title: Acute and long-term effects of tissue culture on contractile reactivity in renal arteries of the rat.
abstract: To evaluate long-term effects of contractile and mitogenic stimuli on the contractile reactivity of arterial smooth muscle, we measured the incorporation of the thymidine analogue 5-bromo-2'-deoxyuridine (BrdUrd) and mechanical responses in arterial segments that had been maintained in tissue culture. The experiments were performed on renal arteries that had been isolated from adult rats, chemically sympathectomized, mechanically denuded from endothelium and mounted under distension. Exposure of arterial segments for up to 3 weeks to culture medium supplemented with fetal calf serum resulted in the following consecutive changes: a strong acute contraction, selective pharmacological changes that included decreased contractile responses to phenylephrine and vasopressin and increased relaxing responses to isoproterenol, increased incorporation of BrdUrd, a progressive fall in contractile responses to all vasoconstrictor stimuli, and an increase in excitability. Serum-free medium resulted in a much smaller acute arterial contraction, induced less incorporation of BrdUrd, accelerated the occurrence of hyperexcitability, but did not affect early pharmacological changes or the subsequent fall in overall arterial contractility with tissue culture. Dialysis of the serum or addition of ketanserin abolished the contractile effect of serum but did not affect the incorporation of BrdUrd or the loss of contractility with tissue culture. Addition of serotonin to serum-free culture medium mimicked the contractile response to serum but not the stimulation of BrdUrd incorporation. These data indicate that tissue culture alters the properties of the arterial wall, that contraction does not underlie the proliferative response of arterial smooth muscle to serum-derived mitogens in vitro, and that stimulation of DNA synthesis does in itself not lead to selective changes in arterial contractility.
authors: Department of Pharmacology, University of Limburg, Maastricht, The Netherlands.
authors: J G
authors: JG
authors: De Mey
authors: M P
authors: MP
authors: Uitendaal
authors: H C
authors: HC
authors: Boonen
authors: M J
authors: MJ
authors: Vrijdag
authors: M J
authors: MJ
authors: Daemen
authors: H A
authors: HA
authors: Struyker-Boudier
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D001973:Bromodeoxyuridine; D002417:Cattle / Q000097:blood; D003470:Culture Media; D046508:Culture Techniques; D004247:DNA / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000502:physiology; D008297:Male; D051381:Rats; D011921:Rats, Inbred WKY; D012077:Renal Artery / Q000378:metabolism / Q000502:physiology*; D012701:Serotonin / Q000502:physiology; D013997:Time Factors; D014661:Vasoconstriction
chemical_list: D003470:Culture Media; D012701:Serotonin; D004247:DNA; D001973:Bromodeoxyuridine