The extracellular matrix of Volvox carteri: molecular structure of the cellular compartment.

The extracellular matrix (ECM) of Volvox contains insoluble fibrous layers that surround individual cells at a distance to form contiguous cellular compartments. Using immunological techniques, we identified a sulfated surface glycoprotein (SSG 185) as the monomeric precursor of this substructure within the ECM. The primary structure of the SSG 185 poly-peptide chain has been derived from cDNA and genomic DNA. A central domain of the protein, 80 amino acid residues long, consists almost exclusively of hydroxyproline residues. The chemical structure of the highly sulfated polysaccharide covalently attached to SSG 185 has been determined by permethylation analysis. As revealed by EM, SSG 185 is a rod-shaped molecule with a 21-nm-long polysaccharide strand protruding from its central region. The chemical nature of the cross-links between SSG 185 monomers is discussed.

The highly efficient sex-inducing pheromone system of Volvox.

The green alga Volvox is one of the simplest multicellular organisms and is capable of both asexual and sexual reproduction. Sexual development is initiated by a glycoprotein pheromone that acts at a concentration below 10(-16) M. The extracellular matrix (ECM) appears to play a key role in signal amplification: several ECM proteins contain a domain with homology to the sex-inducing pheromone.

[The cystic duct stump after laparoscopic cholecystectomy]. / Der Zystikusstumpf nach laparoskopischer Cholezystektomie.

The aim of this study was to evaluate the length of cystic-duct stumps after laparoscopic cholecystectomy. 113 patients underwent intravenous cholangiography 2 to 3 months postoperatively, whereby a cystic-duct remnant of up to 1 cm was found in 34.5% (n = 39) and between 1 and 2 cm in 36.3% (n = 41). In 24.8% (n = 28) a stump measuring 2 to 3 cm was registered and in 4.4% (n = 5) the cystic-duct remnant was more than 3 cm. Possible consequences with regard to the development of the postcholecystectomy syndrome are discussed. In our experience laparoscopic cholangiography seems to be a useful procedure to detect cysticolithiasis and as preliminary measure to undertaking the correct therapeutic steps. Adherence of this strategy might avoid possible later complications by elimination of their principal cause.

Sulfation of a cell surface glycoprotein correlates with the developmental program during embryogenesis of Volvox carteri.

A sulfated cell surface glycoprotein with an apparent molecular weight of 185,000 is synthesized in the multicellular organism Volvox only during the limited period of embryogenesis. The lifetime of sulfate residues on this glycoprotein is very short (half-life about 20 min). Production of this sulfated glycoprotein sharply decreases to a minimum shortly before the onset of the differentiating cell cleavage--e.g., in asexual development, before the 32-cell embryo divides. It is demonstrated that the sulfated glycoprotein behaves in many respects as would the hypothetical cell surface component postulated by a recently published model [Sumper, M. (1979) FEBS Lett. 107, 241-246], which proposes an explanation for the cell-counting mechanism and the spatial control of differentiation that is operative in Volvox embryogenesis.

A novel glycosphingolipid that may participate in embryo inversion in Volvox carteri.

Mouse monoclonal antibody 4-I-244 detects a developmentally regulated antigen in embryos of Volvox carteri and inhibits specifically the morphogenetic process of inversion (the process by which the embryo turns inside out). Antigen 4-I-244 was chemically characterized as a complex phytosphingolipid containing the neutral sugars xylose, galactose, and glucose as well as inositol and phosphate.

Development-dependent modification of the extracellular matrix by a sulphated glycoprotein in Volvox carteri.

We report the chemical characterization of the highly sulphated glycoprotein SSG 185 from Volvox carteri. SSG 185 is a hydroxyproline-containing, extracellular glycoprotein. The sulphate residues are clustered within the parent saccharide structure of SSG 185, since on mercaptolysis all the sulphate residues are recovered in a small saccharide fragment containing mannose, arabinose and sulphate (in a molar ratio of 112). SSG 185 is a short-lived molecule, serving as a precursor for a high mol. wt. component of the extracellular matrix. Synthesis of SSG 185 is developmentally controlled. Different SSG 185 variants, with unknown modifications in the sulphated saccharide fragment, are synthesized at different developmental stages or under the influence of the sexual inducer. These modifications remain conserved in the aggregated state of SSG 185, indicating the development-dependent modification of the extracellular matrix.

Self-assembly and cross-linking of Volvox extracellular matrix glycoproteins are specifically inhibited by Ellman's reagent.

A major impediment to the biochemical characterization of extracellular matrices from algae (as well as higher plants) is the extensive covalent cross-linking that exists in the matrix, rendering most components insoluble and resistant to conventional extraction procedures. In the multicellular green alga Volvox, biogenesis of the extracellular matrix (ECM) is initiated immediately after the process of embryonic inversion. At this stage of development, the sulfhydryl reagent 5, 5'-dithio-bis(2-nitrobenzoic acid), known as Ellman's reagent, interferes in a highly specific manner with ECM biogenesis. Treated post-inversion embryos are no longer able to assemble an intact ECM and consequently dissociate into a suspension of single cells. Dissociated cells remain viable and continue to secrete ECM proteins into the growth medium, as documented by the identification of several members of the pherophorin family. Cross-linked ECM polymers such as sulfated surface glycoprotein 185 remain in a soluble state. Thus, treatment with Ellman's reagent opens a simple approach for the isolation and characterization of otherwise inaccessible monomeric precursors.

A novel extensin that may organize extracellular matrix biogenesis in Volvox carteri.

ISG is a sulphated, extracellular glycoprotein synthesized for only a few minutes in inverting Volvox embryos and inverting sperm cell packets. This control operates at the level of transcription. ISG has been characterized by studies of protein chemistry and electron microscopy. The primary structure of ISG has been derived from genomic DNA and cDNA. ISG is composed of a globular and a rod-shaped domain. The rod-shaped domain represents a member of the extensin family with numerous repeats of Ser-(Hyp)4-6 motifs. A synthetic decapeptide matching the C-terminal sequence is able to disaggregate the organism into individual cells. Immunofluorescence microscopy localizes ISG within the boundary zone of the ECM.

Differential targeting of closely related ECM glycoproteins: the pherophorin family from Volvox.

The alga Volvox carteri represents one of the simplest multicellular organisms. Its extracellular matrix (ECM) is modified under developmental control, e.g. under the influence of the sex-inducing pheromone that triggers development of males and females at a concentration below 10(-16) M. A novel ECM glycoprotein (pherophorin-S) synthesized in response to this pheromone was identified and characterized. Although being a typical member of the pherophorins, which are identified by a C-terminal domain with sequence homology to the sex-inducing pheromone, pherophorin-S exhibits a completely novel set of properties. In contrast to the other members of the family, which are found as part of the insoluble ECM structures of the cellular zone, pherophorin-S is targeted to the cell-free interior of the spherical organism and remains in a soluble state. A main structural difference is the presence of a polyhydroxyproline spacer in pherophorin-S that is linked to a saccharide containing a phosphodiester bridge between two arabinose residues. Sequence comparisons indicate that the self-assembling proteins that create the main parts of the complex Volvox ECM have evolved from a common ancestral gene.

How a sex pheromone might act at a concentration below 10(-16) M.

The sex-inducing pheromone of Volvox carteri is a glycoprotein that triggers development of males and females at a concentration below 10(-16) M. Evidence is presented for the existence of a novel mechanism of signal amplification operating within the extracellular matrix of this multicellular organism. A family of 70 kDa matrix glycoproteins denoted pherophorins bear a C-terminal domain being homologous to the sex-inducing pheromone. Under the influence of the pheromone, this domain is liberated by highly specific proteolysis.

Intraperitoneal cholelithiasis after laparoscopic cholecystectomy--behavior of 'lost' concrements and their role in abscess formation.

UNLABELLED: In two experimental studies we sought preliminary information about the behavior of concrements lost in the peritoneal cavity during laparoscopic cholecystectomy. MATERIALS AND METHODS: In study 1, human gallstones were analyzed using X-ray diffraction, classified in three groups and examined with an ultramicroscope; then they were implanted in the peritoneal cavity of rats. After 8 weeks or 6 months, the animals were sacrificed and the concrements analyzed again as before. The tissues surrounding the calculi were also examined histologically. In study 2, human gallstones were examined with regard to bacterial contamination on the surface or in the middle of the calculi. The cholesterol content was analyzed, and the stones were divided into three groups and implantated in the rats as in the first study. After 8 weeks, the animals were sacrificed and areas with identifiable tissue reactions were examined histologically and microbiologically. RESULTS: The concrements lost their crystalline formation without any relation to their former cholesterol content, as shown by X-ray diffraction as well as ultramicroscopy. Mineralogically, these changes are a certain sign of structural dissolution. Cholesterol stones only caused abscess formations in association with gram-negative bowel germs. Sterile pigment concrements often led to a mesenchymal reaction such as granulomas. Contaminated pigment stones also resulted in extensive abscess formations.