Abstract
Caveolae are flask-shaped membrane invaginations present in most mammalian cells. They are distinguished by the presence of a striated coat composed of the protein, caveolin. Caveolae have been implicated in numerous cellular processes, including potocytosis in which caveolae are hypothesized to co-localize with folate receptor α and participate in folate uptake. Our laboratory has recently localized folate receptor α to the basolateral surface of the retinal pigment epithelium (RPE). It is present also in many other cells of the retina. In the present study, we asked whether caveolae were present in the RPE, and if so, whether their pattern of distribution was similar to folate receptor α. We also examined the distribution pattern of caveolin-1, which can be a marker of caveolae. Extensive electron microscopical analysis revealed caveolae associated with endothelial cells. However, none were detected in intact or cultured RPE. Laser scanning confocal microscopical analysis of intact RPE localized caveolin-1 to the apical and basal surfaces, a distribution unlike folate receptor α. Western analysis confirmed the presence of caveolin-1 in cultured RPE cells and laser scanning confocal microscopy localized the protein to the basal plasma membrane of the RPE, a distribution like that of folate receptor α. This distribution was confirmed by electron microscopic immunolocalization. The lack of caveolae in the RPE suggests that these structures may not be essential for folate internalization in the RPE.
Similar content being viewed by others
References
Anderson RG, Kamen BA, Rothberg KG, Lacey SW (1992) Potocytosis: sequestration and transport of small molecules by caveolae. Science 255: 410–411.
Anderson RGW (1993) Caveolae: Where incoming and outgoing messengers meet. Proc Natl Acad Sci USA 90: 10909–10913.
Bretscher MS, Whytock S (1977) Membrane-associated vesicles in fibroblasts. J Ultrastruct Res 61: 215–217.
Chancy CD, Kekuda R, Huang W, Prasad PD, Kuhnel, J-M, Sirotnak, FM, Roon P, Ganapathy V, Smith SB (2000) Expression and differential polarization of the reduced-folate transporter-1 and the folate receptor α in mammalian retinal pigment epithelium. J Biol Chem 275: 20676–20684.
Dunn KC, Aotaki-Keen E, Putkey FR, Hjelmeland LM (1996) ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. Exp Eye Res 62: 155–169.
Dunn KC, Marmorstein AD, Bonilha VL, Rodriquez-Boulan E, Giordano F, Hjelmeland LM (1998) Use of ARPE-19 cell line as a model of RPE polarity; basolateral secretion of FGF5. Invest Ophthalmol Vis Sci 39: 2744–2749.
Feng Y, Venema VJ, Venema RC, Tsai N, Behzadian MA, Caldwell RB (1999) VEG-F-induced permeability increase is mediated by caveolae. Invest Ophthalmol Vis Sci 40: 157–167.
Finnemann SC, Marmorstein AD, Neill JM, Rodriquez-Boulan E (1997) Identification of the retinal pigment epithelium protein RET-PE2 as CE-9/OX-47, a member of the immunoglobin superfamily. Invest Ophthalmol Vis Sci 38: 2366–2374.
Gardiner TA, Archer DB (1986a) Does unidirectional vesicular transport occur in retinal vessels? Brit J Ophthalmol 70: 249–254.
Gardiner TA, Archer DB (1986b) Endocytosis in the retinal and choroidal microcirculation. Brit J Ophthalmol 70: 361–372.
Gundersen D, Orlowski J, Rodriquez-Boulan E (1991) Apical polarity of Na,K-ATPase in retinal pigment epithelium is linked to a reversal of the ankyrin-fodrin submembrane cytoskeleton. J Cell Biol 112: 863–872.
Heth CA, Yankauckas MA, Adamian M, Edwards RB (1987) Characterization of retinal pigment epithelial cells cultured on microporous filters. Curr Eye Res 6: 1007–1019.
Huang W, Prasad PD, Kekuda R, Leibach FH, Ganapathy V (1997) Characterization of the N5-methyltetrahydrofolate uptake in cultured human retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 38:1578–1587.
Islam M, Akhtar RA (2000) Epidermal growth factor stimulates phospholipase cgamma1 in cultured rabbit corneal epithelial cells. Exp Eye Res 70: 261–269.
Kleinman HK, Cannon FB, Laurie GW, Hassell Jr, Aumailley M, Terronova VP, Martin GR, Dubois-Dalcq M (1985) Biological activities of laminin. J Cell Biochem 27: 317–325.
Kono T, Sorgente N, Patterson R, Ryan SJ (1983) Fibronectin and laminin distribution in bovine eyes. Jpn J Ophthalmol 27: 496–505.
Lacey SW, Sanders SM, Rothberg KG, Anderson RGW, Kamen BA (1989) Complementary DNA for the folate binding protein correctly predicts anchoring to the membrane by glycosyl-phosphatidylinositol. J Clin Invest 84: 715–720.
Laemmli UK (1970) Cleavage of structural properties during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Lee HC, Shoda R, Krall JA, Foster JD, Selhub J, Rosenberry TL (1992) Folate binding protein from kidney brush border membranes contains components characteristic of a glycoinositol phospholipid anchor. Biochemistry 31: 3236–3243.
Lowry OH, Rosebrough NJ, Farr AL, Randal RJ (1951) Protein measurements with Folin phenol reagent. J Biol Chem 193: 265–275.
Mayor S, Rothberg KG, Maxfield FR (1994) Sequestration of GPIanchored proteins in caveolae triggered by cross-linking. Science 264: 1948–1951.
Mcgarvey ML, Baron Van Evercooren A, Kleinman HK, Dubois-Dalcq M (1984) Synthesis and effects of basement membrane components in cultured rat schwann cells. Develop Biol 105: 18–28.
Miller SS, Steinberg RH, Oakley Jr. B (1978) The electrogenic sodium pump of the frog retinal pigment epithelium. J Membr Biol 44: 259–279.
Montesano R, Roth J, Robert A, Orci L (1982) Non-coated membrane invaginations are involved in binding and internalization of cholera and tetanus toxin. Nature 296: 651–653.
Okami T, Yamamoto A, Omori K, Takada T, Uyama M, Tashiro Y (1990) Immunocytochemical localization of Na+K+-ATPase in rat retinal pigment epithelial cells. J Histochem Cytochem 38: 1267–1275.
Ostwald TJ, Steinberg RH (1980) Localization of frog retinal pigment epithelium Na+-K+ ATPase. Exp Eye Res 31: 351–360.
Palade GE (1953) Fine structure of blood capillaries. J Appl Phys 24: 1424.
Palade GE, Bruns RR (1968) Structural modulations of plasmalemmal vesicles. J Cell Biol 37: 633–649.
Peters KR, Carley WW, Palade GE (1985) Endothelial plasmalemmal vesicles have a characteristic striped bipolar surface structure. J Cell Biol 101: 2233–2238.
Quinn RH, Miller SS (1992) Ion transport mechanisms in native human retinal pigment epithelium. Invest Ophthalmol Vis Sci 33: 3513–3527.
Ratnam M, Marquardt H, Duhring JL, Freisheim JH (1989) Homologous membrane folate binding proteins in human placenta: cloning and sequence of a cDNA. Biochemistry 28: 8249–8254.
Ratnam M, Freisheim JH (1992) Proteins involved in the transport of folates and antifolates by normal and neoplastic cells. In: Folate Metabolism in Health and Disease. (edited by Picriano MF), pp. 91–120. New York: Wiley-Liss.
Rothberg KG, Ying Y, Kolhouse JF, Kamen BA, Anderson RGW (1990) The glycophospholipid-linked folate receptor internalizes folate without entering the clathrin-coated pit endocytic pathway. J Cell Biol 110: 637–649. 158 C.C. Bridges et al.
Rothberg KG, Heuser JE, Donzell WC, Ying Y, Glenney JR, Anderson RGW (1992) Caveolin, a protein component of caveolae membrane coats. Cell 68: 673–682.
Scheiffele P, Verkade P, Fra AM, Virta H, Simons K, Ikonen E (1998) Caveolin-1 and-2 in the exocytic pathway of MDCK cells. J Cell Biol 140: 795–806.
Schlegel A, Volonte D, Engelman JA, Galbiati F, Mehta P, Zhang X-L, Scherer PE, Lisanti MP (1998) Crowded little caves: structure and function of caveolae. Cell Signal 10: 456–463.
Sirotnak FM, Tolner B (1999) Carrier-mediated membrane transport of folates in mammalian cells. Ann Rev Nutr 19: 91–122.
Smart EJ, Foster DC, Ying YS, Kamen BA (1994) Protein kinase C activators inhibit receptor-mediated potocytosis by preventing internalization of caveolae. J Cell Biol 124: 307–313.
Smith SB, Kekuda R, Gu X, Chancy C, Conway SJ, Ganapathy V (1999a) Expression of folate receptor a in the mammalian retinal pigment epithelium and retina. Invest Ophthalmol Vis Sci 40: 840–848.
Smith SB, Huang W, Chancy C, Ganapathy V (1999b) Regulation of the reduced-folate transporter by nitric oxide in cultured human retinal pigment epithelial cells. Biochem Biophys Res Comm 257: 279–283.
Somlyo AP, Devine CE, Somlyo AB, North SR (1971) Sarcoplasmic reticulum and the temperature-dependent contraction of smooth muscle in calcium-free solutions. J Cell Biol 51: 722–741.
Sramek S, Wallow IHS, Bindley C, Sterken G (1985) Fibronectin in the rat eye. Invest Ophthalmol Vis Sci Suppl 26:330.
Sugrue SP, Hay ED (1981) Response of basal epithelial cell surfaces and cytoskeleton to solubilized extracellular matrix molecules. J Cell Biol 91: 45–54.
Sun, X-L, Antony AC (1996) Identification of an 18-base cis-element in the 5_-untranslated region of human folate receptor-α mRNA which specifically binds 46-kDa cystosolic (trans-factor) proteins. J Invest Med 44: 203A.
Timpl R, Rhode H, Robey PG, Rennard SI, Foidart J-M, Martin GR (1979) Laminin – a glycoprotein from basement membranes. J Biol Chem 254: 9933–9937.
Turksen K, Aubin JE, Sodek J, Kalnins VI (1985) Localization of laminin, type IV collagen, fibronectin, and heparan sulfate proteoglycan in chick retinal pigment epithelium basement membrane during embryonic development. J Histochem Cytochem 33: 665–671.
Verma RS, Gullapalli S, Antony AC (1992) Evidence that the hydrophobicity of isolated, in situ, and de novo-synthesized native human placental folate receptors is a function of glycosyl-phosphatidylinositol anchoring to membranes. J Biol Chem 267: 4119–27.
Vladovsky I, Gospodarowicz D (1981) Respective roles of laminin and fibronectin in adhesion of human carcinoma and sarcoma cells. Nature 289: 304–306.
Vogel U, Sandvig K, Van Deurs B (1998) Expression of caveolin-1 and polarized formation of invaginated caveolae in Caco-2 and MDCK II cells. J Cell Sci 111: 825–832.
Wu M, Fan J, Gunning W, Ratnam M (1997) Clustering of GPI-anchored folate receptor independent of both cross-linking and association with caveolae. J Memb Biol 159: 137–147.
Yamada E (1955) The fine structure of the gall bladder epithelium of the mouse. J Biophys Biochem Cytol 1: 455–458.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bridges, C.C., El-Sherbeny, A., Roon, P. et al. A Comparison of Caveolae and Caveolin-1 to Folate Receptor α in Retina and Retinal Pigment Epithelium. Histochem J 33, 149–158 (2001). https://doi.org/10.1023/A:1017991925821
Issue Date:
DOI: https://doi.org/10.1023/A:1017991925821