Cell Biology & Signaling

Goal of this 3 day workshop is to give an introduction into the crossroads of growth factor signalling and mechanobiology in physiology, pathology and regeneration. Theoretical aspects of the workshop will cover basic concepts in cell-sensation of mechanical cues, integration of extracellular matrix biology and external mechanical cues into intracellular signalling (Prof. Petra Knaus, Dr. Christian Hiepen, Dr. Maria Reichenbach) and coupling of mechanobiology with chromatin and gene transcription (Prof. Sigmar Stricker and Prof. Petra Knaus). Practical aspects will highlight technologies to apply and measure cellular forces, to investigate on the cells force sensing machinery, it’s subcellular localization (Dr. Christian Hiepen, Dr. Maria Reichenbach) and in-silico prediction of genes that underly mechano-dependent regulation (Jerome Jatzlau). The students will review and present primary research articles and review that provide state of art data and concepts in mechanobiology and signal transduction. This workshop is particularly suited for students who want to work at the translational interface of physics, bioinformatics and cell biology.

MONDAY, 30.09.2019

9:00          Welcome (P. Knaus)

9:30          General introduction into BMP/TGFβ receptors and growth factors + context in regenerative medicine (P. Knaus)

11:00        General introduction into cell mechanics and the concept of mechano-biology (C. Hiepen/M. Reichenbach)

12:30        lunch break

13:30        Literature seminar: Reading and summarizing a primary-research article/ review on topic of Mechano and ECM biology, Students will work in groups

                 of two and are allocated 1-2 state of arte article from high impact journal to highlight the focus by joint presentation

                 Focus topic #1: Technologies to investigate cell mechanics (AFM, TFM, nanopillars, optical tweezers, tension-sensors, technologies to apply

                 mechanical forces to cells)

                 Focus topic #2: Focus: Discrete cellular compartments to sense cell mechanics and how they work (Integrins, Focal adhesions, Cell-junctions,

                 primary cilia, the actin cytoskeleton as mechano-sensory structure)

                 Focus topic #3: Chromatin regulation by cell mechanics, nuclear actin cap, LINC complex, mechano-sensitive transcription factors

                 (C. Hiepen/M. Reichenbach)

15:00        coffee break

15:30        Presentation of article (15' each), 10' discussion

17:00        question-time/ closing remarks

17:30 - 18:00 END

TUESDAY, 01.10.2019

9:00          Introduction into experiment A: “Quantitative cell migration on different substrate stiffness” (C. Hiepen)

10:00        Introduction into experiment B: “Application of fluid shear stress and cytoskeletal adaptation” (M. Reichenbach)

11:00        setup life cell imaging microscopy, setup Fluid shear stress devices

12:00        start devices

12:30        lunch break

13:30        Lecture on mechanics in interstitial cells and stem cell niches (S. Stricker)

14:30        Q&A time

15:00        coffee break

15:30        short discussion on next steps

16:00        unmount devices

16:30        collect data experiment A

17:00        Fix cells experiment B

17:30        perm. block. add primary Ab O.N.

19:00        END

WEDNESDAY, 02.10.2019

9:00          wash/ add secondary Ab

10:30        Phalloidin/DAPI/ mount

12:00        microscopy samples

13:00        lunch break

14:00        Hands-on: In-silico analysis: How to investigate mechano-sensitive gene regulation (J. Jatzlau)

16:00        Discussion on experiments (C. Hiepen, M. Reichenbach, J. Jatzlau)

17:30        END