Osteopontin expression in spontaneously developed neointima in fowl (Gallus gallus)

Rhonda J. Kuykindoll, Hiroko Nishimura, Donald Thomason, Satoru Nishimoto

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Fowl show spontaneous elevation of blood pressure and neointimal plaque formation in the abdominal aorta at young ages. A similar neointima can be induced by a balloon-catheter-induced endothelium injury to the fowl aorta. Both spontaneously developed and injury-induced vascular lesions exhibit subendothelial hyperplasia consisting of neointimal cells with a synthetic phenotype and abundant extracellular matrix. The role of the extracellular matrix in the formation of neointima is not known. In this study, we investigated whether osteopontin, an adhesive glycoprotein present in the extracellular matrix, is expressed in aortic smooth muscle tissue of the fowl abdominal aorta, in spontaneously developed neointimal plaques and in the aortic smooth muscle underlying neointimal plaques. Crude protein extracted from isolated aortic smooth muscle tissues and neointimal plaques was fractionated by SDS-polyacrylamide gel electrophoresis and analyzed by immunoblotting with rabbit anti-fowl osteopontin (provided by Dr L. C. Gerstenfeld, Boston University) or anti-alpha smooth muscle actin antibodies. The anti-fowl osteopontin antibody predominantly recognized a 66-70kDa protein band in neointimal plaques that co-migrated with the osteopontin phosphoprotein from chick bone. In contrast, intact aortic smooth muscle and the smooth muscle underlying neointimal plaques equally expressed three proteins (66-70 kDa, approximately 50 kDa and approximately 43 kDa) recognized by the anti-osteopontin antibody. Anti-alpha smooth muscle actin antibody recognized a 43kDa protein band, and the expression of alpha smooth muscle actin was higher in aortic smooth muscle than in neointimal plaques. Osteopontin mRNA expression was examined using reverse transcription-polymerase chain reaction (RT-PCR) of total RNA from vascular tissues with specific primers constructed on the basis of the reported fowl osteopontin nucleotide sequence. The PCR products from intact aortic smooth muscle and neointimal plaques correspond to the product from recombinant plasmid cDNA (a gift from Dr L. C. Gerstenfeld) transcribed in vitro. These results suggest that osteopontin is synthesized in intact aortic smooth muscle and neointimal plaques in fowl and that unmetabolized approximately 66kDa osteopontin protein is a predominant form in the neointima, indicating that osteopontin protein may be actively synthesized in the neointima.

Original languageEnglish (US)
Pages (from-to)273-282
Number of pages10
JournalJournal of Experimental Biology
Volume203
Issue number2
StatePublished - Jan 1 2000

Fingerprint

osteopontin
Neointima
Gallus gallus
Osteopontin
smooth muscle
Smooth Muscle
Chickens
muscle
chickens
antibody
protein
aorta
extracellular matrix
actin
Extracellular Matrix
Actins
Proteins
antibodies
Abdominal Aorta
muscle tissues

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Physiology
  • Ecology, Evolution, Behavior and Systematics
  • Aquatic Science
  • Molecular Biology
  • Animal Science and Zoology
  • Insect Science

Cite this

Osteopontin expression in spontaneously developed neointima in fowl (Gallus gallus). / Kuykindoll, Rhonda J.; Nishimura, Hiroko; Thomason, Donald; Nishimoto, Satoru.

In: Journal of Experimental Biology, Vol. 203, No. 2, 01.01.2000, p. 273-282.

Research output: Contribution to journalArticle

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