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Mouse ß₆ Integrin Sequence, Pattern of Expression, and Role in Kidney Development

LOIS J. AREND^*, ANN M. SMART and JOSIE P. BRIGGS

^* Department of Pathology, University of Michigan, Ann Arbor, Michigan
National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland.

Correspondence to Dr. Lois J. Arend, Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, 601 Elmwood Avenue, Box 626, Rochester, NY 14642. Phone: 716-273-4062; Fax: 716-756-4468; E-mail: lois_arend{at}urmc.rochester.edu

Abstract. Integrins mediate cell-cell and cell-extracellular matrix interactions and play key roles in development. ß₆ integrin expression has been demonstrated in human fetal kidney at a higher level than in the adult, making ß₆ integrin a marker of interest for the study of development of the nephron. The aims of this study were to determine the cDNA sequence for the mouse ß₆ integrin and to characterize ß₆ integrin expression in the developing mouse kidney. Two embryonic mouse kidney cDNA libraries were screened, and the coding region was sequenced. The mouse ß₆ nucleotide coding region sequence shows 82% nucleotide identity to the human sequence. The putative amino acid sequence has 89.5% identity to human ß₆ integrin and contains many conserved domains. By reverse transcription-PCR, ß₆ integrin mRNA expression is very low at 11 d of gestation in the mouse, increases dramatically by E14 and E17 (20-fold, normalized for increases in ß actin), and plateaus by 2 wk of age. ß₆ integrin expression is induced 15- to 20-fold after 5 d in metanephric explant culture. Reverse transcription-PCR of adult rat microdissected nephron segments demonstrates ß₆ integrin mRNA expression in proximal tubule, cortical thick ascending limb, distal nephron segments (inner and outer medullary collecting ducts), and macula densa—containing segments. Lectin-peroxidase and in situ colocalization studies demonstrated expression of ß₆ integrin mRNA in developing proximal tubules and thick ascending limb. Culture of mouse metanephric kidneys with antisense oligonucleotides to ß₆ integrin resulted in inhibition of ureteric bud branching and complete lack of mesenchyme condensation. These studies demonstrate a high homology between the human and mouse ß₆ integrin sequence, a different pattern of expression in the developing mouse kidney compared with the primate kidney, and abnormal metanephric development in culture in the absence of ß₆ integrin. These findings suggest an important role for ß₆ integrin in normal development of the mouse kidney.

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Copyright © 2008 by the American Society of Nephrology. Online ISSN: 1533-3450 Print ISSN: 1046-6673