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During the development of peripheral nerves, Schwann cells are induced to form myelin sheaths round the larger axons. This process involves a complex series of events and the nature of the molecular signals that regulate and control myelin formation in Schwann cells is not well understood. Our previous experiments on rat Schwann cells in vitro, using serum-free defined medium, showed that a myelinrelated protein phenotype could be induced in early postnatal Schwann cells in culture by elevation of intracellular cyclic AMP levels in the absence of growth factors, conditions under which the cells are not dividing. Cells with this phenotype expressed the major myelin glycoprotein Po and expression of p75 NGF receptor, N-CAM, GFAP and A5E3 proteins was down-regulated. These changes are all characteristics associated with myelination in vivo. In contrast, when cyclic AMP levels were elevated in the presence of serum, suppression of cyclic AMP-induced differentiation resulted and DNA synthesis was induced. In this paper, we have used this model system and extended our analysis to explore the relationship between defined growth factors and suppression of myelination. We have used pure recombinant growth factors normally present in peripheral nerves, i.e. FGF1 and FGF2 and TGFβ1, TGFβ2, and TGFβ3 and shown that, like serum, they can strongly suppress the forskolin-mediated induction of the Po gene, both at the level of mRNA and protein synthesis. For both growth factor families, the suppression of Po gene expression is dose-dependent and takes place in serum-starved cells that are mitotically quiescent. In the case of FGF2, however, even more complete suppression is obtained when the cells are simultaneously allowed to enter the cell cycle by inclusion of high concentrations of insulin in the culture medium. The present results raise the possibility that, in addition to the positive axonal signals that are usually envisaged to control the onset of myelination, growth factors present in the nerve may exert negative regulatory signals during development and thus help control the time of onset and the rate of myelination in peripheral nerves.

Using immunocytochemical methods we describe the distribution of different TGF β isoforms and the effects of excess retinoic acid on their expression during early mouse embryogenesis ( days of development). In normal embryos at 9 days, intracellular TGF β1 is expressed most intensely in neuroepithelium and cardiac myocardium whereas extracellular TGF β1 is expressed in mesenchymal cells and in the endocardium of the heart. At later stages, intracellular TGF β1 becomes very restricted to the myocardium and to a limited number of head mesenchymal cells; extracellular TGF β1 continues to be expressed widely in cells of mesenchymal origin, particularly in head and trunk mesenchyme, and also in endocardium. TGF β1 is widely expressed at all stages investigated while TGF β3 is not expressed strongly in any tissue at the stages examined. Exposure of early neural plate stage embryos to retinoic acid caused reduced expression of TGF β1 and TGF β2 proteins but had no effect on TGF β1. Intracellular TGF β1 expression was reduced in all tissues except in the myocardium, while extracellular TGF β1 was specifically reduced in neuroepithelium and cranial neural crest cells at early stages. TGF β1 was reduced in all embryonic tissues. The down-regulation of intracellular TGF β1 was observed up to 48 hours after initial exposure to retinoic acid while some downregulation of TGF β1 was still seen up to 60 hours after initial exposure. TGF βs are known to modulate the expression of various extracellular matrix molecules involved in cell growth, differentiation and morphogenesis. The interaction between retinoic acid and TGF β is discussed in relation to morphogenesis.

We have examined by Northern analysis and in situ hybridisation the expression of TGF β1 β2and β3 during mouse embryogenesis. TGF β1 is expressed predominantly in the mesodermal components of the embryo e.g. the hematopoietic cells of both fetal liver and the hemopoietic islands of the yolk sac, the mesenchymal tissues of several internal organs and in ossifying bone tissues. The strongest TGF β2 signals were found in early facial mesenchyme and in some endodermal and ectodermal epithelial cell layers e.g., lung and cochlea epithelia. TGF β3 was strongest in prevertebral tissue, in some mesothelia and in lung epithelia. All three isoforms were expressed in bone tissues but showed distinct patterns of expression both spatially and temporally. In the root sheath of the whisker follicle, TGF β1 β2 and β3 were expressed simultaneously. We discuss the implication of these results in regard to known regulatory elements of the TGF β genes and their receptors.

We have studied the expression of genes encoding transforming growth factors (TGFs) beta1, beta2 and beta3 during development of the secondary palate in the mouse from 11.5 to 15.5 days postcoitum using in situ hybridisation. The RNA detected at the earliest developmental stage is TGF beta3, which is localised in the epithelial component of the vertical palatal shelf. This expression continues in the horizontal palatal shelf, predominantly in the medial edge epithelium, and is lost as the epithelial seam disrupts, soon after palatal shelf fusion. TGF betal RNA is expressed with the same epithelial pattern as TGF beta3, but is not detectable until the horizontal palatal shelf stage. TGF beta2 RNA is localised to the palatal mesenchyme underlying the medial edge epithelia in the horizontal shelves and in the early postfusion palate. The temporal and spatial distribution of TGF betal, beta! and beta3 RNAs in the developing palate, together with a knowledge of in vitro TGF beta biological activities, suggests an important role for TGF beta isoforms in this developmental pro cess.