1. Positional information: fields, boundaries, and gradients Development requires a dramatic increase in the amount of information contained within the organism. The "new" information is contained in the genome, and is gradually translated into cellular processes. The principal ways in which this happens is by (1) subdivision of larger fields of cells into smaller fields, and (2) specifying the "address" of each cell within the field. This is a recursive process that requires translation of gradients of gene expression into sharp boundaries, and initiation of new gradients by these boundaries

2. Specification of cell fates Positional cues (Pattern formation): Cell fate is determined by its spatial position within a morphogenetic field during a critical time period Historical cues (Cell lineage): Cell fate is determined by inherited molecules or gene expression states Both mechanisms are required for cell specification, and often act simultaneously.

3. Pattern formationY X Cell fate Cue 1 Cue 2 Morphogenetic (progenitor) field is a region or a group of cells that show no overt differentiation, but that is “destined” to give rise to a particular organ or structure. Morphogenetic fields are to some extent autonomous and integrated.

4. Primary and secondary fields Y X etc.

5. Morphogen gradients A morphogen is a (usually) secreted molecule that induces cell fate decisions in recipient cells in a concentration-dependent manner Requires: Spatially restricted production Long-range distribution (passive or active) Reception and interpretation Interpretation is context-dependent Most animal morphogens belong to a small number of well-conserved and widely distributed families

6. Morphogen gradient Source Position Concentration Positional information Positional information may include both scalar and vector components (distance and direction)

7. Gradients and cell polarity

8. Wolpert's "French flag" model Single gradient

9. Wolpert's "French flag" model Double gradient

10. Signal transduction

11. Positional information is translated into the activation and repression of target genes Responses to morphogen gradients: activation / repression of target genes; cell proliferation and growth; morphogenetic movements Responses are context-specific

12. Translating boundary into gradient hh en Hh ptc smo ci dpp ci Boundary of Engrailed expression serves as the source of Hedgehog gradient Hedgehog activates expression of a second morphogen, Dpp, which establishes a bidirectional gradient

13. Translating gradient into boundary

14. Threshold responses to the Dorsal morphogen gradient sna sog zen

15. Threshold responses to the Dpp morphogen gradient

16. (Lost in dpp / - ) hnt ush

17. msh ind vnd After the gradient: Refining position-specific cell fates

18. Cowden and Levine 2003 After the gradient: Refining position-specific cell fates

19. Short-range (contact-mediated) signaling Notch signaling

20. Sensory organ precursor lineage in Drosophila

21. Interplay of position and lineage

22. Context-dependent action of morphogen gradients Anterior-posterior Dorso-ventral Same morphogen, different targets, different responses sna sog zen

23. "Selector genes" provide the context in which positional information is interpreted Act as digital switches that “toggle” between distinct fates Can be induced by morphogens or other selector genes Form multi-layer hierarchies

24. Distal-less Dpp EGFR Wg Target genes, including other selectors and signals Signaling pathways activate selector genes… … and vice versa

25. Types of selector genes Organ Spatial region Cell / tissue type

26. HOX genes and axial patterning

27. Combinatorial specification of cell fates

28. Combinatorial control of cell fates Signal 1Signal 2Selector ASelector B Target Gene XTarget Gene Y Target Gene Z Cell fate  Cell fate  Cell fate A relatively small “toolkit” of signals and selector genes can specify a wide range of cell fates by a combinatorial mechanism

29. Control of gene expression by selector genes and signaling pathways Different signal/selector combinations define different cell fates and gene expression domains

30. Precise spatial control of cell fates Culi and Modolell 1998

31. Precise spatial control of cell fates Garcia-Garcia et al 1999

32. Precise spatial control of cell fates Renaud and Simpson 2002

33. Combinatorial control by overlapping selectors Selector A Selector B Selector C Cell fate 1 Cell fate 2 Cell fate 3 Cell fate 4 Cell fate 5

34. Axial patterning by overlapping regional determinants Multiple positions along the Proximo-Distal axis are defined by selector genes expressed in overlapping concentric domains

35. Axial patterning by overlapping regional determinants Kojima 2004

36. Progressive regionalization of morphogenetic fields

37. Combinations of regional identities establish new regional identities

38. Regulatory hierarchies in development

39. Genes and molecules that control animal development are widely conserved Signaling pathways HedgehogNotch Dpp/ TGF  Ras/ raf Wingless/ WntJak/ Stat Selector genes HOX genes eyeless/ Pax6(eye development) Distal-less(appendages) tinman(heart)