RESUMEN
Two series of small polyatomic ions, HxCO+ and HxN2(+) (x = 1, 2, 3), were systematically characterized using three correlated theoretical techniques: density functional theory using the B3LYP functional, spin-restricted second-order perturbation theory, and singles + doubles coupled cluster theory with perturbative triples. On the basis of thermodynamic data, the existence of these ions in inductively coupled plasma mass spectrometry (ICP-MS) experiments is not surprising since the ions are predicted to be considerably more stable than their corresponding dissociation products (by 30-170 kcal/mol). While each pair of isoelectronic ions exhibit very similar thermodynamic and kinetic characteristics, there are significant differences within each series. While the mechanism for dissociation of the larger ions occurs through hydrogen abstraction, the triatomic ions (HCO+ and HN2(+)) appear to dissociate by proton abstraction. These differing mechanisms help to explain large differences in the abundances of HN2(+) and HCO+ observed in ICP-MS experiments.
RESUMEN
We have shown previously that components of the extracellular matrix (ECM) modulate neuronal development. Here, we searched for additional ECM elements that might play roles in retinal histogenesis and identified a secreted glycoprotein that is heavily expressed in the retina. This molecule, named by others Wnt Inhibitory Factor-1 (WIF-1), is expressed during and after the period of rod photoreceptor morphogenesis in the mouse. We show that a potential WIF-1 ligand, Wnt4, as well as a potential Wnt4 receptor, fzd4, and a potential Wnt4 coreceptor, LRP6, are expressed in the region of, and at the time of, rod photoreceptor genesis. WIF-1 and Wnt4 are coexpressed during retinal development and bind to each other; therefore, they are likely to interact during rod production. WIF-1 protein inhibits rod production, and anti-WIF-1 antibodies increase rod production; in contrast, Wnt4 promotes rod production. Together, these data suggest that WIF-1 and Wnt4, both components of the ECM, regulate mammalian photoreceptor development.
Asunto(s)
Proteínas Portadoras/metabolismo , Matriz Extracelular/metabolismo , Neuronas/metabolismo , Proteínas Represoras/metabolismo , Retina/crecimiento & desarrollo , Retina/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Animales , Anticuerpos/farmacología , Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/genética , Diferenciación Celular/genética , Células Cultivadas , Proteínas de la Matriz Extracelular , Receptores Frizzled , Regulación del Desarrollo de la Expresión Génica/genética , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intercelular , Péptidos y Proteínas de Señalización Intracelular , Proteína-6 Relacionada a Receptor de Lipoproteína de Baja Densidad , Ratones , Neuronas/citología , Unión Proteica/fisiología , Proteínas/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , ARN Mensajero/metabolismo , Receptores de Superficie Celular , Receptores Acoplados a Proteínas G , Receptores de LDL/metabolismo , Proteínas Represoras/antagonistas & inhibidores , Proteínas Represoras/genética , Retina/citología , Células Fotorreceptoras Retinianas Bastones/citología , Células Fotorreceptoras Retinianas Bastones/metabolismo , Regulación hacia Arriba/genética , Proteínas Wnt , Proteína Wnt4RESUMEN
Beta-ig is a secretory protein embodied by fasciclin I-like repeats containing sequences that might bind integrins and glycosaminoglycans in vivo. Expression of Beta-ig is responsive to Transforming Growth Factor-beta and the protein is found to be associated with extracellular matrix (ECM) molecules, implicating Beta-ig as an ECM adhesive protein of developmental processes. The spatiotemporal distribution of Beta-ig during various stages of murine development was examined and its ability to support adhesion of various cell types assessed. In situ hybridization of mouse embryos (E12.5-E18.5) indicated a prominent, distinct expression pattern for Beta-ig message in connective tissue. Beta-ig transcripts were abundantly expressed during mesenchymal cell condensation in areas of axial, craniofacial and appendicular primordial cartilage from E12.5-E14.5. Beginning at E15.5, Beta-ig transcripts appeared in collagen-rich tissues, including dura mater and corneal stroma. During E16.5-E18.5, Beta-ig transcripts were observed in proliferating chondrocytes and areas of endochondral ossification in joint and articular cartilage formation. Connective tissues expressed Beta-ig transcripts within the nasal septum and surrounding cartilage primordia, and in the pericardium, optic cup, kidney, ovary, esophagus, diaphragm, bronchi, trachea and corneal epithelium, and during cardiac valve formation. These patterns of expression indicate that Beta-ig may be involved in tissue morphogenesis. Cells derived from mesenchyme attached onto a substratum comprised of purified recombinant Beta-ig. Taken together, the results indicate that Beta-ig is expressed principally in collagen-rich tissues where it may interact with cells and ECM molecules, perhaps playing a role in tissue morphogenesis.
Asunto(s)
Proteínas de la Matriz Extracelular/metabolismo , Proteínas de la Matriz Extracelular/fisiología , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta/fisiología , Secuencia de Aminoácidos , Animales , Adhesión Celular/fisiología , Moléculas de Adhesión Celular/fisiología , Células Cultivadas , Condrocitos/metabolismo , Condrocitos/fisiología , Desarrollo Embrionario y Fetal , Matriz Extracelular/metabolismo , Regulación del Desarrollo de la Expresión Génica , Mesodermo/metabolismo , Ratones , Datos de Secuencia Molecular , Especificidad de ÓrganosRESUMEN
Molecules of the extracellular matrix (ECM) play important roles in the development and maintenance of myotendinous junctions (MTJs), specialized regions of muscle to bone union. In this report we provide evidence that skeletal muscle cells synthesize the collagen- and fibronectin-binding ECM protein betaIG-H3 and that betaIG-H3 is localized to MTJs. In situ hybridization experiments revealed that during E16.5-E18.5 of murine development, betaIG-H3 RNA transcripts were expressed where developing skeletal muscle fibers contact primordial cartilage and bone. Immunohistochemical analysis verified that the betaIG-H3 protein itself localized distinctively at MTJs, and ultrastructural analysis suggested that betaIG-H3 associates with extracellular fibers and the surface of cells. In vitro, recombinant betaIG-H3 functioned as an adhesion substratum for skeletal muscle cells. Adhesion was significantly reduced by anti-integrin alpha7 and beta1 antibodies, suggesting that betaIG-H3 binds to skeletal muscle cells via alpha7beta1 integrin. Localization of betaIG-H3 to the termini of skeletal muscle fibers and the binding of betaIG-H3 to cells and to molecules of the ECM suggests that betaIG-H3 may play an organizational and structural role in developing MTJs, linking skeletal muscle to components of the ECM.