Pusch, Michael

[["dc.bibliographiccitation.firstpage","785"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","792"],["dc.bibliographiccitation.volume","75"],["dc.contributor.author","Pusch, Michael"],["dc.contributor.author","Magrassi, Raffaella"],["dc.contributor.author","Wollnik, Bernd"],["dc.contributor.author","Conti, Franco"],["dc.date.accessioned","2017-09-07T11:54:25Z"],["dc.date.available","2017-09-07T11:54:25Z"],["dc.date.issued","1998"],["dc.description.abstract","The voltage-gated potassium channel protein KvLQT1 (Wang et al., 1996. Nature Genet. 12:17–23) is believed to underlie the delayed rectifier potassium current of cardiac muscle together with the small membrane protein minK (also named IsK) as an essential auxiliary subunit (Barhanin et al., 1996. Nature. 384:78–80; Sanguinetti et al., 1996. Nature. 384:80–83). Using the Xenopus oocyte expression system, we analyzed in detail the gating characteristics of homomeric KvLQT1 channels and of heteromeric KvLQT1/minK channels using two-electrode voltage-clamp recordings. Activation of homomeric KvLQT1 at positive voltages is accompanied by an inactivation process that is revealed by a transient increase in conductance after membrane repolarization to negative values. We studied the recovery from inactivation and the deactivation of the channels during tail repolarizations at −120 mV after conditioning pulses of variable amplitude and duration. Most measurements were made in high extracellular potassium to increase the size of inward tail currents. However, experiments in normal low-potassium solutions showed that, in contrast to classical C-type inactivation, the inactivation of KvLQT1 is independent of extracellular potassium. At +40 mV inactivation develops with a delay of 100 ms. At the same potential, the activation estimated from the amplitude of the late exponential decay of the tail currents follows a less sigmoidal time course, with a late time constant of 300 ms. Inactivation of KvLQT1 is not complete, even at the most positive voltages. The delayed, voltage-dependent onset and the incompleteness of inactivation suggest a sequential gating scheme containing at least two open states and ending with an inactivating step that is voltage independent. In coexpression experiments of KvLQT1 with minK, inactivation seems to be largely absent, although biphasic tails are also observed that could be related to similar phenomena."],["dc.identifier.doi","10.1016/s0006-3495(98)77568-x"],["dc.identifier.gro","3145177"],["dc.identifier.pii","S000634959877568X"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2884"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","0006-3495"],["dc.title","Activation and Inactivation of Homomeric KvLQT1 Potassium Channels"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1621"],["dc.bibliographiccitation.issue","8"],["dc.bibliographiccitation.journal","Journal of Bone and Mineral Research"],["dc.bibliographiccitation.lastpage","1635"],["dc.bibliographiccitation.volume","36"],["dc.contributor.affiliation","Rössler, Uta; 1\r\nBIH Center for Regenerative Therapies (BCRT)\r\nCharité ‐ Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt‐Universität zu Berlin, and Berlin Institute of Health\r\nBerlin Germany"],["dc.contributor.affiliation","Stelzer, Nina; 1\r\nBIH Center for Regenerative Therapies (BCRT)\r\nCharité ‐ Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt‐Universität zu Berlin, and Berlin Institute of Health\r\nBerlin Germany"],["dc.contributor.affiliation","Bose, Shroddha; 6\r\nInstitute of Chemistry and Biochemistry\r\nFreie Universität Berlin\r\nBerlin Germany"],["dc.contributor.affiliation","Kopp, Johannes; 1\r\nBIH Center for Regenerative Therapies (BCRT)\r\nCharité ‐ Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt‐Universität zu Berlin, and Berlin Institute of Health\r\nBerlin Germany"],["dc.contributor.affiliation","Søe, Kent; 8\r\nClinical Cell Biology, Department of Pathology\r\nOdense University Hospital\r\nOdense C Denmark"],["dc.contributor.affiliation","Cyganek, Lukas; 11\r\nStem Cell Unit, Clinic for Cardiology and Pneumology\r\nUniversity Medical Center Göttingen\r\nGöttingen Germany"],["dc.contributor.affiliation","Zifarelli, Giovanni; 13\r\nIstituto di Biofisica, CNR\r\nGenoa Italy"],["dc.contributor.affiliation","Ali, Salaheddine; 1\r\nBIH Center for Regenerative Therapies (BCRT)\r\nCharité ‐ Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt‐Universität zu Berlin, and Berlin Institute of Health\r\nBerlin Germany"],["dc.contributor.affiliation","von der Hagen, Maja; 14\r\nAbteilung Neuropädiatrie, Medizinische Fakultät Carl Gustav Carus\r\nTechnische Universität Dresden\r\nDresden Germany"],["dc.contributor.affiliation","Strässler, Elisabeth Tamara; 15\r\nDepartment of Cardiology\r\nCharité ‐ Universitätsmedizin Berlin, Campus Benjamin Franklin\r\nBerlin Germany"],["dc.contributor.affiliation","Hahn, Gabriele; 17\r\nInstitut und Poliklinik für Radiologische Diagnostik\r\nMedizinische Fakultät Carl Gustav Carus Technische Universität Dresden\r\nDresden Germany"],["dc.contributor.affiliation","Pusch, Michael; 13\r\nIstituto di Biofisica, CNR\r\nGenoa Italy"],["dc.contributor.affiliation","Stauber, Tobias; 6\r\nInstitute of Chemistry and Biochemistry\r\nFreie Universität Berlin\r\nBerlin Germany"],["dc.contributor.affiliation","Izsvák, Zsuzsanna; 19\r\nMax‐Delbrück‐Center for Molecular Medicine (MDC), Helmholtz Association\r\nBerlin Germany"],["dc.contributor.affiliation","Gossen, Manfred; 20\r\nBerlin‐Brandenburg Center for Regenerative Therapies\r\nCharité Virchow Campus\r\nBerlin Germany"],["dc.contributor.affiliation","Stachelscheid, Harald; 22\r\nCharité ‐ Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt‐Universität zu Berlin, and Berlin Institute of Health\r\nBerlin Germany"],["dc.contributor.author","Rössler, Uta"],["dc.contributor.author","Hennig, Anna Floriane"],["dc.contributor.author","Stelzer, Nina"],["dc.contributor.author","Bose, Shroddha"],["dc.contributor.author","Kopp, Johannes"],["dc.contributor.author","Søe, Kent"],["dc.contributor.author","Cyganek, Lukas"],["dc.contributor.author","Zifarelli, Giovanni"],["dc.contributor.author","Ali, Salaheddine"],["dc.contributor.author","Kornak, Uwe"],["dc.contributor.author","Pusch, Michael"],["dc.contributor.author","von der Hagen, Maja"],["dc.contributor.author","Strässler, Elisabeth Tamara"],["dc.contributor.author","Hahn, Gabriele"],["dc.contributor.author","Stauber, Tobias"],["dc.contributor.author","Izsvák, Zsuzsanna"],["dc.contributor.author","Gossen, Manfred"],["dc.contributor.author","Stachelscheid, Harald"],["dc.date.accessioned","2021-06-01T09:41:55Z"],["dc.date.available","2021-06-01T09:41:55Z"],["dc.date.issued","2021"],["dc.date.updated","2022-02-09T13:20:40Z"],["dc.description.abstract","ABSTRACT Human induced pluripotent stem cells (hiPSCs) hold great potential for modeling human diseases and the development of innovative therapeutic approaches. Here, we report on a novel, simplified differentiation method for forming functional osteoclasts from hiPSCs. The three‐step protocol starts with embryoid body formation, followed by hematopoietic specification, and finally osteoclast differentiation. We observed continuous production of monocyte‐like cells over a period of up to 9 weeks, generating sufficient material for several osteoclast differentiations. The analysis of stage‐specific gene and surface marker expression proved mesodermal priming, the presence of monocyte‐like cells, and of terminally differentiated multinucleated osteoclasts, able to form resorption pits and trenches on bone and dentine in vitro. In comparison to peripheral blood mononuclear cell (PBMC)‐derived osteoclasts hiPSC‐derived osteoclasts were larger and contained a higher number of nuclei. Detailed functional studies on the resorption behavior of hiPSC‐osteoclasts indicated a trend towards forming more trenches than pits and an increase in pseudoresorption. We used hiPSCs from an autosomal recessive osteopetrosis (ARO) patient (BIHi002‐A, ARO hiPSCs) with compound heterozygous missense mutations p.(G292E) and p.(R403Q) in CLCN7, coding for the Cl−/H+‐exchanger ClC‐7, for functional investigations. The patient's leading clinical feature was a brain malformation due to defective neuronal migration. Mutant ClC‐7 displayed residual expression and retained lysosomal co‐localization with OSTM1, the gene coding for the osteopetrosis‐associated transmembrane protein 1, but only ClC‐7 harboring the mutation p.(R403Q) gave strongly reduced ion currents. An increased autophagic flux in spite of unchanged lysosomal pH was evident in undifferentiated ARO hiPSCs. ARO hiPSC‐derived osteoclasts showed an increased size compared to hiPSCs of healthy donors. They were not able to resorb bone, underlining a loss‐of‐function effect of the mutations. In summary, we developed a highly reproducible, straightforward hiPSC‐osteoclast differentiation protocol. We demonstrated that osteoclasts differentiated from ARO hiPSCs can be used as a disease model for ARO and potentially also other osteoclast‐related diseases. © 2021 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR)."],["dc.description.sponsorship","Berlin Institute of Health"],["dc.description.sponsorship","BCRT crossfield project GenoPro"],["dc.description.sponsorship","BIH Center for Regenerative Therapies"],["dc.description.sponsorship","European Community's Seventh Framework Programme"],["dc.description.sponsorship","Deutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/501100001659"],["dc.identifier.doi","10.1002/jbmr.4322"],["dc.identifier.pmid","33905594"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/85075"],["dc.identifier.url","https://sfb1002.med.uni-goettingen.de/production/literature/publications/394"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-425"],["dc.relation","SFB 1002: Modulatorische Einheiten bei Herzinsuffizienz"],["dc.relation","SFB 1002 | S01: In vivo und in vitro Krankheitsmodelle"],["dc.relation.eissn","1523-4681"],["dc.relation.issn","0884-0431"],["dc.relation.workinggroup","RG Cyganek (Stem Cell Unit)"],["dc.rights","This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made."],["dc.title","Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7 ‐related osteopetrosis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.subtype","original_ja"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","313"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Bioscience Reports"],["dc.bibliographiccitation.lastpage","321"],["dc.bibliographiccitation.volume","7"],["dc.contributor.author","Penner, Reinhold"],["dc.contributor.author","Pusch, Michael"],["dc.contributor.author","Neher, Erwin"],["dc.date.accessioned","2022-03-01T11:43:50Z"],["dc.date.available","2022-03-01T11:43:50Z"],["dc.date.issued","1987"],["dc.description.abstract","Transient increases of intracellular calcium and exocytotic activity of rat peritoneal mast cells following stimulation with compound 48/80 were monitored using the Ca-indicator dye fura-2 and the capacitance measurement technique. It is known that mast cells very rapidly lose their secretory response towards antigenic or compound 48/80-induced stimulation in the whole-cell recording configuration of the patch-clamp technique due to “washout” of signal mediators. In contrast, we found that calcium transients remained unaffected by intracellular dialysis for as long as 10 min. The fast “washout” phenomenon of exocytosis could be overcome by supplementing the pipette filling solution with guanosinetriphosphate (GTP) indicating a major role for GTP-binding proteins in secretion. The restoration of exocytosis was transient and decayed within three minutes, suggesting diffusional escape of one or several other cytoplasmic substances involved in stimulus-secretion coupling. Quantitative aspects of this process and the implications of its differential effects on Ca-transients versus secretion are discussed."],["dc.description.abstract","Transient increases of intracellular calcium and exocytotic activity of rat peritoneal mast cells following stimulation with compound 48/80 were monitored using the Ca-indicator dye fura-2 and the capacitance measurement technique. It is known that mast cells very rapidly lose their secretory response towards antigenic or compound 48/80-induced stimulation in the whole-cell recording configuration of the patch-clamp technique due to “washout” of signal mediators. In contrast, we found that calcium transients remained unaffected by intracellular dialysis for as long as 10 min. The fast “washout” phenomenon of exocytosis could be overcome by supplementing the pipette filling solution with guanosinetriphosphate (GTP) indicating a major role for GTP-binding proteins in secretion. The restoration of exocytosis was transient and decayed within three minutes, suggesting diffusional escape of one or several other cytoplasmic substances involved in stimulus-secretion coupling. Quantitative aspects of this process and the implications of its differential effects on Ca-transients versus secretion are discussed."],["dc.identifier.doi","10.1007/BF01121453"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/102855"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.relation.eissn","1573-4935"],["dc.relation.issn","0144-8463"],["dc.title","Washout phenomena in dialyzed mast cells allow discrimination of different steps in stimulus-secretion coupling"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","19"],["dc.bibliographiccitation.issue","Suppl."],["dc.bibliographiccitation.journal","The Journal of Physiology"],["dc.bibliographiccitation.lastpage","25"],["dc.bibliographiccitation.volume","482"],["dc.contributor.author","Jentsch, Thomas J."],["dc.contributor.author","Günther, Willy"],["dc.contributor.author","Pusch, Michael"],["dc.contributor.author","Schwappach, Blanche"],["dc.date.accessioned","2017-09-07T11:54:18Z"],["dc.date.available","2017-09-07T11:54:18Z"],["dc.date.issued","2014"],["dc.description.abstract","We review the properties of ClC chloride channels, members of an expanding gene family originally discovered by the cloning of the ClC‐0 chloride channel from Torpedo electric organ. There are at least nine different ClC genes in mammals, several of which seem to be expressed ubiquitously, while others are expressed in a highly specific manner (e.g. the muscle‐specific ClC‐1 channel and the kidney‐specific ClC‐K channels). The newly cloned rat ClC‐4 is strongly expressed in liver and brain, but also in heart, muscle, kidney and spleen. ClC chloride channels are structurally unrelated to other channel proteins and have twelve putative transmembrane domains. They function as multimers with probably four subunits. Functional characterization is most advanced with ClC‐0, ClC‐1 (mutations which cause myotonia) and ClC‐2, a swelling‐activated chloride channel. Many of the new ClC family members cannot yet be expressed functionally."],["dc.identifier.doi","10.1113/jphysiol.1995.sp020560"],["dc.identifier.gro","3145141"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2844"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","0022-3751"],["dc.title","Properties of voltage-gated chloride channels of the ClC gene family"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","805"],["dc.bibliographiccitation.issue","5"],["dc.bibliographiccitation.journal","Human Molecular Genetics"],["dc.bibliographiccitation.lastpage","811"],["dc.bibliographiccitation.volume","6"],["dc.contributor.author","Wollnik, Bernd"],["dc.contributor.author","Kubisch, C."],["dc.contributor.author","Steinmeyer, K"],["dc.contributor.author","Pusch, Michael"],["dc.date.accessioned","2017-09-07T11:50:58Z"],["dc.date.available","2017-09-07T11:50:58Z"],["dc.date.issued","1997"],["dc.description.abstract","Mutations in the muscular voltage-dependent Cl- channel, CIC-1, lead to recessive and dominant myotonia, Here we analyse the effects of one dominant (G200R) and three recessive (Y150C, Y261C, and M485V) mutations after functional expression in Xenopus oocytes, Glycine 200 is a highly conserved amino acid located in a conserved stretch in the putatively cytoplasmic loop between domains D2 and D3, Similar to several other dominant mutations the amino acid exchange G200R leads to a drastic shift by similar to 65 mV of the open probability curve to more positive voltages, As explored by co-expression studies, the shift is intermediate in heteromeric mutant/WT channels, Open channel properties such as single channel conductance, rectification or ion selectivity are not changed, Thus we identified a new region of the CIC-1 protein in which mutations can lead to drastic shifts of the voltage dependence, The recessive mutation M485V, which is located in a conserved region at the beginning of domain D10, leads to a drastic reduction of the single channel conductance from 1.5 pS for WT to similar to 0.3 pS, In addition, the mutant is strongly inwardly rectifying and deactivates incompletely at negative voltages, ion-selectivity, however, is unchanged, These electrophysiological properties fully explain the recessive phenotype of the mutation and identify a new region of the protein that is involved in ion permeation and gating of the CIC-1 channel, The other two recessive mutations (Y150C and Y261C) had been found in a compound heterozygous patient, Surprisingly, expression of these mutants in oocytes yielded currents indistinguishable from WT CIC-1 when explored by two-electrode voltage clamp recording and patch clamping (either singly or both mutations co-expressed), Other mechanisms that are not faithfully represented by the Xenopus expression system must therefore be responsible for the myotonic symptoms associated with these mutations."],["dc.identifier.doi","10.1093/hmg/6.5.805"],["dc.identifier.gro","3144604"],["dc.identifier.isi","A1997WX60800021"],["dc.identifier.pmid","9158157"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2247"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0964-6906"],["dc.title","Identification of functionally important regions of the muscular chloride channel CIC-1 by analysis of recessive and dominant myotonic mutations"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","204"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Pflügers Archiv European Journal of Physiology"],["dc.bibliographiccitation.lastpage","211"],["dc.bibliographiccitation.volume","411"],["dc.contributor.author","Pusch, Michael"],["dc.contributor.author","Neher, Erwin"],["dc.date.accessioned","2022-03-01T11:47:17Z"],["dc.date.available","2022-03-01T11:47:17Z"],["dc.date.issued","1988"],["dc.identifier.doi","10.1007/BF00582316"],["dc.identifier.pii","BF00582316"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/103979"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.relation.eissn","1432-2013"],["dc.relation.issn","0031-6768"],["dc.title","Rates of diffusional exchange between small cells and a measuring patch pipette"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]