@article{BogenBenderLoeweetal.2015, author = {Bogen, Oliver and Bender, Olaf and Loewe, Jana and Blenau, Wolfgang and Thevis, Beatrice and Schroeder, Wolfgang and Margolis, Richard U. and Levine, Jon D. and Hucho, Ferdinand}, title = {Neuronally produced versican V2 renders C-fiber nociceptors IB4-positive}, series = {Journal of neurochemistry}, volume = {134}, journal = {Journal of neurochemistry}, number = {1}, publisher = {Wiley-Blackwell}, address = {Hoboken}, issn = {0022-3042}, doi = {10.1111/jnc.13113}, pages = {147 -- 155}, year = {2015}, abstract = {A subpopulation of nociceptors, the glial cell line-derived neurotrophic factor (GDNF)-dependent, non-peptidergic C-fibers, expresses a cell-surface glycoconjugate that can be selectively labeled with isolectin B4 (IB4), a homotetrameric plant lectin from Griffonia simplicifolia. We show that versican is an IB4-binding molecule in rat dorsal root ganglion neurons. Using reverse transcriptase polymerase chain reaction (RT-PCR), insitu hybridization and immunofluorescence experiments on rat lumbar dorsal root ganglion, we provide the first demonstration that versican is produced by neurons. In addition, by probing Western blots with splice variant-specific antibodies we show that the IB4-binding versican contains only the glycosaminoglycan alpha domain. Our data support V2 as the versican isoform that renders this subpopulation of nociceptors IB4-positive (+). A subset of nociceptors, the GDNF-dependent non-peptidergic C-fibers can be characterized by its reactivity for isolectin B4 (IB4), a plant lectin from Griffonia simplicifolia. We have previously demonstrated that versican V2 binds IB4 in a Ca2+-dependent manner. However, given that versican is thought to be the product of glial cells, it was questionable whether versican V2 can be accountable for the IB4-reactivity of this subset of nociceptors. The results presented here prove - for the first time - a neuronal origin of versican and suggest that versican V2 is the molecule that renders GDNF-dependent non-peptidergic C-fibers IB4-positive.}, language = {en} } @article{WalzBaumannKrachetal.2006, author = {Walz, Bernd and Baumann, Otto and Krach, Christian and Baumann, Arnd and Blenau, Wolfgang}, title = {The aminergic control of cockroach salivary glands}, year = {2006}, abstract = {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}, language = {en} } @article{HauserCazzamaliWilliamsonetal.2006, author = {Hauser, Frank and Cazzamali, Giuseppe and Williamson, Michael and Blenau, Wolfgang and Grimmelikhuijzen, CJ.}, title = {A review of neurohormone GPCRs present in the fruitfly "Drosophila melanogaster" and the honey bee "Apis mellifera"}, issn = {0301-0082}, doi = {10.1016/j.pneurobio.2006.07.005}, year = {2006}, abstract = {G protein-coupled receptor (GPCR) genes are large gene families in every animal, sometimes making up to 1-2\% of the animal's genome. Of all insect GPCRs, the neurohormone (neuropeptide, protein hormone, biogenic amine) GPCRs are especially important, because they, together with their ligands, occupy a high hierarchic position in the physiology of insects and steer crucial processes such as development, reproduction, and behavior. In this paper, we give a review of our current knowledge on Drosophila melanogaster GPCRs and use this information to annotate the neurohormone GPCR genes present in the recently sequenced genome from the honey bee Apis mellifera. We found 35 neuropeptide receptor genes in the honey bee (44 in Drosophila) and two genes, coding for leucine-rich repeats-containing protein hormone GPCRs (4 in Drosophila). In addition, the honey bee has 19 biogenic amine receptor genes (21 in Drosophila). The larger numbers of neurohormone receptors in Drosophila are probably due to gene duplications that occurred during recent evolution of the fly. Our analyses also yielded the likely ligands for 40 of the 56 honey bee neurohormone GPCRs identified in this study. In addition, we made some interesting observations on neurohormone GPCR evolution and the evolution and co-evolution of their ligands. For neuropeptide and protein hormone GPCRs, there appears to be a general co-evolution between receptors and their ligands. This is in contrast to biogenic amine GPCRs, where evolutionarily unrelated GPCRs often bind to the same biogenic amine, suggesting frequent ligand exchanges ("ligand hops") during GPCR evolution.}, language = {en} } @article{BlenauBaumann2005, author = {Blenau, Wolfgang and Baumann, Arnd}, title = {Molecular characterization of the ebony gene from the American cockroach, Periplaneta americana}, year = {2005}, abstract = {Biogenic amines are an important class of primary messengers in the central (CNS) and peripheral nervous systems and in peripheral organs. These substances regulate and modulate many physiological and behavioral processes. Various inactivation mechanisms for these substances exist to terminate biogenic amine-mediated signal transduction. In vertebrates, the enzymes monoamine oxidase and/or catechol-O-methyl-transferase are involved in these processes. In insects, however, in which both enzymes are low in abundance or absent, biogenic amines are inactivated mainly by N- acetylation or O-sulphation. In Droso-philo, beta-alanyl conjugation mediated by the Ebony protein has recently been shown to be a novel and alternative pathway for biogenic amine inactivation. Here, we report the cloning of ebony cDNA (Peaebony) from a brain-specific cDNA library of the cockroach Periplaneta americana. The open reading frame encodes a protein of 860 amino acid residues (PeaEbony). The PeaEbony polypeptide shares homology to Ebony sequences from Anopheles gambiae, Apis mellifera, and Drosophila melonogaster. In addition, PeaEbony exhibits sequence similarity to a family of microbial non-ribosomal peptide synthetases. The mRNA encoding PeaEbony is highly expressed in the cockroach brain and to a lesser extent in the salivary glands. PeaEbony is, therefore, probably involved in the inactivation of various biogenic amines through beta-alanyl conjugation in the cockroach CNS. Since the salivary glands in Periplaneta are innervated by dopaminergic and serotonergic neurons, PeaEbony probably also biochemically modifies dopamine and serotonin in these acinar glands. Arch. Insect Biochem. (c) 2005 Wiley-Liss, Inc}, language = {en} } @article{BogenBenderSchlenstedtetal.2005, author = {Bogen, Oliver and Bender, Olaf and Schlenstedt, Jana and Goswami, C. and Schroder, W. and Blenau, Wolfgang and Gillen, C. and Dreger, M. and Hucho, F.}, title = {Neuronal expression of the IB4-binding versican}, issn = {0022-3042}, year = {2005}, language = {en} } @article{RietdorfBlenauWalz2005, author = {Rietdorf, Katja and Blenau, Wolfgang and Walz, Bernd}, title = {Protein secretion in cockroach salivary glands requires an increase in intracellular cAMP and Ca2+ concentrations}, issn = {0022-1910}, year = {2005}, abstract = {The salivary glands in the cockroach Periplaneta americana secrete protein-containing saliva when stimulated by serotonin (5-HT) and protein-free saliva upon dopamine stimulation. In order to obtain information concerning the signalling pathways involved in 5-HT-induced protein secretion, we have determined the protein content of saliva secreted after experimental manipulations that potentially elevate intracellular Ca2+ and cyclic nucleotide concentrations in isolated glands. We have found that 5-HT stimulates the rate of protein secretion in a dose-dependent manner (threshold: 3 x 10(-8) M; EC50 1.5 x 10(-6) M). The maximal rate of 5-HT-induced protein secretion was 2.2 +/- 0.2 mu g/min. Increasing intracellular Ca2+ or cAMP by bath application of ionomycin (5 mu M), db cAMP (10 mM), forskolin (100 mu M) or IBMX (100 mu M), respectively, stimulated protein secretion at significantly lower rates, whereas db cGMP (1 mM) did not activate protein secretion. The high rates and the kinetics of 5-HT-induced protein secretion could only be mimicked by either applying forskolin together with IBMX (with or without ionomycin) or by applying IBMX together with ionomycin. Our measurements suggest that 5-HT-induced protein secretion is mediated by an elevation of [cAMP](i) and that Ca2+ may function as a co-agonist and augment the rate of protein secretion. (c) 2005 Elsevier Ltd. All rights reserved}, language = {en} } @article{MustardKurshanHamiltonetal.2005, author = {Mustard, J. A. and Kurshan, P. T. and Hamilton, Ingrid S. and Blenau, Wolfgang and Mercer, Alison R.}, title = {Developmental expression of a tyramine receptor gene in the brain of the honey bee, Apis mellifera}, year = {2005}, language = {en} } @article{Blenau2005, author = {Blenau, Wolfgang}, title = {Cellular actions of biogenic amines}, year = {2005}, language = {en} } @article{MargWalzBlenau2004, author = {Marg, S. and Walz, Bernd and Blenau, Wolfgang}, title = {The effects of dopamine receptor agonists and antagonists on the secretory rate of cockroach (Periplaneta americana) salivary glands}, issn = {0022-1910}, year = {2004}, abstract = {The acinar salivary glands of the cockroach, Periplaneta americana, are innervated by dopaminergic and serotonergic nerve fibers. Serotonin stimulates the secretion of protein-rich saliva, whereas dopamine causes the production of protein-free saliva. This suggests that dopamine acts selectively on ion-transporting peripheral cells within the acini and the duct cells, and that serotonin acts on the protein-producing central cells of the acini. We have investigated the pharmacology of the dopamine-induced secretory activity of the salivary gland of Periplaneta americana by testing several dopamine receptor agonists and antagonists. The effects of dopamine can be mimicked by the non-selective dopamine receptor agonist 6,7-ADTN and, less effectively, by the vertebrate D1 receptor-selective agonist chloro-APB. The vertebrate D1 receptor-selective agonist SKF 38393 and vertebrate D2 receptor-selective agonist R(-)- TNPA were ineffective. R(+)-Lisuride induces a secretory response with a slower onset and a lower maximal response compared with dopamine-induced secretion. However, lisuride-stimulated glands continue secreting saliva, even after lisuride-washout. Dopamine-induced secretions can be blocked by the vertebrate dopamine receptor antagonists cis(Z)- flupenthixol, chlorpromazine, and S(+)-butaclamol. Our pharmacological data do not unequivocally indicate whether the dopamine receptors on the Periplaneta salivary glands belong to the D1 or D2 subfamily of dopamine receptors, but we can confirm that the pharmacology of invertebrate dopamine receptors is remarkably different from that of their vertebrate counterparts. (C) 2004 Elsevier Ltd. All rights reserved}, language = {en} } @article{ThammBalfanzScheineretal.2010, author = {Thamm, Markus and Balfanz, Sabine and Scheiner, Richarda and Baumann, Arnd and Blenau, Wolfgang}, title = {Characterization of the 5-HT1A receptor of the honeybee (Apis mellifera) and involvement of serotonin in phototactic behavior}, issn = {1420-682X}, doi = {10.1007/s00018-010-0350-6}, year = {2010}, abstract = {Serotonin plays a key role in modulating various physiological and behavioral processes in both protostomes and deuterostomes. The vast majority of serotonin receptors belong to the superfamily of G-protein-coupled receptors. We report the cloning of a cDNA from the honeybee (Am5-ht1A) sharing high similarity with members of the 5-HT1 receptor class. Activation of Am5-HT1A by serotonin inhibited the production of cAMP in a dose-dependent manner (EC50 = 16.9 nM). Am5-HT1A was highly expressed in brain regions known to be involved in visual information processing. Using in vivo pharmacology, we could demonstrate that Am5-HT1A receptor ligands had a strong impact on the phototactic behavior of individual bees. The data presented here mark the first comprehensive study-from gene to behavior-of a 5-HT1A receptor in the honeybee, paving the way for the eventual elucidation of additional roles of this receptor subtype in the physiology and behavior of this social insect.}, language = {en} } @article{VerlindenVleugelsMarchaletal.2010, author = {Verlinden, Heleen and Vleugels, Rut and Marchal, Elisabeth and Badisco, Liesbeth and Pfl{\"u}ger, Hans-Joachim and Blenau, Wolfgang and Vanden Broeck, Jozef}, title = {The role of octopamine in locusts and other arthropods}, issn = {0022-1910}, doi = {10.1016/j.jinsphys.2010.05.018}, year = {2010}, abstract = {The biogenic amine octopamine and its biological precursor tyramine are thought to be the invertebrate functional homologues of the vertebrate adrenergic transmitters. Octopamine functions as a neuromodulator, neurotransmitter and neurohormone in insect nervous systems and prompts the whole organism to "dynamic action". A growing number of studies suggest a prominent role for octopamine in modulating multiple physiological and behavioural processes in invertebrates, as for example the phase transition in Schistocerca gregaria. Both octopamine and tyramine exert their effects by binding to specific receptor proteins that belong to the superfamily of G protein-coupled receptors. Since these receptors do not appear to be present in vertebrates, they may present very suitable and specific insecticide and acaricide targets. (C) 2010 Elsevier Ltd. All rights reserved.}, language = {en} } @article{VerlindenVleugelsMarchaletal.2010, author = {Verlinden, Heleen and Vleugels, Rut and Marchal, Elisabeth and Badisco, Liesbeth and Tobback, Julie and Pfl{\"u}ger, Hans-Joachim and Blenau, Wolfgang and Vanden Broeck, Jozef}, title = {The cloning, phylogenetic relationship and distribution pattern of two new putative GPCR-type octopamine receptors in the desert locust (Schistocerca gregaria)}, issn = {0022-1910}, doi = {10.1016/j.jinsphys.2010.03.003}, year = {2010}, abstract = {The biogenic amine octopamine functions as a neuromodulator, neurotransmitter and neurohormone in insect nervous systems. It plays a prominent role in modulating multiple physiological and behavioural processes in invertebrates. Octopamine exerts its effects by binding to specific receptor proteins that belong to the superfamily of G protein-coupled receptors. We found two partial sequences of putative octopamine receptors in the desert locust Schistocerca gregaria (SgOct alpha R and SgOct beta R) and investigated their transcript levels in males and females of both phases and during the transition between long-term solitarious and gregarious locusts. The transcript levels of SgOctaR are the highest in the central nervous system, whereas those of SgOct beta R are the highest in the flight muscles, followed by the central nervous system. Both SgOct alpha R and SgOct beta R show higher transcript levels in long-term gregarious locusts as compared to solitarious ones. The rise of SgOct beta R transcript levels already appears during the first 4 h of gregarisation, during which also the behavioural changes take place.}, language = {en} } @article{ThammRolkeJordanetal.2013, author = {Thamm, Markus and Rolke, Daniel and Jordan, Nadine and Balfanz, Sabine and Schiffer, Christian and Baumann, Arnd and Blenau, Wolfgang}, title = {Function and distribution of 5-HT2 receptors in the honeybee (apis mellifera)}, series = {PLoS one}, volume = {8}, journal = {PLoS one}, number = {12}, publisher = {PLoS}, address = {San Fransisco}, issn = {1932-6203}, doi = {10.1371/journal.pone.0082407}, pages = {12}, year = {2013}, abstract = {Background: Serotonin plays a pivotal role in regulating and modulating physiological and behavioral processes in both vertebrates and invertebrates. In the honeybee (Apis mellifera), serotonin has been implicated in division of labor, visual processing, and learning processes. Here, we present the cloning, heterologous expression, and detailed functional and pharmacological characterization of two honeybee 5-HT2 receptors. Methods: Honeybee 5-HT2 receptor cDNAs were amplified from brain cDNA. Recombinant cell lines were established constitutively expressing receptor variants. Pharmacological properties of the receptors were investigated by Ca2+ imaging experiments. Quantitative PCR was applied to explore the expression patterns of receptor mRNAs. Results: The honeybee 5-HT2 receptor class consists of two subtypes, Am5-HT2 alpha and Am5-HT2 beta. Each receptor gene also gives rise to alternatively spliced mRNAs that possibly code for truncated receptors. Only activation of the full-length receptors with serotonin caused an increase in the intracellular Ca2+ concentration. The effect was mimicked by the agonists 5-methoxytryptamine and 8-OH-DPAT at low micromolar concentrations. Receptor activities were blocked by established 5-HT receptor antagonists such as clozapine, methiothepin, or mianserin. High transcript numbers were detected in exocrine glands suggesting that 5-HT2 receptors participate in secretory processes in the honeybee. Conclusions: This study marks the first molecular and pharmacological characterization of two 5-HT2 receptor subtypes in the same insect species. The results presented should facilitate further attempts to unravel central and peripheral effects of serotonin mediated by these receptors.}, language = {en} } @article{ReimThammRolkeetal.2013, author = {Reim, Tina and Thamm, Markus and Rolke, Daniel and Blenau, Wolfgang and Scheiner, Ricarda}, title = {Suitability of three common reference genes for quantitative real-time PCR in honey bees}, series = {Apidologie : a quality journal in bee science}, volume = {44}, journal = {Apidologie : a quality journal in bee science}, number = {3}, publisher = {Springer}, address = {Paris}, issn = {0044-8435}, doi = {10.1007/s13592-012-0184-3}, pages = {342 -- 350}, year = {2013}, abstract = {Honey bees are important model organisms for neurobiology, because they display a large array of behaviors. To link behavior with individual gene function, quantitative polymerase chain reaction is frequently used. Comparing gene expression of different individuals requires data normalization using adequate reference genes. These should ideally be expressed stably throughout lifetime. Unfortunately, this is frequently not the case. We studied how well three commonly used reference genes are suited for this purpose and measured gene expression in the brains of honey bees differing in age and social role. Although rpl32 is used most frequently, it only remains stable in expression between newly emerged bees, nurse-aged bees, and pollen foragers but shows a peak at the age of 12 days. The genes gapdh and ef1 alpha-f1, in contrast, are expressed stably in the brain throughout all age groups except newly emerged bees. According to stability software, gapdh was expressed most stably, followed by rpl32 and ef1 alpha-f1.}, language = {en} } @misc{TroppmannBalfanzKrachetal.2014, author = {Troppmann, Britta and Balfanz, Sabine and Krach, Christian and Baumann, Arnd and Blenau, Wolfgang}, title = {Characterization of an Invertebrate-Type Dopamine Receptor of the American Cockroach, Periplaneta americana}, series = {International journal of molecular sciences}, volume = {15}, journal = {International journal of molecular sciences}, number = {1}, publisher = {MDPI}, address = {Basel}, issn = {1422-0067}, doi = {10.3390/ijms15010629}, pages = {629 -- 653}, year = {2014}, abstract = {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.}, language = {en} } @article{ScheweBlenauWalz2012, author = {Schewe, Bettina and Blenau, Wolfgang and Walz, Bernd}, title = {Intracellular pH regulation in unstimulated Calliphora salivary glands is Na+ dependent and requires V-ATPase activity}, series = {The journal of experimental biology}, volume = {215}, journal = {The journal of experimental biology}, number = {8}, publisher = {Company of Biologists Limited}, address = {Cambridge}, issn = {0022-0949}, doi = {10.1242/jeb.063172}, pages = {1337 -- 1345}, year = {2012}, abstract = {Salivary gland cells of the blowfly Calliphora vicina have a vacuolar-type H+-ATPase (V-ATPase) that lies in their apical membrane and energizes the secretion of a KCl-rich primary saliva upon stimulation with serotonin (5-hydroxytryptamine). Whether and to what extent V-ATPase contributes to intracellular pH (pH(i)) regulation in unstimulated gland cells is unknown. We used the fluorescent dye BCECF to study intracellular pH(i) regulation microfluorometrically and show that: (1) under resting conditions, the application of Na+-free physiological saline induces an intracellular alkalinization attributable to the inhibition of the activity of a Na+-dependent glutamate transporter; (2) the maintenance of resting pHi is Na+, Cl-, concanamycin A and DIDS sensitive; (3) recovery from an intracellular acid load is Na+ sensitive and requires V-ATPase activity; (4) the Na+/H+ antiporter is not involved in pHi recovery after a NH4Cl prepulse; and (5) at least one Na+-dependent transporter and the V-ATPase maintain recovery from an intracellular acid load. Thus, under resting conditions, the V-ATPase and at least one Na+-dependent transporter maintain normal pH(i) values of pH.7.5. We have also detected the presence of a Na+-dependent glutamate transporter, which seems to act as an acid loader. Despite this not being a common pH(i)-regulating transporter, its activity affects steady-state pH(i) in C. vicina salivary gland cells.}, language = {en} } @article{FrenchSimcockRolkeetal.2014, author = {French, Alice S. and Simcock, Kerry L. and Rolke, Daniel and Gartside, Sarah E. and Blenau, Wolfgang and Wright, Geraldine A.}, title = {The role of serotonin in feeding and gut contractions in the honeybee}, series = {Journal of insect physiology}, volume = {61}, journal = {Journal of insect physiology}, publisher = {Elsevier}, address = {Oxford}, issn = {0022-1910}, doi = {10.1016/j.jinsphys.2013.12.005}, pages = {8 -- 15}, year = {2014}, language = {en} } @article{HeindorffBlenauWalzetal.2012, author = {Heindorff, Kristoffer and Blenau, Wolfgang and Walz, Bernd and Baumann, Otto}, title = {Characterization of a Ca2+/calmodulin-dependent AC1 adenylyl cyclase in a non-neuronal tissue, the blowfly salivary gland}, series = {Cell calcium}, volume = {52}, journal = {Cell calcium}, number = {2}, publisher = {Churchill Livingstone}, address = {Edinburgh}, issn = {0143-4160}, doi = {10.1016/j.ceca.2012.04.016}, pages = {103 -- 112}, year = {2012}, abstract = {Crosstalk between intracellular signalling pathways is a functionally important and widespread phenomenon in cell physiology across phyla. In the salivary gland of the blowfly, serotonin induces fluid secretion via parallel activation of both the InsP(3)/Ca2+ and the cAMP/PKA signalling pathways, which interact on multiple levels. We have determined the molecular identity of a link between both pathways that mediates a Ca2+-dependent rise of intracellular cAMP. Whereas hydrolysis of cAMP via phosphodiesterases is largely independent of Ca2+, cAMP synthesis by adenylyl cyclases (AC) is potentiated in a Ca2+/calmodulin (Ca2+/CaM)-dependent manner. The existence of a Ca2+/CaM-dependent AC is supported by physiological data and a molecular approach. We have cloned Cv rutabaga cDNA, encoding the first blowfly AC, and confirmed its expression in the salivary gland via reverse transcription followed by polymerase chain reaction. The putative gene product of Cv rutabaga is a Ca2+/CaM-dependent type I AC and shows highest homology to Rutabaga from Drosophila. Thus, a Ca2+/CaM-dependent AC serves as a link between the InsP(3)/Ca2+ and the cAMP/PKA signalling pathways in the salivary gland of the blowfly and might be important for the amplification and optimization of the secretory response.}, language = {en} } @article{RoeserJordanBalfanzetal.2012, author = {R{\"o}ser, Claudia and Jordan, Nadine and Balfanz, Sabine and Baumann, Arnd and Walz, Bernd and Baumann, Otto and Blenau, Wolfgang}, title = {Molecular and pharmacological characterization of serotonin 5-HT2 alpha and 5-HT7 receptors in the salivary glands of the blowfly calliphora vicina}, series = {PLoS one}, volume = {7}, journal = {PLoS one}, number = {11}, publisher = {PLoS}, address = {San Fransisco}, issn = {1932-6203}, doi = {10.1371/journal.pone.0049459}, pages = {13}, year = {2012}, abstract = {Secretion in blowfly (Calliphora vicina) salivary glands is stimulated by the biogenic amine serotonin (5-hydroxytryptamine, 5-HT), which activates both inositol 1,4,5-trisphosphate (InsP(3))/Ca2+ and cyclic adenosine 3',5'-monophosphate (cAMP) signalling pathways in the secretory cells. In order to characterize the signal-inducing 5-HT receptors, we cloned two cDNAs (Cv5-ht2 alpha, Cv5-ht7) that share high similarity with mammalian 5-HT2 and 5-HT7 receptor genes, respectively. RT-PCR demonstrated that both receptors are expressed in the salivary glands and brain. Stimulation of Cv5-ht2 alpha-transfected mammalian cells with 5-HT elevates cytosolic [Ca2+] in a dose-dependent manner (EC50 = 24 nM). In Cv5-ht7-transfected cells, 5-HT produces a dose-dependent increase in [cAMP](i) (EC50 = 4 nM). We studied the pharmacological profile for both receptors. Substances that appear to act as specific ligands of either Cv5-HT2 alpha or Cv5-HT7 in the heterologous expression system were also tested in intact blowfly salivary gland preparations. We observed that 5-methoxytryptamine (100 nM) activates only the Cv(5)-HT2 alpha receptor, 5-carboxamidotryptamine (300 nM) activates only the Cv5-HT7 receptor, and clozapine (1 mu M) antagonizes the effects of 5-HT via Cv5-HT7 in blowfly salivary glands, providing means for the selective activation of each of the two 5-HT receptor subtypes. This study represents the first comprehensive molecular and pharmacological characterization of two 5-HT receptors in the blowfly and permits the analysis of the physiological role of these receptors, even when co-expressed in cells, and of the modes of interaction between the Ca2+- and cAMP-signalling cascades. Citation: Roser C, Jordan N, Balfanz S, Baumann A, Walz B, et al. (2012) Molecular and Pharmacological Characterization of Serotonin 5-HT2a and 5-HT7 Receptors in the Salivary Glands of the Blowfly Calliphora vicina.}, language = {en} } @misc{BlenauThamm2011, author = {Blenau, Wolfgang and Thamm, Markus}, title = {Distribution of serotonin (5-HT) and its receptors in the insect brain with focus on the mushroom bodies lessons from Drosophila melanogaster and Apis mellifera}, series = {Arthropod structure \& development}, volume = {40}, journal = {Arthropod structure \& development}, number = {5}, publisher = {Elsevier}, address = {Oxford}, issn = {1467-8039}, doi = {10.1016/j.asd.2011.01.004}, pages = {381 -- 394}, year = {2011}, abstract = {The biogenic amine serotonin (5-hydroxytryptamine, 5-HT) plays a key role in regulating and modulating various physiological and behavioral processes in both protostomes and deuterostomes. The specific functions of serotonin are mediated by its binding to and subsequent activation of membrane receptors. The vast majority of these receptors belong to the superfamily of G-protein-coupled receptors. We report here the in vivo expression pattern of a recently characterized 5-HT(1) receptor of the honeybee Apis mellifera (Am5-HT(1A)) in the mushroom bodies. In addition, we summarize current knowledge on the distribution of serotonin and serotonin receptor subtypes in the brain and specifically in the mushroom bodies of the fruit fly Drosophila melanogaster and the honeybee. Functional studies in these two species have shown that serotonergic signaling participates in various behaviors including aggression, sleep, circadian rhythms, responses to visual stimuli, and associative learning. The molecular, pharmacological, and functional properties of identified 5-HT receptor subtypes from A. mellifera and D. melanogaster will also be summarized in this review.}, language = {en} }