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Immuno-histochemical expression of a1, a2 and a3 integrin subunits during angiogenesis in vitro H. Suda1, Y. Asami1, E. Murata1, K. Fujita1 and M. Akita2 1Department of Anatomy, Saitama Medical School, Saitama, Japan and 2Division of Morphological Science, Biomedical Research Center, Saitama Medical School, Saitama, Japan Offprint requests to: Prof. Dr. M. Akita, Division of Morphological Science, Biomedical Research Center, Saitama Medical School, 38 Moroyama, Iruma-gun, Saitama 350-0495, Japan. Fax: +81-49-295-5998. e-mail: makita@saitama-med.ac.jp
Summary. Aortic
explants were obtained from mouse fetuses and cultured in collagen
gels. Immuno-fluorescence microscopy, antibodies (anti a1, a2 and a3 integrin subunits) were used. Fibroblastic cells
migrated from the aortic explant after one day of cultivation.
The migrating cells located in the peripheral part of the aortic
explant were positive for a1 and a2 integrin subunit antibodies. Immuno-fluorescence-positive
staining for the a3
integrin subunit antibody was clearly seen in the migrating cells
located near the aortic explant and surrounding tube-like structures.
In an immuno-electron microscope study performed by pre-embedding
immuno labeling, gold particles associated with the a3 integrin subunit
were found to reside on the membranes of the cells surrounding
the capillary-like tubes. Two synthetic peptides, GRGDSP (Gly-Arg-Gly-Asp-Ser-Pro)
and KDGEA (Lys-Asp-Gly-Glu-Ala), were added to the growth medium
to study their effects on cell migration. KDGEA, a compound containing
the recognition sequence for a2ß1 integrin, decreased cell migration, while
GRGDSP exhibited no effect. Key words: Integrin,
a subunit, Angiogenesis, Culture Immuno-histochemistry |