Eileen Lynd-Balta

The organization of midbrain projections to the striatum in the primate: sensorimotor-related striatum versus ventral striatum.

In order to examine the organization of nigrostriatal projections in the primate, the retrograde tracers Lucifer Yellow conjugated to dextran amines and horseradish peroxidase conjugated to wheatgerm agglutinin were injected into different regions of the dorsolateral and ventral striatum. Based on the topography of cortical inputs to the striatum, the dorsolateral striatum is associated with the motor system, and the ventral striatum is related to the limbic system. Our results indicate that although midbrain neurons projecting to the ventral and dorsolateral striatum are mostly separate, there are neurons projecting to these different striatal territories that overlap in the medial substantia nigra. The dopaminergic neurons of the ventral mesencephalon can be subdivided into dorsal and ventral tiers that include the cells of the ventral tegmental area, the substantia nigra pars compacta, and the retrorubral area. Neurons projecting to the ventral striatum are found in both the dorsal and ventral tiers. A large number of neurons occupying the medial densocellular zone of the ventral tier are labeled following injections into different regions of the ventral striatum. Neurons projecting to the sensorimotor-related striatum are derived almost exclusively from the ventral tier. Many of these neurons are located very ventrally in the substantia nigra, where clusters of neurons invade the pars reticulata. In addition, labeled neurons are found throughout the mediolateral extent of the densocellular zone of the pars compacta. Notably, neurons are labeled in the medial densocellular zone following injections into the dorsolateral and ventral striatum. Mesencephalic neurons projecting to different striatal territories are distinct in that dorsal tier neurons mainly innervate the ventral striatum, whereas the ventral columns of neurons in the ventral tier innervate the sensorimotor-related striatum. Thus, the dopaminergic regulation of the sensorimotor-related striatum and the ventral striatum may be different. However, a subgroup of dopaminergic neurons in the medial densocellular zone projects to both striatal territories. Such divergent projections may allow the substantia nigra to serve as a link, connecting different striatal territories, via their connections with the substantia nigra.

The organization of midbrain projections to the ventral striatum in the primate

Because the dopaminergic neurons of the midbrain form a continuum, boundaries between the ventral tegmental area, substantia nigra pars compacta, and retrorubral area are difficult to distinguish in the primate. Therefore, dopaminergic neurons have been subdivided into more readily discernible dorsal and ventral tiers. The projections from these dorsal and ventral tier neurons of the ventral mesencephalon to the ventral striatum were labeled by injections of horseradish peroxidase conjugated to wheatgerm agglutinin and Lucifer Yellow conjugated to dextran amines into different regions of the nucleus accumbens, the ventral caudate nucleus, and the rostral, ventral putamen in the primate. Neurons projecting to the ventral striatum are not topographically organized in the ventral mesencephalon. Retrogradely labeled neurons are found in the medial densocellular zone of the ventral tier following injections into all regions of the ventral striatum except the ventromedial shell region of the nucleus accumbens. These medial nigral neurons have diverging projections throughout the mediolateral extent of the ventral striatum. In addition, neurons of the dorsal tier project to all ventral striatal regions examined. Notably, neurons projecting to the shell region of the nucleus accumbens are limited to the dorsal tier, throughout the rostrocaudal extent of the substantia nigra. Both dorsal and ventral tier neurons innervate the ventral striatum. Not only do neurons of the ventral tegmental area project to the ventral striatum, but also many of the pars compacta. The projections to the shell region of the nucleus accumbens are more restricted, suggesting that the dopaminergic regulation of this accumbens subterritory is distinct from the rest of the ventral striatum.

P2X7 receptor immunoreactive profile confined to resting and activated microglia in the epileptic brain

The purpose of this study was to identify the CNS cellular constituent immunoreactive for specific P2X7 receptor antiserum in the kainate-induced seizure and non-seizure rat brain. Analysis of P2X7 immunocytochemistry (ICC) revealed small immunoreactive cells with processes showing distinct morphological changes as seizures progressed in time. These morphological changes were reminiscent of reactive glia during CNS injury. In order to determine the identity of this non-neuronal cellular constituent, we employed dual ICC techniques using sequential antibody incubations and reacted the sections with contrasting chromagens. Specific glial markers tested in the series included Iba1 (microglia), COX-1 (microglia), and GFAP (astroglia). Results of this study revealed distinct colocalization when sections immunostained for P2X7 were dual immunostained with antisera specific for microglia (Iba1, COX-1). In contrast, no colocalization was evident when sections were dual immunostained with P2X7 and GFAP, an astrocytic marker. In the latter experiment, dual ICC revealed two distinct cell populations with contrasting color demonstrating a population of distinct GFAP immunopositive cells and a population of distinct P2X7 immunopositive cells. We conclude that P2X7 antiserum used in this study is specific for and identifies microglia in rat and that there exists a timeline of progressive changes in microglia morphology that can be demonstrated following kainate-induced seizures. In addition, the morphological changes in microglia following seizure induction that can be identified with P2X7 antisera or with antisera specific for microglia suggest a neuroinflammatory milieu in areas of CNS seizure activity.

Integrative Aspects of Basal Ganglia Circuitry

The basal ganglia is a set of interconnected subcortical structures that influence cortical activity through corticobasal ganglia loops. Information is thought to flow from the cortex via the striatum to the pallidum and substantia nigra, from the pallidum and the substantia nigra to the thalamus, and finally back to the cortex. Based on some of the pathways and physiology of the basal ganglia sensorimotor circuit in the primate, several theories concerning the organization of the primate basal ganglia have been suggested. A current one proposes that the frontal cortex and the basal ganglia are arranged in parallel, functionally segregated circuits (Alexander et al., 1990). In this concept, each individual circuit has a specific, functionally distinct region of the frontal cortex as a nodal point, and projects to anatomically distinct sectors of the striatum, the pallidum, the substantia nigra, and the thalamus. From there, the thalamocortical pathway completes the loop to cortex. In addition to the sensorimotor circuit, others have been proposed that involve distinct regions of the basal ganglia associated with particular cortical regions including the dorsolateral prefrontal cortex, the orbitofrontal cortex, and the cingulate cortex.

Distribution of AMPA receptor subunits in the hippocampal formation of temporal lobe epilepsy patients.

The immunocytochemical distribution of the AMPA-selective receptor subunits GluR1 and GluR2/3 were mapped in the human hippocampal formation obtained from surgery for medically intractable temporal lobe epilepsy. GluR2/3 immunoreactivity was detected in all principal cell types of the hippocampal formation, including hilar neurons, granule cells of the dentate gyrus, and pyramidal cells of the cornu ammonis fields and subiculum. GluR2/3 immunostaining typically filled the cell bodies and processes of neurons. A comparison of GluR2/3 immunoreactivity in a sclerotic specimen versus a non-sclerotic specimen demonstrated a profound loss of staining, specifically in the areas where neuronal dropout was occurring, including CA1, CA3 and the hilus. An analysis of GluR1 immunoreactivity in non-sclerotic specimens revealed that it was predominantly localized to cellular processes throughout the cornu ammonis fields, with a sparse staining of the dentate gyrus outer molecular layer and little to no staining of the dentate gyrus inner molecular layer. Similar to the GluR2/3-immunostained patterns, GluR1 immunoreactivity was lost in the cornu ammonis fields of sclerotic hippocampal specimens, corresponding to patterns of neuronal dropout. Our most compelling finding was a unique extensive pattern of GluR1 and Glu2/3 immunoreactivity throughout the molecular layers of the dentate gyrus of severely compromised hippocampi. The altered staining of GluR1 and GluR2/3 complements some of the patterns of axonal sprouting already described for the dentate gyrus, with a conjecture that their anatomy and distribution pattern underlies to some degree the reorganization of the sclerotic hippocampus. A combination of enhanced glutamatergic transmission and changes in neuropeptides that modulate hippocampal circuitry could greatly affect the degree of excitability in the hippocampal formation. The alterations of GluR1 and GluR2/3 immunoreactivity in the dentate gyrus add another component to the concept of reorganization in the epileptic sclerotic hippocampus.

Developing Information Literacy Skills Early in an Undergraduate Curriculum

Several core competencies related to information literacy have been identified by the Association of College and Research Libraries. Students must learn to gather relevant information and communicate their findings effectively. The collaborative activity described here, which could easily be adapted for other disciplines, introduces first-semester freshmen to the standards of professional scientific writing, the different forms of publication, search strategies to effectively find information using a relevant database, and plagiarism. Analysis of our pre- and post-activity assessment demonstrates that students gain both confidence and knowledge on several important skills as a result of this activity. Providing content-relevant information literacy experience lays the foundation for students to be successful consumers of information.

Enhanced cyclooxygenase-2 expression in olfactory-limbic forebrain following kainate-induced seizures

Cyclooxygenase-2 is expressed at low levels in a subset of neurons in CNS and is rapidly induced by a multiplicity of factors including seizure activity. A putative relationship exists between cyclooxygenase-2 induction and glutamatergic neurotransmission. Cyclooxygenase-1 is constitutively expressed in glial cells and has been specifically linked to microglia. In this study we evaluated cyclooxygenase-2 protein immunocytochemically and found markedly enhanced immunostaining primarily in olfactory-limbic regions at 2, 6 and 24 h following kainate-induced status epilepticus. Impressive enhanced cyclooxygenase-2 immunoreactivity was localized in anterior olfactory nucleus, tenia tecta, nucleus of the lateral olfactory tract, piriform cortex, lateral and basolateral amygdala, orbital frontal cortex, nucleus accumbens (shell) and associated areas of ventral striatum, entorhinal cortex, dentate gyrus granule cells and hilar neurons, hippocampal CA subfields and subiculum. Alternate sections were processed for dual immunocytochemical analysis utilizing c-Fos and cyclooxygenase-2 antiserum to examine the possibility that the neuronal induction of cyclooxygenase-2 was associated with seizure activity. Neurons that showed a timeline of cyclooxygenase-2 upregulation were found to possess c-Fos immunopositive nuclei. Additional results from all seizure groups showed cyclooxygenase-1 induction in microglia, which was confirmed by Western blot analysis of hippocampus. Western blot and real-time quantitative RT-PCR analysis showed significant upregulation of cyclooxygenase-2 expression, confirming its induction in neurons. These data indicate that cyclooxygenase-2 induction in a neuronal network can be a useful marker for pathways associated with seizure activity.

NF-κB transcription factor subunits in rat brain: Colocalization of p65 and α-MSH

Abstract The subunit proteins p50 and p65 of the transcription factor NF-κB and the I-κB inhibitory protein were immunocytochemically identified and mapped in rat brain. The p65 subunit was localized to the cytoplasm of neurons in the lateral hypothalamus and colocalized with α-MSH in neurons identified as the α-2 component of the α-MSH system. The p50 subunit protein was distributed throughout the neocortex, basal ganglia, thalamic, and hypothalamic nuclei, and certain nuclei of the pons and medulla. The I-κB protein, which is necessary for the cytoplasmic sequenstration of the NF-κB transcription factor complex, was identified specifically in regions of limbic, hypothalamic, and autonomic nuclei.

AMPA receptor alterations precede mossy fiber sprouting in young children with temporal lobe epilepsy

Following neurological injury early in life numerous events, including excitotoxicity, neural degeneration, gliosis, neosynaptogenesis, and circuitry reorganization, may alone or in concert contribute to hyperexcitability and recurrent seizures in temporal lobe epilepsy. Our studies provide new evidence regarding the temporal sequence of key elements of hippocampal reorganization, mossy fiber sprouting and glutamate receptor subunit up-regulation, in a subset of young temporal lobe epileptic patients. Without evidence of mossy fiber sprouting, the youngest age group (3-10 years old) of mesial temporal lobe epileptic patients demonstrated enhanced glutamate receptor subunit profiles, suggesting that the dendritic change precedes axonal sprouting. However, sclerotic hippocampal specimens from epileptic patients ages 12-15 years old had the characteristic features of glutamate receptor up-regulation and mossy fiber sprouting first identified in the adult, indicating that reconstructed circuits appear early in the course of the disease. Non-sclerotic hippocampal specimens from lesion associated temporal lobe epileptic patients of all age groups showed minimal cell loss, sparse staining of glutamate receptor subunits in the dentate gyrus, and little or no mossy fiber sprouting. These compelling findings suggest a progressive sequence of events in the reorganization of the dentate gyrus of sclerotic hippocampal specimens. We suggest that cell loss and up-regulation of glutamate receptor subunits appear early in temporal lobe epilepsy and contribute to the synaptic plasticity that may facilitate the subsequent sprouting of mossy fiber collaterals which compound an already precipitous state of decline. The combination of pre-synaptic and post-synaptic changes serves as a potential substrate for hyperexcitability.

Adrenocorticotropic Hormone Immunoreactivity in the Hippocampal Formation of Temporal Lobe Epilepsy Patients

Summary: Purpose: We wished to identify immunocytochemically the distribution of proopiomelanocortin‐related peptides in the hippocampal formation of patients with epilepsy.