“Wall-papering” and elaborate nest architecture in the ponerine antHarpegnathos saltator

Summary Excavation of 18 nests ofHarpegnathos saltator from southern India revealed an unusually complex architecture for a ponerine ant. The inhabited chambers are not deep in the ground. The uppermost chamber is protected by a thick vaulted roof, on the outside of which is an intervening space serving as isolation from the surrounding soil. In large colonies, the vaulted roof is extended into a shell which encloses several superimposed chambers. Little openings, which may be encircled by moulded flanges, occur in the upper region of the shell. The inside of the chambers is partly or completely lined with strips of empty cocoons. A refuse chamber is always found deeper than the inhabited chambers; live dipteran larvae (family Milichiidae) are typically present. These elaborate nests represent a large energetic investment, and we speculate therefore that nest emigration is unlikely in this species. Consequently, colony fission may never occur, unlike other ants where gamergates reproduce.     

ISOLATION AND CHARACTERIZATION OF PHASE-SPECIFIC COMPLEMENTARY DNAs FROM SPOROPHYTES AND GAMETOPHYTES OF PORPHYRA PURPUREA (RHODOPHYTA) USING SUBTRACTED COMPLEMENTARY DNA LIBRARIES1

The red alga Porphyra purpurea (Roth) C. Agardh has a life cycle that alternates between shell-boring, filamentous sporophytes and free-living, foliose gametophytes. The significant morphological differences between these two phases suggest that many genes should be developmentally regulated and expressed in a phase-specific manner. In this study, we prepared and screened subtracted complementary DNA (cDNA) libraries specific for the sporophyte and gametophyte of P. purpurea. This involved the construction of cDNA libraries from each phase, followed by the removal of common clones through subtractive hybridization. Sampling of the subtracted libraries indicated that 8–10% of the recombinant colonies in each library were specific for the appropriate phase. Of 20 putative phase-specific cDNAs selected from each subtracted library, eight unique clones were obtained for the sporophyte and seven for the gametophyte. After confirming their phase-specificities by hybridization to gametophyte and sporophyte messenger RNA, these 15 phase-specific cDNAs were sequenced, and the deduced amino acid sequences were used to search protein databanks. Two proteins encoded by the sporophyte-specific cDNAs and two by the gametophyte-specific cDNAs were identified by their similarity to databank entries.     

α2-Adrenoceptor-Mediated Inhibition of Electrically Evoked [3H]Noradrenaline Release from Chick Sympathetic Neurons: Role of Cyclic AMP

Abstract: This study explores the role of cyclic AMP in electrically evoked [³H]noradrenaline release and in the α₂-adrenergic modulation of this release in chick sympathetic neurons. Along with an increase in stimulation-evoked tritium overflow, applications of forskolin enhanced the formation of intracellular cyclic AMP. Both effects of forskolin were potentiated by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine. The forskolin-induced increase in overflow was abolished by the Rp-diastereomer of cyclic AMP-thioate, an antagonist at cyclic AMP-dependent protein kinases, and 1,9-dideoxy-forskolin, an inactive analogue at adenylyl cyclase, had no effect on the evoked overflow. A 24-h pretreatment with either cholera toxin or forskolin reduced the subsequent forskolin-induced accumulation of cyclic AMP and inhibited the stimulation-evoked release. Basal cyclic AMP production, however, remained unaltered after forskolin treatment and was enhanced after 24 h of cholera toxin exposure. The α₂-adrenergic agonist bromoxidine did not affect the formation of cyclic AMP stimulated by forskolin but reduced electrically evoked release. However, effects of bromoxidine on ³H overflow were attenuated by forskolin as well as by 8-bromo-cyclic AMP. Effects of bromoxidine on [³H]noradrenaline release were paralleled by an inhibition of voltage-activated Ca²⁺ currents, primarily through a delayed time course of current activation. This effect was abolished when either forskolin or 8-bromo-cyclic AMP was included in the pipette solution. Both substances, however, failed to affect Ca²⁺ currents in the absence of bromoxidine. These results suggest that the signaling cascade of the α₂-adrenergic inhibition of noradrenaline release involves voltage-activated Ca²⁺ channels but not cyclic AMP. Elevated levels of cyclic AMP, however, antagonize this α₂-adrenergic reduction, apparently through a disinhibition of Ca²⁺ channels.     

Purification and partial characterization of a glutamyl-tRNA synthetase from the unicellular green algaScenedesmus obliquus,mutant C-2A′

The synthesis of 5-aminolevulinic acid commences with the ligation of glutamate to a specific tRNA^Glu by a glutamyl-tRNA synthetase (E.C. 6.1.1.17) (Huang et al., 1984, Science 225, 1482–1484). The synthetase from the yellow pigment mutant C-2A of the unicellular green alga Scenedesmus obliquus was purified by sequential column chromatography on Sephacryl S-300, Blue Sepharose, phosphocellulose P11 and by fast protein liquid chromatography (FPLC) on Mono Q. After denaturing sodium dodecylsulfate (SDS)-gel electrophoresis the purified enzyme preparation revealed a single protein band with a molecular mass of 55 kDa, proving the apparent homogeneity of the glutamyl-tRNA synthetase. A molecular mass of 105 ± 10 kDa was determined for the native protein by chromatography on Sephadex G-150. From these data it can be concluded that the glutamyl-tRNA synthetase from S. obliquus is a homodimer. The purified protein is active within a pH range from 7.0 to 9.0 with a maximum activity at pH 8.0. Kinetics for the binding of glutamate to the tRNA, performed with highly purified enzyme preparations, showed a K _m value of 2.3 M ± 0.3 for glutamate.Abbreviations ALA 5-aminolevulinic acid - FPLC fast protein liquid chromatography - Glu glutamate - Hepes N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - SDS sodium dodecylsulfate - Tricine N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]-glycine This work was supported by a grant of the Deutsche Forschungsgemeinschaft. U.C. Vothknecht is grateful for a Nachwuchs-förderungsstipendium des Landes Hessen. The authors want to thank Ms. B. Böhm, J. Gade and K. Eckhardt for skillful technical assistance. The authors also want to thank Dr. C.G. Kannangara (Carlsberg Institute, Kopenhagen, Denmark) for the donation of tRNA from barley and Dr. D. Jahn (FB Biology/Microbiology, Philipps-University, Marburg, FRG) for the tRNA^Glufrom E. coli.     

Vacuolar ion currents in the primitive green algaEremosphaera viridis: The electrical properties are suggestive of both the Characeae and higher plants

Summary The electrical properties of the vacuolar membrane of the primitive green algaEremosphaera virdis were investigated using the patch-clamp technique. In whole vacuole measurements two types of transport systems with long activation time-constants were identified. The first, showing marked outward rectification, was activated by an increase in the cytosolic calcium concentration. Furthermore, it displayed sensitivity to micromolar concentrations of the anion channel blocker Zn²⁺ and to acidification of the cytosol. In contrast, the second time-activated current component was almost insensitive to changes in cytosolic pH and was blocked by the potassium channel inhibitor TEA. In addition to these slowly activating current components, the vacuolar membrane contained at least two further transport systems, responsible for an instantaneous current. These two current components were distinguished by their different sensitivity to protons, cytosolic calcium, and TEA. Comparing these electrical properties to those observed in vacuoles of higher plants or in cytoplasmic droplets from characean algae, respectively, it seems thatEremosphaera is intermediate, corresponding to the systematic position of this simple green alga.Abbreviations [Ca²⁺]_cyt cytosolic free calcium concentration - EGTA ethyleneglycol-bis(-aminoethylether)N,N,N,N-tetraacetic acid - HEPES N-[2-hydroxyethyl]piperazine-N-[2-ethanesulfonic acid] - I electric current - IRC inward rectifying current - MES 2-[N-morpholino]ethanesulfonic acid - ORC outward rectifying current - pH_cyt cytosolic pH - pH_vac vacuolar pH - P_o open probability - P_x permeability coefficient of ion species X - TEA tetraethylammonium chloride - Tris tris[hydroxymethyl]aminomethane - V voltage     

Comparative sensitivity analysis of stable stage structures and reproductive values

New formulas for deriving the sensitivities of stable stage structures and reproductive values to changes in vital rates are presented. They enable comparison of the sensities to changes of different elements in the projection matrix; in other words, comparison of partial derivatives of the eigenvectors. These kinds of sensitivities can be used in applied problems such as an analysis of the effect of harvesting on the population structure. However, in this paper, we examine the application of the sensitivities in a more general ecological context. We investigate why the stable stage structure of the mustard aphid,Lipaphis erysimi, changes very little in the temperature interval 10–30°C. The sensitivities of the stable stage structure at 15°C and 25°C were derived. The character of the sensitivites were the same in both temperatures although the stage structure was more sensitive to changes at 15°C than at 25°C. The sensitivity analysis also revealed that the temperature variation results in changes in fecundity and developmental rate that have a counteractive effect on the population structure.     

Cytological characterization of isolated sperm cells of perennial ryegrass (Lolium perenne L.)

Summary The cytoplasmic content and the distribution of intramembrane particles (IMPs) of the plasma membrane of isolated sperm cells of perennial ryegrass (Lolium perenne L.) have been characterized using flow cytometry, transmission electron microscopy, confocal scanning laser microscopy and freeze-fracture studies. The isolated haploid sperm cells contain a variety of cell organelles with the exception of microtubules. Proplastids and plastids with starch were observed, although only rarely. Vacuoles containing remnants of organelles and stacked lamellae of endoplasmic reticulum with cytoplasmic inclusions were observed frequently, indicating that autophagy takes place. The number of mitochondria varies from 11 to 26 with an average of 17. Generally, the nucleus has a lobed shape and displays various interphasic stages of chromatin condensation. The analysis of the number of mitochondria and the nuclear state did not show evidence of sperm cell dimorphism. The cytological variability observed, could be explained by differences in developmental stages already present in vivo at the moment of isolation. No correlation between the number of mitochondria and the nuclear cross-sectioned area and/or the condensation state of the chromatin could be found. The density of intramembrane particles of the plasma membrane on the exoplasmic fracture face is more than twice that on the protoplasmic fracture face. That is the opposite of what was found for sporophytic cells of perennial ryegrass. These results are discussed in relation to the potential use of these cells for biotechnology and developmental studies.     

Postpartum anestrus in French beef cattle: an epidemiological study

Postpartum anestrus of lactating beef cows was studied by means of an epidemiological study carried out on 878 lactating beef cows in 60 French herds. The cows calved between October 1992 and March 1993 and were housed 2 mo after calving, when the anestrus status was determined by progesterone radioimmunoassays. Data analysis was performed using a multiple logistic model in order to adjust for confounding and interaction. Fifty-one percent of the primiparous and 23% of the multiparous cows were found to be in anestrus. Factors significantly related to anestrus were parity (primiparous); breed (Charolais); housing type (tie housing); suckling (compared to weaning at birth); and, among those that were under the control of the farmer, calving conditions (manual exploration of the birth canal); body condition score at calving (3 or less, on a 5-point scale); and loss in body condition score after calving (1 point or more within 2 mo). Previous reproductive performance for multiparous cows such as a long calving interval and induced estrus in the previous year also appeared to be related to anestrus.     

Pool sequencing of natural HLA-DR,DQ, and DP ligands reveals detailed peptide motifs,constraints of processing,and general rules

We have approached the problem of MHC class II ligand motifs by pool sequencing natural peptides eluted from HLA-DR, DQ, and DP molecules. The results indicate surprisingly clear patterns, although not quite as clear as with natural class I ligands. The most striking feature is a highly dominant Proline at position 2. We interpret this to be a consequence of aminopeptidase N-like activity in processing. Another general aspect is the existence of three to four hydrophobic or aromatic anchors, whereby the first and the last are separated by five to eight residues. The peptide motifs for HLA-DR1, DR5, DQ7, and DPw4 are allele-specific and differ by spacing and occupancy of anchors. The anchors tend to be flanked by clusters of charged residues, and small residues, especially Ala, are frequent in the motif centers. These detailed motifs allow one to interpret most previous (DR-) motifs as fitting one or more of the anchors or conserved clusters. The relative motif symmetry suggests the possibility of bidirectional binding of peptides in the class II groove.