RESUMEN
Pax genes are a group of critical developmental transcriptional regulators in both invertebrates and vertebrates, characterized by the presence of a paired DNA-binding domain. Pax proteins also often contain an octapeptide motif and a C-terminal homeodomain. The genome of Nasonia vitripennis (Hymenoptera) has recently become available, and analysis of this genome alongside Apis mellifera allowed us to contribute to the phylogeny of this gene family in insects. Nasonia, a parasitic wasp, has independently evolved a similar mode of development to that of the well-studied Drosophila, making it an excellent model system for comparative studies of developmental gene networks. We report the characterization of the seven Nasonia Pax genes. We describe their genomic organization, and the embryonic expression of three of them, and uncover wider conservation of the octapeptide motif than previously described.
Asunto(s)
Redes Reguladoras de Genes/genética , Proteínas de Insectos/genética , Factores de Transcripción Paired Box/genética , Filogenia , Avispas/genética , Secuencia de Aminoácidos , Animales , Abejas/genética , Embrión no Mamífero/metabolismo , Componentes del Gen , Genómica , Hibridación in Situ , Proteínas de Insectos/metabolismo , Datos de Secuencia Molecular , Factores de Transcripción Paired Box/metabolismo , Péptidos/genética , Alineación de Secuencia , Especificidad de la EspecieRESUMEN
Previous studies found that normal human serum (NHS) contains an immunoglobulin G (IgG) antibody that mediates initiation of the classical complement pathway by nonencapsulated Cryptococcus neoformans. The present study used an enzyme-linked immunosorbent assay with whole nonencapsulated yeast cells as solid-phase antigens to demonstrate the presence of high levels of IgG antibody in each of 11 sera from normal adult donors. The IgG antibodies were of the IgG2 subclass. The antibody activity was blocked completely by treatment of serum with isolated yeast glucan. Treatment of serum with mannan or chitin had no effect on antibody levels. Antibody activity was adsorbed completely by treatment of serum with zymosan particles. Adsorption with intact cells of Saccharomyces cerevisiae or Candida albicans had no effect, suggesting that the glucan on S. cerevisiae or C. albicans is not surface exposed. Assessment of the opsonic requirements for phagocytosis of nonencapsulated cryptococci by monocyte-derived human macrophages (MO-M phi) showed that high levels of phagocytosis occurred when yeast cells were opsonized with NHS. Removal of anti-glucan antibody by adsorption with whole nonencapsulated cryptococci did not diminish opsonic activity. Heat-inactivated serum or anti-glucan antibody affinity purified from NHS lacked opsonic activity. Taken together, these results indicate that phagocytosis of nonencapsulated cryptococci by monocyte-derived human macrophages has an obligatory requirement for opsonic ligands of the complement system, with no contribution by the anti-glucan IgG that is found in NHS.
Asunto(s)
Anticuerpos Antifúngicos/inmunología , Cryptococcus neoformans/inmunología , Especificidad de Anticuerpos , Pared Celular/inmunología , Humanos , Isotipos de Inmunoglobulinas/inmunología , Proteínas OpsoninasRESUMEN
The morphology and electrophysiology of the canine distal colon were studied to compare this region with the proximal colon. Many morphological characteristics were similar including the presence of interstitial cells at the submucosal surface of the circular layer. Muscle cells near the submucosal surface had resting membrane potentials (RMPs) of -79 +/- 1 mV, and slow waves were generated in this region. Slow waves had similar waveform characteristics to those of the proximal colon, but rapid oscillations were superimposed on slow waves of some preparations. RMPs and slow waves decreased with distance from the submucosal surface. The latter were not resolvable in the myenteric half of the circular layer. Cells at the myenteric border had RMPs of -49.5 +/- 2 mV and a higher frequency oscillation of 16 min-1. Acetylcholine increased slow-wave amplitude and duration and caused fast oscillations on the plateau phase of slow waves. Isolated circular myocytes were studied with the patch-clamp technique. Cells from the submucosal border displayed voltage-dependent inward and outward currents. With outward currents blocked, the inward current was composed of two components. Nifedipine (10(-6) M) blocked a portion of the inward current but left a substantial transient component. The effect of nifedipine correlated with its effects on tissues, suggesting that two components of Ca2+ current participate in slow waves. These studies describe numerous similarities in the structure and activity of the proximal and distal portions of the colon but also show some potentially important differences between these regions.