The polarity of the membrane skeleton in retinal pigment epithelial cells of developing chicken embryos and in primary culture.
We studied the morphogenesis and the membrane skeleton in the retinal pigment epithelium during chicken embryogenesis and in culture, by using immunofluorescence and electron microscopy. During embryogenesis two distinct membrane skeletal structures were formed, an apical and a basolateral one. The former was seen in the apical surface already in the 10-day-old embryos. It was comprised of ankyrin and alpha-fodrin and showed a codistribution with Na+,K(+)-ATPase and an as yet uncharacterized cadherin-like molecule. The basolateral membrane skeleton was seen in the lateral walls already in the 10-day-old embryos, and later, between the 13th and 17th embryonic days, it also appeared at the basal membrane, coincidentally with the formation of the basal infoldings. It consisted of ankyrin and alpha-fodrin, but did not codistribute with any of the integral membrane proteins studied (Na+,K(+)-ATPase and cadherins). In culture, the retinal pigment epithelial cells retained their polarized morphology. Compared with the situation in vivo, however, there was a distinct translocation of the membrane skeletal components fodrin and ankyrin from the apical surface to the lateral walls, accompanied by a similar redistribution of Na+,K(+)-ATPase and the cadherin-like molecule. The results suggest that (1) there is, in the retinal pigment epithelium, an apical Na+,K(+)-ATPase-membrane skeleton structure stabilized by contacts between the retinal pigment epithelium and the neural retina, possibly mediated by a cadherin-like molecule, and that (2) there is another fodrin/ankyrin-based membrane skeleton in the basolateral walls that is important for the maintenance of the extensive folding of these surface areas.
Huotari
V
V
Biocenter Oulu, University of Oulu, Finland.
Sormunen
R
R
Lehto
V P
VP
Eskelinen
S
S
D000818:Animals; D017487:Ankyrins / Q000032:analysis; D015820:Cadherins / Q000032:analysis; D002352:Carrier Proteins / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure*; D016764:Cell Polarity; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology* / Q000254:growth & development; D015151:Immunoblotting; D008840:Microfilament Proteins / Q000032:analysis; D008856:Microscopy, Fluorescence; D010857:Pigment Epithelium of Eye / Q000737:chemistry / Q000166:cytology* / Q000196:embryology; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis; D013049:Spectrin
D017487:Ankyrins; D015820:Cadherins; D002352:Carrier Proteins; D008840:Microfilament Proteins; D000254:Sodium-Potassium-Exchanging ATPase; D013049:Spectrin