Copulatory Mechanism in Holocnemus pluchei and Pholcus opilionoides,With Notes on Male Cheliceral Apophyses and Stridulatory Organs in Pholcidae (Araneae)

Huber, B. A. 1994. Copulatory mechanism in Holocnemus pluchei and Pholcus opilionoides, with notes on male cheliceral apophyses and stridulatory organs in Pholcidae (Araneae).—Acta Zoologica (Stockholm) 76: 291–300. The pholcid spiders Holocnemus pluchei (Scopoli, 1763) and Pholcus opilionoides (Schrank, 1781) are investigated with respect to functional morphology of their genital organs using freeze-fixation of spiders during copula in liquid nitrogen and subsequent preparation of histological serial sections of the copulatory organs in functional contact. Special attention is paid to the mode of male pedipalpal arrestation before copulation, which is achieved in two quite different ways: in Pholcus by contact of the lateral cheliceral apophysis with the pedipalpal trochanter-apophysis, in Holocnemus by locking the pedipalpal trochanter between chelicera and pedipalpal coxa. The condition in Pholcus is considered to be apomorphic and to present a synapomorphy of about a dozen genera for which the name “Pholcus-group” is proposed. The stridulatory apparatus of Holocnemus pluchei is described, its biological significance discussed and an overview of accounts on stridulation in Pholcidae given.     

Serological Characterization of the Three Major Proteins of Vesicular Stomatitis Virus

The three major proteins of vesicular stomatitis virus-Indiana, glycoprotein (G), nucleoprotein (N), and membrane protein (M), were isolated and characterized by means of specific monocomponent antisera. G, N, and M proteins are distinct, nonrelated antigens with specific serological properties. The G protein is the only antigen inducing the formation of virus-neutralizing antibodies and was shown to confer immunity to mice. Specific complement-fixing and precipitating activity was demonstrated for each of the three antisera. The future use of isolated rhabdovirus components and of monospecific antisera is considered for therapeutic and diagnostic purposes as well as for virus strain differentiation and classification work.     

Quantification of mycorrhizal water uptake via high-resolution on-line water content sensors

The benefits of mycorrhizas for host plants are well known for a large number of species. However, experimental evaluations of the hyphal contribution to the total water uptake and the assessment of the bulk flow velocity in the hyphae are so far contradictory. Barley (Hordeum vulgaris L. Scarlet) with the inoculum Glomus intraradices was grown in a split plant-hyphal chamber with a 5 mm air gap. During the preparation of the chambers with a loamy-silt soil, water content sensors were inserted in each of the plant and the hyphal compartments. These sensors allow non-destructive measurements with high resolution. In total, 8 drying periods with a length of several days were applied with repeated watering following each drying period. A clear decline in water content in the hyphal compartment during each drying period supports the ability of hyphae to transfer water into the plant compartment. The difference between the decline in the hyphal compartment with and without arbuscular mycorrhyzal fungi is significant at the p?<?0.000001 level. The direct and indirect hyphal contribution to the total water uptake was estimated to be about 20%. The application of capacitance sensors for water content determination with a special geometry adapted to the plant-hyphal chambers allows the evaluation of the hyphal water flow with high accuracy.     

The synthesis of 15N- and deuterium-substituted, spin-labeled analogues of NAD+ and their use in EPR studies of dehydrogenases

Calcium efflux and EGTA-induced calcium release from an internal platelet membrane fraction have been studied after the oxalate-supported calcium uptake had reached steady state. Increasing external calcium concentrations stimulate the calcium efflux velocity, with an apparent half-maximal stimulation at about 5 microM outside calcium concentration and a maximal velocity of calcium efflux of 4.66 +/- 2.32 nmol X min-1 X mg-1. Moreover, the ratio of the liberated calcium on the loaded calcium seems to be independent of the increasing external calcium concentration. Increasing the calculated internal calcium concentration by varying the oxalate potassium concentration from 10 mM to 1 mM results in an increase of the liberated calcium from the membrane vesicles from 7.4% to 63%, respectively, without changing the calcium efflux velocity. Similar conclusions can be drawn from the observation of results from the calcium efflux and EGTA-induced calcium release methods. Moreover, calcium pump reversal does not seem to be responsible for the calcium efflux or calcium release. All these different points added to the previously described regulation of calcium efflux by the catalytic subunit of cAMP protein kinase suggest us that the mechanism of calcium liberation by the platelet membranes is different from the calcium uptake.     

Phospholipid protection against proteolysis of D-beta-hydroxybutyrate dehydrogenase, a lecithin-requiring enzyme

D-beta-Hydroxybutyrate dehydrogenase is a lipid-requiring enzyme which is localized on the inner face of the mitochondrial inner membrane. The apodehydrogenase, i.e. the purified enzyme devoid of lipid, has been purified from beef heart mitochondria and as such is inactive. It can be reactivated by insertion into phospholipid vesicles containing lecithin. Proteolytic digestion with different proteases has been carried out to obtain insight into the orientation of the enzyme in the membrane and to assess the extent of immersion of the protein into the phospholipid bilayer. Digestion of the apodehydrogenase with either trypsin, chymotrypsin, Staphylococcus aureus protease, thermolysin, carboxypeptidases A and Y, or Pronase (from Streptomyces griseus) leads to loss of activity, as assayed with phospholipid. Limited digestion with carboxypeptidase results in complete inactivation. Of the proteases tested, only Pronase and chymotrypsin cleave and inactivate the enzyme inserted into phospholipid vesicles (enzyme-phospholipid complex). For the enzyme-phospholipid complex, the loss of activity with Pronase digestion follows a single exponential decay to less than 10% of the initial activity. With chymotrypsin digestion, the staining intensity of the original approximately 31,500-dalton polypeptide decreases more rapidly than the loss of enzymic activity. The enzyme-phospholipid complex, after limited cleavage with chymotrypsin, retains enzymic activity and resonance energy transfer from protein to bound NADH and an approximately 26,000-dalton polypeptide is observed. Phospholipid alters the cleavage pattern with both chymotrypsin and Pronase, and the rate of inactivation of the enzyme-phospholipid complex is slowed in the presence of NAD(H). Moreover, the rate of inactivation of the apodehydrogenase with chymotrypsin is diminished approximately 3-fold in the presence of NAD+. Digestion of submitochondrial vesicles with either trypsin, chymotrypsin, or Pronase rapidly inactivates D-beta-hydroxybutyrate dehydrogenase; the addition of NAD+ or NADH, together with dithiothreitol and increased salt (to 50 mM), decreases the rate of inactivation, and with trypsin, virtually eliminates inactivation.(ABSTRACT TRUNCATED AT 400 WORDS)