ebook img

Retinoids: Cell Differentiation and Clinical Applications Part B PDF

488 Pages·1990·11.622 MB·English
Save to my drive
Quick download
Download
Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.

Preview Retinoids: Cell Differentiation and Clinical Applications Part B

Methods in Enzymology Volume 190 Retinoids Part B Cell Dlrerentiation and Clinical Applications EDITED BY Lester Packer DEPARTMENT OF MOLECULAR AND CELL BIOLOGY UNIVERSITY OF CALIFORNIA, BERKELEY BERKELEY. CALIFOKNIA Editorial Advisory Board Frank Chytil Leonard M&tone Dewitt Goodman Concetta Nicotra Maria A. Livrea James A. Olson Stanley S. Shapiro AP 0 ACADEMIC PRESS, INC. Harcourl Brace Jovanovich, Publishers San Diego New York Boston London Sydney Tokyo Toronto Preface Spectacular progress and unprecedented interest in the field of retinoids prompted us to consider this topic for two volumes in the Methods ni Enzymology series: Volume 189, Retinoids, Part A: Molecular and Meta- bolic Aspects and Volume 190, Retinoids, Part B: Cell Differentiation and Clinical Applications. From a historical perspective we know that studies in the 1930s showed that vitamin A (retinol) and retinal had a role in the visual process. It was also recognized that some link between vitamin A and cancer incidence existed. Several decades ago it was discovered that retinoic acid had a dramatic effect on the chemically induced DMBA mouse skin carcinogen- esis model in which enormous reductions in the tumor burden were ob- served. This led to the realization that retinoids had important effects on cell differentiation. This resulted almost immediately in the synthesis and evaluation of new retinoids. Indeed, the effects of retinoids on cell differ- entiation appear to be more universal and of greater importance than their light-dependent role in vision and microbial energy transduction. Progress has been rapid, and the importance of accurate methodology for this field is imperative to its further development. The importance of methodology applies to the use of retinoids in basic research in molecular, cellular, and developmental biology, and in clinical medicine. In medicine, applications have been mainly to cancer and in dermatology to the treat- ment of skin diseases and skin aging. As new retinoids are being tested in biological models and in clinical medicine, interest in the nutrition and pharmacology of retinoids has arisen. Moreover, the beneficial effects of retinoids in pharmacological treatment have led to a recognition of the "double-edged sword" of toxicity (teratogenicity). In Section I of this volume, Cell Differentiation, the effects of retinoids in various cell differentiation systems are covered. Many new systems in which retinoids exhibit their effects have been employed. Both normal diploid cells and cell lines ni vitro have been used, and the methods and systems employed are presented. In addition, tissue and organ culture are important areas for retinoid methodology. The effects of retinoids as mor- phogens and teratology agents are also included. In Sections II, Nutrition, Tissue and Immune Status, and Antioxidant Action, and III, Pharmacoki- netics, Pharmacology, and Toxicology, nutritional and pharmacological methods are presented. Retinoids in the treatment of skin disease and in cancer chemotherapy are probably the most important areas in which methodological developments have occurred. New methodology has also iiix xiv ECAFERP revealed the antioxidant activity of retinoids, and since any antioxidant may also be a pro-oxidant such considerations may be important for clinical pharmacology and therapeutics. Volume 981 covers structure and analysis, receptors, transport, and binding proteins, and enzymology and metabolism. I am very grateful to the Advisory Board--Frank Chytil, DeWitt Goodman, Mafia A. Livrea, Leonard Milstone, Concetta Nicotra, James A. Olson, and Stanley S. Shapiro m for their unique input, advice, counsel, and encouragement in the planning and organization of this volume. In most instances, I met with every member of the board on one or more occasions to discuss the topics and to identify the most important contribu- tors. Indeed, we found almost universal acceptance, and virtually no one turned down our invitation to contribute to this volume. In fact it was somewhat autocatalytic in that many contributors, realizing the timeliness and significance of having all of the methods dealing with retinoids in- cluded, made suggestions for additional contributions which were evalu- ated by the board. In a few instances we may have been somewhat over- zealous, and more than one article on a method has been included. We do apologize for this slight redundancy for the sake of completeness. RETSEL REKCAP Contributors to Volume 190 elcitrA numbers are in sesehtnerml gniwollof the names of .;notubirtnoc snoitallfffA listed are current. SERGIO ADAMO (9), Department of Experi- ROSERT J. ALOTEPAC (37), Research Labo- mental Medicine, University of L'Aquila, ratories, The R. W. Johnson Pharmaceu- 67100 L ,aliuqA" Italy tical Research Institute, Raritan, New BRAD AMOS (23), Department of Tumor Bi- Jersey 08869 ology, M. D. Anderson Cancer Center, WERONA L. DARROW (12), Department of University of Texas, Houston, Texas Molecular Pharmacology, State University 77030 of New York at Stony Brook, Stony Brook, EVA ANDERSSON (18), Department of Der- New York 11794 matology, University Hospital, S-581 85 LinkOping, Sweden WILLEM J. DE GRIP (1), Center of Eye Re- search Nijmegen, University of Nijmegen, ADRIANNE BENDICH (27), Department of 6500 HB Nijmegen, The Netherlands Clinical Nutrition, Hoffmann-La Roche Inc., Nutley, New Jersey 07110 Luioi M. DE LUCA (9, 10), National Cancer TROND BERG (6), Institute for Nutrition Re- Institute, Bethesda, Maryland 20892 search, University of Oslo, N-0316 Oslo ,3 SAMOHT I. DORAN (35), Preclinical Re- Norway search, Roche Dermatologics, Hoffmann- RUNE FFOHMOLB (6), Institute for Nutrition La Roche Inc., Nutley, New Jersey 07110 Research, University of Oslo, N-0316 Oslo ,3 Norway R. TRELHE (3 l), Department of Dermatology and Venereology, University Medical NEVETS A. BOOK (43), Health and Welfare Center Steglitz, The Free University of Agency, State of California, Sacramento, Berlin, Berlin, Federal Republic of Ger- California 95814 many D. LUCILLE BREDBERG (17), Department of Ophthalmology, University of Washing- GREGOR EICHELE (21), Department of Cel- ton, Seattle, Washington 98195 lular and Molecular Physiology, Harvard ERODOEHT R. BREITMAN (13), Laboratory Medical School, Boston, Massachusetts 02115 of Biological Chemistry, Division of Cancer Treatment, National Cancer Insti- DENISE A. FAHERTY (27), Department of tute, National Institutes of Health, Be- Immunopharmacology, Hoffmann-La thesda, Maryland 20892 Roche Inc., Nutley, New Jersey 07110 C. D. B. BRIDGES (15), Department of Bio- SHAO-LING FONG (15), Departments of Oph- logical Sciences, Purdue University, West thalmology and Biochemistry and Molecu- Lafayette, Indiana 47907 lar Biology, Indiana University, Indianap- ECNATSNOC E. FFOHREKCNIRB (19), Depart- olis, Indiana 46202 ment of Medicine and Biochemistry, Dart- mouth Medical School, Hanover, New ELAINE V. FUCHS (2), Department of Molec- Hampshire 03756 ular Genetics and Cell Biology, Howard Hughes Medical Institute, University of ADRIAAN BROUWER (5), TNO Institute for Chicago, Chicago, Illinois 60637 Experimental Gerontology, 2280 HV Rijs- wijk, The Netherlands GERARD J. GENDIMENICO (37), Research GRAEME F. BRYCE (38), Roche Dermatolo- Laboratories, The R. W. Johnson Phar- gics, Hoffmann-La Roche Inc., Nutley, maceutical Research Institute, Raritan, New Jersey 07110 New Jersey 08869 ix X CONTRIBUTORS TO VOLUME 190 TERAGRAM A. GEORGE (4), Cell Biology ,hcnarB National Institutes of Health, Be- ,noitceS Laboratory of Pulmonary -ohtaP ,adseht Maryland 29802 ,ygoloib National Institute of -nemnorivnE SEMAJ YELRUH (34), Business ,tnempoleveD tal Health Sciences, Research Triangle Roche ,scigolotamreD Hoffmann-La Roche Park, North aniloraC 27709 Inc., Nutley, New Jersey 01170 EGRO~G J. GIUDlCE (2), Department of -reD NOTNA M. NETTEJ (4), Cell Biology ,noitceS ,ygolotam Medical College of ,nisnocsiW Laboratory of Pulmonary ,ygoloibohtaP Milwaukee, Wisconsin 62235 National Institute of Environmental H. KCINLLOG (31), Department of -lotamreD Health ,secneicS Research Triangle Park, ogy and ,ygoloereneV University Medical North aniloraC 27709 Center Steglitz, The Free University of ECILA F. KARL (7), Department of -mehcoiB Berlin, Berlin, Federal Republic of -reG istry, Washington State ,ytisrevinU Pull- many man, Washington 46199 ENNAOJ REMLAB GREEN (32), Nutrition -eD IMEGIHS KATO (9), St. Marianna ytisrevinU partment, Pennsylvania State ,ytisrevinU School of Medicine, Miyamae-Ku, Kawa- ytisrevinU Park, Pennsylvania 20861 sam 213, Japan LEAHCIM H. GREEN (32), Nutrition -trapeD ECNERET I~ALEY (36), Department of -nilC ment, Pennsylvania State ,ytisrevinU -inU ical Biochemistry, Cambridge ,ytisrevinU versity Park, Pennsylvania 20861 Addenbrooke's Hospital, Cambridge CB2 HPESOJ F. GRIPPO (16), Department of 2QR, England ygolocixoT and ,ygolohtaP Hoffmann-La SAERDNA RELTSIK (44, 45, 46), Clinical Re- Roche Inc., Nutley, New Jersey 01170 ,hcraes .F Hoffmann-La Roche Ltd., CH- LEAHCIM D. DLOWSIRG (7), Department of 4002 Basel, Switzerland Biochemistry, Washington State -revinU ENIARROL H. NAMGILK (40), Department of sity, Pullman, Washington 46199 ,ygolotamreD University of ainavlysnneP GARY L. GROVE (39), KGL's Skin Study School of ,enicideM ,aihpledalihP -lysnneP ,retneC Broomall, Pennsylvania 80091 vania 40191 MARY Jo GROVE (39), KGL's Skin Study DICK L. KNOOK (5), TNO Institute for Ex- ,retneC Broomall, Pennsylvania 80091 perimental ,ygolotnoreG 2280 HV Rijs- ENIARROL J. GUDAS (14), Department of wijk, The sdnalrehteN lacigoloiB Chemistry and Molecular -rahP ARDNEVED M. RAHHCOK (33), Department ,ygolocam Harvard Medical School and of Anatomy, Thomas Jefferson ,ytisrevinU Dana-Farber Cancer Institute, Boston, ,aihpledalihP Pennsylvania 70191 Massachusetts 51120 DAFNA LOTAN (11), Department of Tumor HENK F. J. SKIRDNEH (5), TNO Institute for ,ygoloiB M. .D Anderson Cancer ,retneC Experimental ,ygolotnoreG 2280 HV Rijs- University of Texas, Houston, Texas wijk, The sdnalrehteN 03077 MIDORI USTAMARIH (29), Department of NEBUER LOTAN (11, 23), Department of ,yrtsimehcorueN Institute for -loiborueN Tumor ,ygoloiB M. .D Anderson Cancer ,ygo Okayama University Medical ,loohcS Center, University of Texas, Houston, Okayama ,007 Japan Texas 03077 W. BRIAN DRAWOH (44, 45, 46), La Jolla MLOCLAM MADEN (20), Anatomy and Cancer Research Foundation, La Jolla, Human Biology Group, Kings ,egelloC ainrofilaC 73029 London WC2R 2LS, England AISEERF L. HUANG (10), National Institute ECNERWAL J. TTENRAM (30), Department of of Child Health and Human ,tnempoleveD ,yrtsimehcoiB tlibrednaV ytisrevinU Medi- ygolonircodnE and noitcudorpeR hcraeseR cal ,retneC Nashville, Tennessee 23273 CONTRIBUTORS TO VOLUME 190 xi JOhN L. McGuw.~ (37), Research Laborato- Berlin, Berlin, Federal Republic of Ger- ries, The R. W. Johnson Pharmaceutical many Research Institute, Raritan, New Jersey OLA ROLLMAN (18), Department of Derma- 08869 tology, University Hospital, S-751 85 Upp- RAJENDRA G. MEHTA (42), Laboratory of sala, Sweden Pathophysiology, HT Research Institute, DENNIS R. POOR (10), tnemtrapeD of Cell Chicago, Illinois 60616 Biology, Baylor College of Medicine, SEMAJ A. MEZICK (37), Research Laborato- Houston, Texas 77030 ries, The R. W. Johnson Pharmaceutical MARVIN E. ROSENTHALE (37), Research Research Institute, Raritan, New Jersey Laboratories, The R. W. Johnson Phar- 08869 maceutical Research Institute, Raritan, DRANOEL M. MILSTONE (8), Department of New Jersey 08869 Dermatology, Yale University School of Medicine, New Haven, Connecticut 06510 A. ENIRAHTAC SSOR (28), tnemtrapeD of Physiology and Biochemistry, Division of RICHARD C. MOON (42), Laboratory of Nutrition, Medical College of Pennsylva- Pathophysiology, HT Research Institute, nia, Philadelphia, Pennsylvania 19129 Chicago, Illinois 60616 JOHN C. SAARI (17), Department of Ophthal- HEINZ NAU (47), Institute of Toxicology and mology, University of Washington, Seattle, Embryopharmacology, The Free Univer- Washington 98195 sity of Berlin, D-IO00 Berlin ,33 Federal Republic of Germany RETEP G. SKCAS (11), Department of Tumor NEELHTAK M. NAUSS (28), Health Effects Biology, M. D, Anderson Cancer Center, Institute, Cambridge, Massachusetts University of Texas, Houston, Texas 02139 77030 C. E. ORFANOS (3 l), Department of Derma- ROTCIV M. NYZSYKOMAS (30), Department tology and Venereology, University Medi- of Pharmaceutical Chemistry, School of cal Center Steglitz, The Free University of Pharmacy, University of California, San Berlin, Berlin, Federal Republic of Ger- Francisco, San Francisco, California many 94143 RETSE, REKCAP (29), Department of Molec- YELNATS S. SHAPIRO (34, 35, 38), Preclini- ular and Cell Biology, University of Cali- cal Research, Roche Dermatologics, fornia, Berkeley, Berkeley, California Hoffmann-La Roche Inc., Nutley, New 94720 Jersey 07110 MARIA CENOP (3), Department of Dermatol- MICHAEL I. NAMREHS (16), Department of ogy, University Hospital Leiden, 2300 RC Cell Biology, Hoffmann-La Roche Inc., Leiden, The Netherlands Nutley, New Jersey 07110 JAMES I. REARICK (4), Department of Bio- HTENNEK L. SIMPSON (25), Food Science chemistry, Kirksville College of Osteo- and Nutrition Research Center, West pathic Medicine, Kirksville, Missouri Kingston, Rhode Island 02892 63501 RICHARD J. RICKLES (12), Department of JOHN EDGAR SMITH (24), Nutrition Depart- Biochemistry and Molecular Biology, Har- ment, Pennsylvania State University, Uni- vard University, Cambridge, Massachu- versity Park, Pennsylvania 16802 setts 02138 DAVID L. MUCOTS (20), Department of Biol- G. RINCK (31), Department of Dermatology ogy, Indiana University-Purdue University and Venereology, University Medical at Indianapolis, Indianapolis, Indiana Center Steglitz, The Free University of 46205 xii CONTRIBUTORS TO VOLUME 190 SIDNEY STRICKLAND (12), Department of ,yrtsimehc Medical College of ,nisnocsiW raluceloM ,ygolocamrahP State ytisrevinU Milwaukee, Wisconsin 62235 of New York at Stony Brook, Stony ,koorB ARABRAB A. DOOWREDNU (26), National New York 49711 Eye Institute, National Institutes of KAMPE TEELMANN (41), Pharmaceutical Health, Bethesda, Maryland 29802 ,noisiviD .F Hoffmann-La Roche Ltd., SREDNA TSIUQLHAV (18, 22), Department of 2004-HC Basel, Switzerland ,ygolotamreD University Hospital S-581 ANITSIRHC THALLER (21), Department of 58 ,gnipOkniL Sweden ralulleC and Molecular ,ygoloisyhP Har- HARAS WEDDEN (21), Department of Anat- vard Medical School Boston, Massachu- omy, University Medical School, Edin- setts 51120 hgrub EH8 ,GA9 Scotland ADRIAN M. TIMMERS (1), Department of ARIJ MIEHREEW (3), Department of -amreD ,ygolomlahthpO University of ,adirolF ,ygolot University Hospital Leiden, 2300 ,ellivseniaG Florida 01623 RC Leiden, The sdnalrehteN HANS TORM.~ (18, 22), Department of -reD CALVIN C. WILLHITE (43), Toxic secnatsbuS ,ygolotam University Hospital, S-581 58 Control Program, Department of Health ,gnipOkniL Sweden Services, State of California, Berkeley, YLLAS S. TWINING (28), Department of -oiB ainrofilaC 01749 1 SHORT-TERM INCUBATION OF ISOLATED RPE CELLS 3 1 Analysis of the Visual Cycle by Short-Term Incubation of Isolated Retinal Pigment Epithelial Cells By ADRIAN M. SREMMIT and MELLIW J. DE GRIP Introduction The recycling of retinoids in retina and retinal pigment epithelium (RPE), effectuating the regeneration of bleached visual pigment to the photoactive state, is called the visual cycle. The trigger reaction for the process of vision as well as the visual cycle is photoinduced isomerization (11-cis to all- trans) of the chromophore of vertebrate visual pigments) Major parts of the pathway of the visual cycle remained obscure for decades. Only re- cently has insight been obtained into some of the long-standing enigmas: which type of retinoid is isomerized where and how and transported in what form to the outer segments to eventually regenerate the visual pig- merit, rhodopsin. 2 This regeneration pathway is initiated in the RPE where all.trans-refinol is enzymatically isomerized to 11-cis-retinol followed later by conversion to 11-cis-retinaldehyde. The isomerization reaction is driven by the free energy of hydrolysis of the retinyl ester? Retinoid metabolism in the RPE, with respect to the visual cycle, encompasses several steps: uptake of retinol, intracellular transport, acyla- tion, isomerization, oxidation, and secretion of retinoid. In order to study such a complex set of metabolic pathways in RPE cells, the development of a reliable in vitro system would be highly desirable. Investigations on the multifaceted retinoid metabolism in RPE cells in vitro require isolated RPE cells in which the complexity of cellular organization is preserved to a high extent and which are viable and metabolically active? In order to meet these requirements of physiological fitness, we optimized the isola- tion of bovine RPE cells and the in vitro incubation conditions by applying a variety of criteria. These criteria included morphology (ultrastructure of the cells), viability (exclusion of viability stains and retention of small cellular proteins), and metabolic activity (energy charge). Here we describe an approach to study retinoid metabolism in isolated bovine RPE cells during short-term in vitro incubation. t G. Wald, Nature (London) 219, 800 (1968). 2 p. S. Bemstein, C. W. Law, and R. R. Rando, .corP Natl. Acad. Sci. U.S.A. 84, 1849 (1987). 3 R. R. Rando, J. Canada, P. S. Deigner, and C. W. Law, Invest. Ophthalmol. Visual Sci. 30, 331 (1989). 4 A. M. M. Timmers, W. J. De Grip, and F. J. M. Daemen, in "Proceedings on Retinal Proteins" (N. G. Abdulaev and Y. A. Ovchinnikov, eds.), p. 381. VNU Science Press, Utrecht, 1987. thgirypoC © 0991 yb ¢im~l~A ~ .cnI SDOHTEM NI ,YGOLOMYZNE .LOV 091 AllfishtsofreProducfionin~ay mrof t~xl. 4 LAMRON SLLEC 1 Isolation of Viable Bovine Retinal Pigment Epithelial Cells Principle Isolation of intact RPE cells requires release of cells from their intimate contact with both retina and BrOchs membrane. Strong reduction of the attachment of RPE to retina and BrOchs membrane is achieved by perfu- sion of the intact bovine eye through the central ophthalmic artery with a divalent cation-free isotonic salt buffer (perfusion buffer; Ca ,+2 Mg2+-free Hanks-EDTA s) kept at 0 6.° A high recovery of RPE cells which are not contaminated by either red blood cells or rod outer segments is obtained after 12-18 min ofperfusion. The yield of 1-2 × 601 RPE cells per bovine eye represents 20-40% of the total RPE cell population. Furthermore, over 80% of the cells exclude didansylcystine (viability stain), 6 and 85% of the cellular retinol-binding protein (CRBP) is retained in the cells as assayed with the Lipidex 1000 binding assay.7 The ultrastructure of the isolated cells is very well preserved (Fig. .)1 The yield, purity, and integrity of this RPE cell population meet the high standards for in vitro studies. Procedure Bovine eyes are enudeated 20 min after the death of the animal; the optic nerve is kept at least 2 cm long. The eyes are immediately transported in a fight-tight container to the laboratory, where excess fat and muscle are trimmed off under dim red light. The eyes are wrapped in aluminum foil, leaving the optic nerve accessible, and kept in a dark container at 8-10 °. The central ophthalmic artery is cannulated with a blunt 21-gauge needle connected to a reservoir of perfusion buffer (137.8 mM NaCI; 5.4 mM KC1; 0.3 mM Na,zHPO4; 0.4 mM KH2PO4; 2 mM EDTA; 5.5 mM glucose; 10 mM HEPES, pH 7.4) positioned 100- 120 cm above the eye (Fig. 2). The central ophthalmic artery, which supplies the entire eye and runs along the optic nerve, can be readily identified by its translucent white color and the blood clot at its end. Further differentiation from fat tissue is achieved by pulling it gently with forceps; this ruptures fat tissue but not the artery. The artery is grasped with two small forceps (5SA, Technical Tools, Rotterdam), pulled over the blunt needle, and tied with a suture. The wrapped eye is peffused with ice-cold peffusion buffer for 13- 71 min at a flow rate of 0.5-1 ml/min. Routinely, eyes are processed within 2 hr 5 j. Heller and P. Jones, Exp. Eye Res. 30, 481 (1980). 6 A. M. M. Timmers, E. A. Dratz, W. J. De Grip, and F. J. M. Daemen, Invest. Ophthalmol. Visual Sci. 25, 1013 (1984). 7 A. M. M. Timmers, W. A. H. M. van G-roningen-Luyben, F. J. M. Daemen, and W. J. De Grip, .J LipidRes. 27, 979 (1986).

See more

The list of books you might like

Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.