The expression of megalin (gp330) and LRP diverges during F9 cell differentiation

RP Czekay, RA Orlando, L Woodward… - Journal of Cell …, 1995 - journals.biologists.com
RP Czekay, RA Orlando, L Woodward, ED Adamson, MG Farquhar
Journal of Cell Science, 1995journals.biologists.com
The receptor-associated protein, RAP, is a chaperonin-like molecule that binds to two
members of the low density lipoprotein receptor (LDLR) superfamily–megalin (gp330) and
the LDL receptor-related protein (LRP). In F9 embryonal carcinoma cells, expression of RAP
mRNA increases when differentiation is induced with retinoic acid and dibutyryl-cyclic AMP.
We have investigated the expression of megalin and LRP and their interaction with RAP in
F9 cells using biochemical and immunocytochemical methods. Both receptors are …
Abstract
The receptor-associated protein, RAP, is a chaperonin-like molecule that binds to two members of the low density lipoprotein receptor (LDLR) superfamily – megalin (gp330) and the LDL receptor-related protein (LRP). In F9 embryonal carcinoma cells, expression of RAP mRNA increases when differentiation is induced with retinoic acid and dibutyryl-cyclic AMP. We have investigated the expression of megalin and LRP and their interaction with RAP in F9 cells using biochemical and immunocytochemical methods. Both receptors are expressed in uninduced F9 cells, but only megalin co-precipitates with RAP. When F9 cells were induced to differentiate into parietal endoderm, the expression of megalin was dramatically increased. The expression of megalin exceeded that of LRP and RAP by an order of magnitude and both receptors co-precipitated with RAP. By immunoelectron microscopy, megalin and LRP were localized to clathrin-coated pits at the cell surface in both undifferentiated and differentiated F9 cells, whereas RAP was found mainly in the ER. A sizeable pool of LRP was also detected in the ER. When F9 cells were grown in suspension in the presence of RA and induced to develop into embryoid bodies, the expression of megalin and LRP segregated into different cell types: megalin was found in the outer epithelial layer and LRP in the stem cells of the inner core. Our results demonstrate that F9 cells induced to differentiate in monolayer culture express megalin, LRP and RAP, and RAP is capable of interacting simultaneously with both receptors. In embryoid bodies the expression of megalin and LRP diverges, and only megalin is expressed in the outer epithelial layer. The high levels of expression and cell specificity of megalin and LRP suggest important roles for these receptors in early embryonic development.
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