ABSTRACT
We have investigated the distribution of PEP-19, a neuron-specific protein, in the adult human brain. Immunohistochemistry for PEP-19 appears to define the basal ganglia and related structures. The strongest immunoreactivity is seen in the caudate nucleus and putamen, each of which showed both cell body and neuropil PEP-19 immunoreactivity. The substantia nigra and both segments of the globus pallidus showed PEP-19 immunoreactivity only in the neuropil. Cell bodies and dendrites of the thalamic nuclei ventralis lateralis and ventralis anterioralis were less strongly immunoreactive. Cerebellar Purkinje cells and their dendrites were immunoreactive, as were the presubiculum/subiculum regions and dentate gyrus granule cells of the hippocampus. The CA zones of the hippocampus were not immunoreactive. Preliminary data from immunoblotting experiments indicate that PEP-19 immunoreactivity is significantly reduced in cerebellum in Alzheimer's disease. While there were no apparent alterations of immunoreactivity in Down's syndrome or in Parkinson's disease, immunohistochemical analysis showed a massive loss of PEP-19 immunoreactivity in the caudate nucleus, putamen, globus pallidus and substantia nigra in Huntington's disease. These results show that PEP-19, a neuron-specific, calmodulin-binding protein, is distributed in specific areas of the adult human brain. The reduction in PEP-19 immunoreactivity in Alzheimer's disease and Huntington's disease suggests that PEP-19 may play a role in the pathophysiology of these diseases through a mechanism of calcium/calmodulin disregulation. This may be especially apparent in Huntington's disease where the distribution of the product of the abnormal gene, huntingtin, alone is not sufficient to explain the pattern of pathology. Abnormal huntingtin associates more strongly with calmodulin than does normal huntingtin [Bao et al. (1996) Proc. natn. Acad. Sci. U.S.A., 93, 5037-5042] suggesting a disruption of calmodulin-mediated intracellular mechanism(s), very likely involving PEP-19.
Subject(s)
Basal Ganglia/metabolism , Brain Chemistry/physiology , Huntington Disease/metabolism , Nerve Tissue Proteins/metabolism , Adult , Aged , Aged, 80 and over , Animals , Calmodulin-Binding Proteins , Chromosome Mapping , Chromosomes, Human, Pair 21/genetics , Chromosomes, Human, Pair 21/physiology , Down Syndrome/metabolism , Female , Humans , Immunoblotting , Immunohistochemistry , Male , Middle Aged , Nerve Tissue Proteins/deficiency , Nerve Tissue Proteins/genetics , RatsABSTRACT
IL-1 beta is one of the cytokines known to affect astroglial cells in normal brain development, brain injury and neurodegenerative diseases. IL-1 beta causes astrocytes to become more reactive, alter the expression and release of molecules and in some cases to proliferate. We have investigated the mitogenic effect and signal transduction pathway induced by IL-1 beta in U373 cells, a human astrocytoma cell-line. Recombinant human IL-1 beta induced mitogenesis of U373 cells in a dose-dependent fashion as assessed by tritiated thymidine incorporation. The following signal transduction mechanisms, reported to be induced in other systems by IL-1 beta, were investigated in U373 cells: (1) activation of phosphatidylcholine-specific phospholipase C as assayed by incorporation of tritiated choline into cellular phospholipids, (2) production of diacylglycerol, a lipid second messenger, (3) activation of sphingomyelinase, and (4) activation of mitogen-activated protein kinase (MAPK). Of these, IL-1 beta activated only MAPK. In cultured rat astrocytes, IL-1 beta caused activation of MAPK without inducing proliferation.
