PHA-L, injections of the median raphe nucleus demonstrated a reduction of raphe axons in the hippocampal formation after PCA. In rats treated with PCA, raphe axons labeled with PHA-L also appeared to have fewer boutons than raphe axons labeled in control cases. The density of PHA-L containing axons in the hippocampal formation of rats injected 3 days and 6 weeks after PCA was less than control but there was no difference between the experimental groups. Based upon the results from synaptosomal uptake and anterograde tracing experiments, we feel that compensatory proliferation of 5-HT axons does not occur within 6 weeks of PCA-induced damage to the 5-HT plexus of the hippocampal formation. The data derived from the retrograde tracing experiment are thought to reflect reduced uptake and transport of WGA-HRP as an acute effect of PCA.

Reorganization of the area dentata serotoninergic plexus after lesions of the median raphe nucleus.

Serotoninergic projections from the dorsal and median raphe nuclei to the area dentata of the hippocampal formation terminate mainly in the molecular layer and hilus, respectively. Consequently, a reduction in the density of the hilar serotoninergic plexus is seen by immunocytochemistry 2 weeks after lesions of the median raphe nucleus. Hippocampal serotonin concentration and serotonin high affinity uptake are also significantly reduced. Six weeks after lesion, surviving serotoninergic axons form a dense band in the inner molecular layer of the dorsal area dentata, a region that usually contains a sparse serotoninergic plexus. Moreover, serotoninergic fibers transverse the molecular layer and pass through the granule cell layer to reinnervate the hilus. Serotonin concentration and high affinity uptake have recovered to near normal levels by 6 weeks postlesion. Changes in the anatomical distribution of the area dentata serotoninergic plexus have not been reported in cases in which serotoninergic sprouting follows axotomy of serotoninergic projections. Thus direct lesions of serotoninergic neurons can produce a homotypic compensatory response that is qualitatively different from that generated by axotomy. The mechanistic basis for this reorganization is unclear, but the apparent extension of serotoninergic axon collaterals toward the hilus suggests that the denervated hilar neuropil is guiding reinnervation. Finally, anatomical evidence from animals studied 10 weeks postlesion suggests that the compensatory proliferation of serotoninergic axons observed 6 weeks after median raphe lesion is a transient event.

Evidence for decreased transport of tryptophan hydroxylase in Alzheimer's disease.

Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the synthesis of serotonin and a specific marker for serotonergic neurons. These neurons are affected in Alzheimer's disease (AD) in several ways: serotonin is decreased in axon terminals, serotonin neurons accumulate neurofibrillary protein, and these neurons are lost in AD brains. One subcellular mechanism which may underlie degeneration of neurons in AD is decreased axonal transport with accumulation of enzymes and their potentially toxic metabolites in the cell body. To determine whether there is a defect in axonal transport in serotonin neurons in AD we measured TPH activity, serotonin and its oxidative metabolite 5-hydroxyindoleacetic acid (5-HIAA) in dorsal raphe cell bodies from Alzheimer and control cases. TPH activity is increased 4.7-fold in raphe neuron cell bodies in Alzheimer brains. Serotonin and 5-HIAA are increased by 4.0- and 2.0-fold, respectively in Alzheimer compared to control raphe cell bodies. In contrast, in synaptic terminals of the amygdala 5-HT and 5-HIAA were decreased by 41% and 50%, respectively in the same AD cases. We propose that the accumulation of TPH and its products in the raphe perikarya in AD results from a diminished transport of TPH to axon terminals. The accumulation of oxidative metabolites of serotonin may contribute to the degeneration of serotonergic neurons in AD.

Intersubnuclear connections within the rat trigeminal brainstem complex.

Prior intracellular recording and labeling experiments have documented local-circuit and projection neurons in the spinal trigeminal (V) nucleus with axons that arborize in more rostral and caudal spinal trigeminal subnuclei and nucleus principalis. Anterograde tracing studies were therefore carried out to assess the origin, extent, distribution, and morphology of such intersubnuclear axons in the rat trigeminal brainstem nuclear complex (TBNC). Phaseolus vulgaris leucoagglutinin (PHA-L) was used as the anterograde marker because of its high sensitivity and the morphological detail provided. Injections restricted to TBNC subnucleus caudalis resulted in dense terminal labeling in each of the more rostral ipsilateral subnuclei. Subnucleus interpolaris projected ipsilaterally and heavily to magnocellular portions of subnucleus caudalis, as well as subnucleus oralis and nucleus principalis. Nucleus principalis, on the other hand, had only a sparse projection to each of the caudal ipsilateral subnuclei. Intersubnuclear axons most frequently traveled in the deep bundles within the TBNC, the V spinal tract, and the reticular formation. They gave rise to a number of circumscribed, highly branched arbors with many boutons of the terminal and en passant types. Retrograde single- or multiple-labeling experiments assessed the cells giving rise to TBNC intersubnuclear collaterals. Horseradish peroxidase (HRP) and/or fluorescent tracer injections into the thalamus, colliculus, cerebellum, nucleus principalis, and/or subnucleus caudalis revealed large numbers of neurons in subnuclei caudalis, interpolaris, and oralis projecting to the region of nucleus principalis. Cells projecting to more caudal spinal trigeminal regions were most numerous in subnuclei interpolaris and oralis. Some cells in lamina V of subnucleus caudalis and in subnuclei interpolaris and oralis projected to thalamus and/or colliculus, as well as other TBNC subnuclei. Such collateral projections were rare in nucleus principalis and more superficial laminae of subnucleus caudalis. TBNC cells labeled by cerebellar injections were not double-labeled by tracer injections into the thalamus, colliculus, or TBNC. These findings lend generality to currently available data obtained with intracellular recording and HRP labeling methods, and suggest that most intersubnuclear axons originate in TBNC local-circuit neurons, though some originate in cells that project to midbrain and/or diencephalon.

Principalis- or parabrachial-projecting spinal trigeminal neurons do not stain for GABA or GAD.

Retrograde transport and immunohistochemical double-labeling methods (Weinberg et al., 1985) were used to assess the distribution and projection status of spinal trigeminal (SpV) neurons that stain positively for glutamic acid decarboxylase (GAD) or gamma-aminobutyric acid (GABA). Large bilateral injections of diamidino yellow into the rostral and lateral pons, inclusive of V nucleus principalis and the parabrachial nucleus, retrogradely labeled large numbers of cells in each SpV subnucleus. Many cells in SpV subnuclei caudalis, interpolaris, and oralis also exhibited GABA immunoreactivity; the largest numbers were in caudalis and the smallest numbers were in oralis. However, none of the GABA- or GAD-immunoreactive SpV cells were double-labeled with diamidino yellow, though some reticular neurons displayed both GABA and the retrograde tracer. This negative result refutes a previously offered hypothesis that SpV local-circuit neurons with principalis collaterals are GABA-ergic (Jacquin et al., 1989b). These data also indicate that parabrachial-projecting SpV neurons are not GABA-ergic.

Development and lesion induced reorganization of the cortical representation of the rat's body surface as revealed by immunocytochemistry for serotonin.

Immunocytochemistry with an antiserum directed against serotonin (5-HT) was used to assess the development of the representation of the body surface in the rat's primary somatosensory cortex (S-I). Within 1 hour of birth (P-O), 5-HT-positive fibers were present in the marginal zone, the cortical plate, and developing layers V and VI. Immunoreactivity in the marginal zone consisted of a thin band of coarse fibers oriented parallel to the pia. Only a small number of isolated fibers were visible in the cortical plate. A denser network of both coarse and fine fibers could be seen in presumptive layers V and VI. By the first hour of P-I, 5-HT-positive axons in the deeper cortical plate were organized into a crude representation of the rat's body surface. At this age, aggregates of fibers corresponding to the head, lower jaw, trunk, and forepaw could be clearly distinguished. These regions of dense 5-HT immunoreactivity consisted primarily of fine caliber axons that had invaded the lower part of the cortical plate. Dense aggregates of fine caliber axons were also visible in developing layers V and VI. Coarse 5-HT-positive fibers were visible in all layers, but they did not appear to contribute to the pattern that corresponded to the body surface. By the first hour of P-2, the map of the body surface in S-I was more refined and a row-related organization of 5-HT-immunoreactive fibers was visible in the portion of the cortex representing the vibrissa pad. The laminar distributions of coarse and fine caliber serotoninergic axons at this age were essentially the same as on P-I. By P-2.5 (60 hours after birth), patches of 5-HT-positive fibers corresponding to individual vibrissa follicles were clearly evident. These consisted of dense aggregates of fine caliber axons that were centered in presumptive layer IV, but which also extended above and below this lamina. Over the next 3 days, the pattern continued to mature. By P-4, dense 5-HT labelling was also visible in the secondary somatosensory cortex (S-II). By the beginning of P-5, clusters of fibers corresponding to more rostral facial hairs and individual digits within the forepaw representation could also be discerned. By P-12, the differential distribution of 5-HT fibers in S-I was no longer visible. Thus, immunocytochemistry for serotonin showed a representation in S-I homeomorphic with the body surface prior to the age at which it can be discerned with other methods thought to reveal thalamocortical axons. Transection of the infraorbital nerve (ION) on the day of birth altered the organization of the vibrissal representation in the contralateral cortex from the earliest age at which it could be detected by 5-HT immunocytochemistry in normal animals. However, the departure from the normal organization was gradual. Row-related organization was clearly visible in the cortices of rats sacrificed on P-3, but not in those of rats that were killed on P-5. These results suggested that the organization of the 5-HT innervation of the cortex may be guided by thalamic afferents and further that some aspects of this guidance persist, albeit temporarily, after ION transection on P-0. The 5-HT immunoreactivity that we observed in the developing somatosensory cortex was not contained in thalamocortical axons. Unilateral electrocautery of the ventrobasal thalamus on P-4 did not reduce the density or alter the pattern of the 5-HT innervation of the cortex in rats that were examined on P-6.

Patterns of resting discharge in neurons of the raccoon main cuneate nucleus.

1. The presence and pattern of resting discharge were examined in 100 single neurons of the raccoon main cuneate nucleus (MCN). Of these, 66 were activated, either antidromically or synaptically, by electrical stimulation of the contralateral thalamic ventrobasal complex (VB), and 34 were activated by stimulation of the ipsilateral cerebellum (CB). 2. Forty-one percent of VB-activated neurons displayed a resting discharge, whereas 32% of CB-activated neurons did. Most neurons activated from VB and showing a resting discharge fired in bursts of 2-5 spikes, whereas those activated from CB and showing a resting discharge generally fired as single, irregularly spaced spikes, with occasional bursts in some neurons. 3. All neurons antidromically activated from VB were histologically localized within the clusters region of the MCN, whereas those antidromically activated from CB were confined to its polymorphic region. Neurons synaptically activated from either VB or CB were located in either of these regions. 4. Differences in the proportions of neurons displaying a resting discharge did not vary significantly as a function of type of preparation: methoxyflurane anesthesia, pentobarbital sodium anesthesia, decerebrate (the latter CB-activated only). 5. Although the sample sizes were too small to demonstrate statistical significance, neurons exhibiting a resting discharge were more likely to show a bursting pattern in methoxyflurane-anesthetized preparations than were neurons in pentobarbital sodium-anesthetized preparations. 6. The probability of having no resting discharge, firing in bursts, or firing in single spikes was not related to cutaneous submodality [rapidly adapting (RA), slowly adapting (SA), Pacinian (Pc)], or to receptive field (RF) locus (glabrous versus hairy skin). 7. The overall mean rate of firing (11.8 Hz) was not significantly different for bursting versus nonbursting neurons. 8. In bursting neurons, median interspike intervals (ISIs) varied between 1.3 and 2.3 ms. Most bursting neurons also had a range of short or minimal interburst intervals (MIBIs), characteristic for each neuron, whose medians varied from neuron to neuron between 34 and 90 ms. Distributions of within-burst ISIs and MIBIs had comparable coefficients of variation, varying between 0.031 and 0.223. 9. The application of a mechanical stimulus to a neuron's peripheral RF led to a decrease in interburst intervals, accompanied, depending upon the unit, by either an increase or a decrease in the number of spikes per burst. 10. Results are discussed in terms of the functional significance of resting discharge, including bursting, and possible roles in somatosensory information

Evidence for retrograde degeneration of epinephrine neurons in Alzheimer's disease.

Alzheimer's disease (AD) is associated with a progressive loss of locus ceruleus neurons. These noradrenergic neurons receive a major afferent projection from epinephrine neurons in epinephrine cell groups in the brainstem. The epinephrine neurons have a specific enzymatic marker, phenylethanolamine N-methyltransferase (PNMT), which allows them to be identified chemically and immunohistochemically. We have previously reported a decrease in PNMT in brains of patients with AD. We now report that the decrease in PNMT activity in projections to the locus ceruleus is not due to the loss of epinephrine neurons, although up to 33% of these neurons are atrophic. The decrease in presynaptic PNMT does, however, correlate with the loss of postsynaptic locus ceruleus neurons in brains from AD patients. The percentage of degenerating neurons in the epinephrine nuclei also correlates significantly with the amount of loss of locus ceruleus neurons in AD. In addition, there is a 55% decrease in mitogen activity, a nonspecific measure of growth or maintenance factors, in dialysed locus ceruleus extracts from the AD patients compared to those from control subjects. The mitogen activity in the locus ceruleus was significantly correlated with PNMT activity and with the density of locus ceruleus neurons in all cases examined. These findings provide evidence for the hypothesis that retrograde degeneration is a mechanism of neuronal degeneration in AD and suggest that trophic factors may play a role in this process.

Peripheral sympathetic ingrowth can alter metabolic activity within the hippocampal formation.

After cholinergic denervation of the hippocampal formation, peripheral sympathetic fibers arising from the superior cervical ganglia grow into the dentate gyrus and CA3 region; the functional significance of sympathetic ingrowth into the hippocampal formation is unknown. Utilizing electrical stimulation of the preganglionic trunk in combination with 2-deoxy-D-[3H]-glucose fine-grained autoradiograms, we demonstrated an alteration of hippocampal glucose metabolism, suggesting that this neuronal rearrangement makes functional connections within the hippocampus.