Molecular analysis of the role of caspase-8 during liver regeneration and tumorigenesis [Elektronische Ressource] / vorgelegt von Julia Freimuth

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• Publié par: rheinisch-westfalischen_technischen_hochschule_-rwth-_aachen
• Publié le: 01 janvier 2009
• Nombre de lectures: 19
• Langue: Deutsch
• Poids de l'ouvrage: 9 Mo

Extrait

Molecular Analysis of the Role of Caspase-8
during Liver Regeneration and Tumorigenesis



Von der Fakultät für Mathematik, Informatik und Naturwissenschaften
der RWTH Aachen University zur Erlangung des akademischen Grades
eines Doktors der Naturwissenschaften genehmigte Dissertation

vorgelegt von

Diplom-Biologin
Julia Freimuth
Bremen


Berichter:
Privatdozent Dr. rer nat Christian Liedtke
Universitätsprofessor Dipl. Ing. Dr. Werner Baumgartner

Tag der mündlichen Prüfung:
28. Oktober 2009


Diese Dissertation ist auf den Internetseiten der Hochschulbibliothek online verfügbar.

















"Who believes to be has ceased to come to be"
- Sokrates -




















Teile dieser Arbeit wurden bereits vorab veröffentlicht oder zur Publikation
eingereicht:
1. Beraza N, Malato Y, Sander LE, Al-Masaoudi M, Freimuth J, Riethmacher D,
Gores G, Roskams T, Liedtke C, Trautwein C (2009). Hepatocyte specific
NEMO deletion promotes NK/NKT cell dependent liver damage. J Exp Med
206, 1727-1737
2. Freimuth J, Riethmacher D, Trautwein C and Liedtke C (2009): Depletion of
Caspase-8 in mice modulates TNF-induced complex formation and cell cycle
signaling in hepatocytes following partial hepatectomy. Vortrag, 44.
Jahrestagung der EASL, Kopenhagen, Dänemark
3. Liedtke C, Freimuth J, Streetz KL, Beraza N, Lambertz D, Gassler N,
Riethmacher D, and Trautwein C (2008): Depletion of caspase-8 protects from
Fas- and LPS-mediated liver injury but not from Concanavalin A induced
hepatitis in mice. Journal of Hepatology 48, S18 Vortrag, 43. Jahrestagung der
EASL, Mailand, Italien
4. Freimuth J, Riethmacher D, Trautwein C, Liedtke C (2008). Inaktivierung von
Caspase–8 beeinflusst die Proliferation von Hepatozyten nach partieller
Hepatektomie in der Maus. Z Gastroenterol 46, 112. Poster, 24.
Jahrestagung der GASL, Frankfurt.
5. Liedtke C, Freimuth J, Streetz KL, Beraza N, Lambertz D, Gaßler N,
Riethmacher D, Trautwein C (2008). Charakterisierung von hepatozyten-
spezifischen Caspase–8 knockout Mäusen in experimentellen
Leberschädigungsmodellen. Z Gastroenterol 46, 108. Poster, 24.
Jahrestagung der GASL, Frankfurt.
6. Freimuth J, Riethmacher D, Trautwein C and Liedtke C (2007). Inactivation of
caspase-8 affects hepatocyte proliferation after partial hepatectomy in mice.
Hepatology 46. Poster, 58. Jahrestagung der AASLD, Boston, MA, USA
7. Liedtke C, Freimuth J, Streetz KL, Beraza N, Lambertz D, Riethmacher D and
Trautwein C (2007). Analysis of hepatocyte-specific caspase-8 knockout mice
in experimental models of liver injury. Hepatology 46. Vortrag, 58.
Jahrestagung der AASLD, Boston, MA, USA
8. Freimuth J, Riethmacher D, Trautwein C, and Liedtke C (2007). Hepatocyte-
specific Depletion of Caspase-8 Accelerates the Onset of Liver Regeneration

in Mice. Journal of Hepatology 46, S21-S21, Vortrag, 41. Jahrestagung der
EASL, Barcelona, Spanien

Table of content I

Table of content
Table of content.......................................................................................I
1 Introduction .........................................................................................1
1.1 The liver ........................................................................................................... 1
1.1.1 Anatomy and Physiology of the Liver ......................................................... 1
1.2 Apoptosis ........................................................................................................ 2
1.2.1 The significance of apoptosis ..................................................................... 2
1.2.2 Morphological features of apoptosis........................................................... 2
1.2.3 Apoptotic signaling cascades 4
1.3 TNF-  signaling pathways mediate inflammatory or apoptotic signals..... 6
1.3.1 TNF-R1 and Tumor-Necrosis-Factor-  (TNF) ............................................ 6
1.3.1.1 Signaling complexes for cell survival or apoptosis............................... 8
1.3.1.1.1 NF- B in pro-survival signaling after TNF-induced stimulation............. 8
1.3.1.1.2 JNK mediated pathway in TNF-induced signaling ................................ 9
1.4 Caspases ......................................................................................................... 9
1.4.1 Classification of Caspases ......................................................................... 9
1.4.2 Activation and Regulation of Caspase-8................................................... 11
1.5 Regulation of Liver Regeneration................................................................ 12
1.5.1 The Cell Cycle .......................................................................................... 12
1.5.2 Liver regeneration after Partial Hepatectomy (PH)................................... 14
1.6 Hepatocellular carcinoma (HCC) ................................................................. 15
1.7 Magnetic Resonance Imaging (MRI)............................................................ 16
1.8 The Cre/ loxP technology for generating conditional knockout mice...... 18
1.8.1 Establishing of a conditional Caspase-8 knockout mouse........................ 18
2 Aim of the present study ..................................................................20
3 Material and Methods........................................................................21
3.1 Material .......................................................................................................... 21
3.1.1 Chemicals................................................................................................. 21
3.1.1.1 Radiochemicals.................................................................................. 23
3.1.2 Instruments and equipment ...................................................................... 23
3.1.3 Consumables ........................................................................................... 24
3.1.3.1. General material ............................................................................... 24 Table of content II

3.1.3.2 Material for animal experiments......................................................... 24
3.1.3.3 Material for cell culture....................................................................... 25
3.1.4 Antibodies................................................................................................. 25
3.1.4.1 Primary antibodies ............................................................................. 25
3.1.4.1.1 Primary antibodies.............................................................................. 25
3.1.4.1.2 Supershift antibodies .......................................................................... 26
3.1.4.2 Secondary antibodies ........................................................................ 26
3.1.4.2.1 Secondary antibodies HRP-labeled.................................................... 26
3.1.4.2.2 Secondary antibodies for fluorescence microscopy............................ 26
3.1.5 Enzymes................................................................................................... 27
3.1.6 Analytical chemicals, reagents and kits 27
3.2 Methods ......................................................................................................... 28
3.2.1 Animal experimental models .................................................................... 28
3.2.1.1 Housing and breeding of mice ........................................................... 28
3.2.1.2 Used mouse strains ........................................................................... 28
3.2.1.2.1 Hepatocyte-specific conditional Caspase-8 knockout mice ................ 28
3.2.1.2.2 Hepatocyte-specific conditional Caspase-8/NEMO double knockout
mice................................................................................................................... 29
tg3.2.1.2.3 Caspase-8 c-myc .............................................................................. 29
3.2.1.3 Genotyping of genetically modified mice............................................ 29
3.2.1.4 Application techniques for drugs and other substances in mice......... 29
3.2.1.4.1 Intraperitoneal (i.p.) injection .............................................................. 29
3.2.1.4.2 Intravenious (i.v.) injection.................................................................. 29
3.2.1.5 Blood and tissue sampling procedures .............................................. 30
3.2.1.5.1 Taking of blood samples..................................................................... 30
3.2.1.5.2 Extraction and sectioning of complete livers....................................... 30
3.2.1.6 Partial Hepatectomy (PH) 31
3.2.1.7 Experimental protocol for Magnetic Resonance Imaging (MRI) in mice
.................................................................................................................