Metabotropic glutamate receptors in mechanisms of plasticity of the central nervous system in the honeybee <Emphasis Type="Italic">Apis mellifera</Emphasis>

Localization of metabotropic glutamate receptors (MGR) in head ganglion of honeybee Apis mellifera, and mechanisms of participation of activated MGR in CNS plasticity are investigated by means of complex approach using immunochemical, electrophysiological and behavioral methods. Influence of MGR activation on GABAergic system and inotropic glutamate receptors (IGR) of AMPA- and NMDA-subtypes is studied. MGR are revealed in lateral and medial calices of mushroom bodies. The inhibiting influence of MGR on AMPA- and NMDA receptors is shown using method of conditioned reflex. Previous activation of MGR neutralizes the inhibiting effect of GABA. Modulating role of heterogeneous MGR population in mechanisms of CNS plasticity on the level of glutamatergic synapse, and at interaction with GABAergic system is discussed.     

The Genus Syringa: Molecular Markers of Species and Cultivars

RAPD analysis was carried out with 22 accessions of the genus Syringa, including six species, one interspecific hybrid, and 15 cultivars. In total, 512 polymorphic fragments were detected; species-specific and cultivar-specific markers were identified. For the first time, genetic polymorphism and genome similarity coefficients were estimated and phylogenetic relationships were established for the genus Syringa.     

RAPD analysis of genome polymorphism in the family Lemnaceae

The multilocus RAPD analysis of intergeneric, inter- and intraspecific nuclear genome polymorphism was used for the first time to assess intergeneric, interspecific, and intraspecific polymorphism in Lemnaceae growing on the territory of Russia. The origin of the chosen accessions overlapped with the natural range of duckweeds in Russia. Seventy-five Lemnaceae accessions representing eight species (L. minor, L. gibba, L. turionifera, L. japonica. L. trisulca, L. aequinoctialis, S. polyrhiza, and L. punctata) from three genera (Lemna, Spirodela, and Landoltia), were analyzed. The highest variability levels were revealed in L. minor accessions (0.03-0.20). Species L. trisulca and S. polyrhiza were characterized by values of genetic distance 0.01-0.18 and 0.03-0.16, respectively. The lowest polymorphism levels were detected for L. turionifera (0.01-0.11). The dendrogram based on RAPD data showed that L. aequinoctialis was the most genetically distant species of the genus Lemna. Accessions of species L. turionifera and L. japonica, as well as L. minor and L. gibba, did not form separate species-specific subclusters; rather, they fell into clusters with L. japonica/L. turionifera and L. minor/L. gibba. Accessions of the genera Spirodela and Landoltia formed two separate clusters combined into one group.     

L-Glutamate in formation of long-term memory in the honeybee <Emphasis Type="Italic">Apis mellifera</Emphasis>

Participation of metabotropic glutamate receptors (mGluR) in associative learning of the honeybee Apis mellifera was studied in behavioral and electrophysiological experiments with use of pharmacological analysis. By the method of conditioned reflexes, effects of systemic injections of mGluR agonists and antagonists were studied on retention of elaborated conditioned reflexes in short-term and long-term memory. Injection of aminocyclopentandicarbonic acid (ACPD), ibotenate, and phosphoserine stimulated the memory in 3 h after the single learning procedure. The long- term memory was inhibited by mGluR antagonists — methylcarboxyphenylglycine and amino- phosphonopropionate as well as by non-competitive antagonists of mGluR₁(CPCCOEt) and of mGluR₅ (SIB 1757). Electrophysiological experiments demonstrated changes of characteristics of the action potential recorded from mushroom bodxies in response to stimulation of antennal lobes and during injections of mGluR III phosphoserine and the mGluR₁ antagonist CPCCOEt. The obtained data allow suggesting participation of different groups of metabotropic glutamate receptors in the process of the honeybee associative learning.Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 40, No. 6, 2004, pp. 539–545.     

Biotechnological synthesis of ribavirin. Effect of ribavirin and its various combinations on the reproduction of Vaccinia virus

The biotechnological method of synthesis of ribavirin, vidarabin, and 6-azauridine by the use of immobilized recombinant enzymatic preparations of nucleoside phosphorylase was improved. The effect of ribavirin and its combinations with the other synthesized nucleosides on the reproduction of Vaccinia virus was studied using cultures of Vero cells. The combination of ribavirin and vidarabin was shown to provide an antiviral effect at lesser concentrations than when these compounds were taken separately. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 6; see also http://www.maik.ru.     

Regulation and localization of CAS substrate domain tyrosine phosphorylation

Crk-associated substrate (CAS) is a tyrosine kinase substrate implicated in integrin control of cell behavior. Phosphorylation, by Src family kinases, of multiple tyrosine residues in the CAS substrate domain (SD) is a major integrin signaling event that promotes cell motility. In this study, novel phosphospecific antibodies directed against CAS SD phosphotyrosine sites ("pCAS" antibodies) were characterized and employed to investigate the cellular regulation and localization of CAS SD tyrosine phosphorylation. An analysis of CAS and focal adhesion kinase (FAK) variants expressed in CAS- and FAK-deficient cell lines, respectively, indicated that CAS SD tyrosine phosphorylation is substantially achieved by Src family kinases brought into association with CAS through two distinct mechanisms: direct binding to the CAS Src-binding domain and indirect association through a FAK bridge. Cell immunostaining with pCAS antibodies revealed that CAS SD tyrosine phosphorylation occurs exclusively at sites of integrin adhesion including both nascent focal complexes formed at the edges of extending lamellipodia as well as mature focal adhesions underlying the cell body. These findings further document a role for FAK as an important upstream regulator of CAS SD tyrosine phosphorylation and implicate CAS-mediated signaling events in promoting membrane protrusion/lamellipodium extension during cell motility.     

Absence of the effect of opioid peptides on muscarine receptors in the frog vestibular apparatus

We studied the effects of opioid peptide leu-enkephaline, a specific antagonist of acetylcholine receptors atropine, and non-selective opiate antagonist naloxone on synaptic transmission and responses evoked by acetylcholine in semicircular organs of the frog. A decrease in frequency of acetylcholine (0.1-5.0 microM) responses under leu-enkephaline (10 nM) id not differ from the frequency decline induced by leu-enkephaline alone. Atropine (1 microM) left the response to leu-enkephaline intact while blocking the excitatory effect of acetylcholine. No modification of the acetylcholine response under leu-enkephaline was observed in the presence of naloxone (1 microM). The findings suggest that no interaction exists between the acetylcholine-mediated excitatory action on resting activity in the isolated semicircular canal preparation and the suppressive action of leu-enkephaline.     

Genetic diversity and phylogenetic relationships of the genus Lycopersicon (Tourn.) Mill. as revealed by inter-simple sequence repeat (ISSR) analysis

Inter-simple sequence repeat (ISSR) analysis was for the first time used to study the genetic diversity and phylogenetic relationships in 54 wild accessions and cultivars of the genus Lycopersicon. Analysis involved 14 ISSR primers homologous to microsatellite repeats and containing additional selective anchor nucleotides. In total, 318 ISSR fragments were amplified for the wild and cultivated tomato genomes. The interspecific polymorphism revealed with the ISSR primers was 95.6%. Species-specific ISSR fragments were detected for each tomato species. The highest number (more than 20) of species-specific fragments were obtained for L. esculentum sensu lato, although the intraspecific variation of ISSR patterns was low. UPGMA cluster analysis was used to construct a dendrogram and to estimate the genetic distances between the species of the genus Lycopersicon; between populations of L. peruvianum, L. pimpinellifolium, and L. esculentum; and between tomato cultivars. The ISSR-based phylogeny was generally consistent with Lycopersicon taxonomy based on morphological and molecular evidence, suggesting the applicability of ISSR analysis for genotyping and phylogenetic studies in tomato.     

Organization of fusicoccin receptor in higher plant plasma membrane: relationship between affinity and molecular mass

Higher plant plasma membranes carry receptors of different affinity for the phytotoxin fusicoccin. Reception of fusicoccin involves proteins belonging to the highly conserved 14-3-3 family, but the complete structure of the fusicoccin receptor (FCR) is unknown. Using radiation inactivation analysis, we estimated the molecular masses of low-affinity and high-affinity FCR at 63 +/- 7 and 130 +/- 15 kD, respectively. The dose dependences of receptor inactivation indicate that microsomal specimens contain "silent" FCRs of 420 +/- 90 kD in amounts commensurate with that of the active FCRs. Both low- and high-affinity FCRs are inactivated by hydrolytic enzymes from the outer surface of the plasma membrane, and impairment of protoplast integrity causes an irreversible transition of the low-affinity binding site into the high-affinity one. A scheme is proposed for the organization of different types of FCR in the plasma membrane, implying that the membrane affinity for fusicoccin reflects the interaction between proteins in the FCR complex.