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Tubular Organs
From Uv et al, 2003
Drosophila Tracheal System
•Simple structure
•Powerful genetics
•Easy observation
Embryonic Tracheal Development
From Uv et al, 2003
I
II
III
IV
Drosophila Trachea Has Four Types of Tubes
Drosophila Trachea Has Four Types of Tubes
From Uv et al, 2003
4-5 µM 1 µM 0.5 µM
2-5 cells compose the lumen circumference
single tube-shaped cellswith AJ encircle the lumen
doughnut shaped cells with no AJ
protrusions from single cells without AJ
1 µM or 4-5 µM
Steps of Tracheal Tube Formation
• Tracheal sac formation Specification of tracheal cells (tracheal placodes) Invagination of the tracheal placode
• Branch identity determination• General branch outgrowth via FGF signaling• Branch-specific outgrowth is controlled by regional signals in addtition to
FGF signaling 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
Specification of Tracheal Cells
From Affolter and Shilo, 2000
Trh = trachealess (bHLH PAS transcription factor)Tgo=Tango (bHLH Pas transcription factor, binding partner for Trh)Vvl/Dfr = Ventral veinless-Drifter (POU domain transcription regulator)
Invagination of Tracheal Placode
EGFR = epidermal growth factor receptorSpitz=EGFR ligandRho = Rhomboid (EGF pathway activator)
From Affolter and Shilo, 2000
Model of Tracheal Cell Invagination
Trh, Vvl
Rho (EGF pathway)
Apical Actin Enrichment
Localized Apical Cell Constriction
Ordered Cell invagination
Brodu V and Casanova J 2006
Steps of Tracheal Tube Formation
• Tracheal sac formation Specification of tracheal cells (tracheal placodes) Invagination of the tracheal placode
• Branch identity determination• General branch outgrowth via FGF signaling• Branch-specific outgrowth is controlled by regional signals in addtition to
FGF signaling 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
DB = dorsal branch
DTa/p = dorsal trunk (anterior/posterior)
VB = visceral branch
SB = spiracular branch
LTa/p = lateral trunk (anterior/posterior)LTp is also called GB or ganglionic branch
Branches of the Drosophila Trachea
From Cabernard et al, 2004
From Affolter M 2002
Determination of branch identity
EGF=Epidermal growth factor : GB and DTWg= wigless: DTDpp = Decapentaplegic (Transforming growth factor β-like): DB, LT
EGF αPS1 integrin
Steps of Tracheal Tube Formation
• Tracheal sac formation Specification of tracheal cells (tracheal placodes) Invagination of the tracheal placode
• Branch identity determination• General branch outgrowth via FGF signaling• Branch-specific outgrowth is controlled by regional signals in addtition to
FGF signaling 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
FGF Signaling Directs Primary Branch Outgrowth
From Cabernard et al, 2004
bnl/FGF = blueBtl/FGFR in Trachea cells = brown
Primary Branching Requires Bnl/Btl Signaling
From Cabernard et al, 2004green = actin-GFP (WT)Red = cells lacking Btl/FGFr
Steps in Tracheal Branching Morphogenesis
Ghabrial A, Krasnow M
Model of the FGF signaling Pathway in the Control of Cell Migration in Drosophila.
Bnl: BranchlessBtl: BreathlessDof: Downstream of FGF, StumpsCsw: Corkscrew(phosphatase)HSPG: heparan sulfate proteoglycan
Petit V, Affolter M
Steps of Tracheal Tube Formation
• Tracheal sac formation Specification of tracheal cells (tracheal placodes) Invagination of the tracheal placode
• Branch identity determination• General branch outgrowth via FGF signaling• Branch-specific outgrowth is controlled by regional signals in addtition to
FGF signaling 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
Dpp Signaling is Required for Dorsal Branch Migration in Addition to FGF Signaling
Affolter M 2002
Dpp (ligand)Pnt (receptor)
From Englund et al, 2002
Branch Migration Requires Slit/Robo Signaling
blue= tracheal lumenBrown =DSRF (GB marker)
From Cabernard et al, 2004
Branch Elongation via Cell Rearrangements
Type I Type II Type IIType I
AJ Remodeling during Intercalation Process
From Ribeiro C et al 2003
Steps of Tracheal Tube Formation
• Tracheal sac formation Specification of tracheal cells (tracheal placodes) Invagination of the tracheal placode
• Branch identity determination• General branch outgrowth via FGF signaling• Branch-specific outgrowth is controlled by regional signals in addtition to
FGF signaling 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
From Uv et al, 2003
Formation of Specialized Tracheal Cells
Fusion Process
E-Cadherin Membrane
Cytoskeleton Lumen
Migration Contact Adhesion Invagination Lumen Formation
Dysfusion is Expressed in Tracheal Fusion Cells
DT
DB
LT
btl-lacZ Dys
Dysfusion is Required for Fusion Process
btl-gal4, UAS-actin-GFP btl-gal4, UAS-actin-GFP ; dys
Wild-type dys mutant
dys Misexpression Causes Inhibition of Migration
DysMab 2A12
Wild type
btl-gal4; UAS-dys
Dysfusion Regulates Downstream Targets with Various Functions
• Shotgun and CG13196 (cell adhesion)↑• members only (protein trafficking) ↑• CG15252 (unknown function) ↑• Trachealess (tracheal migration) ↓
Steps of Tracheal Tube Formation
• Tracheal sac formation Specification of tracheal cells (tracheal placodes) Invagination of the tracheal placode
• Branch identity determination• General branch outgrowth via FGF signaling• Branch-specific outgrowth is controlled by regional signals in addtition to
FGF signaling 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
From Uv et al, 2003
I
II
III
IV
VB that has ramified to form dozens of fine terminal branches on the gut
Larval Tracheal System
Ghabrial and Krasnow MA 2003
Terminal branch expansion in response to Hypoxia induced branchless
Jarecki J, Johnson E and Krasnow MA 1999 Type IV tube
Model for Patterning of Terminal Branching by Bnl
Jarecki J, Johnson E and Krasnow MA 1999
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