Formation of cation channels in planar lipid bilayers by brefeldin A.

Brefeldin A (BFA) is a novel agent with the unique property of effecting a rapid increase of Golgi cisternae volume and subsequent loss of a recognizable Golgi apparatus in treated cells. Although a receptor-mediated mechanism has been proposed, the molecular basis of BFA action remains unknown (Lippincott-Schwartz, J., Glickman, J., Donaldson, J. G., Robbins, J., Kreis, T. E., Seamon, K. B., Sheetz, M. P., and Klausner, R. D. (1991) J. Cell Biol. 112, 567-577). Since a variety of ionophores distort Golgi architecture by initially causing osmotic swelling of the cisternae (Mollenhauer, H. H., Morre, D. J., and Rowe, L. D. (1990) Biochim. Biophys. Acta 1031, 225-246), Golgi membrane permeabilization by BFA seemed possible. We examined the effects of BFA on the conductance of planar lipid bilayers bathed in several aqueous salt solutions. Addition of BFA (1 microgram/ml) quickly augmented alkali cation conductance (K+ greater than Na+ much greater than Li+) but not anion conductance of the bilayer. Lower concentrations (1 ng/ml) indicated that BFA formed discrete, cation-selective channels in these bilayers. Given that Golgi cisternae volume increases immediately upon treatment with BFA, these findings suggest that alteration of ion gradients or Golgi membrane potential followed by an influx of water may be the mechanism by which BFA initiates disruption of Golgi structural integrity. Subsequent functional perturbations may then ensue either as a consequence of these initial structural changes or by a combination of several distinct mechanisms.     

Uptake and utilization of CDP-choline in primary brain cell cultures from fetal brain

The utilization of double-labeled CDP-choline by cultured brain cells has been studied. CDP-choline is demonstrated to be rapidly hydrolysed into CMP and choline phosphate. The fragments, or their hydrolysis products, penetrate into the cells and are utilized for lipid synthesis. At short times after the isotope administration a rapid labeling of phosphatidylcholine was detected, when cells were incubated with CDP-choline. The same was not seen when cells were incubated with labeled choline. From these observations it can be inferred that either CDP-choline can penetrate the cell membrane or that some mechanism involving CDP-choline and leading to phospholipid synthesis can work at the external surface of the plasma membranes.     

The brush cells of the common bile duct of the rat

Summary Two different fixative procedures (immersion and perfusion) and four different fixative solutions were used in order to obtain the best preservation of the brush cells of the common bile duct of the rat. The results indicate that only perfusion fixation through the common bile duct is suitable, independent of the fixative solutions and their osmolarity.Numerous brush cells were seen in the proximal and distal regions of the common bile duct. In these locations, they could be implicated in a registration and/or regulation of intraluminal pressure variations.     

Fast-white and fast-red isomyosins in guinea pig muscles

Myosins isolated from guinea pig fast-white Tensor Fasciae Latae (TFL) and fast-red Masseter (MASS) muscles displayed similar light chains pattern as revealed by two-dimen-sional gel electrophoresis. In contrast, the heavy chain stru$\text{c}$ ture was found to be different as shown by electrophoretic peptide maps obtained by treating myosins with different proteases.     

The recently established diatom Coscinodiscus wailesii (Coscinodiscales, Bacillariophyta) in Brazilian waters. I: Remarks on morphology and distribution

Coscinodiscus wailesii Gran et Angst is a large centric diatom (280–500 μm diameter) from marine phytoplankton, characterized by a cylindrical frustule with flat valvar surface, two marginal rings of rimoportulae on the mantle, and two macrorimoportulae. Cells from cultured and natural populations collected in Paranaguá Bay, Paraná, southern Brazil were observed under light and scanning electron microscopes to verify the populations’ correct identity and morphology. In both populations, a typical central rosette or a hyaline area was found in the valvar center. The species’ distribution in Brazilian waters was revised, and a discussion on possible vectors of transport was made. Blooms of the species occur sporadically in the coast of Paraná, seeming to affect the local trophic chain.     

Electrophoretic Polymorphisms and Their Taxonomic Implications in Callitrichini (Primates, Platyrrhini)

Five hundred forty-three blood samples from 15populations of the four genera of callitrichin primateswere studied electrophoretically. Polymorphism andgenetic distances were estimated for 20 loci, 13 of which were polymorphic. The lion tamarin(Leontopithecus) studied here exhibited theleast variability for these loci, while the monospecificCebuella showed the most. The genetic distancesobserved between Callithrix andCebuella genera support previous evidenceindicating a close taxonomic relationship between them.Genetic distance values obtained in this study alsosupport the synonimization of the kuhli form with Callithrix jacchuspenicillata.     

Genomic studies on killer yeasts belonging to the genusPichia

Twenty-four species belonging to the genusPichia were investigated using restriction fragment length polymorphism (RFLP) and Southern blot hybridization of their genomic DNA.Saccharomyces cerevisiae, Kluyveromyces lactis, Williopsis mrakii andCandida albicans were also included in this study. The RFLP patterns were obtained from digestion of yeast DNA with several restriction endonuclease enzymes, and showed various bands with different mobility; in most isolates, the more deeply stained bands were species-specific. This observation was confirmed by the results obtained from Southern blot hybridization of theEcoRI andXhoI RFLP patterns withP. anomala UCSC 25F DNA, digested with the same enzymes, used as probes. These bands are likely to be ribosomal DNA as shown by hybridization of digested DNA from unrelated yeast species (S. cerevisiae, K. lactis andC. albicans). However, one hybridized band, located at 3.9–4.1 Kb, seems to be peculiar to thePichia species. Our study confirms the usefulness of molecular tools in studying genetic relatedness among yeasts.     

Alterations in the morphology of glycoconjugate molecules caused by histochemical procedures: Comparison of renal glomeruli and articular cartilage

Summary The fine structure of the glycoconjugate molecules was investigated in the glomerular capillary wall of the rat kidney fixed by vascular perfusion, and in the human and rat articular cartilage fixed by immersion. Kidney and cartilage were either prefixed in aldehyde alone (group a), or with the addition of Alcian Blue 8 GX (group b), or Alcian Blue and 0.3m MgCl₂ (group c), or Acridine Orange at a low (0.01%) and high (0.1%) concentration (group d). The specimens were postfixed either in OsO₄ phosphate or cacodylate, with the exception of some of the samples in group a, for which a solution of potassium ferrocyanide-reduced OsO₄ was used (group e). All samples were conventionally dehydrated and embedded in Epon. In addition, some of the tissue samples in group c were cryoprotected, frozen in liquid Freon (–150° C) or in nitrogen slush (–210° C), both postfixed and dehydrated by cryosubstitution, and embedded in Epon (group f).The present investigations demonstrate that some well known extracellular structures such as the laminae rarae of the glomerular basement membrane or the interfibrillar matrix of the articular cartilage can be considerably altered in their morphology by the histological procedures applied. Whereas the precipitated glycoconjugates, as seen after staining with cationic dyes or reduced OsO₄ and conventional dehydration, can easily be recognized, the superposition of the extended molecules, as preserved by freezing and substitution, prevents their demonstration in native conformation.Preliminary reports on some of these findings were given at the Symposium on Morphological Sciences, Rome 1988 (Realeet al., 1989), at the XI Meeting of the Federation of European Connective Tissue Societies, Amsterdam 1988 (Brandes & Reale, 1988) and at the Symposium Recent Advances in Hereditary Nephritis, Milan, July 1989 (Reale & Luciano, 1990).