TY - JOUR A1 - Stuckas, Heiko A1 - Messerschmidt, Katrin A1 - Putzler, Sascha A1 - Baumann, Otto A1 - Schenk, Jörg A. A1 - Tiedemann, Ralph A1 - Micheel, Burkhard T1 - Detection and characterization of gamete-specific molecules in Mytilus edulis using selective antibody production N2 - The mussel Mytilus edulis can be used as model to study the molecular basis of reproductive isolation because this species maintains its species integrity, despite of hybridizing in zones of contact with the closely related species M. trossulus or M. galloprovincialis. This study uses selective antibody production by means of hybridoma technology to identify molecules which are involved in sperm function of M. edulis. Fragmented sperm were injected into mice and 25 hybridoma cell clones were established to obtain monoclonal antibodies (mAb). Five clones were identified producing mAb targeting molecules putatively involved in sperm function based on enzyme immunoassays, dot and Western blotting as well as immunostaining of tissue sections. Specific localization of these mAb targets on sperm and partly also in somatic tissue suggests that all five antibodies bind to different molecules. The targets of the mAb obtained from clone G26-AG8 were identified using mass spectrometry (nano-LC-ESI-MS/MS) as M6 and M7 lysin. These acrosomal proteins have egg vitelline lyses function and are highly similar (76%) which explains the cross reactivity of mAb G26- AG8. Furthermore, M7 lysin was recently shown to be under strong positive selection suggesting a role in interspecific reproductive isolation. This study shows that M6 and M7 lysin are not only found in the sperm acrosome but also in male somatic tissue of the mantle and the posterior adductor muscle, while being completely absent in females. The monoclonal antibody G26-AG8 described here will allow elucidating M7/M6 lysin function in somatic and gonad tissue of adult and developing animals. Y1 - 2009 UR - http://www3.interscience.wiley.com/cgi-bin/jhome/37692 U6 - https://doi.org/10.1002/Mrd.20916 SN - 1040-452X ER - TY - JOUR A1 - Baumann, Otto A1 - Bauer, Alexandra T1 - Development of apical membrane organization and V-ATPase regulation in blowfly salivary glands JF - The journal of experimental biology N2 - Secretory cells in blowfly salivary gland are specialized via morphological and physiological attributes in order to serve their main function, i.e. the transport of solutes at a high rate in response to a hormonal stimulus, namely serotonin (5-HT). This study examines the way that 5-HT-insensitive precursor cells differentiate into morphologically complex 5-HT-responsive secretory cells. By means of immunofluorescence microscopy, immunoblotting and measurements of the transepithelial potential changes, we show the following. (1) The apical membrane of the secretory cells becomes organized into an elaborate system of canaliculi and is folded into pleats during the last pupal day and the first day of adulthood. (2) The structural reorganization of the apical membrane is accompanied by an enrichment of actin filaments and phosphorylated ERM protein (phospho-moesin) at this membrane domain and by the deployment of the membrane-integral part of vacuolar-type H+-ATPase (V-ATPase). These findings suggest a role for phospho-moesin, a linker between actin filaments and membrane components, in apical membrane morphogenesis. (3) The assembly and activation of V-ATPase can be induced immediately after eclosion by way of 8-CPT-cAMP, a membrane-permeant cAMP analogue. (4) 5-HT, however, produces the assembly and activation of V-ATPase only in flies aged for at least 2 h after eclosion, indicating that, at eclosion, the 5-HT receptor/adenylyl cyclase/cAMP signalling pathway is inoperative upstream of cAMP. (5) 5-HT activates both the Ca2+ signalling pathway and the cAMP signalling cascade in fully differentiated secretory cells. However, the functionality of these signalling cascades does not seem to be established in a tightly coordinated manner during cell differentation. KW - secretory cell KW - moesin KW - morphogenesis KW - serotonin KW - vacuolar ATPase KW - cAMP Y1 - 2013 U6 - https://doi.org/10.1242/jeb.077420 SN - 0022-0949 VL - 216 IS - 7 SP - 1225 EP - 1234 PB - Company of Biologists Limited CY - Cambridge ER - TY - JOUR A1 - Baumann, Otto A1 - Salvaterra, Paul M. A1 - Takeyasu, Kunio T1 - Developmental changes in beta-subunit composition of Na,K-ATPase in the Drosophila eye N2 - The Drosophila genome contains at least three loci for the Na,K-ATPase beta-subunit; however, only the protein products of nrv1 and nrv2 have been characterized hitherto. Here, we provide evidence that nrv3 also encodes for a functional Na,K-ATPase beta-subunit, as its protein product co-precipitates with the Na,K-ATPase alpha-subunit. Nrv3 expression in adult flies is restricted to the nervous system in which Nrv3 is enriched in selective types of sensory cells. Because Nrv3 expression is especially prominent in the compound eye, we have analyzed the subcellular and developmental distribution of Nrv3 within the visual cells and related this distribution to those of the alpha-subunit and of the beta-subunits Nrv1 and Nrv2. Prospective visual cells express Nrv2 in the third larval instar stage and during the first half of pupal development. During the last third of pupal life, Nrv3 gradually replaces Nrv2 as the Na,K-ATPase beta-subunit in the photoreceptor cells. Adult photoreceptors express Nrv3 as their major beta-subunit; the visual cells R1-R6 co-express Nrv2 at a low level, whereas R7 and R8 co-express Nrv1. Notably, beta-subunits do not co- distribute exactly with the alpha-subunit at some developmental stages, supporting the concept that the alpha-subunit and beta-subunit can exist in the plasma membrane without being engaged in alpha/beta heterodimers. The non-visual cells within the compound eye express almost exclusively Nrv2, which segregates together with the alpha-subunit to septate junctions throughout development. Y1 - 2010 UR - http://www.springerlink.com/content/100524 U6 - https://doi.org/10.1007/s00441-010-0948-x SN - 0302-766X ER - TY - JOUR A1 - Schulz, Irene A1 - Baumann, Otto A1 - Samereier, Matthias A1 - Zoglmeier, Christine A1 - Gräf, Ralph T1 - Dictyostelium Sun1 is a dynamic membrane protein of both nuclear membranes and required for centrosomal association with clustered centromeres N2 - Centrosomal attachment to nuclei is crucial for proper mitosis and nuclear positioning in various organisms, and generally involves Sun-family proteins located at the inner nuclear envelope. There is still no common scheme for the outer nuclear membrane proteins interacting with Sun I in centrosome/nucleus attachment. Here we propose a model in which Sun1 mediates a physical link between centrosomes and clustered centromeres through both nuclear membranes in Dictyostelium. For the first time we provide a detailed microscopic analysis of the centrosomal and nuclear envelope localization of endogenous Dictyostelium Sun1 during interphase and mitosis. By immunogold electron microscopy we show that Sun1 is a resident of both nuclear membranes. Disruption of Sun1 function by overexpression of full-length GFP-Sun1 or a GFP-Sun-domain deletion construct revealed not only the established function in centrosome/nucleus attachment and maintenance of ploidy, but also a requirement of Sun1 for the association of the centromere cluster with the centrosome. Live-cell imaging visualized the occurrence of mitotic defects, and demonstrated the requirement of microtubules for dynamic distance changes between centrosomes and nuclei. FRAP analysis revealed at least two populations of Sun1, with an immobile fraction associated with the centrosome, and a mobile fraction in the nuclear envelope. Y1 - 2009 UR - http://www.sciencedirect.com/science/journal/01719335 U6 - https://doi.org/10.1016/j.ejcb.2009.06.003 SN - 0171-9335 ER - TY - JOUR A1 - Dubreuil, R. R. A1 - Grushko, T. A1 - Baumann, Otto T1 - Differential effects of a labial mutation on the development, structure, and function of stomach acid-secreting cells in Drosophila larvae and adults Y1 - 2001 ER - TY - JOUR A1 - Baumann, Otto T1 - Disruption of actin filaments causes redistribution of ryanodine receptor Ca2+ channels in honeybee photoreceptor cells Y1 - 2001 ER - TY - JOUR A1 - Zimmermann, Bernhard A1 - Dames, Petra A1 - Walz, Bernd A1 - Baumann, Otto T1 - Distribution and serotonin-induced activation of vacuolar-type H+-ATPase in the salivary glands of the blowfly Calliphora vicina Y1 - 2003 ER - TY - JOUR A1 - Baumann, Otto T1 - Distribution of Na+, K+-ATPase in photoreceptor cells of insects. Y1 - 1997 ER - TY - JOUR A1 - Baumann, Otto T1 - Distribution of nonmuscle myosin-II in honeybee photoreceptor cells and its possible role in maintaining compound eye architecture Y1 - 2001 ER - TY - JOUR A1 - Baumann, Otto T1 - Distribution of ryanodine receptor Ca2+ channels in insect photoreceptor cells Y1 - 2000 ER -