The spatial and temporal relationships between cytoplasmic filaments and the morphogenesis of the intestinal brush border were examined by transmission electron microscopy of normally developing tissue and of tissue exposed to a variety of experimental conditions in organ culture. Distinct stages in the development of the brush border were identified: (1) Irregular projections of the apical plasma membrane that contain a network of microfilaments are converted to uniform projections filled with a core bundle of straight microfilaments (7-11d of incubation). (2) Rootlets form by an elongation or aggregation of filaments (11-15d). (3) The terminal web forms first as a network of short filaments just below the apical plasma membrane, then secondarily stratifies into two layers (19d of incubation to 3d posthatching). (4) Core filaments elongate as microvilli achieve their maturity (21d of incubation to 5d posthatching). Microvillus formation was not perturbed by culturing 9d tissue in high concentrations of Ca++ or Mg++, either with or without the ionophore, A23187. Rootlet formation was stimulated by high Mg++, with or without A23187, and, for reasons unknown, by ethanol. Terminal web formation was not stimulated by Mg++ or Ca++, but the integrity of the terminal web was lost when 21d embryonic tissue was cultured with EGTA or cytochalasin B. After stratification, the terminal web could not be disrupted by EGTA, but instead was aggregated to the center of the apical end of the cell.
The beta-crystallins are a major constituent of the mammalian lens, where they associate into dimers, tetramers and higher order aggregates. Appropriate association of lens crystallins is important for lens transparency. To examine the associative properties of betaB2-crystallin, we have expressed mouse betaB2-crystallin using a baculovirus system. Recombinant mouse betaB2-crystallin has an estimated monomer molecular weight of 24 kDa by SDS-PAGE, appropriate immunoreactivity and appropriate secondary structure as assessed by circular dichroism analysis. The recombinant betaB2-crystallin associates into a homodimer with a weight average molecular mass of 39 kDa. The betaB2-crystallin homodimer has an estimated Kd of 5 x 10(-6) M, slightly greater than that of betaA3-crystallin, 0.8 x 10(-6) M. When recombinant betaB2-crystallin is combined with recombinant betaA3-crystallin, a heterodimer is formed within 10 min of incubation at room temperature. When equilibrium is reached in 4-6 h, approximately half of each crystallin associates into heterodimers. Subunit exchange between betaB2-crystallin and betaA3-crystallin occurs readily in the absence of any denaturing agents. Thus, rbetaA3-rbetaB2 heterodimer formation can occur under conditions similar to those found in the eye lens.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.