Strategische personeelsplanning Vergunningverlening en Handhaving door Margreet Schotman
Hans Schotman , Leena Karhinen , and Catherine Rabouille
description
Transcript of Hans Schotman , Leena Karhinen , and Catherine Rabouille
![Page 1: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/1.jpg)
dGRASP-Mediated Noncanonical Integrin Secretion
Is Required for Drosophila Epithelial Remodeling
Hans Schotman, Leena Karhinen, and Catherine Rabouille
Developmental Cell 14, 171–182, February 2008
Presented by Ian M. Parnham & Steven de Maat, supervised by Dr. Catherine Rabouille
![Page 2: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/2.jpg)
Oogenesis
• Divided in to early (stages 1–6), mid- (stages 7–10a), and late (stages 10b–14) oogenesis.
• During oogenesis, the 15 nurse cells, the oocyte, and a surrounding monolayer of somatically derived follicle cells together comprise the egg chamber
2
![Page 3: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/3.jpg)
Follicle Epithelium (Stage 10)
3
• BC: Border cells• NC: Nurse cells• CC: Centripetal
Cells• CFC: Cylindrical
Follicular Cells
• In stage 10A the Follicular cells migrate, covering the oocyte and changing there shape from cubical to columnar.
• In stage 10B follicle cells completely cover oocyte separating nurse cells from the oocyte, elongating there shape
![Page 4: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/4.jpg)
What are we going to look at?
4
Immunofluoresence (IF) Immunoelectron microscopy (IEM)
ZOC: zone of contactFigure 1
![Page 5: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/5.jpg)
Observations
• Architecture of epithelial tissue depends intercellular junctions and adhesion
• Adhesion with extracellular matrix components at the basal side is mediated by integrins and integral plasma membrane αβ heterodimers
• In the wing imaginal discs of Dropsophila larvae, where αPS1βPS(PS1) and αPS2βPS (PS2) maintain integrity of the developing wing.
5
![Page 6: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/6.jpg)
dGRASP• dGRASP has been considered bona fide
Golgi proteins.
• Involved in formation and maintenance of the Golgi
• GrhA (GRASP65/55 homologue) has been shown to mediate Golgi independent secretion of AcbA in Dictyostelium.
6
![Page 7: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/7.jpg)
Where does dGRASP localize to, during Oocyte development?
7Figure 1
![Page 8: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/8.jpg)
Aim
Investigate the role of dGRASP in follicular epithelium
surrounding the developing oocyte
8
![Page 9: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/9.jpg)
Do dGRASP binding partners also translocate towards the open ZOC during oocyte development?
9Figure 1
![Page 10: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/10.jpg)
Where does the newly localized dGRASP come from?
10Figure 2
![Page 11: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/11.jpg)
Do dgrasp RNA & dGRASP translocate to the same place?
11Figure 2
![Page 12: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/12.jpg)
Recap
• Stage 10A to 10B: formation of open ZOC
• Stage 10B dGRASP localizes to the open ZOC, almost no co-localisation with Golgi
• Stage 10B dgrasp mRNA also localizes to the open ZOC
![Page 13: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/13.jpg)
What does dGRASP do at the open ZOC?
13
Wild Type – Stage 10B
dgrasp100.2 – Stage 10B
dGRASP is important in maintaining follicular epithelium integrity
Figure 3
![Page 14: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/14.jpg)
How are integrins deposited during the progression from stage 10A -> 10B
14
Integrin αPS1 Is deposited at the open ZOC in stage 10B
Figure 3
![Page 15: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/15.jpg)
Is Integrin αPS1 also deposited at the open ZOC of dgrasp100.2 in stage 10B?
15Figure 4
![Page 16: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/16.jpg)
Is Integrin αPS1 also deposited at the open ZOC of dgrasp100.2 in stage 10B?
16Figure 4
![Page 17: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/17.jpg)
Is βPS also effected by dgrasp100.2?
17Figure 4
![Page 18: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/18.jpg)
Recap• dGRASP mutant leads to disrupted follicular
epithelium• αPS1 is deposited at the open ZOC• αPS1 deposition at the open ZOC is dependent on
dGRASP• βPS and other adherens junction proteins were
dGRASP independent
• How does dGRASP enable αPS1 deposition?
![Page 19: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/19.jpg)
Is αPS1 deposition Golgi dependent?
19Figure 6
![Page 20: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/20.jpg)
Is αPS1 deposition dSyntaxin-5 dependent?
Figure 6
![Page 21: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/21.jpg)
Conclusion
• In stage 10B dGRASP localizes to the open ZOC, almost no co-localisation with Golgi
• dGRASP mutant leads to disrupted follicular epithelium, by interfering with αPS1 deposition at the open ZOC
• dGRASP mediated αPS1 deposition is Golgi independent
21
![Page 22: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/22.jpg)
Discussion
22
Is αPS1 newly synthesized at the open ZOC?
Figure 5
![Page 23: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/23.jpg)
Discussion
23
Is αPS1 newly synthesized at the open ZOC?
Figure 5
![Page 24: Hans Schotman , Leena Karhinen , and Catherine Rabouille](https://reader036.fdocuments.net/reader036/viewer/2022070422/568165f7550346895dd91f68/html5/thumbnails/24.jpg)
The End
24