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Cloning of cDNA for the α Subunit of Mouse Insulin-Like Growth Factor I Receptor and the Role of the Receptor in Metanephric Development
Jun Wada, Zheng Z. Liu, Keith Alvares, Anil Kumar, Elisabeth Wallner, Hirofumi Makino and Yashpal S. Kanwar
Proceedings of the National Academy of Sciences of the United States of America
Vol. 90, No. 21 (Nov. 1, 1993), pp. 10360-10364
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/2363463
Page Count: 5
You can always find the topics here!Topics: Kidneys, Complementary DNA, Receptors, Pregnancy, Glycoproteins, Proteoglycans, In situ hybridization, Morphogenesis, Somatomedins, Nucleotide sequences
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Various growth factors influence mammalian development by binding to specific cell surface receptors. These interactions are followed by a series of intracellular transductional events leading to a wide variety of biological effects. To establish the role of insulin-like growth factor I receptor (IGF-IR) in renal development, cDNA for the α subunit of the mouse IGF-IR was isolated, characterized, and used in expression studies and antisense experiments in a metanephric organ culture system. A 989-bp insert, encoding the signal peptide and 299 amino acids, isolated from a newborn mouse kidney cDNA library had 99% and 91% homology with the nucleotide sequences encoding the rat and the human IGF-IR, respectively. An ≈11-kb message was readily detected by Northern blot analysis of RNA from the developing kidney at day 13 of gestation, and it declined during the subsequent embryonal and neonatal periods. In situ hybridization revealed high levels of message over the ureteric bud and its branches. A lower level of message was seen in the neonatal kidney, confined mainly to the tubules. Antisense oligodeoxynucleotide-treated metanephric kidneys were reduced in size and had a decreased population of nephrons with marked disorganization of ureteric bud branches. Immunofluorescence studies indicated an arrest of IGF-IR translation after antisense exposure. Immunoprecipitation studies showed a marked decrease in the biosynthesis of various extracellular matrix proteins that serve as regulators of morphogenesis. These studies suggest that the nucleotide sequence encoding the α subunit of mouse IGF-IR is highly conserved and that the receptor might play an essential role in the organogenesis of the kidney.
Proceedings of the National Academy of Sciences of the United States of America © 1993 National Academy of Sciences