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Laboratory of Pediatrics (M.v.K., C.A.H.G., N.F.J.D., D.J.L.-K., S.L.S.D., J.W.v.N.), Subdivision Molecular Endocrinology, and Department of Pathology (E.C.Z.), Erasmus University Rotterdam, Postbus 1738, 3000 DR Rotterdam, The Netherlands; and Department of Neuroscience and Cell Biology (A.G.P.S., S.L.B., J.E.P.), University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854
Address all correspondence and requests for reprints to: Dr. J. W. van Neck, Erasmus University Rotterdam, Laboratory of Pediatrics, Room Ee15.00, Postbus 1738, 3000 DR Rotterdam, The Netherlands. E-mail: vanneck{at}kgk.fgg.eur.nl
The insulin-like growth factor (IGF) system is an important regulator of fetal growth and differentiation. IGF bioavailability is modulated by IGF binding proteins (IGFBPs). We have generated six different antisera, directed to synthetic peptide fragments of mouse IGFBP-1 through -6. The specificity of the produced antisera was demonstrated by enzyme-linked immunosorbent assay, Western blotting, and by immunohistochemistry on sections of mouse embryos of 13.5 days post coitum. Specificity for the IGFBP-2 through -6 antisera also was confirmed immunohistochemically in liver and lung of corresponding gene deletion (knock-out) mutant mice and wild-type litter mates.
Immunohistochemistry and messenger RNA (mRNA) in situ hybridization on sections of mouse embryos of 13.5 days post coitum revealed tissue-specific expression patterns for the six IGFBPs. The only site of IGFBP-1 protein and mRNA production was the liver. IGFBP-2, -4, and -5 protein and mRNA were detected in various organs and tissues. IGFBP-3 and -6 protein and mRNA levels were low. In several tissues, such as lung, liver, kidney, and tongue, more than one IGFBP (protein and mRNA) could be detected. Differences between mRNA and protein localization were extensive for IGFBP-3, -5, and -6, suggesting that these IGFBPs are secreted and transported.
These results confirm the different spatial localization of the IGFBPs, on the mRNA and protein level. The overlapping mRNA and protein localization for IGFBP-2 and -4, on the other hand, may indicate that these IGFBPs also function in an auto- or paracrine manner.
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