G-actin pool and actin messenger RNA during development of the apical processes of the retinal pigment epithelial cells of the chick.
It has been suggested that during development an increase in the pool of G-actin may drive the elongation of actin-containing processes which occur in several types of epithelial cells. The apical processes of chick retinal pigment epithelial (RPE) cells elongate during the last 7 days of embryonic life (E15-E21) reaching lengths of 20 microns or more by hatching (E21). F-actin bundles form the cores of these processes. We followed the elongation by measuring F-actin in the cells and cytoskeletons. In correlation with this, we studied by DNAse assay the levels of monomeric actin in supernatants of cell extracts from E13, before elongation starts, to E17, when elongation is well underway. Total F-actin increased 1.9-fold over this time period and cytoskeletal actin increased 2.5-fold. In supernatants from extracts of E13 RPE the monomeric actin concentration was 51 +/- 0.5 micrograms/ml. From estimates of cell volume we calculated the cellular monomeric actin concentration at E13 as at least 510 micrograms/ml (13 microM). We compared this with monomeric actin levels in extracts from RPE at E15 and E17. Allowing for the estimated increase in cell volume, our data show little overall change in cellular monomeric actin concentration at these times. Changes in the level of actin mRNA were measured over the same time period. Normalized to equal RNA, we found a twofold increase in beta actin mRNA and a four- to fivefold increase in message for gamma actin at E17 as compared to E13. In summary, we show that (1) there is a substantial pool of monomeric actin in these epithelial cells before elongation starts; (2) process elongation is not associated with a significant change in the size of this pool; and (3) process elongation is associated with a significant increase in actin mRNA.
Nachmias
V T
VT
Department of Anatomy, School of Medicine, University of Pennsylvania, Philadelphia 19104-6058.
Philp
N
N
Momoyama
Y
Y
Choi
J K
JK
D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D002642:Chick Embryo; D003851:Deoxyribonucleases / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008871:Microvilli / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D014746:Vimentin / Q000032:analysis
D000199:Actins; D012333:RNA, Messenger; D014746:Vimentin; D003851:Deoxyribonucleases