0000000000016859
AUTHOR
Ulrich Hoeger
Nucleoside uptake in male germ cells of the polychaeteNereis virens
Summary Uptake of inosine and guanosine was measured in male germ ceils of the polychaete Nereis virens at different stages of development. In spermatogonia I (spg I) and spermatid stages, total inosine uptake at 12°C and ambient concentrations of 100 μmol/1 was relatively low (10–50 nmol·ml of packed cell volume pcv−l·h−1). A rapid increase (150–300 nmol·ml of pcv-1·h−1) was found during transition from spg I to spg II with a subsequent decline to low values (10–30 nmol·ml of pcv−1·h−1) in spermatocyte and spermatid stages. This transient increase may be related to the proliferative activity of spg I stages leading to spg II stages, which increases the demand of purine precursors for nucle…
Metabolite supply in oocytes of Nereis virens: role of nucleosides
Gamete development in Nereis virens and other nereidid polychaetes is organized in a simple way. Follicular tissues are absent and the germ cells develop floating freely in the coelomic fluid, which thus serves as the vehicle for the supply of substances required for oocyte growth. This overview focusses on the role of exogenous purine nucleosides for the growing oocytes. Eleocytes, a coelomic cell type which is proliferated in large amounts at the beginning of sexual maturation, supply purine nucleosides to support nucleic acid synthesis in the oocytes. Eleocytes can store large amounts of purine nucleotides (up to 50 µ mol ml−1 cell vol.) in the form of AMP and ADP. During oogenesis, thes…
Hydrolytic enzymes in the coelomic cells of the polychaeteNereis virens during sexual maturation
Nereis virens were collected between April 1989 and April 1990 at Yerseke, Oosterscheldt Bay, The Netherlands. Activities of the hydrolytic enzymes arylsulfatase, acid phosphatase, N-acetylglucosaminidase, aminopeptidase, and carboxylesterase were investigated in coelomic cells (elaeocytes) of individuals at different stages of maturation as determined by the time course of oocyte growth. On a protein-content basis, up to ten-fold higher specific activities were present in the elaeocytes compared to the body-wall tissue. Acid phosphatase, N-acetylglucosaminidase, leucine aminopeptidase, and long- and medium-chain esterase increased continuously from the beginning to the later stage of sexua…
Glutathionyl-biliverdin IXα, a new heme catabolite in a marine annelid: Sex and cell specific accumulation.
Abstract We have isolated and characterized the green pigment accumulating in coelomic cells (eleocytes) of the common clam worm Nereis virens by means of RP-HPLC and ESI-tandem mass spectrometry. This pigment is a novel biliverdin-glutathione conjugate in which the glutathione is linked to the biliverdin-backbone via a thioether bond. The yolk precursor vitellogenin, a female-specific high-density lipoprotein ( ρ = 1179 kg/m 3 ) with a native molecular mass of ∼500 kDa and a subunit mass of ∼150 kDa, is capable of transporting this pigment as well as heme. The sex-independent large discoidal lipoprotein present in the coelomic fluid, which is sequestered by the eleocytes, could also be sh…
One precursor, three apolipoproteins: The relationship between two crustacean lipoproteins, the large discoidal lipoprotein and the high density lipoprotein/β-glucan binding protein
The novel discoidal lipoprotein (dLp) recently detected in the crayfish, differs from other crustacean lipoproteins in its large size, apoprotein composition and high lipid binding capacity, We identified the dLp sequence by transcriptome analyses of the hepatopancreas and mass spectrometry. Further de novo assembly of the NGS data followed by BLAST searches using the sequence of the high density lipoprotein/1-glucan binding protein (HDL-BGBP) of Astacus leptodactylus as query revealed a putative precursor molecule with an open reading frame of 14.7 kb and a deduced primary structure of 4889 amino acids. The presence of an N-terminal lipid bind- ing domain and a DUF 1943 domain suggests the…
Nucleotide pool changes in coelomic cells (eleocytes) of the polychaete Nereis virens during sexual maturation
Eleocytes (a type of coelomic cell) of the polychaete Nereis virens can store large amounts of adenine nucleotides at certain times. Since eleocytes have specific functions related to gametogenesis, we tested whether the presence of these large nucleotide stores in eleocytes is specific to gender or related to specific events during gametogenesis. Nucleotide pools in eleocytes isolated at different stages of sexual maturation from N. virens were analysed using high-performance liquid chromatography. Eleocytes from immature and male animals had extremely high concentrations of both AMP and ADP (each > 10 µmol/ml of packed cell volume). In eleocytes from male animals, the high nucleotide stor…
Transmission electron microscopical studies on some haemolymph proteins from the marine polychaete Nereis virens.
Abstract The hexagonal bilayer haemoglobin molecule from Nereis virens has been investigated in a comparative study using several different negative stain electron microscopical specimen preparations (i.e. by conventional adsorption to continuous carbon support films, by the negative staining-carbon film technique and by negative staining across the holes of holey carbon support films with air-drying and rapid freezing/cryo-negative staining). The benefits and limitations of these different approaches are indicated, with the overall conclusion that negative staining with ammonium molybdate across holes creates the best possibilities for molecular imaging, and also has the potential for the …
Annelid Coelomic Fluid Proteins
The coelomic cavity is part of the main body plan of annelids. This fluid filled space takes up a considerable volume of the body and serves as an important site of exchange of both metabolites and proteins. In addition to low molecular substances such as amino acids and glucose and lactate, the coelomic fluid contains different proteins that can arise through release from adjacent tissues (intestine) or from secretion by coelomic cells. In this chapter, we will review the current knowledge about the proteins in the annelid coelomic fluid. Given the number of more than 20,000 extant annelid species, existing studies are confined to a relatively few species. Most studies on the oligochaetes …
Adenylate storage, metabolism and utilization in coelomic cells of the polychaeteNereis virens (Annelida, polychaeta)
Eleocytes are specialized coelomic cells in nereid annelids which assume a central role during germ cell development. They may contain extremely high concentrations of both adenosine monophosphate (AMP) and adenosine diphosphate (ADP) (each >10 μmol/ml of cell vol.), whereas the adenosine triphosphate (ATP) content is comparatively low (0.8 μmol/ml cell vol.).31P nuclear magnetic, resonance (NMR) studies of living eleocytes suggest the compartmentalization of both AMP and ADP in the large acidic vacuole characteristic for this cell type. Eleocytes are thus capable of storing high concentrations of ADP and AMP without inhibiting energy metabolism, by sequestering these compounds in a separat…
Lipid accumulation and metabolism in polychaete spermatogenesis: Role of the large discoidal lipoprotein
In most oviparous animals, lipoprotein-mediated lipid transport plays an important role in the nutrient supply for the oocyte. In male gametes, lipids are used as energy substrates in spermatozoa but nothing is yet known about their origin and metabolism throughout spermatogenesis. The lipid profiles analyzed from different stages of male germ cell development in the marine annelid Nereis virens were found to undergo a dramatic change from primary triacylglycerides at the beginning of germ cell development to cholesterol and phospholipids at the end of development as demonstrated by HPLC with evaporative light scattering detection and mass spectrometry. The uptake of a large discoidal lipop…
Extreme nucleoside concentrations in a marine annelid: a novel nucleoside storing cell in the polychaete Nereis virens.
A new type of free coelomic cell was isolated from the coelomic fluid of the polychaete Nereis virens. This cell type was present only during sexual maturation and only in males with fully developed spermatozoa. In vivo acridine orange staining and electron microscopy showed that this cell type consists mainly of a large acidic vacuole with a very low proportion of cytoplasm. This cell type stores extremely high concentration of purine nucleosides reaching concentrations of >700 millimolar on a cell weight basis. The nucleoside concentrations are the highest reported from any living cell so far. Inosine (280-711 micromol/g cell mass), guanosine (8-109 micromol/g), deoxyinosine (3-26 micromo…
Crustacean Hemolymph Lipoproteins
Lipoproteins mediate the transport of apolar lipids in the hydrophilic environment of physiological fluids such as the vertebrate blood and the arthropod hemolymph. In this overview, we will focus on the hemolymph lipoproteins in Crustacea that have received most attention during the last years: the high density lipoprotein/β-glucan binding proteins (HDL-BGBPs), the vitellogenins (VGs), the clotting proteins (CPs) and the more recently discovered large discoidal lipoproteins (dLPs). VGs are female specific lipoproteins which supply both proteins and lipids as storage material for the oocyte for later use by the developing embryo. Unusual within the invertebrates, the crustacean yolk protein…
Metabolic enzymes in coelomic cells (eleocytes) of the polychaete Nereis virens: sex specific changes during sexual maturation
The activities of some enzymes of the intermediary metabolism and the content of soluble protein and carbohydrate (glycogen plus free glucose) were measured in one type of coelomic cells (eleocytes) of the polychaete Nereis virens. Specimens used in this study were collected between 1989 and 1991 in Oosterscheldt Bay, The Netherlands, and divided into six different stages of sexual maturation as determined by the mean oocyte volume. In both sexes, the soluble protein content in eleocytes of immature individuals (11 mg ml−1 cell vol) increased three-fold. In prespawning N. virens the soluble protein content decreased to less than 2 mg protein ml−1 cell vol in females but not in males. In bot…
Lipoprotein-induced phenoloxidase-activity in tarantula hemocyanin.
Phenoloxidases play vital roles in invertebrate innate immune reactions, wound closure and sclerotization processes in arthropods. In chelicerates, where phenoloxidases are lacking, phenoloxidase-activity can be induced in the oxygen carrier hemocyanin in vitro by proteolytic cleavage, incubation with the artificial inducer SDS, or lipids. The role of protein-protein interaction has up to now received little attention. This is remarkable, as lipoproteins - complexes of proteins and lipids - are present at high concentrations in arthropod hemolymph. We characterized the three lipoproteins present in tarantula hemolymph, two high-density lipoproteins and one very high-density lipoprotein, and…
The Pandinus imperator haemolymph lipoprotein, an unusual phosphatidylserine carrying lipoprotein.
The haemolymph lipoprotein of the scorpion, Pandinus imperator was isolated and characterised. Contrary to the lipoproteins of insects and the discoidal HDL-lipoproteins of a crayfish and polychaete, the Pandinus lipoprotein consists of three instead of two apoproteins (apoPiLp I = 230 kDa, apoPiLp II = 130 kDa and apoPiLp III = 120 kDa). The apolipoproteins are arranged in varying stoichiometries as judged by cross-linking experiments. In lipoprotein samples from individual animals, the two smaller subunits occurred in a 1:1 stoichiometry, while the relative amount of the 230 kDa peptide varied. The lipoprotein is a slightly heart-shaped HDL with a diameter of approximately 15 nm. It is pr…
A discoidal lipoprotein from the coelomic fluid of the polychaete Nereis virens.
A discoidal lipoprotein was isolated from the coelomic fluid of the polychaete, Nereis virens, by density gradient centrifugation. The lipoprotein was present in both sexes and moved as a uniform band in an agarose gel. The average diameter of the lipoprotein particles determined by electron microscopy was 42 nm with a thickness of 10 nm. SDS electrophoresis showed two apoprotein subunits with molecular masses of 247 and 85 kDa, respectively. In lectin blots, both apoproteins were reactive with Concanavalin A indicating the presence of N-glycans. The small subunit was also reactive with peanut lectin, indicating additional O-glycosylation. The total lipid content was 48% and consisted mainl…