GATA-6: a zinc finger transcription factor that is expressed in multiple cell lineages derived from lateral mesoderm.

Members of the GATA family of zinc finger transcription factors play important roles in the development of several mesodermally derived cell lineages. In the studies described in this report, we have isolated and functionally characterized the murine GATA-6 cDNA and protein and defined the temporal and spatial patterns of GATA-6 gene expression during mammalian development. The GATA-6 and -4 proteins share high-level amino acid sequence identity over a proline-rich region at the amino terminus of the protein that is not conserved in other GATA family members. GATA-6 binds to a functionally important nuclear protein binding site within the cardiac-specific cardiac troponin C (cTnC) transcriptional enhancer. Moreover, the cTnC promoter enhancer can be transactivated by overexpression of GATA-6 in noncardiac muscle cells. During early murine embryonic development, the patterns of GATA-6 and -4 gene expression are similar, with expression of GATA-6 restricted to the precardiac mesoderm, the embryonic heart tube, and the primitive gut. However, coincident with the onset of vasculogenesis and development of the respiratory and urogenital tracts, only the GATA-6 gene is expressed in arterial smooth muscle cells, the developing bronchi, and the urogenital ridge and bladder. These data are consistent with a model in which GATA-6 functions in concert with GATA-4 to direct tissue-specific gene expression during formation of the mammalian heart and gastrointestinal tract, but performs a unique function in programming lineage-restricted gene expression in the arterial system, the bladder, and the embryonic lung.

SM22 alpha, a marker of adult smooth muscle, is expressed in multiple myogenic lineages during embryogenesis.

SM22 alpha is a calponin-related protein that is expressed specifically in adult smooth muscle. To begin to define the mechanisms that regulate the establishment of the smooth muscle lineage, we analyzed the expression pattern of the SM22 alpha gene during mouse embryogenesis. In situ hybridization demonstrated that SM22 alpha transcripts were first expressed in vascular smooth muscle cells at about embryonic day (E) 9.5 and thereafter continued to be expressed in all smooth muscle cells into adulthood. In contrast to its smooth muscle specificity in adult tissues, SM22 alpha was expressed transiently in the heart between E8.0 and E12.5 and in skeletal muscle cells in the myotomal compartment of the somites between E9.5 and E12.5. The expression of SM22 alpha in smooth muscle cells, as well as early cardiac and skeletal muscle cells, suggests that there may be commonalities between the regulatory programs that direct muscle-specific gene expression in these three myogenic cell types.

[The symmetrical relation of intestines, the habenular nuclei and the intestinal venous system in dorsal-doral and ventral-ventral grown parabiontic larva of mountain salamanders (Triturus alpestris)]. / Die Symmetrieverhältnisse der Eingeweide, der Habenulakerne und des Darmvenensystems bei dorsal-dorsal und ventral-ventral verwachsenen Parabiose-larven des Bergmolches (Triturus alpestris).

Embryos of the newt Triturus alpestris were fused in their dorsal regions (DD-parabionts) of their ventral regions (VV-parabionts), while the longitudinal axes were in equal direction. In these parabionts situs and symmetry of the gut, the heart, the habenular nuclei and the vitelline vein were studied. The fusion of the embryos of the DD-parabionts was performed in three developmental stages (after Harrison): phase of neurulation (14-18, N-parabionts), late phase of neurulation (19-22, E-parabionts) and tail bud stage (23-27, S-parabionts). VV-parabionts were only fused in phase of neurulation (14-18). The external development of the parabionts in most cases was normally. DD-parabionts showed a more or less pronounced deformation of their longitudinal axes. In DD-parabionts all organs developed separately in nearly all cases. In VV-parabionts a partial fusion of the heart was sometimes observed, while fusion of large part of gut occurred regularly. In DD-parabionts complete and incomplete inversions of all organs and the vitelline vein are extremely abundant in the N-parabionts, much lower in E-parabionts and nearly absent in S-parabionts. The differences are statistically of high significance. The "pair situs" symmetries of N-parabionts (DD-parabionts) show always a strong dominance of a transindividual organ- and vitellin vein-symmetry. In VV-parabionts (N-parabionts) the tendency for inversion of the asymmetric organs is statistically significant lower as in DD-parabionts. In regard to this observation there is a preference for an asymmetric "pair situs" in the heart, habenular nuclei and vitelline vein system. In "polarisied" DD-parabionts ("left" as well as "right" parabionts) the asymmetric organs as well as the vitelline vein system inversions were more often observed in the "left" parabiont compared with the "right" parabiont in N- and E-parabionts. These right-left differences partial are statistically significant (P < 0.05). The organ situs-correlation as well as the situs-correlation between vitelline vein and gut or heart, respectively, are mostly positive (concordant). The frequency of dominance of "left" organ inversions argues against a morphogenetic "left dominance" of the amphibian embryo. The determination of organ-asymmetries is irreversible only after the end of neurulation in tail-bud-stage. The results suggest the efficiency of a "symmetry-factor", which determines the organ symmetry during neurulaphase. The differences between DD- and VV-parabionts are statistically significant and point to a dorso-ventral polarity of the embryo, with a maximal morphogenetic potency in the dorsal region.

Furosemide differentially relaxes airway and vascular smooth muscle in fetal, newborn, and adult guinea pigs.

Furosemide, an inhibitor of Cl-dependent Na+,K+ cotransport, is the most frequently used diuretic in newborns. Recently, furosemide was also demonstrated to decrease bronchial hyper-responsiveness in adults, although little is known about the direct effect of furosemide on smooth muscle of immature animals. This in vitro study was designed to determine the action of furosemide on airway and vascular smooth muscle during ontogeny. Extrathoracic trachea (ET), main stem bronchi, main pulmonary artery, and thoracic aorta ring segments from fetal, newborn, and adult Hartley albino guinea pigs were suspended in HEPES solution for measurement of isometric tension. Furosemide (30 or 300 microM) was administered after preconstriction with an ED35-70 concentration of histamine or acetylcholine for airway and ED40-100 concentration of norepinephrine for vessels. Furosemide (30 microM) caused significant relaxation of airway smooth muscle at all ages. After histamine-induced preconstriction, fetal airway segments exhibited greatest relaxation (183 +/- 28%), with newborn airway demonstrating 123 +/- 15% relaxation and modest relaxation seen in adults (40 +/- 4%). This pattern was similar for both ET and bronchus and appeared greater for histamine compared with ACh preconstriction. Epithelial removal slightly enhanced relaxation. Furosemide also relaxed pulmonary artery segments, but at a 10-fold higher concentration. In striking contrast to the pattern seen in airway, adult pulmonary artery relaxed more than newborn and newborn, more than fetus. Cyclooxygenase blockade and endothelium removal did not change pulmonary artery relaxation. Furosemide did not significantly relax aorta after NE preconstriction. Taken together, these results suggest that furosemide may be more effective in relaxing airway compared with vascular smooth muscle, and the ontogeny of these responses indicates a greater efficacy and selectivity in airways of immature animals.

Heparin inhibits proliferation of fetal vascular smooth muscle cells in the absence of platelet-derived growth factor.

Proliferation of smooth muscle cells from the pulmonary arteries and aortas of fetal calves is inhibited by heparin in vitro. This effect is reversible and dose dependent. Comparisons with effects of other polysaccharides indicate that only extensively sulfated polysaccharides inhibit proliferation of smooth muscle cells but that specific structural features of heparin are required to achieve maximum effect. Heparin-Sepharose chromatography of medium containing fetal calf serum reduces the ability of that medium to promote growth of smooth muscle cells from fetal pulmonary arteries, suggesting that heparin may remove soluble growth factors in serum. However, inhibition of fetal pulmonary artery smooth muscle cell proliferation by heparin is identical in media supplemented either with serum prepared from fetal calf plasma, in which platelet-derived growth factor (PDGF) is not detectable, or with fetal calf serum, which contains relatively abundant PDGF (114 pg/ml). Thus, inhibition of fetal pulmonary artery smooth muscle cell proliferation by heparin is not mediated solely by decreased availability or activity of exogenous PDGF. These studies suggest that morphogenesis of the smooth muscle investment of the pulmonary arteries could be regulated by local production of heparin-like inhibitors of smooth muscle cell growth.