Localization of IGF-I, IGF-I receptor, and IGFBP-2 in developing Umbrina cirrosa (Pisces: Osteichthyes)

In this study, the distribution of IGF-I, IGF type I receptor (IGF-IR), and IGF-binding protein 2 (IGFBP-2) was investigated during larval and post-larval developmental stages of the shi drum (Umbrina cirrosa) by immunohistochemistry using antisera raised against Sparus aurata IGF-I and IGF-IR, and against mouse IGFBP-2. Immunoreactivity of the mitogenic marker PCNA (proliferating cell nuclear antigen) was used for assessment of cellular proliferation. Distribution of IGF-I mRNA was studied by in situ hybridization. IGF-I immunoreactivity was detected in liver and developing intestine already in 1-5 day post-hatching larvae. From day 11, immunostaining in the intestine was evident in the enterocytes of the anterior intestine and in the apical zone of the epithelium of developing posterior intestine. Positive reaction with IGF-I antibody was also detected in chondrocytes, in the epithelium of the skin, gills and in the central nervous system (CNS), and lateral muscle. At hatching IGF-IR immunoreactivity was already detectable in developing CNS, notochord, and skin. From day 6 immunostaining was evident in the olfactory epithelium, in eyes and from day 11 in the developing olfactory bulbs and CNS. Positive reaction with IGF-IR antibody was also detected in chondrocytes, in the epithelium of the skin, gills, heart, and in the lateral muscle. Immunoreactive IGFBP-2, as detected by anti-mouse IGFBP-2 antiserum, exhibited generally a similar distribution pattern to that of IGF-I and IGF-IR. In situ hybridization, which has been performed by using riboprobes from S. aurata cDNA, revealed IGF-I mRNA in skeletal musculature, liver, and CNS. These data strongly suggest a role for the IGF system during development and growth of U. cirrosa.