Intra-nuclear localization of two envelope proteins, gB and gD, of herpes simplex virus.

The envelopes of herpes simplex virus (HSV) particles are acquired from the inner nuclear membrane (INM) of the infected cell and virus-coded glycoproteins are present in the envelope of mature virions. Our ultrastructural study examined the process of virus envelopment and the targeting of two major viral glycoproteins, gB and gD, to the INM in HSV-infected human embryonic fibroblasts. It was shown that envelopment and transport of virus particles from the nucleus is facilitated by the formation of a dynamic tubulo-reticulum arising from the INM.

A negative contrast stain for ultra-thin frozen sections.

Ultra-thin frozen sections are ideal substrates for immunolabelling in high resolution electron microscopy. However, visualization of subcellular structures is inferior to that obtained with corresponding plastic sections. Although negative staining is generally effective and even superior to positive staining, the accumulated stain is often too heavy, obscuring morphology and markers used for immunocytochemical localization of antigens.

Introduction of a gonadotropin receptor expression plasmid into immortalized granulosa cells leads to reconstitution of hormone-dependent steroidogenesis.

We have recently succeeded in immortalizing rat granulosa cells by co-transfection with SV-40 DNA and the Ha-ras oncogene. These cells lost their response to gonadotropins, but expressed the cytochrome P450scc mitochondrial system enzymes and produced progesterone and 20 alpha-hydroxy-4-pregnan-3-one (20 alpha-OH-P) upon cAMP stimulation (Suh, B. S.

A 150 Kilodalton Cell Surface Protein Is Induced by Salt in the Halotolerant Green Alga Dunaliella salina.

Dunaliella salina is an extremely halotolerant, unicellular, green alga lacking a rigid cell wall. Osmotic adaptation to high salinities is based on the accumulation of glycerol. To uncover other functions responsible for halotolerance, protein profiles of algae continuously grown in different salinities were compared.

A major stylar matrix polypeptide (sp41) is a member of the pathogenesis-related proteins superclass.

A novel stylar-specific glycosylated protein, sp41, was characterized. Sp41 constitutes greater than 12% of the transmitting tract tissue soluble proteins and is mainly localized in the extracellular matrix. Two cDNA clones corresponding to sp41 mRNA were isolated and sequenced.

Induction and mitochondrial localization of cytochrome P450scc system enzymes in normal and transformed ovarian granulosa cells.

After ovulation of an oocyte, granulosa cells of the ovarian follicle differentiate into luteal cells and become a major factor dedicated to the synthesis of the steroid hormone progesterone. We recently established granulosa cell lines by cotransfection of granulosa cells with SV-40 and Ha-ras oncogene. In these cells progesterone secretion can be induced by cAMP as in normal rat granulosa cells.

Ultracryotomy for the study of unfixed membranes.

Plasma membranes from chromaffin cells of bovine adrenal medullae and from chicken macrophages were isolated on a urografin density gradient, frozen and sectioned without previous chemical fixation. Their receptor binding sites were localized by specific labelling. The sections were then post-fixed in the presence of K2Cr2O7 to produce positive staining of the membrane proteins.

Immunocytochemical localization of the mannose receptor on ultrathin cryosections of chicken macrophages.

Emulphogene-solubilized chicken macrophages were used for the isolation of the mannose receptor by affinity chromatography on mannose-sepharose. From 5 X 10(9) cells 1 microgram protein was obtained, which was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) into 2 bands with an approximate molecular weight of 130 and 170 kDa. The agglutinating activity was assayed with mannan-coated M.

Electron Microscope Characterization of Carbohydrate Residues on the Body Wall of Xiphinema index.

The location of carbohydrate moieties on the outer cuticle of Xiphinema index was examined by electron microcopy using several different reagents: a) The periodic acid-thiosemicarbazide-silver proteinate reaction was used as a general stain for carbohydrates. In sectioned material it stained the canal system and deeper layers of the cuticle as well as the outer surface, b) Cationized ferritin at pH 2.5, which identifies carboxyl and sulfate groups, was used to identify sialic acid residues and also labelled parts of the canal system, c) Ferritin-goat anti rabbit IgG coupled to a DNP ligand was used to label either sialyl or galactosyl/N-acetyl-D-galactosaminyl residues, d) Ferritin hydrazide, a new reagent, was used for the ultrastructural localization of glyco-conjugates.

Binding of mannosylated ferritin to chicken bone marrow macrophages.

Ferritin conjugated to diazo p-aminophenylmannoside and mannan was used for ultrastructural visualization of binding and endocytosis via the mannose receptor. Conjugates were bound by live macrophages but not by glutaraldehyde-fixed cells. Binding was inhibited by 0.

Caenorhabditis elegans: characters of negatively charged groups on the cuticle and intestine.

Partial characterization of carboxyl, sulfate, and phosphate groups on the Caenorhabditis elegans cuticle and intestinal microvilli was achieved by en face labeling of floating cryosections at two pH levels and specific blockage of sulfate groups by Alcian blue. All negatively charged groups on the cuticle and intestinal microvilli labeled heavily at pH 7.2-7.

Partial Characterization of the Cuticle Surface of Meloidogyne javanica Females.

Negative charges on the outer cuticular surface of Meloidogyne javanica females were visualized with electron microscope labelling techniques. Evidence is presented that the electronegative charge is not borne on neuraminic acid. Ruthenium red staining indicated acid mucopolysaccharides on the outer surface.

Transplantable granulocytic leukemia in strain 13 guinea pigs.

The occurrence of a granulocytic leukemia in 1 of 40 female strain 13/N guinea pigs given N-nitroso-N-butylurea continuously in their drinking water for 21 weeks is reported here. This leukemia has been successfully transplanted in this guinea pig strain for 13 transplant generations by i.p.

Fine structure of the esophagus of Pratylenchus penetrans.

The fine structure of the esophagus of Pratylenchus penetrans is described. The gland lobe is syncytial and contains two types of nuclei: three large nuclei with little chromatin, and more numerous smaller nuclei with large amounts of chromatin. Some of the smaller nuclei are associated only with glandular tissue, whereas others are part of nerve ceils within the esophagus.

Protein and glycoprotein electrophoretic patterns of enriched fractions of primary and secondary granules from guinea pig polymorphonuclear leukocytes.

The postnuclear supernatant fraction of sucrose homogenates of guinea pig polymorphonuclear leukocytes (PMNL) was subjected to differential centrifugation to obtain a total particulate fraction, a particle-free supernatant fraction, highly enriched fractions of primary and secondary granules, and a membrane-rich fraction. The various fractions were solubilized in buffer containing sodium dodecyl sulfate (SDS) and analyzed for protein and glycoproteincomponents by SDS -polyacrylamide gel electrophoresis. The major glycoprotein components of the postnuclear supernatant fraction were found mainly associated with the enriched fraction of secondary granules and, to a lesser extent, with the membrane-rich fraction.