Klonieren und Charakterisieren von P/Q-Typ-Calciumkanälen für Mikroskopie an lebenden Zellen

Juha, Martin

dc.contributor.advisor	Neher, Erwin Prof. Dr.	de
dc.contributor.author	Juha, Martin	de
dc.date.accessioned	2013-09-03T13:58:36Z
dc.date.available	2013-09-13T22:50:04Z
dc.date.issued	2013-09-03	de
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-0001-BB49-0
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-4030
dc.language.iso	deu	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/3.0/
dc.subject.ddc	610	de
dc.title	Klonieren und Charakterisieren von P/Q-Typ-Calciumkanälen für Mikroskopie an lebenden Zellen	de
dc.type	doctoralThesis	de
dc.title.translated	Cloning and characterization of P/Q-type calcium channels for live cell imaging	de
dc.contributor.referee	Wouters, Fred Prof. Dr.	de
dc.date.examination	2013-09-03	de
dc.description.abstracteng	In all excitable cells voltage-dependent calcium channels can be found. They mediate the entry of calcium into cells following depolarization. Calcium contributes to a wide variety of cellular response like muscle contraction, neurotransmitter release, gene expression and also synaptogenesis. For the neurotransmitter release vesicles are needed and the release is supported from a variety of different proteins. Research in the field of neurotransmitter release revealed the probable existence of two different vesicle populations within the RRP which differ in their release probability, the velocity of recruitment and their fusion competence. These different characteristics might be caused by a significant heterogeneity concerning the distance between the vesicle and the controlling calcium channels. To approximate these presumptions and to specify the exact location of P/Q-type calcium channels a specific labeling of the α1-subunit of these channels was performed. The labeling technique included the covalent labeling of the α1-subunit using a short 11- residue -long ACP-ybbR-Tag and an adenoviral expression system under the requirements of live cell imaging. Besides this, experiments using the STED-microscope were carried out. Although the specific labeling of P/Q-type calcium channels could neither be successfully performed in HEK293 cells, nor in hippocampal neurons nor by using the STED-microscope, it could be demonstrated that the ACP labeling procedure works in general using the chosen methods. A specific membrane labeling was performed. It could also be proven that the inserted tag did not disturb the function of the calcium channel.	de
dc.contributor.coReferee	Oppermann, Martin Prof. Dr.	de
dc.subject.ger	Calciumkanäle	de
dc.subject.ger	Vesikelpool	de
dc.subject.ger	Transmitterfreisetzung	de
dc.subject.ger	Calciumkanal-bedingte Erkrankungen	de
dc.subject.ger	Färbemethoden	de
dc.subject.ger	schnell-freisetzende Vesikel	de
dc.subject.ger	langsam-freisetzende Vesikel	de
dc.subject.eng	calcium channels	de
dc.subject.eng	vesicle pool	de
dc.subject.eng	ACP-labeling	de
dc.subject.eng	ybbR	de
dc.subject.eng	transmitter release	de
dc.subject.eng	SNARE	de
dc.subject.eng	calcium channelopathies	de
dc.subject.eng	labeling methods	de
dc.subject.eng	slow-releasing vesicles	de
dc.subject.eng	fast-releasing vesicles	de
dc.subject.eng	calcium cluster	de
dc.subject.eng	live cell imaging	de
dc.identifier.urn	urn:nbn:de:gbv:7-11858/00-1735-0000-0001-BB49-0-4
dc.affiliation.institute	Medizinische Fakultät	de
dc.subject.gokfull	Physiologie / Pathophysiologie - Allgemein- und Gesamtdarstellungen (PPN619875283)	de
dc.subject.gokfull	Methoden und Techniken in der Medizin (PPN619875143)	de
dc.subject.gokfull	Physik / Biopyhsik / Biomedizinische Technik - Allgemein- und Gesamtdarstellungen (PPN619875100)	de
dc.subject.gokfull	Biochemie / Physiologische Chemie / Pathobiochemie - Allgemein- und Gesamtdarstellungen (PPN619875313)	de
dc.description.embargoed	2013-09-13	de
dc.identifier.ppn	766821927

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