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Sebastian Wabnig, Jana Fiona Liewald, Szi-Chieh Yu, Alexander Gottschalk
Synaptic vesicles (SVs) undergo a cycle of biogenesis and membrane fusion to release transmitter, followed by recycling. How exocytosis and endocytosis are coupled is intensively investigated. We describe an all-optical method for identification of neurotransmission genes that can directly distinguish SV recycling factors in C. elegans, by motoneuron photostimulation and muscular RCaMP Ca2+ imaging. We verified our approach on mutants affecting synaptic transmission. Mutation of genes affecting SV recycling (unc-26 synaptojanin, unc-41 stonin, unc-57 endophilin, itsn-1 intersectin, snt-1 synaptotagmin) showed a distinct 'signature' of muscle Ca2+ dynamics, induced by cholinergic motoneuron photostimulation, i...
2015: PloS One
Jasper Akerboom, Nicole Carreras Calderón, Lin Tian, Sebastian Wabnig, Matthias Prigge, Johan Tolö, Andrew Gordus, Michael B Orger, Kristen E Severi, John J Macklin, Ronak Patel, Stefan R Pulver, Trevor J Wardill, Elisabeth Fischer, Christina Schüler, Tsai-Wen Chen, Karen S Sarkisyan, Jonathan S Marvin, Cornelia I Bargmann, Douglas S Kim, Sebastian Kügler, Leon Lagnado, Peter Hegemann, Alexander Gottschalk, Eric R Schreiter, Loren L Looger
Genetically encoded calcium indicators (GECIs) are powerful tools for systems neuroscience. Here we describe red, single-wavelength GECIs, "RCaMPs," engineered from circular permutation of the thermostable red fluorescent protein mRuby. High-resolution crystal structures of mRuby, the red sensor RCaMP, and the recently published red GECI R-GECO1 give insight into the chromophore environments of the Ca(2+)-bound state of the sensors and the engineered protein domain interfaces of the different indicators. We characterized the biophysical properties and performance of RCaMP sensors in vitro and in vivo in Caenorhabditis elegans, Drosophila larvae, and larval zebrafish...
2013: Frontiers in Molecular Neuroscience
S Marzorati, R Melzi, R Nano, B Antonioli, V Di Carlo, L Piemonti, F Bertuzzi
Islet transplantation is a new approach to treat type 1 diabetic patients. Despite its great potential and progressively increasing success rate, islet engraftment still represents an unsolved problem. Only part of the transplanted beta-cell mass survives after infusion due to hypoxia and inflammatory reactions, principally mediated by macrophages. We have demonstrated that human islets release monocyte chemoattractant protein-1 (MCP-1), one of the most powerful macrophage chemokines, which may impair the fate of a transplant...
April 2004: Transplantation Proceedings
I D Grozdova, E V Niupenko, E V Sveshnikova
Immunochemical analysis of the cAMP-dependent protein kinase regulatory subunit type II was performed with the use of two rabbit antisera elicited to a free R-subunit from pig brain and to a RcAMP complex. Quantitative precipitation of the homogeneous antigen revealed six determinants on the R-molecule. Of these at least one is localized in the R-fragment (37 kD), the others--in the N-terminal part of the R-molecule. The antigenic determinants seem to be remoted from the cAMP-binding centers, since the attachment of the affinity purified antibody Fab-fragments to the R-subunit did not influence the cAMP-binding activity of the latter...
July 1990: Biokhimii︠a︡
P O Schwille, C Bornhof, R Thun, D Scholz, R Bötticher, W Schellerer, A Sigel
In three groups (n = 12 each) of male controls (22--43 years), patients with recurring calcium urolithiasis (21--36 years) and hyperparathyroidism (HPT; 17--71 years) proven by surgery renal cyclic adenosine monophosphate (RcAMP), fractional tubular phosphate reabsorption and serum parathyroid hormone (PTH) were measured during endogenous creatinine clearance. RcAMP (muMol/g creatinine) was: controls 1.48 +/- SEM 0.27; stone formers 2.037 +/- 0.343 (not significantly different); HPT 6.234 +/- 0.454 (p less than 0...
June 15, 1978: Klinische Wochenschrift
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