Acetylcholine Receptor Targets on Cortical Pyramidal Neurones as Targets for Alzheimer's Therapy

Experimental lesions using the retrogradely transported toxin, volkensin, have been used in conjunction with autoradiography to investigate the cellular localization of 5–HT_1A, muscarinic M₁and nicotinic receptors. Selective destruction of neocortical pyramidal neurones forming the corticostriatal or corticocortical pathways was achieved by intrastriatal or intracortical injection of volkensin. Selective destruction of layer V corticostriatal neurones was accompanied by loss of binding in the cortex to 5–HT_1Aand muscarinic M₁receptors, and an upregulation of [³H] nicotine binding contralateral to the pyramidal cell loss. Destruction of corticocortical neurones was accompanied by loss of binding to muscarinic and nicotinic receptors. The presence of these cholinoceptors on corticocortical neurones was confirmed by recording carbachol-induced depolarizations from a novel cortical brain slice preparation. It is proposed that cholinoceptors represent a consistent marker for neocortical pyramidal cells, and as such are viable targets for the continuing development of therapies designed to ameliorate the cortical hypoactivity observed in Alzheimer's disease. Ligands for these receptors may also be suitable for positron emission tomography to assess pyramidal neurone numbers in suspected Alzheimer's disease.     

NMDA receptors assessed by autoradiography with [3H]L-689,560 are present but not enriched on corticofugal-projecting pyramidal neurones.

Experimental lesions followed by binding of [3H]4-trans-2-carboxy-5,7-dichloro-4-phenylamino-carbonylamino-1,2 ,3,4- tetrahydroquinoline ([3H]L-689,560, a novel ligand that binds to the glycine modulatory site), [3H]glycine and [3H]glutamate (N-methyl-D-aspartate (NMDA) sensitive) to cryostat sections and quantitative autoradiography were used to investigate the cellular localization of the NMDA receptor complex in the neocortex of the rat. The lesions were produced by intrastriatal injections of either volkensin (2 and 6 ng) or ricin (10 ng): both are suicide transport agents but only the former is retrogradely transported in the CNS. The binding of [3H]L-689,560 was significantly reduced in rats receiving 2 or 6 ng volkensin in deep cortical layers of Fr1/Fr2 ipsilateral to the striatal lesion. Similar reductions were also seen in [3H]glycine and [3H]glutamate binding, but only in rats receiving 6 ng volkensin. Quantitative histological analysis had previously revealed a loss of large infragranular pyramidal neurones with sparing of both interneurones and supragranular pyramidal neurones. There were no significant reductions in binding of any ligand in the superficial layers. In cortical areas Par1/Par2, [3H]L-689,560 was also significantly reduced in deep layers but only in rats receiving 6 ng volkensin. Binding was also reduced in the superficial layers by contrast to Fr1/Fr2. [3H]Glycine and [3H]glutamate binding were unaffected in this area. Binding of [3H]L-689,560 was unaffected in any area following intrastriatal ricin injection. The present study indicates that the NMDA receptor complex is present on pyramidal cells forming the corticofugal pathways. This is discussed in terms of the 5-HT1A receptor which is enriched on these cells.     

Changes in Cortical Nicotinic Acetylcholine Receptor Numbers Following Unilateral Destruction of Pyramidal Neurones by Intrastriatal Volkensin Injection

Experimental lesions using the retrogradely transported toxic lectin, volkensin, were used in conjunction with quantitative autoradiography to investigate the cellular localization of nicotinic and adenosine A₁receptors. Lesions were produced by unilateral intrastriatal injection of volkensin, ricin (another toxic lectin but not transported in the central nervous system), quinolinate, and unilateral intrathalamic injection of ibotenate. Volkensin injection significantly reduced the number and mean cell size of large, infragranular pyramidal neurones in cortical areas Fr1/Fr2 (close to the midline) and more laterally in Par1/Par2. Selective destruction of these cells was accompanied by significant increases in the binding of [³H] nicotine in cortical areas contralateral to the lesion. A small but significant reduction in the binding of [³H] 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) to adenosine A₁receptors was observed only in deep layers of Fr1/Fr2 on the side ipsilateral to the lesion. No other toxin consistently changed the binding of either ligand in control animal groups with the exception of [³H] nicotine where small reductions were observed in the middle layers of one thalamic injection group. These data indicate differential plasticity of nicotinic receptors compared with other receptors studied previously using this paradigm. In the light of these findings, nicotinic receptors are discussed as targets for pharmacological manipulation of the activity of pyramidal neurones.     

Selective localization of striatal D1 receptors to striatonigral neurons

A new technique for producing anatomically selective lesions within the brain was used to investigate the cellular localization of the D1 and D2 receptor. The cytotoxic lectin, volkensin, is taken up by nerve terminals and retrogradely transported, killing those neurons projecting to the site of injection. Comparison of D1 and D2 binding following a unilateral volkensin injection into the substantia nigra has demonstrated that striatal D1 binding sites are selectively localized to striatonigral projection neurons.     

Organization of local axon collaterals of efferent projection neurons in rat visual cortex

We have studied the laminar origins of local long-range connections within rat primary visual cortex (area 17), by using retrograde tracing of nerve cell bodies with fluorescent markers. Injections throughout the thickness of cortex produce distinct laminar labeling patterns which indicate that a substantial number of cells in layers 2/3, 5, and 6 have wide local axon collateral arbors, while the local arbors of layer 4 cells are much narrower. Double labeling experiments which combined area 17 injections with injections into different projection targets of area 17 (opposite area 17, area 18a, and area 18b) show that many cortico-cortically projecting cells make widespread projections within area 17. In contrast, the overwhelming majority of subcortically projecting cells have narrow collateral arbors within area 17. Anterograde tracing of local projections within areas 17 with the lectin Phaseolus vulgaris leucoagglutinin shows an extensive system of horizontally running fibers which terminate in distinct 0.15-0.25 mm wide clusters up to 1.8 mm from the injection site. On horizontal sections the termination pattern resembles a closely spaced lattice. The results indicate that cortico-cortically projecting cells provide for long-range interactions between distant points of the visuotopic map, while subcortically projecting cells mediate information within a cortical column. Interestingly, subcortically projecting cells differ functionally from cortico-cortically projecting cells in that they are not orientation selective (Klein et al., Neurosci. 17:57-78, '86; Mangini and Pearlman, J. Comp. Neurol. 193:203-222, '80; Simmons and Pearlman, J. Neurophysiol. 50:838-848, '83). We therefore suggest that cortico-cortically projecting cells with wide collateral arbors are orientation selective and that clustered long-range projections within area 17 connect columns with similar functional specificity.     

A selective lesion of striatonigral neurons decreases presynaptic binding of [H]hemicholinium-3 to striatal interneurons

We have used the suicide transport agent, volkensin, to produce selective lesions of striatal efferent neurons projecting to the substantia nigra in the rat. In order to evaluate potential trans-synaptic effects, we examined cholinergic interneurons intrinsic to the striatum following destruction of striatonigral projection neurons by nigral injection of volkensin. There was no change in the number of large interneurons identified either by Nissl stain or by immunocytochemistry for choline acetyltransferase, indicating that volkensin was not directly toxic to this group of neurons. However, [³H]hemicholinium-3 binding to the choline re-uptake site on the presynaptic cholinergic terminals decreased. No change in [³H]hemicholinium-3 binding was seen after destruction of dopaminergic afferents with 6-hydroxydopamine. Striatonigral afferents to the cholinergic interneurons contain substance P which has been shown to stimulate acetylcholine release. The decrease in [³H]hemicholinium-3 binding may reflect loss of this afferent input. However, striatonigral neurons are an efferent target of the cholinergic interneuron as well, and a presynaptic effect due to loss of target neurons also may contribute.     

Modeccin and volkensin but not abrin are effective suicide transport agents in rat CNS

Suicide transport is a term applied to the technique of producing anatomically selective neural lesions using axonally transported cytotoxins. Because the cytotoxic lectins, abrin, modeccin and volkensin are effective suicide transport agents in the peripheral nervous system, the present study sought to determine if they were effective suicide transport agents in the rat CNS. Toxins were stereotactically pressure microinjected unilaterally into the caudate nucleus of rats. After 2-13 days survival, brain sections were processed for catecholamine histofluorescence or Nissl stained with Cresyl violet. All 3 agents produced extensive necrosis at the caudate injection site. In addition, modeccin and volkensin but not abrin produced destruction of neurons in the ipsilateral substantia nigra and intralaminar thalamus. Histofluorescence confirmed loss of dopaminergic neurons from the ipsilateral substantia nigra after modeccin or volkensin but not abrin injections. These results indicate that modeccin and volkensin are effective suicide transport agents within the rat CNS, presumably due to retrograde axonal transport of the toxins. These agents may prove extremely useful in producing anatomically selective lesions of neurons afferent to a toxin injection site.     

Interhemispheric and subcortical collaterals of medial prefrontal cortical neurons in the rat

Efferent neurons of rat medial prefrontal cortex, projecting to subcortical structures and contralateral homotypical areas, were analyzed using anatomical and electrophysiological methods. Anterograde labelling with radioactive amino acids demonstrated the pathways of these efferents in the rostral part of the brain; terminal fields in contralateral cortical areas and localization of fibers projecting subcortically were particularly examined. The laminar location of cells projecting to the contralateral prefrontal cortex or through the striatum was investigated by means of the retrograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injected in these two structures. Cells innervating the contralateral prefrontal cortex were distributed in layers II–III and V, whereas injection of WGA-HRP in the striatum labelled cells in layer V. The antidromic activation technique was used to identify the cortical neurons which innervate the ipsilateral and contralateral subcortical structures as well as contralateral homotypical cortical areas. Among 743 recorded neurons, 282 neurons were antidromically driven from at least one of the stimulated sites (e.g. right striatum (RS), left striatum (LS), and contralateral prefrontal cortex (L-Cx)). The mean conduction velocities were 0.6 m/s and 0.8 m/s for subcortical and cortical efferents, respectively. The reciprocal collision test provided evidence for the existence of branched axons for 35% of the antidromically activated cells. All the possible branching patterns were found. The results of this study thus demonstrate the existence of single neocortical neurons that send axon collaterals to cortex and subcortical structures.     

NEURONOTOXICITY OF AXONALLY TRANSPORTED TOXIC LECTINS, ABRIN, MODECCIN AND VOLKENSIN IN RAT PERIPHERAL NERVOUS SYSTEM

In an attempt to find new and more useful suicide transport agents, the cytotoxic lectins abrin, modeccin and volkensin were pressure microinjected into peripheral nerves (vagus, hypoglossal and sciatic) of adult rats. After 33 h-5 days survival, the brainstems, spinal cords and corresponding sensory ganglia were examined histologically. All three lectins produced profound chromatolysis, and destruction of sensory and motor neurons projecting axons through the injected nerves. Volkensin and modeccin were significantly more potent than any previously reported suicide transport agent. It is concluded that abrin, modeccin and volkensin are effective, unselective suicide transport agents in the rat peripheral nervous system but none is clearly superior to ricin for making restricted sensory and motor neuron ablations. However, modeccin and volkensin are fundamentally different from any previously reported suicide transport agents with respect to spread within the CNS which destroyed neurons adjacent to those initially taking up and transporting the toxin. Possibly this is due to the different oligosaccharide binding specificity of modeccin and volkensin compared to other suicide transport agents. Modeccin and/or volkensin may prove useful in making lesions of CNS interneurons using the suicide transport strategy.     

Collaterals of corticospinal and pyramidal fibres to the pontine grey demonstrated by a new application of the fluorescent fibre labelling technique

Selective visualization of collaterals of corticospinal and pyramidal fibres to the pons in cat was obtained by retrograde transport of the fluorescent tracer fast blue (FB) through the stem fibres. Unilateral FB injections in the cervical cord and the pyramidal tract respectively produced soft blue fluorescent labelling of pyramidal fibres and of fibres and structures resembling 'terminals' in the pontine grey: contralateral to the spinal injections and ipsilateral to the pyramidal injections. These labelled elements were concluded to represent collaterals of corticospinal and pyramidal fibres because (a) their distribution corresponded to that of the pericruciate corticopontine fibres, (b) their labelling was prevented when the FB injections were preceded by a transection of either the cerebral peduncle or the pyramidal tract which lesions also prevented the FB labelling of the distal parts of the transected axons. Similar findings were obtained when using wheat germ agglutinin-horseradish peroxidase. In other experiments FB-labelling of pyramidal collaterals was combined with retrograde labelling of pontine neurones projecting to the contralateral anterior lobe of the cerebellum using diamidino yellow dihydrochloride as the second tracer. The distributions of the retrogradely labelled neurones and of the pyramidal collaterals in the pontine grey showed an almost complete overlap indicating that these collaterals mainly establish connections with the cerebellar anterior lobe.     

Intrinsic Organization and Connectivity of Intrastriatal Striatal Transplants in Rats as Revealed by DARPP-32 Immunohistochemistry: Specificity of Connections with the Lesioned Host Brain

Intrastriatal grafts of tissue obtained from the striatal or neocortical primordia of rat fetuses have been studied with respect to their intrinsic organization and connectivity using antibodies to DARPP-32 in combination with acetylcholinesterase (AChE) histochemistry, tyrosine hydroxylase (TH) immunocytochemistry, and anterograde and retrograde axonal tracing techniques. The striatal grafts were characterized by distinct patches of DARPP-32-immunoreactive neurons, which were identical to the densely AChE-positive patches stained in adjacent sections from the same specimens. The non-patch areas possessed only few DARPP-32-positive neurons and contained only sparse AChE-positive fibres. The cortical grafts, by contrast, contained no neurons with clear-cut DARPP-32-positivity and they exhibited a sparse, evenly distributed AChE fibre network, similar to that seen in the non-patch areas of the striatal grafts. The host dopaminergic afferents, as revealed by TH immunostaining, had grown selectively into the DARPP-32-positive patches in the striatal grafts, where they formed a dense terminal network around the DARPP-32-positive cell bodies. The non-patch areas, as well as the cortical grafts, received only sparse TH innervation. By contrast, the host cortical afferents, labelled by Phaseolus vulgaris leucoagglutinin from the host frontal cortex, were seen to extend into both the patch and non-patch areas of the striatal grafts. Transplant neurons projecting into the host brain were labelled by Fluoro-Gold injections into the ipsilateral host globus pallidus. These injections labelled large numbers of medium-sized neurons within the striatal grafts and the vast majority of them (over 85%) were confined to the DARPP-32-positive patches. Similar Fluoro-Gold injections labelled only few graft neurons in the cortical grafts. The results indicate that the striatal grafts are composed of a mixture of striatal and non-striatal tissue, and that the striatal graft compartment selectively establishes afferent and efferent connections with the host nigro-pallidal system. These graft connections demonstrate a remarkable specificity in the formation of graft - host connectivity. The results, moreover, suggest that developmental properties of the grafted striatal primordium are retained and expressed in the implanted cell suspension, and that the neuronal systems of the lesioned adult host brain, at least to some extent, remain responsive to growth regulating mechanisms normally operating during ontogenetic development.     

Corticopontine projection in the rat: the distribution of labelled cortical cells after large injections of horseradish peroxidase in the pontine nuclei

The distribution of cortical cells projecting to the pontine nuclei in rats was studied by making large injections of horseradish peroxidase that filled the basilar pons and measuring the density of labelled cells in each cortical area. All retrogradely labelled cells were layer V pyramidal cells. The highest densities of labelled cells were observed in the motor areas. The lowest densities were in temporal association cortex and perirhinal cortex. Visual cortical areas, including the primary visual cortex, provided a major source of pontine projections. The distribution of corticopontine cells within the primary visual cortex was studied in more detail. In all cases the highest density of labelled cells was observed in the region of cortex that represents the nasal visual field. Control injections into brainstem regions adjacent to the pontine nuclei produced a much lower absolute density of retrogradely labelled cortical cells and the distribution of those cells was different from that observed following pontine injections. We conclude that every area of the rat's cerebral cortex projects to the pontine nuclei and that there are consistent variations in the density of the projections both between and within areas.     

Differential synaptic innervation of striatofugal neurones projecting to the internal or external segments of the globus pallidus by thalamic afferents in the squirrel monkey

It is well established that the centromedian nucleus (CM) is the major source of thalamic afferents to the sensorimotor territory of the striatum in monkeys. However, the projection sites of striatal neurones contacted by thalamic afferents still remain to be determined. We therefore carried out an anatomical study aimed at elucidating the hodology of striatal neurones that receive input from the CM in squirrel monkeys. Our approach was to combine the anterograde transport of Phaseolus vulgaris-leucoagglutinin (PHA-L) or biocytin from the CM with the retrograde transport of biotinylated dextran-amine (bio-dex) or PHA-L from the internal (GPi) or external (GPe) segments of the globus pallidus. Following CM injections, rich plexuses of anterogradely labelled, thin varicose fibres aggregated in the form of bands that were confined to the postcommissural region of the putamen. On the other hand, injections into the GPe or GPi led to profuse retrograde labelling of a multitude of medium-sized spiny neurones. In cases where the injections involved the caudoventral two-thirds of the GPe or GPi, the retrogradely labelled striatopallidal cells and the anterogradely labelled thalamostriatal fibres occurred in the sensorimotor territory of the putamen. After injections into either pallidal segments, clusters of retrogradely labelled cells were in register with bands of anterogradely labelled thalamic fibres. However, electron microscopic analysis of striatal regions containing both anterogradely labelled thalamic afferents and retrogradely labelled cells revealed that terminals from the CM frequently form asymmetric synapses with dendritic shafts and spines of striato-GPi cells but rarely with those of striato-GPe cells. In conclusion, our findings demonstrate that thalamic afferents from the CM innervate preferentially striatopallidal neurones projecting to the GPi in monkeys. These results indicate that the striatopallidal neurones contributing to the “direct” and “indirect” output pathways are differentially innervated by thalamic afferents in primates. © 1996 Wiley-Liss, Inc.     

Methamphetamine neurotoxicity: Dissociation of striatal dopamine terminal damage from parietal cortical cell body injury

Methamphetamine (m-AMPH) administration injures both striatal dopaminergic terminals and certain nonmonoaminergic cortical neurons. Fluoro-Jade histochemistry was used to label cortical cells injured by m-AMPH in order to identify factors that contribute to the cortical cell body damage. Rats given four injections of m-AMPH (4 mg/ kg) at 2-h intervals showed hyperthermia (mean = 40.0 ± 0.10°C) and increased behavioral activation relative to animals given saline (SAL). Three days later, m-AMPH-treated animals showed indices of injury to striatal DA terminals (depletion of tyrosine hydroxylase immunoreactivity) and parietal cortical cell bodies (appearance of Fluoro-Jade stained cells). Pretreatment with a dopamine (DA) D1, D2, or N-methyl-D-aspartate (NMDA) receptor antagonist, or administration of m-AMPH in a 4°C environment, prevented or attenuated m-AMPH-induced hyperthermia, behavioral activation, and injury to striatal DA terminals and parietal cortical cell bodies. Animals pretreated with a DA transport inhibitor prior to m-AMPH showed hyperthermia, behavioral activation, and parietal cortical cell body injury, but they did not show striatal DA terminal injury. Pretreatment with a 5HT transport inhibitor failed to prevent m-AMPH-induced damage to striatal DA terminals or parietal cortical cell bodies. Animals given four injections of SAL in a 37°C environment became hyperthermic, but showed no injury to striatal DA terminals or cortical cell bodies. The ability of the DA transport inhibitor to block m-AMPH-induced striatal DA damage, but not cortical injury, and the inability of hyperthermia alone to cause the cortical cell body injury suggests that m-AMPH-induced behavioral activation and hyperthermia may both be necessary for the subsequent parietal cortical cell body damage. Synapse 30:433–445, 1998. © 1998 Wiley-Liss, Inc.     

Critical stages for growth in the development of cortical neurons

In order to study the role of efferent connectivity in the development of CNS neurons, the growth of pyramidal tract neurons within the hamster sensorimotor cortex was studied during normal development and after early postnatal lesions of the pyramidal tract. We first determined, by a combination of Nissl and retrograde HRP techniques, that within the lumbar representation of cortical layer 5B in adult animals two cell populations exist: a large-celled population (40% of the total) projecting to the spinal cord and a small-celled population (60% of the total) projecting intracortically and to targets rostral to the medulla. We could not determine whether large layer 5B cells in the infant sensorimotor cortex also represent the corticospinal population. Nevertheless, measurements of the growth in cross-sectional area of the large cells from 7 days postnatal to adulthood showed that these cells continue to grow until 51 days of age. The most rapid rate of growth occurs between 7 and 14 days, during which time the cross-sectional area of the cell bodies triples, coincident with the arrival of corticospinal axons in the lumbar cord and the beginning of target innervation (Reh and Kalil, '81). The growth of the large neurons in layer 5B was then charted after the pyramidal tract was cut ipsilaterally in the medulla at various postnatal ages. Early lesions of the tract (4-8 days postnatal) interrupt lumbar projection fibers before they establish synapses in the cord. Nevertheless, cortical cell bodies in the lumbar representation continue to grow normally after axotomy until 11 days after birth. At this time, large cells are arrested in development and their cell size remains in the 11-day stage (50% of normal adult large cell size) indefinitely. In contrast, adult lesions of the tract cause a 60% shrinkage of large cells, which in the adult represent corticospinal neurons. No evidence for cortical cell death was found after pyramidal tract lesions at any age. The results of axotomy reveal a turning point in the development of layer 5B cortical neurons. Before the age of 11 days the large cells have an independent program of cell growth that proceeds despite axotomy. After this time, the large cortical neurons appear to require intact axons for further growth and, in the absence of normal connectivity, are arrested in development.     

Morphological differentiation of distinct neuronal classes in embryonic turtle cerebral cortex.

As a starting point for understanding the development of the cerebral cortex in reptiles and for determining how reptilian cortical development compares to that in other vertebrate classes, we studied the appearance and morphological differentiation of cerebral cortical neurons in embryonic turtles. 3H-thymidine birthdate labeling and focal injections of horseradish peroxidase (HRP) in in vitro cortical slices revealed that replicating cells occupy the outer ventricular zone, and subsequently migrate to the ventricular surface where they divide. Postmitotic neurons begin differentiating and elaborating neurites while migrating back through the ventricular zone. On their arrival at the top of the ventricular zone, pyramidal and nonpyramidal neurons can be distinguished morphologically. Cells with multipolar apical dendritic tufts ascending in the marginal zone resemble immature pyramidal neurons. Neurons morphologically similar to these early pyramidal cells were retrogradely labeled by injections of the lipophilic tracer 1,1-dioctadecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (diI) in a known pyramidal cell target, the thalamus. Nonpyramidal neurons, resembling Cajal-Retzius cells, had horizontally oriented long axons and dendrites coursing in the plexiform primordium, the future marginal zone. With further development morphological differences between cell types became accentuated, and pyramidal cell somata were segregated into a single cellular layer flanked by zones containing predominantly nonpyramidal cells. Axon elaboration occurred early in embryonic development, as pyramidal cells sent axonal branches to the septum, thalamus, and cortical targets soon after their generation, and the intracortical axonal plexus became increasingly dense during embryonic life. Over a similar time course the distribution of projecting neurons labeled by thalamic diI injections changed from an initial homogeneous distribution to a preferential location in the superficial half of the cellular layer. Results from this study demonstrate several features of cortical differentiation that are conserved in reptiles and mammals, including similar early morphological differentiation events, the early distinction of principal cell types, and the parallel development of pyramidal and nonpyramidal neurons. The context in which these similar developmental events occur, however, differs profoundly in reptiles and mammals, with differences in the timing and location of neurite elaboration and differences in the appearance and architectonic organization of the cortex. Comparison of cortical developmental patterns between reptiles and mammals shows that similar functional cortical circuits with balanced excitation and inhibition can emerge in diverse cortical structures.     

Suicide retrograde transport of volkensin in cerebellar afferents: direct evidence, neuronal lesions and comparison with ricin

Volkensin and ricin, either free or conjugated with colloidal gold, were injected into the cerebellar cortex of rats. The inferior olive and pontine nuclei were examined to verify the retrograde axonal transport of these two toxins, and the consequent neuronal damage. No evidence was obtained of a retrograde axonal transport of ricin in these pathways. Injection of gold-conjugated volkensin in the cerebellar cortex resulted in retrogradely labelled neurones in the inferior olive after 3 h, and in the pontine nuclei after 6 h. Degenerative changes were very severe in the retrogradely labelled neurones 48 h after the gold-conjugated volkensin injection. In the Nissl-stained material, neuronal degeneration started to be evident in the inferior olive 12 h, and in pontine nuclei 6 h, after volkensin injection. The neuronal degeneration in both the inferior olive and pons increased up to 4 days after the injection. These findings provide direct evidence of the retrograde axonal transport of volkensin in the central nervous system, and of the time course of the consequent degenerative changes in the afferents to the cerebellar cortex.     

Activity of identified cortically projecting and other basal forebrain neurones during large slow waves and cortical activation in anaesthetized rats

To examine the role played by the basal forebrain cholinergic system in cortical activation, neuronal activity was investigated in the globus pallidus and substantia innominata of urethane-anaesthetized rats during large cortical slow waves and spontaneous or elicited low voltage fast activity. An effort was made to identify the neurones by antidromic stimulation from the neocortex (supposedly cholinergic cells) and from the subthalamic nucleus (pallidal cells). Most of the cortically projecting neurones were strongly activated during cortical activation (5-fold increase in firing rate on average), while the discharge rate of pallidal units was increased only slightly (ratio 1.25 on average). In contrast to the cortically projecting cells, some unidentified cells in the substantia innominata fired at a much higher rate during large cortical slow waves as compared to low voltage fast activity. The results are discussed in relation to previous work on the cortically projecting cells and on the mechanisms of cortical activation.     

Pyramidal cell loss in motor cortices in Huntington's disease

Patterns of huntingtin protein aggregation and cortical neuronal loss suggest early involvement of corticostriatal pathways in Huntington's disease. However, theories of pathogenesis of chorea rely on the motor cortices being intact. The motor cortices have not previously been studied at a cellular level in Huntington's disease. We analyzed the neuronal number in the caudate, putamen, and three motor cortical areas in five cases of Huntington's disease and five controls. For each motor cortical region the total neuronal number, number of interneurons, and number of SMI32 immunopositive pyramidal neurons were quantified using previously published techniques and any relationship between cell loss and severity or duration of chorea was examined. The results showed a loss of long projecting SMI32 immunopositive pyramidal neurons in the primary motor cortex with associated morphological changes and suggest a loss of short projecting pyramidal neurons in the premotor cortex. Degeneration in the primary motor cortex correlated with subcortical degeneration. These findings indicate pyramidal cell involvement in Huntington's disease and implicate the degeneration of corticostriatal pathways in the production of chorea.     

The time course of changes in D1 and D2 receptor binding in the striatum following a selective lesion of striatonigral neurons.

The suicide transport agent volkensin was used to produce a selective lesion of striatonigral projection neurons and the time course of changes in binding at striatal D1 and D2 receptors analyzed. Both show a time-dependent decrease with two-thirds of the total change occurring within the first 10 days and a greater decrease in D1 receptor density at all time points. Our results confirm selective localization of D1 receptors to striatonigral neurons and are consistent with localization of some striatal D2 receptors to striatonigral neurons.