Selective Permeability of [<sup>3</sup>H]-D-Mannitol and [<sup>l4</sup>C]-Carboxyl-lnulin Across the Blood-Brain Barrier and Blood-Spinal Cord Barrier in the Rabbit

Prockop, Leon D.; Naidu, K. Akhilender; Binard, Joseph E.; Ransohoff, Joseph

doi:10.1080/10790268.1995.11719399

Cited by 57 publications

(43 citation statements)

References 1 publication

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The BSCB also appears to be more permeable than the BBB in certain sub-regions but is still a tight and highly regulated barrier that protects the spinal cord parenchyma. Studies in vivo have indicated that the BSCB is more permeable than the BBB to small tracers, cytokines and neurotrophins, with lumbar regions of the spinal cord in particular being more permeable [90-94]. Some cytokines and growth factors however, such as IL-1α and granulocyte-macrophage colony-stimulating factor (GMCSF), have been shown to have similar transport across the BSCB as the BBB in vivo [95,96].…”

Section: Discussionmentioning

confidence: 99%

Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitromodels of the rat blood-brain barrier and blood-spinal cord barrier

Watson¹,

Paterson²,

Thom³

et al. 2013

BMC Neurosci

View full text Add to dashboard Cite

BackgroundModelling the blood-CNS barriers of the brain and spinal cord in vitro continues to provide a considerable challenge for research studying the passage of large and small molecules in and out of the central nervous system, both within the context of basic biology and for pharmaceutical drug discovery. Although there has been considerable success over the previous two decades in establishing useful in vitro primary endothelial cell cultures from the blood-CNS barriers, no model fully mimics the high electrical resistance, low paracellular permeability and selective influx/efflux characteristics of the in vivo situation. Furthermore, such primary-derived cultures are typically labour-intensive and generate low yields of cells, limiting scope for experimental work. We thus aimed to establish protocols for the high yield isolation and culture of endothelial cells from both rat brain and spinal cord. Our aim was to optimise in vitro conditions for inducing phenotypic characteristics in these cells that were reminiscent of the in vivo situation, such that they developed into tight endothelial barriers suitable for performing investigative biology and permeability studies.MethodsBrain and spinal cord tissue was taken from the same rats and used to specifically isolate endothelial cells to reconstitute as in vitro blood-CNS barrier models. Isolated endothelial cells were cultured to expand the cellular yield and then passaged onto cell culture inserts for further investigation. Cell culture conditions were optimised using commercially available reagents and the resulting barrier-forming endothelial monolayers were characterised by functional permeability experiments and in vitro phenotyping by immunocytochemistry and western blotting.ResultsUsing a combination of modified handling techniques and cell culture conditions, we have established and optimised a protocol for the in vitro culture of brain and, for the first time in rat, spinal cord endothelial cells. High yields of both CNS endothelial cell types can be obtained, and these can be passaged onto large numbers of cell culture inserts for in vitro permeability studies. The passaged brain and spinal cord endothelial cells are pure and express endothelial markers, tight junction proteins and intracellular transport machinery. Further, both models exhibit tight, functional barrier characteristics that are discriminating against large and small molecules in permeability assays and show functional expression of the pharmaceutically important P-gp efflux transporter.ConclusionsOur techniques allow the provision of high yields of robust sister cultures of endothelial cells that accurately model the blood-CNS barriers in vitro. These models are ideally suited for use in studying the biology of the blood-brain barrier and blood-spinal cord barrier in vitro and for pre-clinical drug discovery.

show abstract

Section: Discussionmentioning

confidence: 99%

Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitromodels of the rat blood-brain barrier and blood-spinal cord barrier

Watson¹,

Paterson²,

Thom³

et al. 2013

BMC Neurosci

View full text Add to dashboard Cite

show abstract

“…In vitro, brain and spinal cord pericytes differ markedly in their potential for tube formation and migration, reflecting more differences between the BBB and BSCB [8]. Such differences in their microenvironment may in turn induce MEC heterogeneity between brain and spinal domains, correlating with a higher BSCB inherent permeability [9, 10]. This increased permeability might result from differences in cell junction protein expression between BBB and BSCB endothelial cells.…”

Section: Introductionmentioning

confidence: 99%

Gene expression comparison reveals distinct basal expression of HOX members and differential TNF-induced response between brain- and spinal cord-derived microvascular endothelial cells

Molino¹,

Jabès²,

Bonnet

et al. 2016

J Neuroinflammation

View full text Add to dashboard Cite

BackgroundThe heterogeneity of endothelial cell types underlies their remarkable ability to sub-specialize and provide specific requirements for a given vascular bed. Here, we compared rat microvascular endothelial cells (MECs) derived from the brain and spinal cord in both basal and inflammatory conditions.MethodsWe used whole rat genome microarrays to compare, at different time points, basal and TNF-α-induced gene expression of rat MECs from in vitro models of the blood-brain barrier (BBB) and blood-spinal cord barrier (BSCB). Validation at both messenger RNA (mRNA) and protein levels was performed on freshly extracted microvessels (MVs) from the brain and spinal cord (BMVs and SCMVs, respectively), as these were considered the closest in vivo tissues to cultured MECs.ResultsMost of the genes encoding adhesion/tight junction molecules and known endothelial markers were similarly expressed in brain and spinal cord MECs (BMECs and SCMECs, respectively). However, one striking finding was the higher expression of several Hox genes, which encode transcription factors involved in positional identity. The differential expression of Hoxa9 and Hoxb7 at the mRNA levels as well as protein levels was confirmed in BMVs and SCMVs. Although the TNF-α response was in general higher in BMECs than in SCMECs at 12 h, the opposite was observed at 48 h. Furthermore, we found that expression of Tnfrsf1a and Tnfrsf1b encoding the TNF receptor super-family member 1a/TNFR1 and 1b/TNFR2, respectively, were constitutively higher in BMVs compared to SCMVs. However, only Tnfrsf1b was induced in SCMECs in response to TNF-α at 24 and 48 h.ConclusionsOur results support a role for HOX members in defining the positional identities of MECs in vivo. Our data also suggest that the delayed transcriptional activation upon TNF-α treatment in SCMECs results from the requirement of the TNF-induced expression of Tnfrsf1b. In contrast, its high basal expression in BMECs might be sufficient to confer an immediate and efficient TNF-α response.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-016-0749-6) contains supplementary material, which is available to authorized users.

show abstract

“…Although the BBB and BSCB are functionally similar, many differences have been identified (Bartanusz et al , 2011). For example, the rodent BSCB has heightened permeability to mannitol and inulin (Daniel et al , 1985; Prockop et al , 1995), interferons (Pan et al , 1997 a ), albumin, sucrose, and tumor necrosis factor α (Pan et al , 1997 b ). The source of these permeability changes is largely unknown, although reduced tight junction protein expression in the spinal cord has been reported (Ge and Pachter, 2006).…”

Section: Introductionmentioning

confidence: 99%

Blood–Spinal Cord Barrier Pericyte Reductions Contribute to Increased Capillary Permeability

Winkler

Sengillo

Bell

et al. 2012

J Cereb Blood Flow Metab

181

195

View full text Add to dashboard Cite

The blood–spinal cord barrier (BSCB) regulates molecular exchange between blood and spinal cord. Pericytes are presumed to be important cellular constituents of the BSCB. However, the regional abundance and vascular functions of spinal cord pericytes have yet to be determined. Utilizing wild-type mice, we show that spinal cord pericyte capillary coverage and number compared with the brain regions are reduced most prominently in the anterior horn. Regional pericyte variations are highly correlated with: (1) increased capillary permeability to 350 Da, 40,000 Da, and 150,000 Da, but not 2,000,000 Da fluorescent vascular tracers in cervical, thoracic, and lumbar regions and (2) diminished endothelial zonula occludens-1 (ZO-1) and occludin tight junction protein expression. Pericyte-deficient mutations (PdgfrβF7/F7 mice) resulted in additional pericyte reductions in spinal cord capillaries leading to overt BSCB disruption to serum proteins, accumulation in motor neurons of cyotoxic thrombin and fibrin and motor neuron loss. Barrier disruption in perciyte-deficient mice coincided with further reductions in ZO-1 and occludin. These data suggest that pericytes contribute to proper function of the BSCB at the capillary level. Regional reductions in spinal cord pericytes may provide a cellular basis for heightened spinal cord barrier capillary permeability and motor neuron loss.

show abstract

Selective Permeability of [³H]-D-Mannitol and [^l4C]-Carboxyl-lnulin Across the Blood-Brain Barrier and Blood-Spinal Cord Barrier in the Rabbit

Cited by 57 publications

References 1 publication

Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitromodels of the rat blood-brain barrier and blood-spinal cord barrier

Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitromodels of the rat blood-brain barrier and blood-spinal cord barrier

Gene expression comparison reveals distinct basal expression of HOX members and differential TNF-induced response between brain- and spinal cord-derived microvascular endothelial cells

Blood–Spinal Cord Barrier Pericyte Reductions Contribute to Increased Capillary Permeability

Contact Info

Product

Resources

About

Selective Permeability of [3H]-D-Mannitol and [l4C]-Carboxyl-lnulin Across the Blood-Brain Barrier and Blood-Spinal Cord Barrier in the Rabbit

Cited by 57 publications

References 1 publication

Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitromodels of the rat blood-brain barrier and blood-spinal cord barrier

Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitromodels of the rat blood-brain barrier and blood-spinal cord barrier

Gene expression comparison reveals distinct basal expression of HOX members and differential TNF-induced response between brain- and spinal cord-derived microvascular endothelial cells

Blood–Spinal Cord Barrier Pericyte Reductions Contribute to Increased Capillary Permeability

Contact Info

Product

Resources

About

Selective Permeability of [³H]-D-Mannitol and [^l4C]-Carboxyl-lnulin Across the Blood-Brain Barrier and Blood-Spinal Cord Barrier in the Rabbit