Functional nucleotide receptor expression and sarcoplasmic reticulum morphology in dedifferentiated porcine coronary smooth muscle cells

Brent J F Hill, Brian R. Wamhoff, Michael Sturek

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The phenotypic dedifferentiation of vascular smooth muscle cells (SMCs) is an early event associated with cell culturing and vascular injury. The purpose of this study was to evaluate the SMC phenotype underlying the functional responsiveness of SMCs to nucleotides in organ culture. Porcine coronary arteries were either used fresh, cold stored (5°C) 4 days, or organ cultured (37°C) 4 days. Fura-2 digital imaging of single SMCs was used to measure the myoplasmic calcium (Cam) response to 10 μM of the following nucleotide receptor agonists: UTP, UDP, ATP, ADP, and 2-MeSATP. In contrast to the nucleotides UDP, ATP, ADP, and 2-MeSATP, the Cam response increased 10-fold and the number of cells that responded to UTP increased 5-fold in SMCs from organ culture compared to SMCs from fresh or cold-stored arteries. Simultaneous imaging of Cam, DNA content, and SR distribution in SMCs from organ culture indicated that the UTP-induced Cam increase occurred exclusively in SMCs that had a dedifferentiated cell phenotype. Three-dimensional image reconstruction of the nucleus and sarcoplasmic reticulum (SR) revealed a novel transnuclear SR distribution that intertwined with the nucleus in fresh SMCs, while in SMCs from organ culture the SR was predominantly perinuclear and cytoplasmic. This study demonstrates that the functional up-regulation of UTP-sensitive receptors and the disappearance of the transnuclear SR distribution are novel features of dedifferentiated coronary SMCs.

Original languageEnglish (US)
Pages (from-to)432-443
Number of pages12
JournalJournal of Vascular Research
Volume38
Issue number5
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Sarcoplasmic Reticulum
Smooth Muscle Myocytes
Swine
Nucleotides
Organ Culture Techniques
Calcium
Uridine Triphosphate
Uridine Diphosphate
Cell Culture Techniques
Adenosine Diphosphate
Adenosine Triphosphate
Phenotype
Computer-Assisted Image Processing
Three-Dimensional Imaging
Fura-2
Vascular System Injuries
Vascular Smooth Muscle
Coronary Vessels
Up-Regulation
Arteries

Keywords

  • Coronary artery
  • Digital imaging
  • Intracellular calcium
  • Nucleus
  • Organ culture
  • P2Y receptors
  • Sarcoplasmic reticulum

ASJC Scopus subject areas

  • Physiology

Cite this

Functional nucleotide receptor expression and sarcoplasmic reticulum morphology in dedifferentiated porcine coronary smooth muscle cells. / Hill, Brent J F; Wamhoff, Brian R.; Sturek, Michael.

In: Journal of Vascular Research, Vol. 38, No. 5, 2001, p. 432-443.

Research output: Contribution to journalArticle

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