44 I ).46 I.1 )Echantillons utilisés, .47 I.1.1.2 )PCR Allèle, p.49 ,
50 II.1.1 )Isolements des glomérules51 II.1.2 )Coloration Immuno-Histo-Fluorescence des glomérules52 II.1.4 )Dosage des protéines glomérulaires52 II.1.5 )Co-précipitation des protéines, 50 II.1 )57 III ) PREPARATION D'UN NOUVEL «APPÂT» Lu/BCAM PAR CLONAGE.........................57 III.1 )Préparation de la matrice, p.60 ,
61 III.2.1 )Préparation de l'insert61 III.2.2 )Préparation des vecteurs N3C, III.2 )Préparation de l'insert des plasmides contenant la partie extracellulaire non mutée (N3C) et mutante, p.65 ,
66 IV.1 )Extraction de plasmides66 IV.2 )Contrôle de la mini-préparation d, ADN : vecteur + insert, p.68 ,
73 VII.1 )Western Blot, 70 VII ), p.74 ,
76 I )76 II )77 III ), 80 III.2 )REVELATION82 III.3 )REVELATION, p.83 ,
87 VI.1 )ANALYSE, p.90 ,
91 VII.1 ), p.92 ,
à l'aide de l'anticorps anti laminine ?5 ultra-purifié (Ac L85 P52) en conditions non réduites et réduites (Figure 36) On obtient le résultat inverse par rapport aux cellules murines bEnd.3. La présence d'une bande à 400kDa en conditions non réduites, mais absente en conditions réduites. Tous ces résultats confirment que nous n'avons pas co-précipité la laminine ,
CD44 is the principal cell surface receptor for hyaluronate, Cell, vol.61, issue.7, pp.1303-1313, 1990. ,
DOI : 10.1016/0092-8674(90)90694-A
A simplified laminin nomenclature, Y and Yurchenco PD. 2005. A simplified laminin nomenclature, pp.326-332 ,
DOI : 10.1016/j.matbio.2005.05.006
The role of laminins in basement membrane function, Journal of Anatomy, vol.193, issue.1, pp.1-21, 1998. ,
DOI : 10.1046/j.1469-7580.1998.19310001.x
Glomerular Endothelial Glycocalyx, Contrib Nephrol, vol.68, pp.104-113, 1988. ,
DOI : 10.1159/000416500
Endothelial characteristics of glomerular capillaries in normal, mercuric chloride-induced, and gentamicin-induced acute renal failure in the rat., Journal of Clinical Investigation, vol.72, issue.1, pp.128-141, 1983. ,
DOI : 10.1172/JCI110950
Molecular structure of the extracellular region of Lutheran blood group glycoprotein and location of the laminin binding site, Blood Cells, Molecules, and Diseases, vol.40, issue.3, pp.446-448, 2008. ,
DOI : 10.1016/j.bcmd.2008.01.004
Hypersensitivity to Transfused Blood, BMJ, vol.2, issue.4411, p.83, 1945. ,
DOI : 10.1136/bmj.2.4411.83
Molecular cloning of the B-CAM cell surface glycoprotein of epithelial cancers: a novel member of the immunoglobulin superfamily, Cancer Res, vol.54, pp.5761-5765, 1994. ,
Aggregation of mononuclear and red blood cells through alpha4beta1 Lu/BCAM interaction in sickle cell disease, Haematologica, 2010. ,
The expected blood-group antibody, anti-Lub, Nature, vol.178, pp.855-856, 1956. ,
Permselectivity of the glomerular capillary wall to macromolecules. I. Theoretical considerations, Biophysical Journal, vol.15, issue.9, pp.861-886, 1975. ,
DOI : 10.1016/S0006-3495(75)85862-0
Red cell and endothelial Lu/BCAM beyond sickle cell disease, Transfusion Clinique et Biologique, vol.15, issue.6, pp.402-405, 2008. ,
DOI : 10.1016/j.tracli.2008.07.011
Ubc9 interacts with Lu/BCAM adhesion glycoproteins and regulates their stability at the membrane of polarized MDCK cells, Biochemical Journal, vol.402, issue.2, pp.311-319, 2007. ,
DOI : 10.1042/BJ20060861
URL : https://hal.archives-ouvertes.fr/hal-00478614
Form and function: The laminin family of heterotrimers, Developmental Dynamics, vol.214, issue.Suppl 1, 2000. ,
DOI : 10.1002/(SICI)1097-0177(200006)218:2<213::AID-DVDY1>3.0.CO;2-R
Expression of red cell surface antigens during erythropoiesis, Vox Sang, vol.78, issue.2, pp.149-153, 2000. ,
Molecular interactions of B-CAM (basal-cell adhesion molecule) and laminin in epithelial skin cancer, Archives of Dermatological Research, vol.13, issue.2, pp.59-66, 2004. ,
DOI : 10.1007/s00403-004-0481-4
Podocyte-Specific Deletion of Integrin-Linked Kinase Results in Severe Glomerular Basement Membrane Alterations and Progressive Glomerulosclerosis, Journal of the American Society of Nephrology, vol.17, issue.5, pp.1334-1344, 2006. ,
DOI : 10.1681/ASN.2005090921
Endothelial Lu/BCAM glycoproteins are novel ligands for red blood cell ??4??1integrin: role in adhesion of sickle red blood cells to endothelial cells, Blood, vol.109, issue.8, pp.3544-3551, 2007. ,
DOI : 10.1182/blood-2006-07-035139
Isoforms of the Lutheran/Basal Cell Adhesion Molecule Glycoprotein Are Differentially Delivered in Polarized Epithelial Cells: MAPPING OF THE BASOLATERAL SORTING SIGNAL TO A CYTOPLASMIC DI-LEUCINE MOTIF, Journal of Biological Chemistry, vol.274, issue.45, pp.31903-31908, 1999. ,
DOI : 10.1074/jbc.274.45.31903
Organisation of the human Lu gene and molecular basis of the Lu(a)/Lu(b) blood group polymorphism, Blood, vol.89, pp.4608-461, 1997. ,
Protein Kinase A-dependent Phosphorylation of Lutheran/Basal Cell Adhesion Molecule Glycoprotein Regulates Cell Adhesion to Laminin ??5, Journal of Biological Chemistry, vol.280, issue.34, pp.30055-3062, 2005. ,
DOI : 10.1074/jbc.M503293200
CD44 may be a laminin receptor of normal and sickle erythrocytes, Blood, vol.86, p.137, 1995. ,
The Endothelium and Thrombosis, Seminars in Thrombosis and Hemostasis, vol.26, issue.05, pp.463-478, 2000. ,
DOI : 10.1055/s-2000-13202
Sickle cell vasoocclusion and rescue in a microfluidic device, Proceedings of the National Academy of Sciences, vol.104, issue.51, pp.20496-20500, 2007. ,
DOI : 10.1073/pnas.0707122105
Endothelial cell membranes contain podocalyxin--the major sialoprotein of visceral glomerular epithelial cells, The Journal of Cell Biology, vol.102, issue.2, pp.484-491, 1986. ,
DOI : 10.1083/jcb.102.2.484
Integrin Structure, Biochemical Society Transactions, vol.28, issue.4, pp.311-339, 2000. ,
DOI : 10.1042/bst0280311
Molecular Dissection of the ??-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10, Journal of Biological Chemistry, vol.279, issue.12, pp.10946-10954, 2004. ,
DOI : 10.1074/jbc.M313626200
The lutheran blood group system, pp.671-694, 1998. ,
Report on group 8 (Lutheran) antibodies, Revue Francaise de Transfusion et Immuno-h??matologie, vol.31, issue.2, pp.433-440, 1988. ,
DOI : 10.1016/S0338-4535(88)80135-1
Molecular cloning, expression, and characterization of podocalyxin-like protein 1 from rabbit as a transmembrane protein of glomerular podocytes and vascular endothelium, J Biol Chem, vol.270, pp.29439-29436, 1995. ,
Laminin ??5 mediates ectopic adhesion of hepatocellular carcinoma through integrins and/or Lutheran/basal cell adhesion molecule, Experimental Cell Research, vol.314, issue.14, pp.2579-2590, 2008. ,
DOI : 10.1016/j.yexcr.2008.05.021
Mesangial cells organize the glomerular capillaries by adhering to the G domain of laminin ??5 in the glomerular basement membrane, The Journal of Cell Biology, vol.129, issue.1, pp.187-196, 2003. ,
DOI : 10.1074/jbc.274.2.728
Identification of the Binding Site for the Lutheran Blood Group Glycoprotein on Laminin alpha 5 through Expression of Chimeric Laminin Chains in Vivo, Journal of Biological Chemistry, vol.277, issue.47, pp.44864-44869, 2002. ,
DOI : 10.1074/jbc.M208731200
Integrin binding specificity of laminin-10/11 : laminin-10/11 are recognized by alpha3 beta1, alpha6 beta1 and alpha6 beta4 Integrins, J Cell Sci, vol.113, pp.869-876, 2000. ,
Endothelium-derived vasoactive factors and the renal vasculature, Am. J. Physiol, vol.260, pp.653-662, 1991. ,
as a Cause of a Mild Haemolytic Disease of the Newborn, Vox Sanguinis, vol.4, issue.6, pp.532-537, 1960. ,
DOI : 10.1111/j.1423-0410.1960.tb04039.x
in Myeloproliferative Disorders, New England Journal of Medicine, vol.352, issue.17, pp.1779-1790, 2005. ,
DOI : 10.1056/NEJMoa051113
Direct interaction between the Lu/B-CAM adhesion glycoproteins and erythroid spectrin+, British Journal of Haematology, vol.69, issue.9, pp.255-264, 2004. ,
DOI : 10.1074/jbc.274.33.23286
2-Aminoethylisothiouronium bromide-treated red cells and the Lutheran antigens Lua and Lub, Transfusion, vol.27, issue.6, pp.505-506, 1987. ,
DOI : 10.1046/j.1537-2995.1987.27688071712.x
Identification of Cell Binding Sites in the Laminin ??5-Chain G Domain, Experimental Cell Research, vol.277, issue.1, pp.95-106, 2002. ,
DOI : 10.1006/excr.2002.5540
The Laminin 511/521 binding site on the Lutheran blood group glycoprotein is located at the flexible junction of Ig domains 2 and 3, Blood, vol.110, issue.9, pp.3398-3406, 2007. ,
DOI : 10.1182/blood-2007-06-094748
Role of Laminin Terminal Globular Domains in Basement Membrane Assembly, Journal of Biological Chemistry, vol.282, issue.29, pp.21437-21447, 2007. ,
DOI : 10.1074/jbc.M702963200
??II-Spectrin Is Critical for Cell Adhesion and Cell Cycle, Journal of Biological Chemistry, vol.284, issue.4, pp.2409-2418, 2009. ,
DOI : 10.1074/jbc.M801324200
Laminins and their roles in mammals, Microscopy Research and Technique, vol.117, issue.5, pp.349-356, 2008. ,
DOI : 10.1002/jemt.20563
LAMININ FUNCTIONS IN TISSUE MORPHOGENESIS, Annual Review of Cell and Developmental Biology, vol.20, issue.1, pp.255-284, 2004. ,
DOI : 10.1146/annurev.cellbio.20.010403.094555
Defective Glomerulogenesis in the Absence of Laminin ??5 Demonstrates a Developmental Role for the Kidney Glomerular Basement Membrane, Developmental Biology, vol.217, issue.2, pp.278-289, 2000. ,
DOI : 10.1006/dbio.1999.9546
Roles for Laminin in Embryogenesis: Exencephaly, Syndactyly, and Placentopathy in Mice Lacking the Laminin ??5 Chain, The Journal of Cell Biology, vol.32, issue.6, pp.1713-1723, 1998. ,
DOI : 10.1016/0955-0674(94)90093-0
Molecular Cloning of a Novel Laminin Chain, ??5, and Widespread Expression in Adult Mouse Tissues, Journal of Biological Chemistry, vol.270, issue.48, pp.28523-28526, 1995. ,
DOI : 10.1074/jbc.270.48.28523
Localization of Lutheran, a novel laminin receptor, in normal, knockout, and transgenic mice suggests an interaction with laminin ?5 in vivo, Developmental Dynamics, vol.274, issue.1, pp.101-114, 2001. ,
DOI : 10.1002/dvdy.1169
A regional localisation for an X-linked suppressor gene (XS) for the Lutheran blood group, Human Genetics, vol.40, issue.2, pp.127-129, 1988. ,
DOI : 10.1007/BF00278180
Role of Rap1 in promoting sickle red blood cell adhesion to laminin via BCAM/LU, Blood, vol.105, issue.8, pp.3322-3329, 2005. ,
DOI : 10.1182/blood-2004-07-2881
Structure and function of podocytes: an update, Anatomy and Embryology, vol.192, issue.5, pp.385-397, 1995. ,
DOI : 10.1007/BF00240371
A Further Example of the Recessive Lu(a-b-) Phenotype, Vox Sanguinis, vol.198, issue.1, pp.66-68, 1975. ,
DOI : 10.1111/j.1423-0410.1975.tb00478.x
Rh-RhAG/ankyrin-R, a new interaction site between the membrane bilayer and the red cell skeleton, is impaired by Rh(null)-associated site mutation, J Biol Chem, vol.278, pp.255526-25536, 2003. ,
Identification of Cell-binding Sites on the Laminin alpha 5 N-terminal Domain by Site-directed Mutagenesis, Journal of Biological Chemistry, vol.276, issue.14, pp.10906-10912, 2001. ,
DOI : 10.1074/jbc.M008743200
Identification of a Major Heparin and Cell Binding Site in the LG4 Module of the Laminin alpha 5 Chain, Journal of Biological Chemistry, vol.275, issue.19, pp.14517-14523, 2000. ,
DOI : 10.1074/jbc.275.19.14517
Lutheran blood group glycoprotein and its newly characterized mouse homologue specifically bind alpha 5 chain-containing human laminin with high affinity, Blood, vol.97, pp.312-320, 2001. ,
Use of domain-deletion mutants to locate Lutheran blood group antigens to each of the five immunoglobulin superfamily domains of the Lutheran glycoprotein : elucidation of the molecular basis of the LuAu(b) polymorphisms, Lu(b) and the Au, pp.4219-4225, 1997. ,
The Lutheran blood group glycoprotein, another member of the immunoglobulin superfamily, is widely expressed in human tissues and is developmentally regulated in human liver., Proceedings of the National Academy of Sciences, vol.92, issue.12, pp.5496-4500, 1995. ,
DOI : 10.1073/pnas.92.12.5496
Evidence that the Lub blood group antigen is located on red cell membrane glycoproteins of 85 and 78 kd, Transfusion, vol.27, issue.1, pp.61-63, 1987. ,
DOI : 10.1046/j.1537-2995.1987.27187121477.x
Laminin isoforms in tumor invasion, angiogenesis and metastasis, Seminars in Cancer Biology, vol.12, issue.3, pp.197-207, 2002. ,
DOI : 10.1016/S1044-579X(02)00023-8
Mouse heart laminin. Purification of native protein and structural comparison with Engelbreth-Holm tumor laminin, J Biol Chem, vol.264, pp.18726-18732, 1989. ,
Genetic inactivation of the laminin ??5 chain receptor Lu/BCAM leads to kidney and intestinal abnormalities in the mouse, AJP: Renal Physiology, vol.294, issue.2, pp.393-406, 2008. ,
DOI : 10.1152/ajprenal.00315.2007
Characterization of a mouse laminin receptor gene homologous to the human blood group Lutheran gene, Immunogenetics, vol.50, issue.5-6, pp.271-277, 1999. ,
DOI : 10.1007/s002510050602
A unique gene encodes spliceoforms of the B-cell adhesion molecule cell surface glycoprotein of epithelial cancer and of the Lutheran blood group glycoprotein, Blood, vol.88, pp.1865-1872, 1996. ,
Sieve plugs in fenestrae of glomerular capillaries-site of the filtration barrier? Cells Tissues Organs, pp.132-138, 2002. ,
Glomerular permeability to proteins. Effects of hemodynamic factors on the distribution of endogenous immunoglobulin G and exogenous catalase in the glomerulus, Lab. Invest, vol.34, pp.415-427, 1976. ,
Domain IVa of laminin ??5 chain is cell-adhesive and binds ??1 and ??V??3 integrins through Arg-Gly-Asp, FEBS Letters, vol.108, issue.2, pp.181-185, 2001. ,
DOI : 10.1016/S0014-5793(01)03167-2
Identification of Endoglycan, a Member of the CD34/Podocalyxin Family of Sialomucins, Journal of Biological Chemistry, vol.275, issue.12, pp.9001-9010, 2000. ,
DOI : 10.1074/jbc.275.12.9001
The biology of the glomerulus: Endothelial cells, Kidney International, vol.45, issue.2, pp.314-319, 1994. ,
DOI : 10.1038/ki.1994.40
Basal-Cell Adhesion Molecule (B-CAM) is Induced in Epithelial Skin Tumors and Inflammatory Epidermis, and is Expressed at Cell???Cell and Cell???Substrate Contact Sites, Journal of Investigative Dermatology, vol.115, issue.6, pp.1047-11053, 2000. ,
DOI : 10.1046/j.1523-1747.2000.00189.x
A human cell-surface glycoprotein that carries Cromer-related blood group antigens on erythrocytes and is also expressed on leucocytes and platelets, Immunology, vol.62, pp.307-313, 1987. ,
Functional Sites in the Laminin Alpha Chains, Connective Tissue Research, vol.2, issue.3, pp.142-152, 2005. ,
DOI : 10.1091/mbc.10.5.1521
The endothelium in glomerular inflammation, Current Opinion in Nephrology and Hypertension, vol.4, issue.3, 1995. ,
DOI : 10.1097/00041552-199505000-00014
Lutheran antigens, CD44-related antigens, and Lutheran regulatory genes, Transfusion Clinique et Biologique, vol.2, issue.4, pp.291-301, 1995. ,
DOI : 10.1016/S1246-7820(05)80095-3
Structure and function of laminin LG modules, Matrix Biology, vol.19, issue.4, pp.309-317, 2000. ,
DOI : 10.1016/S0945-053X(00)00072-X
Supramolecular assembly of basement membranes, BioEssays, vol.217, issue.2, pp.123-132, 1996. ,
DOI : 10.1002/bies.950180208
Laminins: Structure and genetic regulation, Microscopy Research and Technique, vol.271, issue.3, 2000. ,
DOI : 10.1002/1097-0029(20001101)51:3<214::AID-JEMT2>3.0.CO;2-J
Bridging structure with function: Structural, regulatory, and developmental role of laminins, The International Journal of Biochemistry & Cell Biology, vol.40, issue.2, pp.199-214, 2008. ,
DOI : 10.1016/j.biocel.2007.07.015
Basal cell adhesion molecule/lutheran protein. The receptor critical for sickle cell adhesion to laminin., Journal of Clinical Investigation, vol.101, issue.11, pp.2550-2558, 1998. ,
DOI : 10.1172/JCI1204
Blood vessels of human islets of Langerhans are surrounded by a double basement membrane, Diabetologia, vol.48, issue.7, pp.1181-1191, 2008. ,
DOI : 10.1007/s00125-008-0997-9
Increased adhesion to endothelial cells of erythrocytes from patients with polycythemia vera is mediated by laminin ??5 chain and Lu/BCAM, Blood, vol.110, issue.3, pp.894-901, 2007. ,
DOI : 10.1182/blood-2006-10-048298
Lutheran Blood Group Antigen as a Receptor for ??5 Laminins in Gingival Epithelia, Journal of Periodontology, vol.78, issue.9, pp.1810-1818, 2007. ,
DOI : 10.1902/jop.2007.060482
Cloning and Characterization of Human Erythroid Membrane-Associated Protein, Human ERMAP, Genomics, vol.76, issue.1-3, pp.2-4, 2001. ,
DOI : 10.1006/geno.2001.6600
beta1 Integrin and alpha-dystroglycan binding sites are localized to different laminin-G-domain-like (LG) modules within the laminin alpha5 chain G domain, Biochemical Journal, vol.371, issue.2, pp.289-299, 2003. ,
DOI : 10.1042/bj20021500
Molecular architecture of basement membranes, FASEB J, vol.4, pp.1577-1590, 1990. ,
Critical Factors in Basal Cell Adhesion Molecule/Lutheran-mediated Adhesion to Laminin, Journal of Biological Chemistry, vol.274, issue.2, pp.728-734, 1999. ,
DOI : 10.1074/jbc.274.2.728
Hébergement : 1,03? HT/souris soit 3X donc 3,09? HT/test Perfusion de billes : 47,44? HT/souris soit 3X donc 142,32? HT/test Prélèvement (reins) : 26,15? HT/souris soit 3X donc 78, 45? HT/test Coût global de l'ICS : 223.86? HT/expérience SARL MEG Import/Export ,
Paris : 190? HT Coût du transport : 302.69? HT soit 15,13 à 30,27? HT/souris Isolement des glomérules Solution HBSS 1X : 2,47? HT/test Solution de collagénase 60X : 9,30? HT/test Solution DNase 100X : 6, 2? HT /test Coût de l'expérience : 42,97? HT/expérience Immuno-Histo-Fluorescence Solution PBS 1X : 6,63? HT soit 3X donc 1990? HT/test Anticorps Lu : 0,20? HT/test Anti anticorps Lu : 0,18? HT/test Anticorps ?1 laminine, pp.13-020 ,
20? HT/expérience Électrophorèse des protéines Gel 3 à 8% Tris Acétate SDS : 18,10? HT/test Marqueur protéine Plus 3, 50? HT/test Tris Acétate, vol.20, issue.3, p.2733 ,
expérience : 13,14? HT/expérience La spectrométrie de masse Préparation des échantillons : 25? HT soit 6X donc 150? HT/test Mesures, pp.60-63 ,