title: Interstitial cells from rat middle cerebral artery belong to smooth muscle cell type.
abstract: It is now established that non-contractile cells with thin filopodia, also called vascular interstitial cells (VICs), are constitutively present in the media of many, if not all, blood vessels. The aim of this study was to determine the type of cell lineage to which arterial VICs belong using immunocytochemical, and real-time and reverse transcription PCR (RT-PCR). Using RT-PCR, we compared gene expression profiles of single VICs and smooth muscle cells (SMCs) freshly dispersed from rat middle cerebral artery. Both VICs and SMCs expressed the SMC marker, smooth muscle myosin heavy chain (SM-MHC), but did not express fibroblast, pericyte, neuronal, mast cell, endothelial or stem cell markers. Freshly isolated VICs also did not express c-kit, which is the marker for interstitial cells of Cajal in the gastrointestinal tract. Immunocytochemical labelling of contractile proteins showed that VICs and SMCs expressed SM-MHC similarly to the same degree, but VICs in contrast to SMCs had decreased expression of alpha-SM-actin and very low or no expression of calponin. Real-time RT-PCR was consistent with immunocytochemical experiments and showed that VICs had four times lower gene expression of calponin comparing to SMCs, which may explain VICs' inability to contract. VICs had greater expression than SMCs of structural proteins such as non-muscular beta-actin and desmin. The results obtained suggest that VICs represent a subtype of SMCs and may originate from the same precursor as SMCs, but later develop filopodia and a non-contractile cell phenotype.
authors.affiliation: Ion Channels and Cell Signalling Centre, Division of Basic Medical Sciences, St. George's, University of London, London, United Kingdom. m.harhun@sgul.ac.uk
authors.forename: Maksym I
authors.initials: MI
authors.lastname: Harhun
authors.forename: Kinga
authors.initials: K
authors.lastname: Szewczyk
authors.forename: Holger
authors.initials: H
authors.lastname: Laux
authors.forename: Sally A
authors.initials: SA
authors.lastname: Prestwich
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Gordienko
authors.forename: Ray F
authors.initials: RF
authors.lastname: Moss
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Bolton
mesh_terms: D000818:Animals; D003285:Contractile Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D008297:Male; D020768:Middle Cerebral Artery / Q000166:cytology* / Q000648:ultrastructure; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D003285:Contractile Proteins