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Immunogold study of effects of prenatal exposure to lipopolysaccharide and/or valproic acid on the rat blood-brain barrier vessels

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Journal of Neurocytology

Abstract

The involvement of blood microvessels, representing the anatomic site of the blood-brain barrier (BBB), in brain damage induced by prenatal exposure to lipopolysaccharide (LPS) and/or valproic acid (VPA) was studied in four-week-old rats. The immunogold procedure was applied for localization at the ultrastructural level of endogenous albumin and glucose transporter (GLUT-1) in three brain regions: cerebral cortex, cerebellum and hippocampus. Four groups of rats were used: (1) untreated control, (2) prenatally VPA-treated, (3) prenatally LPS-treated, and (4) prenatally LPS- and VPA-treated. The functional state of the BBB was evaluated as follows: (a) by its tightness, i.e., permeability to blood-borne albumin, and (b) by the expression of GLUT-1 in the endothelial cells (ECs). Using morphometry, the labelling density for GLUT-1 was recorded over luminal and abluminal plasma membranes of the ECs, also providing information on their functional polarity. No extensive increase of vascular permeability and/or any considerable dysfunction of the BBB in experimental groups nos. 2 and 3 were observed, although in solitary vascular profiles, increased endocytosis or even transcytosis of albumin by ECs was noted. In experimental group no. 4, some vascular profiles showed scanty leakage (microleakage), manifested by the presence of immunosignals for albumin in the perivascular area. Although some fluctuations in the expression of GLUT-1 occurred in all experimental groups, especially in group no. 3, a most pronounced and significant diminution of the labelling density, in all three regions of the brain, was observed in group no. 4. This finding suggests the synergistic action of prenatally applied LPS and VPA that affects specific transport functions of glucose in the microvascular endothelium. The diminished or disturbed supply of glucose to selected brain regions can be one of the factors leading to previously observed behavioral disturbances in similarly treated rats.

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References

  • ALLEN, G. & COURCHESNE, E. (2003) Differential effects of developmental cerebellar abnormality on cognitive and motor functions in the cerebellum: an fMRI study of autism. American Journal of Psychiatry 160, 262–273.

    Article  PubMed  Google Scholar 

  • BANKS, W. A., KASTIN, A. J., BRENNAN, J. M. & VALLANCE, K. L. (2003) Absorptive endocytosis of HIV-1 gp120 by blood-brain barrier is enhanced by lipopolysaccharide. Experimental Neurology 156, 165–171.

    Article  Google Scholar 

  • BOLZ, S., FARRELL, C. L., DIETZ, K & WOLBURG, H. (1996) Subcellular distribution of glucose transporter (GLUT-1) during development of the blood-brain barrier in rats. Cell and Tissue Research 284, 355–365.

    Article  PubMed  CAS  Google Scholar 

  • CORNFORD, E. M., HYMAN, S. & PARDRIDGE, W. M. (1993) An electron microscopic immunogold analysis of developmental up-regulation of the blood-brain barrier Glut1 glucose transporter. Journal of Cerebral Blood Flow and Metabolism 13, 841–854.

    PubMed  CAS  Google Scholar 

  • CHRISTIANSON, A. L., CHESLER, N. & KROMBERG, J. G. (1994) Fetal valproate syndrome: clinical and neurodevelopmental features in two sibling pairs. Developmental Medicine and Child Neurology 36, 361–369.

    Article  PubMed  CAS  Google Scholar 

  • DERMIETZEL, R., KRAUSE, D., KREMER, M., WANG, C. & STEVENSON, B. (1992) Pattern of glucose transporter (Glut1) expression in embryonic brains is related to maturation of blood-brain barrier tightness. Developmental Biology 193, 152–163.

    CAS  Google Scholar 

  • DE VIVO, D. C., TRIFILETTI, R., JACOBSON, R. I. & HARIK, S. I. (1990) Glucose transporter deficiency causing persistent hypoglycorrhachia: A unique cause of infantile seizures and acquired microcephaly. Annals of Neurology 28, 414–415.

    Google Scholar 

  • DOBROGOWSKA, D. H. & VORBRODT, A. W. (1999) Quantitative immunocytochemical study of blood-brain barrier glucose transporter (Glut-1) in four regions of mouse brain. Journal of Histochemistry and Cytochemistry 47, 1021–1029.

    PubMed  CAS  Google Scholar 

  • DREWES, L. R. (1998) Biology of the blood-brain glucose transporter. In Introduction to the Blood-Brain Barrier (edited by Pardridge, W. M.) pp. 165–174. Cambridge: Cambridge University Press.

    Google Scholar 

  • EL-KHODOR, B., KOZLOWSKI, P. B., RABE, A. & LEE, M. H. (2003) Prenatal valproic acid and lipopolysaccharide synergistically produce behavioral and neuroanatomical changes. Developmental Psychobiology 43, 253.

    Google Scholar 

  • ENGELHARDT, B. & RISAU, W. (1995) Development of the blood-brain barrier. In new concepts of a blood-brain barrier (edited by Greenwood, J., Begley, D.J. & Segal, M.B.) pp.11–31. New York and London: Plenum Press.

    Google Scholar 

  • FARRELL, C. L. & PARDRIDGE, W. M. (1991) Blood-brain barrier glucose transporter is asymmetrically distributed on brain capillary endothelial luminal and abluminal membranes: An electron microscopic immunogold study. Proceedings of the National Academy of Sciences USA 88, 5779–5783.

    Article  CAS  Google Scholar 

  • FATEMI, S. H., EARLE, J., KANODIA, R., KIST, D., EMAMIAN, E. S., PATTERSON, P.H., SHI, L.M. & SIDWELL, R. (2002) Prenatal viral infection leads to pyramidal cell atrophy and macrocephaly in adulthood: Implications for genesis of autism and schizophrenia. Cellular and Molecular Neurobiology 22, 25–33.

    Article  PubMed  Google Scholar 

  • GIBBS, J. P., ADEYEYE, M. C., YANG, Z. & SHEN, D. D. (2004) Valproic acid uptake by bovine brain microvessel endothelial cells: role of active transport. Epilepsy Research 58, 53–66.

    Article  PubMed  CAS  Google Scholar 

  • GORDON, N. & NEWTON, R. W. (2003) Glucose transporter type 1 (GLUT-1) deficiency. Brain Development 25, 477–480.

    Article  PubMed  Google Scholar 

  • HORNIG, M., WEISSENBOCK, H., HORSCROFT, N. & LIPKIN WI (1999) An infection-based model of neurodevelopmental damage. Proceedings of the National Academy of Sciences USA 96, 12102–12107.

    Article  CAS  Google Scholar 

  • HORNIG, M. & LIPKIN, W. I. (2001) Infectious and immune factors in the pathogenesis of neurodevelopmental disorders: Epidemiology, hypotheses, and animal models. Mental Retardation and Developmental Disability Research Review 7, 200–210.

    Article  CAS  Google Scholar 

  • INGRAM, J. L., PECKHAM, S. M., TISDALE, B. & RODIER, P. M. (2000) Prenatal exposure of rats to valproic acid reproduces the cerebellar anomalies associated with autism. Neurotoxicology and Teratology 22, 319–324.

    Article  PubMed  CAS  Google Scholar 

  • KAUFMANN, W. E., COOPER, K. L., MOSTOFSKY, S. H., CAPONE, G. T., KATES, W. R., NEWSCHAFFER, C. J., BUKELIS, I., STUMP, M. H., JANN, A. E. & LANHAM, D. C. (2003) Specificity of cerebellar vermian abnormalities in autism: A quantitative magnetic resonance imaging study. Journal of Children Neurology 18, 463–470.

    Google Scholar 

  • LIPPOLDT, A., KNIESEL, U., LIEBNER, S., KALBACHER, H., KIRSCH, T., WOLBURG, H. & HALLER, H. (2000) Structural alterations of tight junctions are associated with loss of polarity in stroke-prone spontaneously hypertensive rat blood-brain barrier endothelial cells. Brain Research 885, 251–261.

    Article  PubMed  CAS  Google Scholar 

  • MCILVANE, W. J. & CATALDO, M. F. (1996) On the clinical relevance of animal models for the study of human mental retardation. Mental Retardation and Developmental Disability Research Review 2, 188–196.

    Article  Google Scholar 

  • MURATORI, F., CESARI, A. & CASELLA, C. (2001) Autism and cerebellum. An unusual finding with MRI. Panminerva Medicine 43, 311–315.

    CAS  Google Scholar 

  • OMTZIGT, J. G., NAU, H., LOS, F. J., PIJPERS, L. & LINDHOUT, D. (1992) The disposition of valproate and its metabolites in the late first trimester and early second trimester of pregnancy in maternal serum, urine, and amniotic fluid: Effect of dose, co-medication, and the presence of spina bifida. European Journal of Clinical Pharmacology 43, 381–388.

    Article  PubMed  CAS  Google Scholar 

  • PASCUAL, J.M., VAN HEERTUM, R.L., WANG, D., ENGELSTAD, K. & DE VIVO, D.C. (2002) Imaging the metabolic footprint of Glut 1 deficiency on the brain. Annals of Neurology 52, 458–464.

    Article  PubMed  CAS  Google Scholar 

  • PLETNIKOV, M. V., MORAN, T. H. & CARBONE, K. M. (2002) Borna disease virus infection of the neonatal rat: Developmental brain injury model of autism spectrum disorders. Frontiers of Biosciences 7, D593–D607.

    CAS  Google Scholar 

  • SCHNEIDER, M. L., MOORE, C. F. & KRAEMER, G. W. (2001a) Moderate alcohol during pregnancy: Learning and behavior in adolescent rhesus monkeys. Alcohol Clinical and Experimental Research 25, 1383–1392.

    Article  CAS  Google Scholar 

  • SCHNEIDER, T., LABUZ, D. & PRZEWLOCKI, R. (2001b) Nociceptive changes in rats after prenatal exposure to valproic acid. Polish Journal of Pharmacology 53, 531–534.

    CAS  Google Scholar 

  • SOBANIEC-LOTOWSKA, M. E. & SOBANIEC, W. (1996) Morphological features of encephalopathy after chronic administration of the antiepileptic drug valproate to rats. A transmission electron microscopic study of capillaries in the cerebellar cortex. Experimental Toxicology and Pathology 48, 65–75.

    CAS  Google Scholar 

  • TARNAWSKI, J., WEN, G.Y., HEANEY, A., MA, S., EL-KHODOR, B.F., RABE, A., KOZLOWSKI, P.B. & LEE, M.H. (2004) Combined exposure to lipopolysaccharide and valproic acid during early embryonic development in rats. Program No. 359.12. Society for Neuroscience Abstracts Viever/Itinerary Planner, Washington DC.

    Google Scholar 

  • TEMESVÁRI, P., ÁBRAHÁM, C. S., SPEER, C. P., KOVÁCS, J. & MEGYERI, P. (1993) Escherichia coli 0111 B4 lipopolysaccharide given intracisternally induces blood-brain barrier opening during experimental neonatal meningitis in piglets. Pediatric Research 34, 182–186.

    PubMed  Google Scholar 

  • VESZELKA, S., URBÁNYI, Z., PÁZMÁNY, T., NÉMETH, L., OBÁL, I., DUNG, N.T.K., ÁBRAHÁM, C.S., SZABó, G. & DELI, M.A. (2003) Human serum amyloid component attenuates the bacterial lipopolysaccharide-induced increase in blood-brain barrier permeability in mice. Neuroscience Letters 352, 57–60.

    Article  PubMed  CAS  Google Scholar 

  • VORBRODT, A. W. (1993). Morphological evidence of the functional polarization of brain microvascular endothelium. In The Blood-Brain Barrier (edited by Pardridge, W.M.) pp. 137–164. New York: Raven Press.

    Google Scholar 

  • VORBRODT, A. W. (1995) The application of quantitative immunocytochemistry for the evaluation of blood-brain barrier (BBB) to endogenous albumin. In New Concepts of a Blood-Brain Barrier (edited by Greenwood, J., Begley, D.J. & Segal, M. B.) pp. 39–46. New York and London: Plenum Press.

    Google Scholar 

  • VORBRODT, A. W. & DOBROGOWSKA, D. H. (1994) Immunocytochemical evaluation of blood-brain barrier to endogenous albumin in adult, newborn and aged mice. Folia Hictochemica et Cytobiologica 32, 63–70.

    CAS  Google Scholar 

  • VORBRODT, A. W., DOBROGOWSKA, D. H., TARNAWSKI, M. & LOSSINSKY, A. S. (1994) A quantitative immunocytochemical study of the osmotic opening of the blood-brain barrier to endogenous albumin. Journal of Neurocytology 23, 792–800.

    Article  PubMed  CAS  Google Scholar 

  • VORBRODT, A. W., DOBROGOWSKA, D. H., UENO, M. & LOSSINSKY, A.S. (1995) Immunocytochemical studies of protamine-induced blood-brain barrier opening to endogenous albumin. Acta Neuropathologica 89, 491–499.

    PubMed  CAS  Google Scholar 

  • VORBRODT, A. W., DOBROGOWSKA, D. H., TARNAWSKI, M., MEEKER, H. C. & CARP, R. I. (1997) Immunocytochemical evaluation of blood-brain barrier to endogenous albumin in scrapie-infected mice. Acta Neuropathologica 93, 341–348.

    Article  PubMed  CAS  Google Scholar 

  • VORBRODT, A. W., DOBROGOWSKA, D. H., MEEKER, H. C. & CARP, R. I. (1999) Immunogold study of regional differences in the distribution of glucose transporter (GLUT-1) in mouse brain associated with physiological and accelerated aging and scrapie infection. Journal of Neurocytology 28, 711–719.

    Article  PubMed  CAS  Google Scholar 

  • VORBRODT, A. W., DOBROGOWSKA, D. H., KOZLOWSKI, P., TARNAWSKI, M., DUMAS, R. & RABE, A. (2001a) Effect of a single embryonic exposure to alcohol on glucose transporter (Glut-1) distribution in brain vessels of aged mouse. Journal of Neurocytology 30, 167–174.

    Article  CAS  Google Scholar 

  • VORBRODT, A. W., DOBROGOWSKA, D. H., TARNAWSKI, M., MEEKER, H. C. & CARP, R. I. (2001b) Quantitative immunogold study of glucose transporter (GLUT-1) in five brain regions of scrapie-infected mice showing obesity and reduced glucose tolerance. Acta Neuropathologica 102, 278–284.

    CAS  Google Scholar 

  • VORHEES, C. V., WEISENBURGER, W. P. & MINCK, D. R. (2001) Neurobehavioral teratogenic effects of thalidomide in rats. Neurotoxicology and Teratology 23, 255–264.

    Article  PubMed  CAS  Google Scholar 

  • WILLIAMS, G., KING, J., CUNNINGHAM, M., STEPHAN, M., KERR, B. & HERSH, J.H. (2001) Fetal valproate syndrome and autism: Additional evidence of an association. Developmental Medicine and Children Neurology 43, 202–206.

    Article  CAS  Google Scholar 

  • XAIO, H., BANKS, W.A., NIEHOFF, M.L. & MORLEY, J.E. (2001) Effect of LPS on the permeability of the blood-brain barrier to insulin. Brain Research 896, 36–42.

    Article  PubMed  CAS  Google Scholar 

  • XIA, M., WANG, D., PASCUAL, J. M., YANG, H., STINER, S. & DE VIVO, D. C. (2004) Effects of valproic acid on Glut-1 gene expression. Program No. 197.9 Society of Neuroscience Abstract Viewer/Itinerary Planner. Washington, DC.

    Google Scholar 

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  1. M. Tarnawski

    Present address: Lindsley F. Kimball Research Institute of the New York Blood Center, 310 East 67th Street, New York, N.Y., 10021

Authors and Affiliations

  1. New York State Office of Mental Retardation and Developmental Disabilities, Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, New York, 10314, USA

    A. W. Vorbrodt, D. H. Dobrogowska, P. B. Kozlowski, A. Rabe, M. Tarnawski & M. H. Lee

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  1. A. W. Vorbrodt
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  2. D. H. Dobrogowska
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Correspondence to A. W. Vorbrodt.

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Vorbrodt, A.W., Dobrogowska, D.H., Kozlowski, P.B. et al. Immunogold study of effects of prenatal exposure to lipopolysaccharide and/or valproic acid on the rat blood-brain barrier vessels. J Neurocytol 34, 435–446 (2005). https://doi.org/10.1007/s11068-006-8729-x

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  • DOI: https://doi.org/10.1007/s11068-006-8729-x

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