3. MDCK tubulogenesis can be divided into stages with different molecular requirements Requires ERK activation Requires MMP activity From O’Brienet al, 04

4. MDCK tubulogenesis p-EMT requires ERK activation Nuclei=blue Actin = red From O’Brien et al, 04

5. MDCK tubulogenesis redifferentiation Requires MMP activity Nuclei=blue Actin = red From O’Brien et al, 04

6. Cellular Processes involved in Tube Formation 1.Cell-cell adhesion 2.Cell migration 3.Cell shape changes 4.Lumen formation 5.Cell division 6.Cell death

7. Trachea Formation in Drosophila

8. MDCK in collagen forms a polarized cyst - how? Gp135 = apical, red b-catenin = basolateral, green From Zegers et al, 03

9. Drive for three surfaces model - epithelial cells try to establish apical, lateral, and basal domains

10. +dnRAC From Zegers et al, 03 MDCK cyst formation requires Rac for proper orientation of polarity +dnRAC +Laminin

11. Differential Regulation of Junctional Proteins During Tubulogenesis (same paper) 1.Adherens junctions (E-cadherin) move around, and new ones are made 2.ZO-1, a tight junction protein, is between apical and basolateral domains, and during extension is at cell-cell contact points 3.Desmosome components dpI/II are in large intracellular pools during tubulogenesis

12. Steps in Tracheal Branching Morphogenesis Ghabrial A, Krasnow M

13. Model of the FGF signaling Pathway in the Control of Cell Migration in Drosophila. Bnl: Branchless Btl: Breathless Dof: Downstream of FGF, Stumps Csw: Corkscrew(phosphatase) HSPG: heparan sulfate proteoglycan Petit V, Affolter M

14. Steps of Tracheal Tube Formation Tracheal sac formation  Specification of tracheal cells (tracheal placodes)  Invagination of the tracheal placode General branch outgrowth via FGF signaling Branch-specific outgrowth is controlled by regional signals and branch identity genes  Branch identity dentermination  DB migration requires Dpp signaling  GB migration requires slit-robo signaling  Tube elongation through cell intercalation Determination and differentiation of distinct cell types (fusion cell, terminal cell)  Fusion process  Terminal branching

15. Dysfusion is Required for Fusion Process btl-gal4, UAS-actin-GFPbtl-gal4, UAS-actin-GFP ; dys Wild-type dys mutant

16. Dysfusion Regulates Downstream Targets with Various Functions Shotgun and CG13196 (cell adhesion)↑ members only (protein trafficking) ↑ CG15252 (unknown function) ↑ Trachealess (tracheal migration) ↓