@article {68312, title = {A sensitive and specific genetically encoded potassium ion biosensor for in vivo applications across the tree of life.}, journal = {PLoS Biology}, volume = {20}, year = {2022}, month = {2022 Sep 06}, pages = {e3001772}, abstract = {

Potassium ion (K+) plays a critical role as an essential electrolyte in all biological systems. Genetically encoded fluorescent K+ biosensors are promising tools to further improve our understanding of K+-dependent processes under normal and pathological conditions. Here, we report the crystal structure of a previously reported genetically encoded fluorescent K+ biosensor, GINKO1, in the K+-bound state. Using structure-guided optimization and directed evolution, we have engineered an improved K+ biosensor, designated GINKO2, with higher sensitivity and specificity. We have demonstrated the utility of GINKO2 for in vivo detection and imaging of K+ dynamics in multiple model organisms, including bacteria, plants, and mice.

}, issn = {1545-7885}, doi = {10.1371/journal.pbio.3001772}, author = {Wu, Sheng-Yi and Wen, Yurong and Serre, Nelson B C and Laursen, Cathrine Charlotte Heiede and Dietz, Andrea Grost{\o}l and Taylor, Brian R and Drobizhev, Mikhail and Molina, Rosana S and Abhi Aggarwal and Rancic, Vladimir and Becker, Michael and Ballanyi, Klaus and Podgorski, Kaspar and Hirase, Hajime and Nedergaard, Maiken and Fendrych, Maty{\'a}{\v s} and Lemieux, M Joanne and Eberl, Daniel F and Kay, Alan R and Campbell, Robert E and Shen, Yi} } @article {68445, title = {Transmembrane channel-like (tmc) gene regulates Drosophila larval locomotion.}, journal = {Proc Natl Acad Sci U S A}, volume = {113}, year = {2016}, month = {06/2016}, pages = {7243-8}, abstract = {

Drosophila larval locomotion, which entails rhythmic body contractions, is controlled by sensory feedback from proprioceptors. The molecular mechanisms mediating this feedback are little understood. By using genetic knock-in and immunostaining, we found that the Drosophila melanogaster transmembrane channel-like (tmc) gene is expressed in the larval class I and class II dendritic arborization (da) neurons and bipolar dendrite (bd) neurons, both of which are known to provide sensory feedback for larval locomotion. Larvae with knockdown or loss of tmc function displayed reduced crawling speeds, increased head cast frequencies, and enhanced backward locomotion. Expressing Drosophila TMC or mammalian TMC1 and/or TMC2 in the tmc-positive neurons rescued these mutant phenotypes. Bending of the larval body activated the tmc-positive neurons, and in tmc mutants this bending response was impaired. This implicates TMC{\textquoteright}s roles in Drosophila proprioception and the sensory control of larval locomotion. It also provides evidence for a functional conservation between Drosophila and mammalian TMCs.

}, keywords = {Animals, Animals, Genetically Modified, Cell Line, Drosophila melanogaster, Drosophila Proteins, Larva, Locomotion, Membrane Proteins, Mutation, Neurons}, issn = {1091-6490}, doi = {10.1073/pnas.1606537113}, author = {Guo, Yanmeng and Wang, Yuping and Zhang, Wei and Meltzer, Shan and Zanini, Damiano and Yu, Yue and Li, Jiefu and Cheng, Tong and Guo, Zhenhao and Wang, Qingxiu and Jacobs, Julie S and Sharma, Yashoda and Eberl, Daniel F and G{\"o}pfert, Martin C and Jan, Lily Yeh and Jan, Yuh Nung and Wang, Zuoren} } @article {6282, title = {Cbl-associated protein regulates assembly and function of two tension-sensing structures in Drosophila.}, journal = {Development}, volume = {140}, year = {2013}, month = {2013 Feb 1}, pages = {627-38}, abstract = {

Cbl-associated protein (CAP) localizes to focal adhesions and associates with numerous cytoskeletal proteins; however, its physiological roles remain unknown. Here, we demonstrate that Drosophila CAP regulates the organization of two actin-rich structures in Drosophila: muscle attachment sites (MASs), which connect somatic muscles to the body wall; and scolopale cells, which form an integral component of the fly chordotonal organs and mediate mechanosensation. Drosophila CAP mutants exhibit aberrant junctional invaginations and perturbation of the cytoskeletal organization at the MAS. CAP depletion also results in collapse of scolopale cells within chordotonal organs, leading to deficits in larval vibration sensation and adult hearing. We investigate the roles of different CAP protein domains in its recruitment to, and function at, various muscle subcellular compartments. Depletion of the CAP-interacting protein Vinculin results in a marked reduction in CAP levels at MASs, and vinculin mutants partially phenocopy Drosophila CAP mutants. These results show that CAP regulates junctional membrane and cytoskeletal organization at the membrane-cytoskeletal interface of stretch-sensitive structures, and they implicate integrin signaling through a CAP/Vinculin protein complex in stretch-sensitive organ assembly and function.

}, keywords = {Actin Cytoskeleton, Amino Acid, Amino Acid Sequence, Animal Structures, Animals, Binding Sites, Cell Membrane, Cell-Matrix Junctions, Cellular, Cytoskeletal Proteins, Developmental, Drosophila, Electron, Electrophysiological Phenomena, Gene Expression Regulation, Genome, Hearing Disorders, Insect, Integrins, Larva, Mechanotransduction, Microscopy, Multiprotein Complexes, Muscles, Protein Interaction Mapping, Sequence Homology, Signal Transduction, src Homology Domains, Talin, Transmission, Vibration, Vinculin}, issn = {1477-9129}, doi = {10.1242/dev.085100}, author = {Bharadwaj, Rajnish and Roy, Madhuparna and Ohyama, Tomoko and Sivan-Loukianova, Elena and Delannoy, Michael and Lloyd, Thomas E and Zlatic, Marta and Eberl, Daniel F and Kolodkin, Alex L} }