Areas of Interest
The major focus in this laboratory is to elucidate the
mechanisms by which leukocytes and pathogens invade the central
nervous system (CNS). Movement of both soluble and cellular
elements into the CNS is regulated by microvessel endothelial
cells comprising the blood-brain barrier (BBB). It is thus
believed that alterations in the BBB contribute to the
pathogenesis of various neuroinflammatory, neuroinfectious and
neurodegenerative diseases such as multiple sclerosis, AIDS
dementia complex and Alzheimer disease. To evaluate the role
played by the BBB in these disorders, we are employing an in
vitro culture model of the human BBB recently developed in this
laboratory.
Studies in progress in this laboratory include the following:
1.) Analysis of cell-targeted chemokine knockouts (developed in
this laboratory) to determine the role of site-specific release
of chemokines in neuroinflammatory disease.
2.)Assessing how normal aging affects cerebral angiogenesis in
different brain regions, and the role of different exercise
regimens in modifying age-related affects on cerebrovascular
properties.
3.)Using laser capture microdissection (LCM) coupled with gene
array and proteomic platforms to characterize the extent and
nature of heterogeneity along the microvascular tree of the
central nervous system.
4.)Using LCM to probe changes in the blood-brain barrier that
accompany cerebral ischemia.
Lab Rotation Projects
My laboratory is currently performing gene profiling of the
cells comprising the neurovascular unit in the central nervous
system (endothelial cells, astrocytes, and perivascular
microglia/pericytes. Specifically, laser capture microdissection
of these cells in situ is being coupled to quantitative,
real-time PCR and DNA microarray platforms. The objective is to
determine molecular finger prints of the neurovascular unit
throughout the CNS microvascular tree. As different vascular
beds exhibit unique phenotypes, such an approach will be
critical in identifying why particular CNS regions are prone to
diseases with vascular involvement, such as MS, stroke and
Alzheimer's disease.
Selected Publications
Ge, S., Murugesan, N. and Pachter, J.S. Astrocyte- and
endothelial-targeted CCL2 conditional knockout mice: critical
tools for studying the pathogenesis of neuroinflammation. J.
Mol. Neurosci. DOI 10.1007/s12031-009-9197-4.
Murugesan, N., Wu, S.-L., Hancock, W.S. and Pachter, J.S. 2009.
Analysis of mouse brain microvascular endothelium using laser
capture microdissection coupled with proteomics. In, The
Blood-Brain and Other Neural Barriers: Biology and Research
Protocols. Ed. S. Nag. Human Press. (In press).
Macdonald, J., Murugesan, N. and Pachter, J.S. 2008. Validation
of immuno-laser capture microdissection coupled with
quantitative RT-PCR to probe blood-brain barrier gene expression
in situ. J. Neurosci. Method. 174:219-226.
Lu, C., Murugesan, N., Macdonald, J., Wu, S.-L., Pachter, J.S.
and Hancock, W.S. 2008. Analysis of brain microvascular
endothelium using immuno-laser capture microdissection coupled
to a hybrid LTQ-FT-MS proteomics platform. Electrophoresis
29:2689-2695.
Ge, S., Song, L., Serwanski, D.R., Kuziel, W.A. and Pachter, J.S.
2008. Transcellular transport of CCL2 across brain microvascular
endothelial cells. J. Neurochem. 104:1219-1232.
Niciu, M.J., Ma, X.-M., Meskini. R.E., Pachter, J.S., Mains, R.E.
and Eipper, N.A. 2007. Altered ATP7A expression and other
compensatory responses in a murine model of Menkes disease.
Neurobiol. Dis. 27:278-291.
Song, L., Ge, S. and Pachter, J.S. 2007. Caveolin-1 regulates
expression of junction-associated proteins in microvascular
endothelial cells. Blood 109:1515-1523.
Ge, S. and Pachter, J.S. 2006. Isolation and culture of
microvascular endothelial cells from murine spinal cord. J.
Neuroimmunol. 177:209-214.
Kinnecom, K. and Pachter, J.S. 2005. Selective capture of
endothelial cells and perivascular cells from brain microvessels
by laser capture microdissection. Brain Res. Protoc. 16:1-9.
Song, L. and Pachter, J.S. 2005. Technical caveats in
identifying the source of endothelial cells in cultures derived
from brain microvessels. Lab. Invest. 85:1449-1450.
Dzenko, K.A., Song, L., Ge, S., Kuziel, W. and Pachter, J.S.
2005. CCR2 Expression by brain microvascular endothelial cells
is critical for macrophage transendothelial migration. Microvasc.
Res. 70:53-64.
Ge, S., Song, Li. and Pachter, J.S. 2005. Where is the
blood-brain barrierŠ. really? J. Neurosci. Res. 74:421-427.
Ge, S. and Pachter, J.S. 2004. Caveolin-1 knockdown by small
interfering RNA suppresses responses to the chemokine MCP-1 by
human astrocytes. J. Biol. Chem. 278:6688-6695.
Song L, and Pachter JS. 2004. Monocyte chemoattractant
protein-1 alters expression of tight junction-associated
proteins in brain microvascular endothelial cells. Microvasc
Res. Jan;67(1):78-89.
Ge S, and Pachter JS. 2004. Caveolin-1 knockdown by small
interfering RNA suppresses responses to the chemokine monocyte
chemoattractant protein-1 by human astrocytes. J Biol Chem. Feb
20;279(8):6688-95.
Song L, and Pachter JS. 2003. Culture of murine brain
microvascular endothelial cells that maintain expression and
cytoskeletal association of tight junction-associated proteins.
In Vitro Cell Dev Biol Anim. Jul;39(7):313-320.
Pachter JS, de Vries HE, and Fabry Z. 2003. The blood-brain
barrier and its role in immune privilege in the central nervous
system. J Neuropathol Exp Neurol. Jun;62(6):593-604. Review.
Andjelkovic AV, Song L, Dzenko KA, Cong H, and Pachter JS.
2002. Functional expression of CCR2 by human fetal astrocytes. J
Neurosci Res. Oct 15;70(2):219-31.
Dzenko, K.A., Andjelkovic, A.V. and Pachter, J.S. 2001. The
chemokine receptor CCR2 mediates the binding and internalization
of monocyte chemoattractant protein-1 along brain microvessels.
J. Neurosci. 21:9214-9223.
Andjelkovic, A.V., Zochowski, M.R., Morgan, F. and Pachter,
J.S. 2001. Qualitative and quantitative analysis of monocyte
transendothelial migration by confocal microscopy and
three-dimensional image reconstruction. In Vitro Cell. Devel.
Biol. 37:111-120. |
