Annexins I and II show differences in subcellular localization and differentiation-related changes in human epidermal keratinocytes.
The annexins are a family of calcium-dependent phospholipid-binding proteins whose in vitro properties have led to a number of hypotheses suggesting their cellular functions, including membrane fusion in exocytosis and endocytosis. To investigate the topography and possible functions of these proteins we compared the subcellular localization of annexins I, II, IV and VI in skin sections and in cultured epidermal keratinocytes by immunostaining. We found that annexin I staining was in a granular pattern in the monolayer epithelial cells but in an envelope pattern in the stratified keratinocytes. This finding corroborates previous reports that annexin I crosslinks to form cornified envelopes in the mid-epidermis and explains the absence of staining above that level. It is unlikely that this protein is related to exocytosis in the granular layer of the epidermis. In comparison, annexin II staining was also granular and was detected in all nucleated epidermal cells as bands at the cell periphery. However, only annexin II was detected extracellularly among the top layer of cultured cells. The intracellular linear envelope pattern of annexin I and the intercellular pattern of annexin II suggest their interactions with the membrane cytoskeleton in other biological functions. Taken together, both annexins undergo different differentiation-related changes. While methanol fixation enhanced staining of annexin I, it diminished staining of annexin II. Their opposite responses to methanol fixative suggests a different molecular organization of the two annexins with phospholipid in the cell membrane. Annexins IV and VI were predominantly confined to dermal cells including ductal and myoepithelial cells and were not detected in cultured keratinocytes using either cold methanol fixative or prefixation labeling.
Ma
A S
AS
Section of Dermatology, Department of Medicine, The University of Chicago, Illinois 60637, USA.
Ozers
L J
LJ
D017305:Annexin A1 / Q000032:analysis*; D017306:Annexin A2 / Q000032:analysis*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006801:Humans; D007124:Immunoenzyme Techniques; D015603:Keratinocytes / Q000737:chemistry* / Q000166:cytology; D013347:Subcellular Fractions / Q000737:chemistry*
D017305:Annexin A1; D017306:Annexin A2