Fluorescent staining of the actin cytoskeleton in human lymphocytes, monocytes and polymorphonuclear cells using a DNAse 1/anti-DNAse 1 immunoglobulin fluorescein conjugated system.
The actin associated with membrane-enriched extracts of leukocytes can be quantitated by DNAse 1 inhibition. Using this assay, we previously demonstrated that the actin level in monocytes was significantly higher than that in polymorphonuclear, T and B cells respectively. However, the extracellular location of the actin fraction detected by DNAse 1 inhibition (monomeric "G") remained unclear. This study using the DNAse 1/anti DNAse 1 immunoglobulin fluorescein conjugated system demonstrated that G-actin is present primarily in the cortical cell cytoplasm of leukocytes, in confirmation of our previous biochemical findings. Since the solubilized G-actin activities of membrane-rich lymphoid cell fractions, measured by DNAse 1 inhibition, are a reflection of the migratory potential, this immunofluorescent system may permit identification of the leukocytic cell subpopulations that have a potential for active circulation.
Philip
P J
PJ
Unité de Biologie Cellulaire et Tissulaire, Hôpital Pasteur, Nice, France.
Sudaka
I
I
Mely-Goubert
B
B
D000199:Actins / Q000737:chemistry* / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D003599:Cytoskeleton / Q000737:chemistry*; D003850:Deoxyribonuclease I / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D019793:Fluorescein; D005452:Fluoresceins; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D007962:Leukocytes / Q000201:enzymology / Q000378:metabolism*; D008214:Lymphocytes / Q000201:enzymology / Q000378:metabolism; D009000:Monocytes / Q000201:enzymology / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism
D000199:Actins; D005452:Fluoresceins; D007136:Immunoglobulins; D003850:Deoxyribonuclease I; D019793:Fluorescein