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



We 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.



Most 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.



Our 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.
Vect-Horus SAS, Faculté de Médecine - Secteur Nord, 51 Bd Pierre Dramard, 13344, Marseille Cedex 15, France.
Yves
Y
Molino
Vect-Horus SAS, Faculté de Médecine - Secteur Nord, 51 Bd Pierre Dramard, 13344, Marseille Cedex 15, France.
Françoise
F
Jabès
Aix Marseille Univ, CNRS, NICN, Marseille, France.
Amandine
A
Bonnet
Aix Marseille Univ, CNRS, NICN, Marseille, France.
Nicolas
N
Gaudin
Aix Marseille Univ, CNRS, NICN, Marseille, France.
Anne
A
Bernard
Aix Marseille Univ, CNRS, NICN, Marseille, France.
Philippe
P
Benech
Aix Marseille Univ, CNRS, NICN, Marseille, France. michel.khrestchatisky@univ-amu.fr.
Michel
M
Khrestchatisky
D000818:Animals; D001921:Brain / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D046228:Microarray Analysis; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000235:genetics / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D016207:Cytokines; D018398:Homeodomain Proteins; D012333:RNA, Messenger; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha
Endothelial cells; HOX members; Matrix metalloproteinase (MMP); Microvessels; TNF-α response; Vascular beds