@article{BuchnerBlancoRedondoBunzetal.2013, author = {Buchner, Erich and Blanco Redondo, Beatriz and Bunz, Melanie and Halder, Partho and Sadanandappa, Madhumala K. and M{\"u}hlbauer, Barbara and Erwin, Felix and Hofbauer, Alois and Rodrigues, Veronica and VijayRaghavan, K. and Ramaswami, Mani and Rieger, Dirk and Wegener, Christian and F{\"o}rster, Charlotte}, title = {Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the W{\"u}rzburg Hybridoma Library}, series = {PLoS ONE}, journal = {PLoS ONE}, doi = {10.1371/journal.pone.0075420}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-97109}, year = {2013}, abstract = {Several novel synaptic proteins have been identified by monoclonal antibodies (mAbs) of the W{\"u}rzburg hybridoma library generated against homogenized Drosophila brains, e.g. cysteine string protein, synapse-associated protein of 47 kDa, and Bruchpilot. However, at present no routine technique exists to identify the antigens of mAbs of our library that label only a small number of cells in the brain. Yet these antibodies can be used to reproducibly label and thereby identify these cells by immunohistochemical staining. Here we describe the staining patterns in the Drosophila brain for ten mAbs of the W{\"u}rzburg hybridoma library. Besides revealing the neuroanatomical structure and distribution of ten different sets of cells we compare the staining patterns with those of antibodies against known antigens and GFP expression patterns driven by selected Gal4 lines employing regulatory sequences of neuronal genes. We present examples where our antibodies apparently stain the same cells in different Gal4 lines suggesting that the corresponding regulatory sequences can be exploited by the split-Gal4 technique for transgene expression exclusively in these cells. The detection of Gal4 expression in cells labeled by mAbs may also help in the identification of the antigens recognized by the antibodies which then in addition to their value for neuroanatomy will represent important tools for the characterization of the antigens. Implications and future strategies for the identification of the antigens are discussed.}, language = {en} } @article{ReinhardBertoliniSaitoetal.2022, author = {Reinhard, Nils and Bertolini, Enrico and Saito, Aika and Sekiguchi, Manabu and Yoshii, Taishi and Rieger, Dirk and Helfrich-F{\"o}rster, Charlotte}, title = {The lateral posterior clock neurons of Drosophila melanogaster express three neuropeptides and have multiple connections within the circadian clock network and beyond}, series = {Journal of Comparative Neurology}, volume = {530}, journal = {Journal of Comparative Neurology}, number = {9}, doi = {10.1002/cne.25294}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-276456}, pages = {1507 -- 1529}, year = {2022}, abstract = {Drosophila's lateral posterior neurons (LPNs) belong to a small group of circadian clock neurons that is so far not characterized in detail. Thanks to a new highly specific split-Gal4 line, here we describe LPNs' morphology in fine detail, their synaptic connections, daily bimodal expression of neuropeptides, and propose a putative role of this cluster in controlling daily activity and sleep patterns. We found that the three LPNs are heterogeneous. Two of the neurons with similar morphology arborize in the superior medial and lateral protocerebrum and most likely promote sleep. One unique, possibly wakefulness-promoting, neuron with wider arborizations extends from the superior lateral protocerebrum toward the anterior optic tubercle. Both LPN types exhibit manifold connections with the other circadian clock neurons, especially with those that control the flies' morning and evening activity (M- and E-neurons, respectively). In addition, they form synaptic connections with neurons of the mushroom bodies, the fan-shaped body, and with many additional still unidentified neurons. We found that both LPN types rhythmically express three neuropeptides, Allostatin A, Allostatin C, and Diuretic Hormone 31 with maxima in the morning and the evening. The three LPN neuropeptides may, furthermore, signal to the insect hormonal center in the pars intercerebralis and contribute to rhythmic modulation of metabolism, feeding, and reproduction. We discuss our findings in the light of anatomical details gained by the recently published hemibrain of a single female fly on the electron microscopic level and of previous functional studies concerning the LPN.}, language = {en} } @article{ReinhardSchubertBertolinietal.2022, author = {Reinhard, Nils and Schubert, Frank K. and Bertolini, Enrico and Hagedorn, Nicolas and Manoli, Giulia and Sekiguchi, Manabu and Yoshii, Taishi and Rieger, Dirk and Helfrich-F{\"o}rster, Charlotte}, title = {The neuronal circuit of the dorsal circadian clock neurons in Drosophila melanogaster}, series = {Frontiers in Physiology}, volume = {13}, journal = {Frontiers in Physiology}, issn = {1664-042X}, doi = {10.3389/fphys.2022.886432}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-272527}, year = {2022}, abstract = {Drosophila's dorsal clock neurons (DNs) consist of four clusters (DN1as, DN1ps, DN2s, and DN3s) that largely differ in size. While the DN1as and the DN2s encompass only two neurons, the DN1ps consist of ∼15 neurons, and the DN3s comprise ∼40 neurons per brain hemisphere. In comparison to the well-characterized lateral clock neurons (LNs), the neuroanatomy and function of the DNs are still not clear. Over the past decade, numerous studies have addressed their role in the fly's circadian system, leading to several sometimes divergent results. Nonetheless, these studies agreed that the DNs are important to fine-tune activity under light and temperature cycles and play essential roles in linking the output from the LNs to downstream neurons that control sleep and metabolism. Here, we used the Flybow system, specific split-GAL4 lines, trans-Tango, and the recently published fly connectome (called hemibrain) to describe the morphology of the DNs in greater detail, including their synaptic connections to other clock and non-clock neurons. We show that some DN groups are largely heterogenous. While certain DNs are strongly connected with the LNs, others are mainly output neurons that signal to circuits downstream of the clock. Among the latter are mushroom body neurons, central complex neurons, tubercle bulb neurons, neurosecretory cells in the pars intercerebralis, and other still unidentified partners. This heterogeneity of the DNs may explain some of the conflicting results previously found about their functionality. Most importantly, we identify two putative novel communication centers of the clock network: one fiber bundle in the superior lateral protocerebrum running toward the anterior optic tubercle and one fiber hub in the posterior lateral protocerebrum. Both are invaded by several DNs and LNs and might play an instrumental role in the clock network.}, language = {en} } @article{PuetzKramRauhetal.2021, author = {P{\"u}tz, Stephanie M. and Kram, Jette and Rauh, Elisa and Kaiser, Sophie and Toews, Romy and Lueningschroer-Wang, Yi and Rieger, Dirk and Raabe, Thomas}, title = {Loss of p21-activated kinase Mbt/PAK4 causes Parkinson-like symptoms in Drosophila}, series = {Disease Models \& Mechanisms}, volume = {14}, journal = {Disease Models \& Mechanisms}, number = {6}, doi = {10.1242/dmm.047811}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-259222}, pages = {dmm047811}, year = {2021}, abstract = {Parkinson's disease (PD) provokes bradykinesia, resting tremor, rigidity and postural instability, and also non-motor symptoms such as depression, anxiety, sleep and cognitive impairments. Similar phenotypes can be induced in Drosophila melanogaster through modification of PD-relevant genes or the administration of PD inducing toxins. Recent studies correlated deregulation of human p21-activated kinase 4 (PAK4) with PD, leaving open the question of a causative relationship of mutations in this gene for manifestation of PD symptoms. To determine whether flies lacking the PAK4 homolog Mushroom bodies tiny (Mbt) show PD-like phenotypes, we tested for a variety of PD criteria. Here, we demonstrate that mbt mutant flies show PD-like phenotypes including age-dependent movement deficits, reduced life expectancy and fragmented sleep. They also react to a stressful situation with higher immobility, indicating an influence of Mbt on emotional behavior. Loss of Mbt function has a negative effect on the number of dopaminergic protocerebral anterior medial (PAM) neurons, most likely caused by a proliferation defect of neural progenitors. The age-dependent movement deficits are not accompanied by a corresponding further loss of PAM neurons. Previous studies highlighted the importance of a small PAM subgroup for age-dependent PD motor impairments. We show that impaired motor skills are caused by a lack of Mbt in this PAM subgroup. In addition, a broader re-expression of Mbt in PAM neurons improves life expectancy. Conversely, selective Mbt knockout in the same cells shortens lifespan. We conclude that mutations in Mbt/PAK4 can play a causative role in the development of PD phenotypes.}, language = {en} } @article{SchaeblerAmatobiHornetal.2020, author = {Sch{\"a}bler, Stefan and Amatobi, Kelechi M. and Horn, Melanie and Rieger, Dirk and Helfrich‑F{\"o}rster, Charlotte and Mueller, Martin J. and Wegener, Christian and Fekete, Agnes}, title = {Loss of function in the Drosophila clock gene period results in altered intermediary lipid metabolism and increased susceptibility to starvation}, series = {Cellular and Molecular Life Sciences}, volume = {77}, journal = {Cellular and Molecular Life Sciences}, issn = {1420-682X}, doi = {10.1007/s00018-019-03441-6}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232432}, pages = {4939-4956}, year = {2020}, abstract = {The fruit fly Drosophila is a prime model in circadian research, but still little is known about its circadian regulation of metabolism. Daily rhythmicity in levels of several metabolites has been found, but knowledge about hydrophobic metabolites is limited. We here compared metabolite levels including lipids between period\(^{01}\) (per\(^{01}\)) clock mutants and Canton-S wildtype (WT\(_{CS}\)) flies in an isogenic and non-isogenic background using LC-MS. In the non-isogenic background, metabo-lites with differing levels comprised essential amino acids, kynurenines, pterinates, glycero(phospho)lipids, and fatty acid esters. Notably, detectable diacylglycerols (DAG) and acylcarnitines (AC), involved in lipid metabolism, showed lower levels in per\(^{01}\) mutants. Most of these differences disappeared in the isogenic background, yet the level differences for AC as well as DAG were consistent for fly bodies. AC levels were dependent on the time of day in WTCS in phase with food consumption under LD conditions, while DAGs showed weak daily oscillations. Two short-chain ACs continued to cycle even in constant darkness. per\(^{01}\) mutants in LD showed no or very weak diel AC oscillations out of phase with feeding activity. The low levels of DAGs and ACs in per\(^{01}\) did not correlate with lower total food consumption, body mass or weight. Clock mutant flies showed higher sensitivity to starvation independent of their background-dependent activity level. Our results suggest that neither feeding, energy storage nor mobilisation is significantly affected in per\(^{01}\) mutants, but point towards impaired mitochondrial activity, supported by upregulation of the mitochondrial stress marker 4EBP in the clock mutants}, language = {en} } @article{SchlichtingRiegerCusumanoetal.2018, author = {Schlichting, Matthias and Rieger, Dirk and Cusumano, Paola and Grebler, Rudi and Costa, Rodolfo and Mazzotta, Gabriella M. and Helfrich-F{\"o}rster, Charlotte}, title = {Cryptochrome interacts with actin and enhances eye-mediated light sensitivity of the circadian clock in Drosophila melanogaster}, series = {Frontiers in Molecular Neuroscience}, volume = {11}, journal = {Frontiers in Molecular Neuroscience}, number = {238}, doi = {10.3389/fnmol.2018.00238}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-177086}, year = {2018}, abstract = {Cryptochromes (CRYs) are a class of flavoproteins that sense blue light. In animals, CRYs are expressed in the eyes and in the clock neurons that control sleep/wake cycles and are implied in the generation and/or entrainment of circadian rhythmicity. Moreover, CRYs are sensing magnetic fields in insects as well as in humans. Here, we show that in the fruit fly Drosophila melanogaster CRY plays a light-independent role as "assembling" protein in the rhabdomeres of the compound eyes. CRY interacts with actin and appears to increase light sensitivity of the eyes by keeping the "signalplex" of the phototransduction cascade close to the membrane. By this way, CRY also enhances light-responses of the circadian clock.}, language = {en} } @article{HornMitesserHovestadtetal.2019, author = {Horn, Melanie and Mitesser, Oliver and Hovestadt, Thomas and Yoshii, Taishi and Rieger, Dirk and Helfrich-F{\"o}rster, Charlotte}, title = {The circadian clock improves fitness in the fruit fly, Drosophila melanogaster}, series = {Frontiers in Physiology}, volume = {10}, journal = {Frontiers in Physiology}, number = {1374}, issn = {1664-042X}, doi = {10.3389/fphys.2019.01374}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-195738}, year = {2019}, abstract = {It is assumed that a properly timed circadian clock enhances fitness, but only few studies have truly demonstrated this in animals. We raised each of the three classical Drosophila period mutants for >50 generations in the laboratory in competition with wildtype flies. The populations were either kept under a conventional 24-h day or under cycles that matched the mutant's natural cycle, i.e., a 19-h day in the case of pers mutants and a 29-h day for perl mutants. The arrhythmic per0 mutants were grown together with wildtype flies under constant light that renders wildtype flies similar arrhythmic as the mutants. In addition, the mutants had to compete with wildtype flies for two summers in two consecutive years under outdoor conditions. We found that wildtype flies quickly outcompeted the mutant flies under the 24-h laboratory day and under outdoor conditions, but perl mutants persisted and even outnumbered the wildtype flies under the 29-h day in the laboratory. In contrast, pers and per0 mutants did not win against wildtype flies under the 19-h day and constant light, respectively. Our results demonstrate that wildtype flies have a clear fitness advantage in terms of fertility and offspring survival over the period mutants and - as revealed for perl mutants - this advantage appears maximal when the endogenous period resonates with the period of the environment. However, the experiments indicate that perl and pers persist at low frequencies in the population even under the 24-h day. This may be a consequence of a certain mating preference of wildtype and heterozygous females for mutant males and time differences in activity patterns between wildtype and mutants.}, language = {en} }