Early embryonic growth is independent of pituitary growth hormone (GH), since it occurs prior to the differentiation of pituitary somatotrophs. Embryogenesis is therefore thought to be regulated by local growth factors. As GH is now known to be produced in many extrapituitary sites, in which it acts in an autocrine or paracrine manner, the possibility that extra-pituitary GH may participate in embryogenesis and organogenesis was assessed by determining the immunocytochemical presence and location of GH- and GH-receptor (GHR)-like proteins in the peripheral tissues of chick embryos during their 21-day incubation period. Immunoreactive (IR)-GH, detectable by a monoclonal and two polyclonal antibodies for chicken GH, was specifically and ubiquitously present in tissues of 3-day-old embryos. At embryonic day (ED) 5, IR-GH was widespread in ectodermal, mesodermal and endodermal tissues, but it was not present in every cell of each tissue. IR-GH was particularly abundant i! n the neural tube, notochord, limb bud, somites, heart, stomach, liver, kidney, Wolffian duct and the amnion. By ED8, IR-GH was still widespread and was now present in limb bud cartilage, although the heart and liver were no longer GH immunoreactive. GH receptor immunoreactivity was also present in most tissues and cells of ED3-ED8 embryos. These results demonstrate that extrapituitary GH is abundantly present during early embryogenesis, prior to the differentiation of pituitary somatotrophs (at ED12). Since GH- and GHR-like proteins are present in most tissues of the chick embryo, it is proposed that extrapituitary GH may act as a local growth factor during embryonic development.

Growth hormone (GH) gene expression predominantly occurs in the pituitary gland, although it also occurs in many extrapituitary sites, including the brain. The cellular location and ontogeny of neural GH production is, however, largely unknown. This has therefore been determined during chick embryogenesis. In chicks, the brain develops from the neural tube at embryonic day (ED) 3. At this age, the divisions of the brain (the telencephalon, diencephalon, mesencephalon, metencephalon and myelencephalon) have intense GH immunoreactivity (GH-IR) (detected by two polyclonal antibodies and a monoclonal antibody for chicken GH). The otic and optic vesicles were also strongly GH immunoreactive, as were the Vth (semi-lunar), VIIth (facial), VIIIth (acoustic) and IXth (glossopharyngeal) nerve ganglia. This GH-IR was specific for GH and was lost when the antibodies were preabsorbed with recombinant chicken GH. The widespread distribution of GH-IR in the neural tissues of ED 3 embryos was mirrored by the distribution of GH receptor (GHR) immunoreactivity, detected by an antibody raised against the chicken GHR. In ED 6/ED 7 embryos, the neural retina of the eye and the epithelial and lens fiber cells were intensely stained for GH-IR, as was Rathke's pouch and the wall of the diencephalon. In contrast, only a few scattered cells were immunoreactive in the surrounding mesoderm. At ED 14, the GH-IR in the brain was restricted to specific tissues and cells. For instance, immunoreactive cells were present in the molecular and pyramidal layers of the cerebral cortex, in the gray matter of the cerebellum, in the choroid plexus, and in the walls of the ventricles. In summary, GH- and GHR-like proteins are abundant in neural tissues of the chick during the first third of incubation, becoming discretely localized to specific tissues and cells during later incubation. The localization of GH and GHR in these tissues, prior to the ontogeny of plasma GH, suggests autocrine or paracrine roles for GH during early embryogenesis.