@article{GaoNagpalSchneideretal.2015,
  author    = {Gao, Shiqiang and Nagpal, Jatin and Schneider, Martin W. and Kozjak-Pavlovic, Vera and Nagel, Georg and Gottschalk, Alexander},
  title     = {Optogenetic manipulation of cGMP in cells and animals by the tightly light-regulated guanylyl-cyclase opsin CyclOp},
  series = {Nature Communications},
  volume    = {6},
  journal   = {Nature Communications},
  number    = {8046},
  doi       = {10.1038/ncomms9046},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-148197},
  year      = {2015},
  abstract  = {Cyclic GMP (cGMP) signalling regulates multiple biological functions through activation of protein kinase G and cyclic nucleotide-gated (CNG) channels. In sensory neurons, cGMP permits signal modulation, amplification and encoding, before depolarization. Here we implement a guanylyl cyclase rhodopsin from Blastocladiella emersonii as a new optogenetic tool (BeCyclOp), enabling rapid light-triggered cGMP increase in heterologous cells (Xenopus oocytes, HEK293T cells) and in Caenorhabditis elegans. Among five different fungal CyclOps, exhibiting unusual eight transmembrane topologies and cytosolic N-termini, BeCyclOp is the superior optogenetic tool (light/dark activity ratio: 5,000; no cAMP production; turnover (20 °C) ~17 cGMPs\(^{-1}\)). Via co-expressed CNG channels (OLF in oocytes, TAX-2/4 in C. elegans muscle), BeCyclOp photoactivation induces a rapid conductance increase and depolarization at very low light intensities. In O\(_2\)/CO\(_2\) sensory neurons of C. elegans, BeCyclOp activation evokes behavioural responses consistent with their normal sensory function. BeCyclOp therefore enables precise and rapid optogenetic manipulation of cGMP levels in cells and animals.},
  language  = {en}
}
@phdthesis{GeisenhofgebTrinkwalder2019,
  author    = {Geisenhof [geb. Trinkwalder], Michaela},
  title     = {Erforschung des Schicksals des Mittelk{\"o}rpers anhand der ZF1-Methode},
  doi       = {10.25972/OPUS-18219},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-182199},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2019},
  abstract  = {Bei der Teilung einer Zelle werden das Genom und die Zellbestandteile zwischen zwei Tochterzellen aufgeteilt. Dies erfordert verschiedene fein aufeinander abgestimmte Vorg{\"a}nge. Unter anderem ist eine proteinreiche Struktur beteiligt, die 1891 entdeckt wurde: der Mittelk{\"o}rper. In vorliegender Arbeit wurden gezielt gekennzeichnete Mittelk{\"o}rperproteine analysiert und verschiedene Phasen des Transports unterschieden. Es erfolgten erstmals Messungen unter Nutzung der ZF1-Methode. Zudem wird anhand der ZF1-Technik nachgewiesen, dass im Rahmen der Zellteilung die Trennung der interzellul{\"a}ren Br{\"u}cke zu beiden Seiten des Mittelk{\"o}rpers stattfindet, woraufhin dieser nach extrazellul{\"a}r abgegeben wird und {\"u}ber einen der Phagozytose {\"a}hnlichen und von Aktin abh{\"a}ngigen Mechanismus von einer Tochterzelle oder unverwandten Nachbarzelle aufgenommen wird.},
  subject      = {Mitose},
  language  = {de}
}
@article{MarkertSkoruppaYuetal.2020,
  author    = {Markert, Sebastian M. and Skoruppa, Michael and Yu, Bin and Mulcahy, Ben and Zhen, Mai and Gao, Shangbang and Sendtner, Michael and Stigloher, Christian},
  title     = {Overexpression of an ALS-associated FUS mutation in C. elegans disrupts NMJ morphology and leads to defective neuromuscular transmission},
  series = {Biology Open},
  volume    = {9},
  journal   = {Biology Open},
  doi       = {10.1242/bio.055129},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-230662},
  year      = {2020},
  abstract  = {The amyotrophic lateral sclerosis (ALS) neurodegenerative disorder has been associated with multiple genetic lesions, including mutations in the gene for fused in sarcoma (FUS), a nuclear-localized RNA/DNA-binding protein. Neuronal expression of the pathological form of FUS proteins in Caenorhabditis elegans results in mislocalization and aggregation of FUS in the cytoplasm, and leads to impairment of motility. However, the mechanisms by which the mutant FUS disrupts neuronal health and function remain unclear. Here we investigated the impact of ALS-associated FUS on motor neuron health using correlative light and electron microscopy, electron tomography, and electrophysiology. We show that ectopic expression of wild-type or ALS-associated human FUS impairs synaptic vesicle docking at neuromuscular junctions. ALS-associated FUS led to the emergence of a population of large, electron-dense, and filament-filled endosomes. Electrophysiological recording revealed reduced transmission from motor neurons to muscles. Together, these results suggest a pathological effect of ALS-causing FUS at synaptic structure and function organization.},
  language  = {en}
}
@phdthesis{Beer2021,
  author    = {Beer, Katharina Beate},
  title     = {Identification and characterization of TAT-5 interactors that regulate extracellular vesicle budding},
  doi       = {10.25972/OPUS-20672},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-206724},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2021},
  abstract  = {Cells from bacteria to man release extracellular vesicles (EV) such as microvesicles (MV) that carry signaling molecules like morphogens and miRNAs to control intercellular communication during health and disease. MV release also sculpts membranes, e.g. repairing damaged membranes to avoid cell death. HIV viruses also bud from the plasma membrane in a similar fashion. In order to determine the in vivo functions of MVs and regulate their release, we need to understand the mechanisms of MV release by plasma membrane budding (ectocytosis). The conserved phospholipid flippase TAT-5 maintains the asymmetric localization of phosphatidylethanolamine (PE) in the plasma membrane and was the only known inhibitor of ESCRT-mediated ectocytosis in C. elegans. Loss of TAT-5 lipid flipping activity increased the externalization of PE and accumulation of MVs. However, it was unclear how cells control TAT-5 activity to release the right amount of MVs at the right time, since no upstream regulators of TAT-5 were known. To identify conserved TAT-5 regulators we looked for new proteins that inhibit MV release. To do so, we first developed a degradation-based technique to specifically label MVs. We tagged a plasma membrane reporter with the endogenous ZF1 degradation tag (degron) and expressed it in C. elegans embryos. This reporter is protected from degradation inside MVs, but is degraded inside the cell. Thus, the fluorescence is selectively maintained inside MVs, creating the first MV-specific reporter. We identified four MV release inhibitors associated with retrograde recycling, including the class III PI3Kinase VPS-34, Beclin1 homolog BEC-1, DnaJ protein RME-8, and the uncharacterized Dopey homolog PAD-1. We found that VPS-34, BEC-1, RME-8, and redundant sorting nexins are required for the plasma membrane localization of TAT-5, which is important to maintain PE asymmetry and inhibit MV release. Although we confirmed that PAD-1 and the GEF-like protein MON-2 are required for endosomal recycling, they only traffic TAT-5 in the absence of sorting nexin-mediated recycling. Instead, PAD-1 is specifically required for the lipid flipping activity of TAT-5 that inhibits MV release. Thus, our work pinpoints TAT-5 and PE as key regulators of plasma membrane budding, further supporting the model that PE externalization drives ectocytosis. In addition, we uncovered redundant intracellular trafficking pathways, which affect organelle size and revealed new regulators of TAT-5 flippase activity. These newly identified ectocytosis inhibitors provide a toolkit to test the in vivo roles of MVs. In the long term, our work will help to identify the mechanisms that govern MV budding, furthering our understanding of the mechanisms that regulate disease-mediated EV release, membrane sculpting and viral budding.},
  subject      = {Caenorhabditis elegans},
  language  = {en}
}