title: The growth and differentiation of aortal smooth muscle cells after calcitriol treatment are associated with microtubule reorganisation -- an in vitro study.
abstract: The smooth muscle cells (SMCs) of the arterial media play a predominant role in functional and structural alterations of the arterial wall. The transition from the "contractile" to the "synthetic" phenotype appears to be an early critical event in the development of atherosclerotic disease. A number of observations suggest that 1,25(OH)(2)D(3) (calcitriol) is of importance in maintaining normal cardiovascular function through its receptors in cardiac myocytes or aortal SMCs. The present study has focused on the microtubular (MT) network reorganisation after exposure to calcitriol. SMCs isolated by enzymatic digestion from the aortal media of neonatal rats were cultured on glass cover slips. 1 microM of 1,25(OH)(2)D(3) was added to the culture medium every second day. The cytoskeletal features of SMCs after calcitriol were visualised by the immunofluorescence staining of alpha-tubulin. The alterations in alpha-tubulin expression and the distribution of microtubules related to the activities of the vascular smooth muscle cells, namely adhesion, migration, multilayer formation and cell division, were observed. A spindle shape, decreased cell adhesion, low expression of alpha-tubulin and a longitudinally arranged microtubular network manifested the high rate of SMC differentiation in the calcitriol-treated culture. A flat stellate morphology, high expression of alpha-tubulin and a radially distributed three-dimensional microtubular network were observed in the SMCs of the control culture. Destructive changes in the microtubular architecture which altered the cellular shape were evident in SMCs undergoing apoptosis. Cells with apoptotic features were more frequent in calcitriol-exposed culture. In contrast to the regular SMC divisions observed in the control culture, some of the mitotic cells exposed to calcitriol contained broader bipolar, multipolar or disordered spindles. These alterations in the SMCs' microtubular cytoskeleton after calcitriol treatment were concomitant with changes in cell growth, differentiation and apoptosis, and may suggest a similarity to atherosclerotic plaque formation.
authors.affiliation: Laboratory of Electron Microscopy, Medical University, Gdańsk, Poland. ctukaj@amedec.amg.gda.pl
authors.forename: Cecylia
authors.initials: C
authors.lastname: Tukaj
authors.forename: Jerzy
authors.initials: J
authors.lastname: Bohdanowicz
authors.forename: Jolanta
authors.initials: J
authors.lastname: Kubasik-Juraniec
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002117:Calcitriol / Q000494:pharmacology*; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D014404:Tubulin / Q000378:metabolism
chemical_list: D002120:Calcium Channel Agonists; D014404:Tubulin; D002117:Calcitriol