Abstract
In many organs the vascular endothelium forms a barrier which impedes the free diffusion of large molecules. The mechanism by which protein hormones are transported through the endothelial cells to reach their target cells is unknown. We have examined the transport of human chorionic gonadotropin (hCG) in rat testicular microvasculature by electron microscopy and by analysing the transfer of radiolabeled hormone and antibodies. Surprisingly, we have observed that the same receptor molecule which is present in target Leydig cells is also involved in transcytosis through the endothelial cells. The hormone was internalized by coated pits and vesicles on the luminal side of the endothelium. It was then localized in the endosomal compartment and subsequently appeared to be delivered by smooth vesicles into the subendothelial space. Moreover, anti-LH/hCG receptor antibodies were efficiently transported via the same system and delivered into the interstitial space. If generalized, these observations may define a new level of modulation of hormone action and may be of importance for drug targeting into the numerous organs which are responsive to the various protein hormones.
Full Text
The Full Text of this article is available as a PDF (4.3 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bomsel M., Mostov K. Role of heterotrimeric G proteins in membrane traffic. Mol Biol Cell. 1992 Dec;3(12):1317–1328. doi: 10.1091/mbc.3.12.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bukovský A., Chen T. T., Wimalasena J., Caudle M. R. Cellular localization of luteinizing hormone receptor immunoreactivity in the ovaries of immature, gonadotropin-primed and normal cycling rats. Biol Reprod. 1993 Jun;48(6):1367–1382. doi: 10.1095/biolreprod48.6.1367. [DOI] [PubMed] [Google Scholar]
- Casanova J. E., Breitfeld P. P., Ross S. A., Mostov K. E. Phosphorylation of the polymeric immunoglobulin receptor required for its efficient transcytosis. Science. 1990 May 11;248(4956):742–745. doi: 10.1126/science.2110383. [DOI] [PubMed] [Google Scholar]
- Duffy K. R., Pardridge W. M. Blood-brain barrier transcytosis of insulin in developing rabbits. Brain Res. 1987 Sep 8;420(1):32–38. doi: 10.1016/0006-8993(87)90236-8. [DOI] [PubMed] [Google Scholar]
- Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
- Geuze H. J., Slot J. W., Strous G. J., Peppard J., von Figura K., Hasilik A., Schwartz A. L. Intracellular receptor sorting during endocytosis: comparative immunoelectron microscopy of multiple receptors in rat liver. Cell. 1984 May;37(1):195–204. doi: 10.1016/0092-8674(84)90315-5. [DOI] [PubMed] [Google Scholar]
- Ghinea N., Eskenasy M., Simionescu M., Simionescu N. Endothelial albumin binding proteins are membrane-associated components exposed on the cell surface. J Biol Chem. 1989 Mar 25;264(9):4755–4758. [PubMed] [Google Scholar]
- Ghinea N., Fixman A., Alexandru D., Popov D., Hasu M., Ghitescu L., Eskenasy M., Simionescu M., Simionescu N. Identification of albumin-binding proteins in capillary endothelial cells. J Cell Biol. 1988 Jul;107(1):231–239. doi: 10.1083/jcb.107.1.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghinea N., Hasu M. Charge effect on binding, uptake and transport of ferritin through fenestrated endothelium. J Submicrosc Cytol. 1986 Oct;18(4):647–659. [PubMed] [Google Scholar]
- Ghinea N., Simionescu N. Anionized and cationized hemeundecapeptides as probes for cell surface charge and permeability studies: differentiated labeling of endothelial plasmalemmal vesicles. J Cell Biol. 1985 Feb;100(2):606–612. doi: 10.1083/jcb.100.2.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghinea N., Vu Hai M. T., Groyer-Picard M. T., Houllier A., Schoëvaërt D., Milgrom E. Pathways of internalization of the hCG/LH receptor: immunoelectron microscopic studies in Leydig cells and transfected L-cells. J Cell Biol. 1992 Sep;118(6):1347–1358. doi: 10.1083/jcb.118.6.1347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghitescu L., Bendayan M. Transendothelial transport of serum albumin: a quantitative immunocytochemical study. J Cell Biol. 1992 May;117(4):745–755. doi: 10.1083/jcb.117.4.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghitescu L., Fixman A., Simionescu M., Simionescu N. Specific binding sites for albumin restricted to plasmalemmal vesicles of continuous capillary endothelium: receptor-mediated transcytosis. J Cell Biol. 1986 Apr;102(4):1304–1311. doi: 10.1083/jcb.102.4.1304. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldenberg D. M. Monoclonal antibodies in cancer detection and therapy. Am J Med. 1993 Mar;94(3):297–312. doi: 10.1016/0002-9343(93)90062-t. [DOI] [PubMed] [Google Scholar]
- Goud B., McCaffrey M. Small GTP-binding proteins and their role in transport. Curr Opin Cell Biol. 1991 Aug;3(4):626–633. doi: 10.1016/0955-0674(91)90033-u. [DOI] [PubMed] [Google Scholar]
- Harding C., Heuser J., Stahl P. Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes. J Cell Biol. 1983 Aug;97(2):329–339. doi: 10.1083/jcb.97.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hashida R., Anamizu C., Kimura J., Ohkuma S., Yoshida Y., Takano T. Transcellular transport of lipoprotein through arterial endothelial cells in monolayer culture. Cell Struct Funct. 1986 Mar;11(1):31–42. doi: 10.1247/csf.11.31. [DOI] [PubMed] [Google Scholar]
- Herzog V. Transcytosis in thyroid follicle cells. J Cell Biol. 1983 Sep;97(3):607–617. doi: 10.1083/jcb.97.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hipkin R. W., Sánchez-Yagüe J., Ascoli M. Agonist-induced phosphorylation of the luteinizing hormone/chorionic gonadotropin receptor expressed in a stably transfected cell line. Mol Endocrinol. 1993 Jul;7(7):823–832. doi: 10.1210/mend.7.7.8413307. [DOI] [PubMed] [Google Scholar]
- Hopkins C. R. Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells. Cell. 1983 Nov;35(1):321–330. doi: 10.1016/0092-8674(83)90235-0. [DOI] [PubMed] [Google Scholar]
- Hoppe C. A., Connolly T. P., Hubbard A. L. Transcellular transport of polymeric IgA in the rat hepatocyte: biochemical and morphological characterization of the transport pathway. J Cell Biol. 1985 Dec;101(6):2113–2123. doi: 10.1083/jcb.101.6.2113. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jain R. K. Delivery of novel therapeutic agents in tumors: physiological barriers and strategies. J Natl Cancer Inst. 1989 Apr 19;81(8):570–576. doi: 10.1093/jnci/81.8.570. [DOI] [PubMed] [Google Scholar]
- Jefferies W. A., Brandon M. R., Hunt S. V., Williams A. F., Gatter K. C., Mason D. Y. Transferrin receptor on endothelium of brain capillaries. Nature. 1984 Nov 8;312(5990):162–163. doi: 10.1038/312162a0. [DOI] [PubMed] [Google Scholar]
- Kemshead J. T., Coakham H. B., Lashford L. S. Clinical experience of iodine-131 monoclonal antibodies in treating neural tumours. Front Radiat Ther Oncol. 1990;24:166–203. doi: 10.1159/000417782. [DOI] [PubMed] [Google Scholar]
- King G. L., Johnson S. M. Receptor-mediated transport of insulin across endothelial cells. Science. 1985 Mar 29;227(4694):1583–1586. doi: 10.1126/science.3883490. [DOI] [PubMed] [Google Scholar]
- Kishimoto T., Tavassoli M. Transendothelial transport (transcytosis) of iron-transferrin complex in the rat liver. Am J Anat. 1987 Mar;178(3):241–249. doi: 10.1002/aja.1001780305. [DOI] [PubMed] [Google Scholar]
- Larson S. M., Carrasquillo J. A., Reynolds J. C. Radioimmunodetection and radioimmunotherapy. Cancer Invest. 1984;2(5):363–381. doi: 10.3109/07357908409040313. [DOI] [PubMed] [Google Scholar]
- Legrain P., Juy D., Buttin G. Rosette-forming cell assay for detection of antibody-synthesizing hybridomas. Methods Enzymol. 1983;92:175–182. doi: 10.1016/0076-6879(83)92017-7. [DOI] [PubMed] [Google Scholar]
- Loosfelt H., Misrahi M., Atger M., Salesse R., Vu Hai-Luu Thi M. T., Jolivet A., Guiochon-Mantel A., Sar S., Jallal B., Garnier J. Cloning and sequencing of porcine LH-hCG receptor cDNA: variants lacking transmembrane domain. Science. 1989 Aug 4;245(4917):525–528. doi: 10.1126/science.2502844. [DOI] [PubMed] [Google Scholar]
- Loosfelt H., Pichon C., Jolivet A., Misrahi M., Caillou B., Jamous M., Vannier B., Milgrom E. Two-subunit structure of the human thyrotropin receptor. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3765–3769. doi: 10.1073/pnas.89.9.3765. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maratos-Flier E., Kao C. Y., Verdin E. M., King G. L. Receptor-mediated vectorial transcytosis of epidermal growth factor by Madin-Darby canine kidney cells. J Cell Biol. 1987 Oct;105(4):1595–1601. doi: 10.1083/jcb.105.4.1595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melan M. A., Sluder G. Redistribution and differential extraction of soluble proteins in permeabilized cultured cells. Implications for immunofluorescence microscopy. J Cell Sci. 1992 Apr;101(Pt 4):731–743. doi: 10.1242/jcs.101.4.731. [DOI] [PubMed] [Google Scholar]
- Milici A. J., Watrous N. E., Stukenbrok H., Palade G. E. Transcytosis of albumin in capillary endothelium. J Cell Biol. 1987 Dec;105(6 Pt 1):2603–2612. doi: 10.1083/jcb.105.6.2603. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moser D. R., Lowe W. L., Jr, Dake B. L., Booth B. A., Boes M., Clemmons D. R., Bar R. S. Endothelial cells express insulin-like growth factor-binding proteins 2 to 6. Mol Endocrinol. 1992 Nov;6(11):1805–1814. doi: 10.1210/mend.6.11.1282670. [DOI] [PubMed] [Google Scholar]
- Mostov K. E., Kraehenbuhl J. P., Blobel G. Receptor-mediated transcellular transport of immunoglobulin: synthesis of secretory component as multiple and larger transmembrane forms. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7257–7261. doi: 10.1073/pnas.77.12.7257. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mostov K. E., Simister N. E. Transcytosis. Cell. 1985 Dec;43(2 Pt 1):389–390. doi: 10.1016/0092-8674(85)90166-7. [DOI] [PubMed] [Google Scholar]
- Omoto E., Minguell J. J., Tavassoli M. Endothelial transcytosis of iron-transferrin in the liver does not involve endosomal traffic. Pathobiology. 1992;60(5):284–288. doi: 10.1159/000163736. [DOI] [PubMed] [Google Scholar]
- Parham P. Immunology. MHC meets mother's milk. Nature. 1989 Jan 12;337(6203):118–119. doi: 10.1038/337118a0. [DOI] [PubMed] [Google Scholar]
- Powers M. R., Bell D. R. Initial equilibration of albumin and IgG in rabbit hind paw skin and lymph. Microvasc Res. 1990 Sep;40(2):230–245. doi: 10.1016/0026-2862(90)90022-j. [DOI] [PubMed] [Google Scholar]
- Rodewald R., Kraehenbuhl J. P. Receptor-mediated transport of IgG. J Cell Biol. 1984 Jul;99(1 Pt 2):159s–164s. doi: 10.1083/jcb.99.1.159s. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rubin S. C. Monoclonal antibodies in the management of ovarian cancer. A clinical perspective. Cancer. 1993 Feb 15;71(4 Suppl):1602–1612. doi: 10.1002/cncr.2820710427. [DOI] [PubMed] [Google Scholar]
- Salesse R., Dacheux F., Genty N., Garnier J. Dual internalization pathway for lutropin and choriogonadotropin in porcine Leydig cells in primary culture. Biol Cell. 1989;66(3):297–306. [PubMed] [Google Scholar]
- Schnitzer J. E., Carley W. W., Palade G. E. Specific albumin binding to microvascular endothelium in culture. Am J Physiol. 1988 Mar;254(3 Pt 2):H425–H437. doi: 10.1152/ajpheart.1988.254.3.H425. [DOI] [PubMed] [Google Scholar]
- Seddiki T., Delpal S., Ollivier-Bousquet M. Endocytosis and intracellular transport of transferrin across the lactating rabbit mammary epithelial cell. J Histochem Cytochem. 1992 Oct;40(10):1501–1510. doi: 10.1177/40.10.1527373. [DOI] [PubMed] [Google Scholar]
- Siminoski K., Gonnella P., Bernanke J., Owen L., Neutra M., Murphy R. A. Uptake and transepithelial transport of nerve growth factor in suckling rat ileum. J Cell Biol. 1986 Nov;103(5):1979–1990. doi: 10.1083/jcb.103.5.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simionescu N., Siminoescu M., Palade G. E. Permeability of muscle capillaries to small heme-peptides. Evidence for the existence of patent transendothelial channels. J Cell Biol. 1975 Mar;64(3):586–607. doi: 10.1083/jcb.64.3.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simionescu N., Simionescu M., Palade G. E. Differentiated microdomains on the luminal surface of the capillary endothelium. I. Preferential distribution of anionic sites. J Cell Biol. 1981 Sep;90(3):605–613. doi: 10.1083/jcb.90.3.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simionescu N., Simionescu M., Palade G. E. Permeability of intestinal capillaries. Pathway followed by dextrans and glycogens. J Cell Biol. 1972 May;53(2):365–392. doi: 10.1083/jcb.53.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Soda R., Tavassoli M. Transendothelial transport (transcytosis) of iron-transferrin complex in the bone marrow. J Ultrastruct Res. 1984 Jul;88(1):18–29. doi: 10.1016/s0022-5320(84)90178-3. [DOI] [PubMed] [Google Scholar]
- Solari R., Schaerer E., Tallichet C., Braiterman L. T., Hubbard A. L., Kraehenbuhl J. P. Cellular location of the cleavage event of the polymeric immunoglobulin receptor and fate of its anchoring domain in the rat hepatocyte. Biochem J. 1989 Feb 1;257(3):759–768. doi: 10.1042/bj2570759. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vasile E., Simionescu M., Simionescu N. Visualization of the binding, endocytosis, and transcytosis of low-density lipoprotein in the arterial endothelium in situ. J Cell Biol. 1983 Jun;96(6):1677–1689. doi: 10.1083/jcb.96.6.1677. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vuhai-Luuthi M. T., Jolivet A., Jallal B., Salesse R., Bidart J. M., Houllier A., Guiochon-Mantel A., Garnier J., Milgrom E. Monoclonal antibodies against luteinizing hormone receptor. Immunochemical characterization of the receptor. Endocrinology. 1990 Nov;127(5):2090–2098. doi: 10.1210/endo-127-5-2090. [DOI] [PubMed] [Google Scholar]
- Williams S. K. Vesicular transport of proteins by capillary endothelium. Ann N Y Acad Sci. 1983;416:457–467. doi: 10.1111/j.1749-6632.1983.tb35205.x. [DOI] [PubMed] [Google Scholar]
- Yoshikawa T., Pardridge W. M. Biotin delivery to brain with a covalent conjugate of avidin and a monoclonal antibody to the transferrin receptor. J Pharmacol Exp Ther. 1992 Nov;263(2):897–903. [PubMed] [Google Scholar]