Premium
Establishment of minimal requirements for sustained enterocyte migration in a wound healing model
Author(s) -
Pazin Marina Viktoria,
Rudnicki Jean Weston,
Caplan Michael S,
Jilling Tamas
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.687.3
Subject(s) - lamellipodium , cell migration , enterocyte , chemistry , transfection , wound healing , microbiology and biotechnology , cell , biology , biochemistry , small intestine , immunology , gene
Rat small intestinal epithelial cells (IEC6)were monitored by live cell imaging to identify required components for sustained linear migration. Supplementation of HBSS with L‐glutamine, L‐arginine (4mM each), insulin (0.1mg/ml), sodium pyruvate (1mM) and FBS (5%), sustained linear migration for at least two hours at 0.38±0.04 ?m/min. Without sodium pyruvate, migration rate diminished to 0.26±0.02 ?m/min, but remained linear for 2h. Without FBS, cells migrated for 1 h at 0.34±0.07 ?m/min, but slowed to 0.03±0.002 µm/min in the second hour, correlating with severely reduced lamellipodia formation and with the emergence of actin bundles perpendicular to the direction of migration on the leading edge of most cells, as revealed by phalloidin‐AF488 staining of fixed cells. Fluorescence image analysis of IEC6 transfected with AKTPH‐GFP, i.e., a probe for PIP3, indicated a rhythmic accumulation of PIP3 at the leading edges of lamellipodia, which diminished when cells stopped migrating. It is yet to be determined which serum‐derived factors are responsible for the induction of lamellipodia formation and what is the role of PI3 kinase in this process. Supported: NIH HD37581, DK062960.