Daniel S. Zahm

Specificity in the projection patterns of accumbal core and shell in the rat

The efferent projections of the core and shell areas of the nucleus accumbens were studied with a combination of anterograde and retrograde tract-tracing methods, including Phaseolus vulgaris-leucoagglutinin, horseradish peroxidase and fluorescent tracers. Both the core and shell regions project to pallidal areas, i.e. ventral pallidum and entopeduncular nucleus, with a distinct topography in the sense that the core projection is located in the dorsolateral part of ventral pallidum, whereas the shell projects to the medial part of the subcommissural ventral pallidum. Both regions of the accumbens also project to mesencephalon with a bias for the core projection to innervate the substantia nigra-lateral mesencephalic tegmentum, and for the shell projection to reach primarily the ventral tegmental-paramedian tegmentum area. The most pronounced differences between core and shell projections exist in regard to the hypothalamus and extended amygdala. Whereas the core projects primarily to the entopeduncular nucleus including a part that invades the lateral hypothalamus, the shell, in addition, projects diffusely throughout the rostrocaudal extent of the lateral hypothalamus as well as to the extended amygdala, especially its sublenticular part. Both the core and shell of the accumbens have unmistakable striatal characteristics both histologically and in their connectional patterns. The shell, however, has additional features that are reminiscent of the recently described extended amygdala [Alheid G.F. and Heimer L. (1988) Neuroscience 27, 1-39; de Olmos J.S. et al. (1985) In The Rat Nervous System, pp. 223-334]; in fact, the possibility exists that the shell represents a transitional zone that seems to characterize most of the fringes of the striatal complex, where it adjoins the extended amygdala.

Glutamatergic Afferents of the Ventral Tegmental Area in the Rat

Glutamatergic inputs to the ventral tegmental area (VTA), thought crucial to the capacity of the VTA to detect and signal stimulus salience, have been reported to arise in but a few structures. However, the afferent system of the VTA comprises very abundant neurons within a large formation extending from the prefrontal cortex to the caudal brainstem. Neurons in nearly all parts of this continuum may be glutamatergic and equivalently important to VTA function. Thus, we sought to identify the full range of glutamatergic inputs to the VTA by combining retrograde transport of wheat germ agglutinin-bound gold after injections into the VTA with nonisotopic in situ hybridization of the vesicular glutamate transporters (VGLUTs) 1, 2, and 3. We found glutamatergic neurons innervating the VTA in almost all structures projecting there and that a majority of these are subcortical and VGLUT2 mRNA positive. The tremendous convergence of glutamatergic afferents from many brain areas in the VTA suggests that (1) the function of the VTA requires integration of manifold and diverse bits of information and (2) the activity of the VTA reflects the ongoing activities of various combinations of its afferents.

Afferents of the ventral tegmental area in the rat-anatomical substratum for integrative functions

The ventral tegmental area (VTA) is critically important to an organisms capacity to detect rewards and novelty and to enlist appropriate behavioral responses. Although there has been substantial progress concerning information processing at the single cell and molecular levels in the VTA, our knowledge of its overall afferent connections is based principally on the benchmark description by Phillipson ([1979] J. Comp. Neurol. 187:117–144). Given that, since then, the sensitivity of tracing methods and knowledge about the organization of brain structures have increased considerably, we undertook to reevaluate the VTA afferents of the rat. The retrograde tracer Fluoro‐Gold was injected into different parts of the VTA, and labeled neurons were visualized by immunocytochemistry. Retrogradely labeled neurons were not confined to nuclei but rather constituted an elongated formation stretching from the prefrontal cortex rostrally to the medulla oblongata caudally. In the case of descending afferents, this formation was centered on the medial forebrain bundle and the fasciculus retroflexus. The input to the VTA in general was bilateral, with a smaller descending and comparable ascending projection from the contralateral side. Injections of the anterograde tracers Phaseolus vulgaris‐leucoagglutinin or biotinylated dextran amine into selected forebrain structures revealed a surprisingly sparse terminal arborization in the VTA. Furthermore, structures projecting to the VTA innervate other brain areas with similar or greater robustness, which in turn also provide a strong input to the VTA, indicating an anatomical network. Given the importance of the VTA in basic behaviors, this organization might provide a basis for an extraordinary level of afferent integration. J. Comp. Neurol. 490:270–294, 2005.

Functional-anatomical Implications of the Nucleus Accumbens Core and Shell Subterritories

The nucleus accumbens, a major part of the ventral striatum, comprises numerous subterritories and compartments, of which the core and shell appear to be dominant. Shell exhibits greater chemical neuroanatomical diversity than core and is rather directly connected to it by a robust, feed‐forward, striatopallido‐thalamocortico‐striatal pathway. Shell and extended amygdala share afferents, but the two are distinguished by their outputs, strongly toward cortex for shell and descendent toward brain stem effector sites for extended amygdala. Shell responds independently to stimulation by excitatory amino acids and dopamine, which are more mutually permissive in the core. Accordingly, the shell responds to a broad variety of physiological and pharmacological stimuli, including psychomotor and opioid drugs. Whereas locomotion and oro‐facial movements are elicitable from the shell, lesions and blockade of EAA transmission in the core reduce locomotion. It is hypothesized that core‐shell has a feed‐forward functional organization.

The mesopontine rostromedial tegmental nucleus: A structure targeted by the lateral habenula that projects to the ventral tegmental area of Tsai and substantia nigra compacta

Prior studies revealed that aversive stimuli and psychostimulant drugs elicit Fos expression in neurons clustered above and behind the interpeduncular nucleus that project strongly to the ventral tegmental area (VTA) and substantia nigra (SN) compacta (C). Other reports suggest that these neurons modulate responses to aversive stimuli. We now designate the region containing them as the “mesopontine rostromedial tegmental nucleus” (RMTg) and report herein on its neuroanatomy. Dense μ‐opioid receptor and somatostatin immunoreactivity characterize the RMTg, as do neurons projecting to the VTA/SNC that are enriched in GAD67 mRNA. Strong inputs to the RMTg arise in the lateral habenula (LHb) and, to a lesser extent, the SN. Other inputs come from the frontal cortex, ventral striatopallidum, extended amygdala, septum, preoptic region, lateral, paraventricular and posterior hypothalamus, zona incerta, periaqueductal gray, intermediate layers of the contralateral superior colliculus, dorsal raphe, mesencephalic, pontine and medullary reticular formation, and the following nuclei: parafascicular, supramammillary, mammillary, ventral lateral geniculate, deep mesencephalic, red, pedunculopontine and laterodorsal tegmental, cuneiform, parabrachial, and deep cerebellar. The RMTg has meager outputs to the forebrain, mainly to the ventral pallidum, preoptic‐lateral hypothalamic continuum, and midline‐intralaminar thalamus, but much heavier outputs to the brainstem, including, most prominently, the VTA/SNC, as noted above, and to medial tegmentum, pedunculopontine and laterodorsal tegmental nuclei, dorsal raphe, and locus ceruleus and subceruleus. The RMTg may integrate multiple forebrain and brainstem inputs in relation to a dominant LHb input. Its outputs to neuromodulatory projection systems likely converge with direct LHb projections to those structures. J. Comp. Neurol. 513:566–596, 2009.

Morphological differences between projection neurons of the core and shell in the nucleus accumbens of the rat

The somatodendritic morphology of projection neurons in the shell and core of the rat nucleus accumbens was studied. These cells were retrogradely labelled with Fast Blue from the ventral mesencephalon (substantia nigra/ventral tegmental area) and subsequently injected intracellularly with Lucifer Yellow and processed immunocytochemically. Digitized reconstructions revealed that the cell bodies of neurons located throughout the nucleus are small-to-medium in size. Neurons in the shell have significantly fewer dendritic arbours with fewer branch segments, fewer terminal segments, and lower spine densities than those in the core. Values for the same parameters are significantly greater for cells in lateral than in medial parts of the shell but the same for neurons located within and without enkephalin enriched parts of the core, with an exception of spine density being significantly greater in the enkephalin-rich compartment. Calculations based on these data reveal that neurons in the core have as much as 50% more surface area than those in the shell, which suggests that core neurons have a greater potential for collecting synaptic information than have shell cells. Furthermore, the differential distribution and action of various neurochemicals such as dopamine in the shell and core, supports the idea that different morphologies reflect the presence of distinct neuronal circuits in these two territories.

Altered Dendritic Spine Plasticity in Cocaine-Withdrawn Rats

Chronic cocaine treatment is associated with changes in dendritic spines in the nucleus accumbens, but it is unknown whether this neuroplasticity alters the effect of a subsequent cocaine injection on spine morphology and protein content. Three weeks after daily cocaine or saline administration, neurons in the accumbens were filled with the lipophilic dye, DiI. Although daily cocaine pretreatment did not alter spine density compared with daily saline, there was a shift from smaller to larger diameter spines. During the first 2 h after an acute cocaine challenge, a bidirectional change in spine head diameter and increase in spine density was measured in daily cocaine-pretreated animals. In contrast, no change in spine diameter or density was elicited by a cocaine challenge in daily saline animals during the first 2 h after injection. However, spine density was elevated at 6 h after a cocaine challenge in daily saline-pretreated animals. The time-dependent profile of proteins in the postsynaptic density subfraction elicited by a cocaine challenge in daily cocaine-pretreated subjects indicated that the changes in spine diameter and density were associated with a deteriorating actin cytoskeleton and a reduction in glutamate signaling-related proteins. Correspondingly, the amplitude of field potentials in accumbens evoked by stimulating prefrontal cortex was reduced for up to 6 h after acute cocaine in daily cocaine-withdrawn animals. These data indicate that daily cocaine pretreatment dysregulates dendritic spine plasticity elicited by a subsequent cocaine injection.

Specificity in the Projections of Prefrontal and Insular Cortex to Ventral Striatopallidum and the Extended Amygdala

The basal forebrain functional-anatomical macrosystems, ventral striatopallidum, and extended amygdala are innervated by substantially coextensive distributions of neurons in the prefrontal and insular cortex. This suggests two alternative organizational schemes: convergent, in which a given cortical area projects exclusively to only one of these macrosystems and divergent, in which a given cortical area innervates both forebrain macrosystems. To examine the underlying organization and possibly discriminate between these alternatives, rats were injected with two retrograde tracers in different parts of ventral striatopallidum or extended amygdala (homotypic injection pairs) or with one tracer in each macrosystem (heterotypic). The prefrontal and insular cortex was evaluated microscopically for overlap of retrograde labeling and double labeling of neurons. Homotypic injection pairs in the ventral striatum and extended amygdala produced extensive overlap of retrogradely labeled neurons and significant double labeling, suggesting that cortical projections spread broadly within macrosystems. In contrast, heterotypic injection pairs produced significant overlap of retrograde labeling but negligible double labeling, indicating that ventral striatopallidum and extended amygdala receive inputs from separate sets of prefronto- and insular cortical neurons. The caudomedial shell of the nucleus accumbens, a supposed “transition” zone between striatopallidum and extended amygdala, had extended amygdala-like afferents but produced few double-labeled neurons and these only when paired with ventral striatopallidum. The data suggest that a modular organization of the basal forebrain, with postulated independent information processing by the ventral striatopallidal and extended amygdala macrosystems, is reflected in a corresponding segregation of output neurons in the prefrontal and insular cortices.

Differential Foci and Synaptic Organization of the Principal and Spinal Trigeminal Projections to the Thalamus in the Rat

The thalamus is known to receive single‐whisker ‘lemniscal’ inputs from the trigeminal nucleus principalis (Prv) and multiwhisker ‘paralemniscal’ inputs from the spinal trigeminal nucleus (Spv), yet the responses of cells in the thalamic ventroposteromedial nucleus (VPM) are most similar to and contingent upon inputs from PrV. This may reflect a differential termination pattern, density and/or synaptic organization of PrV and SpV projections. This hypothesis was tested in adult rats using anterograde double‐labelling with fluorescent dextrans, horseradish peroxidase (HRP) and choleragenoid, referenced against parvalbumin and calbindin immunoreactivity. The results indicated that Prvs most robust thalamic projection is to the whisker‐related barreloids of VPM. The SpV had robust projections to non‐barreloid thalamic regions, including the VPM ‘shell’ encapsulating the barreloid area, a caudal and ventral region of VPM that lacks barreloids and PrV inputs, the posterior thalamic nucleus, nucleus submedius and zona incerta. Within the barreloid portion of VPM, SpV projections were sparse relative to those from PrV, and most terminal labelling occurred in the peripheral fringes of whisker‐related patches and in inter‐barreloid septae. Thus, PrV and SpV have largely complementary projection foci in the thalamus. Intra‐axonal staining of a small sample of trigeminothalamic axons with whisker or guard hair receptive fields revealed highly localized and somatotopic terminal aggregates in VPM that spanned areas no larger than that of a single barreloid. In the electron microscopic component of this study, HRP transport to the barreloid region of VPM from left SpV and right PrV in the same cases revealed PrV terminals contacting dendrites with a broad range of minor axis diameters (mean ± SD: 1. 51 ± 0. 10 μm). SpV terminals were indistinguishable from those of PrV, but they had a disproportionate number of contacts on narrow dendrites (1. 27 ± 0. 07 μm, P 0. 01). PrV endings were also more likely to contact VPM somata (11. 0 ± 4. 2% of all labelled terminals) than those from SpV (3. 0 ± 1. O%, P 0. 01). Insofar as primary dendrites are thicker than distal dendrites in VPM, these data suggest a differential distribution of PrV and SpV inputs onto VPM cells that may account for their relative efficacies in dictating the responses of VPM cells to whisker stimulation. Multiwhisker receptive fields in VPM may also reflect direct transmission of convergent inputs from PrV.

Neurotoxicity of MAO metabolites of catecholamine neurotransmitters: role in neurodegenerative diseases.

The monoamine oxidase (MAO) metabolites of norepinephrine (NE) or epinephrine (EPI) and of dopamine (DA) are 3,4-dihydroxyphenylglycolaldehyde (DOPEGAL) and 3,4-dihydroxyphenylacetaldehyde (DOPAL), respectively. The toxicity of these catecholamine (CA) MAO metabolites was predicted over 50 years ago. However, until our recent chemical synthesis of these CA aldehyde metabolites, the hypothesis about their toxicity could not be tested. The present paper reviews recent knowledge gained about these compounds. Topics to be reviewed include: chemical synthesis and properties of DOPEGAL and DOPAL; in vitro and in vivo toxicity of CA aldehydes; subcellular mechanisms of toxicity; free radical formation by DOPEGAL versus DOPAL; mechanisms of accumulation of CA aldehydes in Alzheimers disease (AD) and Parkinsons disease (PD) and potential therapeutic targets in Alzheimers disease and Parkinsons disease.