Adénylate-cyclase/Lipase: Induction des récepteurs hormonaux

An adenylate cyclase activity (AC) was found in guinea pig brown adipose tissue (BAT), since the tissue's apparition. This enzymatic activity increased during the development and showed high values at the end of gestation. An increase of AC units per cell was observed, in addition to the cell multiplication. A norepinephrine stimulation of AC activity was observed at the end of gestation: this regulating action disappeared in the first days of extrauterine life. Neither glucagon nor ACTH had any regulating role upon AC activity during fetal and newborn life. The basal lipolytic activity which was observed in BAT of fetuses (61rst day) and neonate dramatically decreased around the 15th day. A potent lipolysis activation by norepinephrine was observed, but only after birth. The correlation observed between these enzymatic activities in presence of norepinephrine seems to indicate that the AC/lipase system was involved in the neonatal thermogenesis of guinea pigs.     

THE MOTOR END-PLATE SPECIFIC FORM OF ACETYLCHOLINESTERASE: APPEARANCE DURING EMBRYOGENESIS AND RE-INNERVATION OF RAT MUSCLE

Abstract– We have solubilized three active molecular forms of AChE from rat muscle and have confirmed the presence of one of these forms (EP form, apparent sedimentation coefficient: 16 s) uniquely at the motor end-plate regions of several skeletal muscles. This form was never detected in smooth muscle extracts. In sternocleidomastoïdian muscle it disappeared after denervation and reappeared after re-innervation in the region where nerve and muscle had come in contact. During the embryonic development of hind leg muscles the EP form appeared on the 14th or 15th day of gestation.
The EP form of muscle AchE appears to be an excellent biochemical marker of the neuromuscular junction.     

Cell-free translation of exogenous mRNA in extracts from dry pea primary axes

Extracts from the primary axes of dry pea (Pisum sativum L.) seeds are able to perform an initiation-dependent translation of exogenous mRNA. SDS polyacrylamide gel electrophoresis of the products synthesized under direction of alfalfa mosaic virus RNA (AMV-RNA) and tobacco mosaic virus RNA (TMV-RNA) shows that the fidelity of translation in this pea system is at least as high as in a wheat embryo cell-free protein synthesizing system. The endogenous messengers are also efficiently translated in extracts from the primary axes of pea seeds. The direct translation of these messengers in a homologous cell-free system may be of interest for a study of the products coded for by the long-lived messengers present in this plant.Abbreviations HEPES N-2-hydroxyethylpiperazine-N-ethanesulfonic acid - SDS sodium dodecyl sulphate - mRNP messenger ribonucleoprotein - AMV-RNA alfalfa mosaic virus RNA - TMV-RNA tobacco mosaic virus RNA - ATA aurin tricarboxylic acid - TCA trichloroacetic acid - S.A. specific activity     

«Noir de Diaminobenzidine»: Une nouvelle méthode histochimique de révélation du fer exogène

Résumé Une nouvelle réaction de mise en évidence histochimique du fer exogène, utilisé comme traceur pour l'étude des connexions dans le système nerveux central, est décrite. Elle comporte la transformation du fer en bleu de Prusse, lequel est utilisé comme catalyseur de l'oxydation de la diaminobenzidine par l'eau oxygénée. La diaminobenzidine oxydée précipite sous forme d'un produit insoluble de couleur brun-noir. Cette réaction présente une sensibilité et un contraste nettement supérieurs à ceux de la réaction classique de Perls.

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Diaminobenzidine black as a new histochemical demonstration of exogenous iron

Summary A new reaction for the histochemical demonstration of exogenous iron, used as a tracer for the study of connectivity in the central nervous system, is described. It consists of, first, the conversion of iron into Prussian blue, which acts secondarily as a catalyst for the oxidation of diaminobenzidine by hydrogen peroxide. The oxidized diaminobenzidine precipitates by polymerization and gives rise to a brown-colored insoluble reaction product. This reaction has a strikingly better sensitivity and contrast than the classical Perls's reaction.

     

Evolution of nuclear rDNA its sequences in the Cladophora albida/sericea clade (Chlorophyta)

Ribosomal DNA ITS sequences were compared among 13 different species and biogeographic isolates from the monophyletic albida/sericea clade in the green algal genus Cladophora. Six distinct ITS sequence types were found, characterized by multiple insertions and deletions and high levels of nucleotide substitution. Conserved domains within the ITS regions indicate the presence of ITS secondary structure. Low transition/transversion ratios among the six types and nearly symmetrical tree-length frequency distributions indicate some saturation, and low phylogenetic signal. Although branching order among five of the six ITS sequence types could not be resolved, estimates of ITS sequence divergence as compared with 18S divergence in a subset of the taxa suggests that the origin of the different ITS types is probably in the mid-Miocene (12 Ma ago) but that biogeographic isolates within a single ITS type (including both Pacific and Atlantic representatives) have probably dispersed on a time scale of thousands rather than millions of years.Correspondence to: J.J. Olsen     

EVOLUTIONARY AND ECOLOGICAL DIFFERENTIATION IN THE PANTROPICAL TO WARM-TEMPERATE SEAWEED DIGENEA SIMPLEX (RHODOPHYTA)1

Genetic differentiation among geographic isolates of the pantropical to warm-temperate red alga Digenea simplex (Wulfen) C. Agardh was investigated using random amplified polymorphic DNA (RAPD) markers, crossing studies, and temperature tolerances experiments. Eleven isolates representing populations from the Caribbean, eastern Atlantic, and Indo-West Pacific were compared. RAPD analysis clearly revealed an Indo-West Pacific group, a Caribbean/Cape Verde Islands group, and a Canary Islands group. Crossing studies showed different levels of inter fertility. In most crosses between Western Australian and Atlantic isolates, no hybrid tetrasporophytes were formed. In crosses between Caribbean and Cape Verde Islands isolates, tetrasporophytes developed, but the viability of tetraspores was reduced. Full sexual compatibility was observed among Cape Verde Islands isolates and among isolates from Bonaire. Temperature tolerance studies indicate that Pacific isolates have a broader temperature survival range than Atlantic isolates, which may be correlated to local temperature extremes. Despite the reduced level of sexual compatibility between Caribbean and Cape Verde Islands isolates, their shared position in the RAPD analysis and similar temperature responses suggest trans-Atlantic dispersal in the near geological past. In addition to their discrete position in the RAPD distance analysis, the Canary Islands isolates were significantly more cold-tolerant than the other Atlantic isolates. This finding is consistent with the hypothesis that the Canary Islands were recolonized from cold-adapted eastern Mediterranean populations after the last Pleistocene glaciation.     

PHYLOGENETIC RELATIONSHIPS OF THE BROWN ALGAL ORDERS ECTOCARPALES, CHORDARIALES, DICTYOSIPHONALES, AND TILOPTERIDALES (PHAEOPHYCEAE) BASED ON RUBISCO LARGE SUBUNIT AND SPACER SEQUENCES

Phylogenetic relationships among 23 species of morphologically simple brown algae belonging to the Ectocarpales sensu stricto , Chordariales, Dictyosiphonales, and Tilopteridales sensu stricto , Phaeophyceae (Fucophyceae), were analyzed using chloroplast-encoded RUBISCO large subunit gene sequences ( rbc L) and the associated RUBISCO spacer sequences. Comparison of the observed and expected sequence divergence at the three codon positions of rbc L showed that the level of mutational saturation within the brown algae is minor. Thus, rbc L is well suited for phylogenetic studies in this group. Unweighted parsimony analyses and a neighbor-joining distance analysis were performed using unambiguously aligned rbc L sequences from the above four orders, one marine raphidophyte and two Tribophyceae (Xantophyceae). Polyphyly of Tilopteridales sensu lato (i.e. including Dictyosiphonales) is verified; we therefore recommend the use of Tilopteridales in the strict sense. The Ectocarpales, Chordariales, and Dictyosiphonales are paraphyletic with respect to each other, forming a highly interwoven clade. A separate parsimony analysis of the RUBISCO spacer as well as a combined rbc L and spacer analysis supported the close relationship among the latter three orders, adding to the evidence that they should be subsumed into the Ectocarpales sensu lato.     

BIOGEOGRAPHY OF CLADOPHOROPSIS MEMBRANACEA (CHLOROPHYTA) BASED ON COMPARISONS OF NUCLEAR rDNA ITS SEQUENCES1

Nucleotides were compared at 988 sites, spanning both internal transcribed spacers (ITS1 and ITS2) of the nuclear ribosomal DNA, among 17 isolates of the green alga Cladophoropsis membranacea (Hofman Bang ex C. Agardh) Boergesen and two isolates of Struvea anastomosans (Harvey) Piccone and Grunow. Collections were made from Bonaire, Curaçao, St. Croix, the Canary Islands, the Cape Verde Islands, Mauritania, Syria, the Red Sea, Okinawa, and Hawaii. Two nucleotide substitutions were found between the sequenced coding regions of C. membranacea and S. anastomosans. Of the 720 nucleotides compared in ITS1 and ITS2, an average of 6.7% sequence divergence was found within C. membranacea and 17.4% between C. membranacea and S. anastomosans. Sequences were analyzed using maximum parsimony. Phylogenetic hypotheses were compared with the biogeographic hypothesis of an east-west Tethyan vicariance. Results of the study allow the identification of widely dispersed biogeographic populations, the identification of an underlying Tethyan imprint, and support for the hypothesis that dispersal has occurred between the Caribbean and the tropical eastern Atlantic. These results demonstrate the usefulness of ITS sequences at the sub-specific level in C. membranacea.