RETINOIC ACID-INDUCED HEPARIN-BINDING PROTEIN EXPRESSION AND LOCALIZATION DURING GASTRULATION, NEURULATION, AND ORGANOGENESIS

Retinoic acid induced heparin-binding protein (RIHB) is a highly basic , soluble polypeptide of the chick embryonic extracellular matrix. We have examined the expression and localization of RIHB during very earl y embryogenesis by in situ hybridization and immunohistochemistry. RIH B mRNA is very weakly detectable above background in the blastodiscs o f unincubated eggs. The expression increases greatly over the first 24 hours of incubation, and is observed throughout the blastodisc in all three of the germ layers following gastrulation. As neurulation occur s, the expression becomes more restricted to certain areas, notably th e ectoderm, the neural folds, and especially the notochord. After the neural tube has formed the expression in the tube itself decreases dra matically, whereas the expression in the head ectoderm and the notocho rd persists. After 72 hours of incubation expression remains relativel y high throughout most of the embryo, with higher levels of expression in regions undergoing organogenesis and lower levels in organs which have already differentiated. RIHB protein is also weakly detectable in unincubated eggs as patches of immunoreactive material between the bl astodisc and the vitelline. After 6 hours of incubation small regions of basement membrane are immunoreactive. RIHB is detected in this matr ix, apparently before even fibronectin. The amount of RIHB protein inc reases dramatically over the first 24 hours of incubation. It is found in basement membrane separating the epiblast from the hypoblast, then later in that separating the ectoderm from the mesoderm, It is also d etected surrounding individual cells, especially of the ectodermal lay er. During neurulation RIHB is observed in the basement membrane surro unding the neural fold and the notochord, and in the lamina separating the ectodermal, mesodermal, and endodermal layers. Eater in developme nt, RIHB is detected in the basement membrane under the epidermis, thr oughout the developing Limbs, and in the lamina of various developing organs, such as the eye, the pulmonary bud, the intestine, and the mes onephros. These results demonstrate that RIHB is highly expressed duri ng the early embryonic period, by all three germ layers, and is an imp ortant and very early component of the embryonic extracellular matrix. Its very broad expression and localization argue for a more general r ole in development than its demonstrated weak neurotrophic activity. ( C) 1994 Wiley-Liss, Inc.