Basic fibroblast growth factor is efficiently released from a cytolsolic storage site through plasma membrane disruptions of endothelial cells

Lakshmi Muthukrishnan, Elizabeth Warder, Paul L McNeil

Research output: Contribution to journalArticle

161 Citations (Scopus)

Abstract

Cells of gut and skin frequently suffer mechanically‐induced plasma membrane disruptions in vivo, and bioactive molecules, including basic fibroblast growth factor (bFGF), could enter and leave cytoplasm through these disruptions. We here provide three lines of evidence that bFGF is released with surprising efficiency through plasma membrane disruptions, resembling those known to occur in vivo, produced by scraping endothelial cells from their culturing substratum. First, 41% of the total of bFGF extractable in 1 M NaCl by freeze‐thaw and sonication was released simply by scraping the endothelial cells. Second, relative to release of lactate dehydrogenase, cells wounded by scraping under conditions promoting >60% cell survival released a significantly larger amount (up to twofold more) of growth promoting activity than did cells uniformly killed and irreversibly permeabilized by scraping in the cold or by freezing and thawing. Last, cells that survived membrane disruptions released, and contained, less bFGF on each subsequent wounding, consistent with release of bFGF through transient (i.e., survivable) membrane disruptions. A polyclonal antibody against bFGF completely neutralized the growth promoting activity released by scraping, confirming that bFGF is released through endothelial cell plasma membrane disruptions. Cell fractionation and immunolocalization, including a novel permeabilization technique for electron microscope immunolocalization, demonstrated a cytosolic location of bFGF. We conclude that many characteristics of bFGF–its broad spectrum of producing and target cell types, cytosolic location, efficient release through biologically and pathologically relevant plasma membrane wounds, and its release from cells that survive membrane wounds–make it a strong candidate as a “wound hormone” for rapidly initiating the cell growth required for routine maintenance of tissue integrity and/or repair after injury.

Original languageEnglish (US)
Pages (from-to)1-16
Number of pages16
JournalJournal of Cellular Physiology
Volume148
Issue number1
DOIs
StatePublished - Jan 1 1991

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Endothelial cells
Fibroblast Growth Factor 2
Cell membranes
Endothelial Cells
Cell Membrane
Membranes
Wounds and Injuries
Cell Fractionation
Sonication
Thawing
Cell growth
Fractionation
L-Lactate Dehydrogenase
Freezing
Cell Survival
Skin
Cytoplasm
Repair
Electron microscopes
Cells

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Basic fibroblast growth factor is efficiently released from a cytolsolic storage site through plasma membrane disruptions of endothelial cells. / Muthukrishnan, Lakshmi; Warder, Elizabeth; McNeil, Paul L.

In: Journal of Cellular Physiology, Vol. 148, No. 1, 01.01.1991, p. 1-16.

Research output: Contribution to journalArticle

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