TY - JOUR A1 - Walz, Bernd A1 - Baumann, Otto A1 - Krach, Christian A1 - Baumann, Arnd A1 - Blenau, Wolfgang T1 - The aminergic control of cockroach salivary glands N2 - The acinar salivary glands of cockroaches receive a dual innervation from the subesophageal ganglion and the stomatogastric nervous system. Acinar cells are surrounded by a plexus of dopaminergic and serotonergic varicose fibers. In addition, seroton-ergic terminals lie deep in the extracellulor spaces between acinar cells. Excitation-secretion coupling in cockroach salivary glands is stimulated by both dopamine and serotonin. These monoamines cause increases in the intracellular concentrations of cAMP and Ca2+. Stimulation of the glands by serotonin results in the production of a protein-rich saliva, whereas stimulation by dopamine results in saliva that is protein-free. Thus, two elementary secretary processes, namely electrolyte/water secretion and protein secretion, are triggered by different aminergic transmitters. Because of its simplicity and experimental accessibility, cockroach salivary glands have been used extensively as a model system to study the cellular actions of biogenic amines and to examine the pharmacological properties of biogenic amine receptors. In this review, we summarize current knowledge concerning the aminergic control of cockroach salivary glands and discuss our efforts to characterize Periplaneta biogenic amine receptors molecularly Y1 - 2006 UR - 1960 = Doi 10.1002/Arch.20128 ER - TY - THES A1 - Krach, Christian T1 - Biogene Aminrezeptoren der Schabe Periplaneta americana : Identifizierung, Charakterisierung und Lokalisierung von PeaTYR1 und PeaDOP2 T1 - Biogenic amine receptors of Periplaneta americana : identification, characterization and localization of PeaTYR1 and PeaDOP2 N2 - Biogene Amine sind eine Substanzklasse, die bei Vertebraten und Invertebraten eine wichtige Komponente des endokrinen Systems darstellen. Sie binden an spezifische Rezeptoren der Gruppe der G-Protein gekoppelten Rezeptoren. In dieser Arbeit wurden zwei neue Rezeptoren aus der Schabe Periplaneta americana kloniert. Durch verschiedene Ansätze konnten zwei vollständige cDNA-Sequenzen isoliert werden. Die Aminosäuresequenzen weisen die größte Ähnlichkeit zu bereits bekannten Tyraminrezeptoren aus Locusta/Bombyx bzw. zu Dopaminrezeptoren aus Apis/Drosophila auf. Entsprechend wurden diese Rezeptoren Pea (P. americana) TYR1 und PeaDOP2 genannt. Deutliche Hinweise auf ihre Funktion lassen sich an den abgeleiteten Aminosäuresequenzen ablesen. Aminosäuren, die wichtig bei der Bildung der Bindungstasche, der Rezeptoraktivierung und der Kopplung eines G-Proteins sind, treten bei beiden Rezeptoren auf. Sequenzalignments stellen die Rezeptoren in die Gruppe anderer Tyraminrezeptoren bzw. der Invertebraten-Typ Dopaminrezeptoren. Das Transkript der beiden Rezeptoren konnte durch RT-PCR in verschiedenen Geweben nachgewiesen werden. Ein Ziel der Arbeit war die Gewinnung eines polyklonalen Antiserums gegen PeaTYR1. Dieses Serum detektiert im Homogenat von Gehirnen mehrere Banden, darunter auch eine mit der kalkulierten Masse von PeaTYR1. Präabsorption des Serums mit dem Peptid, welches zur Reinigung verwendet wurde, zeigt dessen Spezifität. An Gehirnschnitten markiert das Serum große Teile des Protocerebrums aber auch feinere Strukturen der Antennalloben, der optischen Loben und des Zentralkomplexes. Ein weiteres Serum gegen Tyramin führte zu einer Markierung von mehreren Neuronengruppen, welche sich in die optischen Loben und den Zentralkomplex verzweigen. Der αPeaTYR1-CPL3 Antikörper markierte die Plasmamembran von transfizierten HEK293-Zellen. Die Lokalisierung von Rezeptor und Ligand deuten darauf hin, dass Tyramin die optische und olfaktorische Wahrnehmung beeinflussen könnte. N2 - Biogenic amines form a group of substances which play an important role in the endocrine system of invertebrates and vertebrates. They bind to specific receptors of the G-protein coupled type. In this work two new receptors from Periplaneta americana were cloned. By several approaches two full-length cDNA sequences were obtained. The amino acid sequence is very similar to known tyramine receptors of Locusta/Bombyx or dopamine receptors from Apis/Drosophila, respectively. Therefore they were named PeaTYR1 and PeaDOP2. Residues which are important for forming the binding pocket, activation of the receptor or G-protein coupling could be identified in both receptors. Sequence alignments group them together with other tyramine receptors or insect-type dopamine receptors. The corresponding mRNA can be detected in different tissues by RT-PCR. One aim of this work was the preparation of a polyclonal antiserum against PeaTYR1. This serum detects several bands in a brain homogenate; one has the calculated mass of PeaTYR1. Its specificity was proven by praeabsorption. On brain slices the serum labels large regions in the protocerebrum, but also fine structures in the antennal lobes, the optic lobes or the central complex. Another serum against tyramine labels several cell clusters, which ramify within the optic lobes and the central complex. On transfected HEK293 cells the receptor can be detected on the plasma membrane. The localization of receptor and ligand may indicate a role of tyramine in optical/olfactory perception. KW - Rezeptor KW - Tyramin KW - Dopamin KW - Periplaneta americana KW - receptor KW - tyramine KW - dopamine KW - Periplaneta americana Y1 - 2007 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-15561 ER - TY - JOUR A1 - Troppmann, Britta A1 - Balfanz, Sabine A1 - Krach, Christian A1 - Baumann, Arnd A1 - Blenau, Wolfgang T1 - Characterization of an Invertebrate-Type Dopamine Receptor of the American Cockroach, Periplaneta americana JF - International journal of molecular sciences N2 - We have isolated a cDNA coding for a putative invertebrate-type dopamine receptor (Peadop2) from P. americana brain by using a PCR-based strategy. The mRNA is present in samples from brain and salivary glands. We analyzed the distribution of the PeaDOP2 receptor protein with specific affinity-purified polyclonal antibodies. On Western blots, PeaDOP2 was detected in protein samples from brain, subesophageal ganglion, thoracic ganglia, and salivary glands. In immunocytochemical experiments, we detected PeaDOP2 in neurons with their somata being located at the anterior edge of the medulla bilaterally innervating the optic lobes and projecting to the ventro-lateral protocerebrum. In order to determine the functional and pharmacological properties of the cloned receptor, we generated a cell line constitutively expressing PeaDOP2. Activation of PeaDOP2-expressing cells with dopamine induced an increase in intracellular cAMP. In contrast, a C-terminally truncated splice variant of this receptor did not exhibit any functional property by itself. The molecular and pharmacological characterization of the first dopamine receptor from P. americana provides the basis for forthcoming studies focusing on the significance of the dopaminergic system in cockroach behavior and physiology. KW - G-protein-coupled receptor KW - dopamine KW - insect KW - cellular signaling KW - salivary gland KW - biogenic amine Y1 - 2014 U6 - https://doi.org/10.3390/ijms15010629 SN - 1422-0067 VL - 15 IS - 1 SP - 629 EP - 653 PB - MDPI CY - Basel ER - TY - GEN A1 - Blenau, Wolfgang A1 - Rotte, Cathleen A1 - Krach, Christian A1 - Balfanz, Sabine A1 - Baumann, Arnd A1 - Walz, Bernd T1 - Molecular characterization and localization of the first tyramine receptor of the American cockroach (Periplaneta americana) N2 - The phenolamines octopamine and tyramine control, regulate, and modulate many physiological and behavioral processes in invertebrates. Vertebrates possess only small amounts of both substances, and thus, octopamine and tyramine, together with other biogenic amines, are referred to as “trace amines.” Biogenic amines evoke cellular responses by activating G-protein-coupled receptors. We have isolated a complementary DNA (cDNA) that encodes a biogenic amine receptor from the American cockroach Periplaneta americana, viz., Peatyr1, which shares high sequence similarity to members of the invertebrate tyramine-receptor family. The PeaTYR1 receptor was stably expressed in human embryonic kidney (HEK) 293 cells, and its ligand response has been examined. Receptor activation with tyramine reduces adenylyl cyclase activity in a dose-dependent manner (EC50 350 nM). The inhibitory effect of tyramine is abolished by co-incubation with either yohimbine or chlorpromazine. Receptor expression has been investigated by reverse transcription polymerase chain reaction and immunocytochemistry. The mRNA is present in various tissues including brain, salivary glands, midgut, Malpighian tubules, and leg muscles. The effect of tyramine on salivary gland acinar cells has been investigated by intracellular recordings, which have revealed excitatory presynaptic actions of tyramine. This study marks the first comprehensive molecular, pharmacological, and functional characterization of a tyramine receptor in the cockroach. KW - Biogenic amine KW - cellular signaling KW - G-protein-coupled receptor KW - octopamine KW - salivary gland Y1 - 2009 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-44335 ER -