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Expression of Angiopoietin-1, Angiopoietin-2, and the Tie-2 Receptor Tyrosine Kinase during Mouse Kidney Maturation

HAI TAO YUAN^*, CHITRA SURI, GEORGE D. YANCOPOULOS and ADRIAN S. WOOLF^*

^* Nephrourology Unit, Institute of Child Health, University College London Medical School, London, United Kingdom
Regeneron Pharmaceuticals, Inc., Tarrytown, New York.

Correspondence to Dr. Hai Tao Yuan, Nephrourology Unit, Institute of Child Health, University College London Medical School, 30 Guilford Street, London WC1EN 1EH, United Kingdom. Phone: 0171 242 9789; Fax: 0171 916 0011; E-mail: h.yuan{at}ich.ucl.ac.uk

Abstract

Abstract. The Tie-2 receptor tyrosine kinase transduces embryonic onic endothelial differentiation, with Angiopoietin-1 (Ang-1) acting as a stimulatory ligand and Ang-2 postulated to be a naturally occurring inhibitor. Expression of these genes was sought during mouse kidney maturation from the onset of glomerulogenesis (embryonic day 14 [E14]) to the end of nephron formation (2 wk postnatal [P2]), and during medullary maturation into adulthood (P8). Using Northern and slot blotting of RNA extracted from whole organs, these three genes were expressed throughout the experimental period with peak levels at P2 to P3. By in situ hybridization analysis at E18, P1, and P3, Ang-1 mRNA was found to localize to condensing renal mesenchymal cells, proximal tubules, and glomeruli in addition to maturing tubules of the outer medulla. In contrast, Ang-2 transcripts were more spatially restricted, being detected only in differentiating outer medullary tubules and the vasa recta bundle area. Using in situ hybridization and immunohistochemistry, Tie-2 was detected in capillaries of the nephrogenic cortex, glomerular tufts, cortical interstitium, and medulla including vessels in the vasa recta. Using Western blotting of protein extracted from whole organs, Tie-2 protein was detected between E14 and P8 with tyrosine phosphorylated Tie-2 evident from E18. These data are consistent with the hypothesis that Tie-2 has roles in maturation of both glomeruli and vasa rectae.

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