Immunocytochemical evaluation of a cholinergic-specific ganglioside antigen (Chol-1) in the central nervous system of the rat

Summary Previous work from this laboratory has identified gangliosidic surface markers specific for cholinergic neurons. Antibodies to these markers, collectively designated Chol-1, induce complement-mediated lysis of the cholinergic subpopulation of synaptosomes and provide the basis for a new immunocytochemical method for staining cholinergic neurons in rat, guinea pig and human material. The specification and localization of immunocytochemical staining for Chol-1 was investigated in selected areas of the rat central nervous system. The antigen was typically expressed on all neurons previously identified as being cholinergic using monoclonal antibodies to choline acetyltransferase. At spinal levels Chol-1 was present on large and smaller cell bodies in the ventral horn motoneuron area. The preganglionic sympathetic neurons in the thoracic intermediolateral nucleus were also Chol-1-positive. Nerve terminal-like staining was observed in association with stained large Chol-1 positive and smaller unstained Chol-1 negative neurons, and in lamina I and III of the dorsal horn. In the mesencephalon, motoneurons of the oculomotor and trochlear nucleus, as well as neurons within the pedunculopontine tegmental nucleus and the red nucleus were Chol-1-positive. In addition visceromotoneurons of the Edinger-Westphal nucleus were stained with anti Chol-1 antibodies. In the basal forebrain the antibodies gave a positive reaction on well known cholinergic neurons in the vertical and horizontal limbs of the diagonal bands of Broca and the medial forebrain bundle. In agreement with studies using antibodies to choline acetyltransferase, a small sub-population of neostriatal neurons (1–2%) was Chol-1-positive. In the rat retina, both anti-Chol-1 and anti-choline acetyltransferase antibodies gave rise to a nerve terminal-like staining in the same bands within the inner plexiform layer. The anti-Chol-1 antibodies also stain normal and pathological human material and could have a useful application in human neuropathology.     

Age-related changes in glycogen synthase kinase 3beta (GSK3beta) immunoreactivity in the central nervous system of rats

Although glycogen synthase kinase 3beta (GSK3beta) is emerging as a prominent drug target in the treatment of neurodegenerative diseases such as Alzheimer's disease (AD) and stroke, very little is known about age-related changes in GSK3beta expression and GSK3beta phosphorylation. Therefore, we examined age-related changes in immunoreactivities for GSK3beta and phosphorylated GSK3beta (pGSK3beta) in the central nervous system. In aged rats, there were significant increases in GSK3beta immunoreactivity in the cell bodies and processes of pyramidal cells in most cortical regions. GSK3beta immunoreactivity was also significantly increased in the pyramidal layer of CA1-3 regions, and the granule cell layer of dentate gyrus. Age-related increases were prominent in lateral septal nuclei, compared to the medial septal nuclei. Interestingly, both GSK3beta and pGSK3beta was increased in the prefrontal cortex, while GSK3beta and pGSK3beta was differentially localized in the cerebellar cortex. The first demonstration of age-related alterations in immunoreactivities for GSK3beta and pGSK3beta in the basal forebrain area and cholinergic projection targets may provide useful data for investigating the pathogenesis of age-related neurodegenerative diseases including AD.     

Immunohistochemical studies on the cellular localization of GABAA-receptors in explant cultures of rat central nervous system using a monoclonal antibody

Summary Explant cultures of rat spinal cord, brain stem and cerebellum were used to visualize GABA_A-receptors by means of immunohistochemistry. For these studies we have incubated the cultures with the monoclonal antibody bd 17 against the -subunit of the GABA_A/benzodiazepine/chloride channel complex. In spinal cord cultures, many interneurones were immunoreactive whereas only a small number of large neurones, probably motoneurones was specifically stained. In brain stem cultures, groups of large and medium-sized neurones showed immunoreactivity. In cultures of cerebellum, a great number of neurones was specifically stained. Granule cells showed the strongest immunoreactivity whereas other neurones, presumably Purkinje cells and interneurones, were only moderately stained. The immunoreactivity was mainly confined to the cell bodies of the neurones while their processes were only weakly or not stained. In contrast to neurones, no immunoreactivity could be detected on astrocytes.     

Immunohistochemical study on the distribution of insulin-like growth factor-binding protein 4 in the central nervous system of SOD1 transgenic mice as an in vivo model of amyotrophic lateral sclerosis

In the present study, we used the SOD1^G93A mutant transgenic mice as an in vivo model of amyotrophic lateral sclerosis (ALS) and performed immunohistochemical studies to investigate the changes of insulin-like growth factor-binding protein 4 (IGFBP4) in the central nervous system. Decreased expression of IGFBP4 was obvious in the cerebral cortex, hippocampus, cerebellar cortex and inferior olive of SOD1^G93A transgenic mice. In the cerebral cortex, there was a significant decrease in IGFBP4 immunoreactivity in the pyramidal cells. In the hippocampal formation, IGFBP4 immunoreactivity was also decreased in the pyramidal cells of CA1–3 areas and the granule cells of dentate gyrus. In the cerebellar cortex, IGFBP4 immunoreactivity was prominent in the granular layer in wtSOD1 transgenic mice, compared to that in SOD1^G93A transgenic mice. IGFBP4 immunoreactivity was decreased in the inferior olive of SOD1^G93A transgenic mice. This study, showing decreased IGFBP4 in different brain regions of SOD1^G93A transgenic mice, may provide clues to understanding the differential susceptibility of neural structures in ALS, suggesting a role of IGFBP4 in an abnormality of cognitive and/or motor function in ALS. The mechanisms and functional implications of these decreases require elucidation.     

“The developmental and functional logic of neuronal circuits”: commentary on the Kavli Prize in Neuroscience

The first Kavli Prize in Neuroscience recognizes a confluence of career achievements that together provide a fundamental understanding of how brain and spinal cord circuits are assembled during development and function in the adult. The members of the Kavli Neuroscience Prize Committee have decided to reward three scientists (Sten Grillner, Thomas Jessell, and Pasko Rakic) jointly “for discoveries on the developmental and functional logic of neuronal circuits”.