ABSTRACT
ZT3, isolated from a murine muscle cell cDNA library by a low-stringency hybridization, encodes a zinc finger domain containing factor with a transcript of 5.0 kb. A 3' 2.5 kb partial nucleotide sequence contains an ORF of 1.5 kb where 17 canonical C2H2 zinc finger domains organized in tandem were identified. It maps on mouse chromosome 11, close to two mutations which affect skeletal formation. ZT3 expression depends upon differentiation of myogenic cells in culture, since it is upregulated with myogenin and inhibited in scr-transfected C2C12 cells. ZT3 is not expressed in NIH3T3 or C3H10T1/2 fibroblasts, but is induced when fibroblasts are myogenically converted by transfection with the muscle regulatory genes (MRFs). Its expression is also upregulated in the rhabdomyosarcoma cell line RD induced to myogenic differentiation by TPA treatment. In postimplantation embryos, ZT3 is diffusely expressed but higher expression is detectable in the neural tube and encephalic vesicles, in the somites and, at a high level, in the limb buds as they form. During further development ZT3 is expressed in many tissues of neuroectodermal and mesodermal origin, but its expression decreases during fetal development and in the adult it is restricted to skeletal and cardiac muscle and to spleen. This pattern of expression suggests a possible role played by ZT3 in differentiating skeletal muscle. Its expression in other tissues is compatible with the suggestion that members of this class of DNA-binding factors play different roles during post-implantation development and in the adult life.
Subject(s)
DNA-Binding Proteins/biosynthesis , Gene Expression Regulation, Developmental , Muscle Proteins/biosynthesis , Muscle, Skeletal/metabolism , Myocardium/metabolism , 3T3 Cells , Amino Acid Sequence , Animals , Base Sequence , Cell Differentiation , Cell Line , Chromosome Mapping , Crosses, Genetic , DNA, Complementary/genetics , DNA-Binding Proteins/genetics , Fibroblasts/cytology , Fibroblasts/metabolism , Gene Library , Heart/embryology , Heart/growth & development , Kruppel-Like Transcription Factors , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Molecular Sequence Data , Muscle Development , Muscle Proteins/genetics , Muscle, Skeletal/embryology , Muscle, Skeletal/growth & development , Sequence Alignment , Sequence Homology, Amino Acid , Spleen/embryology , Spleen/growth & development , Spleen/metabolismABSTRACT
The expression and distribution of the mRNA coding for galanin precursor, preprogalanin (ppGAL), were analysed in several rat hypothalamic nuclei (periventricular, paraventricular, supraoptic, dorsomedial and arcuate nuclei and the lateral hypothalamic area) during development by an in situ hybridization technique and computer-assisted grain counting over individual cells. ppGAL mRNA (expressed as number of grains/100 microns2) was detectable from postnatal day (PD) 1 in all the nuclei considered, and the amount of transcript per cell was 6-11 times less than in the adult. ppGAL mRNA progressively increased from PD8 to 14 to 21. The level of ppGAL mRNA in all the nuclei at PD21 was about half that in adulthood, except in the dorsomedial nucleus, where the difference was no more than 20%. As an index of the activity of galanin-containing neurons, we measured the basal and K(+)-evoked in vitro release of galanin-like immunoreactivity from hypothalamic slices of PD14, 21 and 90 rats by radioimmunoassay. Basal release of galanin-like immunoreactivity remained at the same level from PD14 to PD90, but the response to KCl (50 mM) stimulation was lower at PD14 (approximately 90%) and PD21 (> 200%) than at PD90 (350%). Basal and K(+)-evoked release was sensitive to tetrodotoxin, indicating a neuronal origin. This study provides the first evidence that the increase in ppGAL mRNA during the ontogeny of hypothalamic nuclei is associated with an increase in galaninergic neuronal function.
Subject(s)
Hypothalamus/metabolism , Neuropeptides/biosynthesis , Peptide Biosynthesis , Age Factors , Animals , Autoradiography , Galanin , Gene Expression , Hypothalamus/growth & development , In Situ Hybridization , In Vitro Techniques , Male , Potassium Compounds/pharmacology , RNA, Messenger/metabolism , Radioimmunoassay , Rats , Rats, Inbred Strains , Tetrodotoxin/pharmacologyABSTRACT
The expression of galanin (GAL) mRNA was determined by in situ hybridization after frontal deafferentation and colchicine treatment in the rat hypothalamus. Frontal deafferentation significantly increased the signal in the paraventricular nucleus (PVN), the supraoptic nucleus (SON), and dorsomedial nucleus (DMN). Colchicine treatment induced a diffuse enhancement of GAL mRNA in hypothalamic nuclei. When the two treatments were combined there was an additivity of GAL mRNA expression in the previous hypothalamic nuclei and also in the arcuate nucleus (AN), where the single treatments did not modify the signal. These results suggest the regulation of GAL mRNA expression mediated by a multineuronal pathway, separate from the colchicine-induced GAL mRNA increase.