Cell identity and positional information

1. Cell identity and positional information

3. How does a neuron find its target?

4. Snail Salivary neuron B4 Snail Salivary Neuron

5. Snail Esophageal Neuron

6. Normal B5 axonal projections C) Normal B4 axonal projections

7. Salivary gland cell Neuron B4 Normal After ET crush After regeneration

8. Axonal Regeneration Neuron B5 is not prohibited from crossing the buccal commissure, But within the ET B5 axons make The proper choices. However Neuron B4 sends axons Indiscriminantly down all ET branches. What does this tell us?

9. There are guidance signals in the ET branches, • two uniquely identified neurons respond differently to these signals. • Either The signals, or the responses to the signals, in a regenerating adult are partially similar and partially different from those in the embryo.

10. Chick embryo- Reversal of spinal cord segments Motor neurons Still reach their proper targets

11. Two big questions: How does a neuron get a proper identity? • How does an axon navigate through an embryo to find a proper target?

12. 1) • Positional information • Gene regulation • 2) • Molecular guidance signals

13. The Central Dogma

14. About 3% to 5% of the total genome is expressed in a given cell at a given time.

15. Regulatory Proteins and Transcription • Proteins called transcription factors function by binding to the promoter and to another region called the enhancer. The enhancer region may be located at a distance from the gene. These transcription factors are necessary for RNA polymerase to attach. Transcription begins when the factors at the promoter region bind with the factors at the enhancer region creating a loop in the DNA.  • In the diagram below, transcription factors are represented by the green rectangle and the red oval.

17. Hundreds of different transcription factors have been discovered; each recognizes and binds with a specific nucleotide sequence in the DNA. A specific combination of transcription factors is necessary to activate a gene. • Transcription factors are regulated by signals produced from other molecules. For example, hormones activate transcription factors and thus enable transcription. Hormones therefore activate certain genes.

18. Blastula Chordin and noggin Expressing Contains siamois (dorsalizing transcription factor) Induced by Wnt growth factor Can induce the Spemann Organizer

19. Twisted Gastrulation (chordin Cofactor) twisted Inhibits dorsaling, Induces ventralizing genes

21. Dorsalizing (neuralizing) & ventralizing (epidermalizing) pathways For epidermal development

22. Dorsalizing (neuralizing) & ventralizing (epidermalizing) pathways For epidermal development

25. In 1822 Geoffroy St. Hillaire suggested that vertebrates were just dorso ventral inversions of invertebrates and represented an inversion of polarity. • This is confirmed by molecular biology.

26. decapentaplegic Chordin Short gastrulation

27. Anterior-Posterior Polarity • There is already polarity in the egg. • Gradients of “morphogens” provide positional information and cascades of diferential activation of transcription factors in different cells.

28. Evidence for two positional information gradients in insect eggs

29. Ligate blastoderm & transplant posterior pole Ligate blastoderm Normal leafhoper embryo Ligate early embryo Hence gradients of positional information sources at each pole. Each turns on one set of genes and represses another. More segments but missing middle 2 partial embryos K. Sander 1975

30. Figure 1   Sander's ligature experiments on the embryo of the leafhopper insect Euscelis. (A) Normal embryo seen in ventral view. The black ball at the bottom represents a cluster of symbiotic bacteria that marks the posterior pole. (B) After ligating the early embryo, partial embryos form, but the head and thoracic segments are missing from both embryos. (C) When ligated later (at the blastoderm stage), more of the missing segments are formed, but the embryos still lack the most central segments. (D) When the posterior pole cytoplasm is transplanted into an embryo ligated at the blastoderm stage, a small but complete embryo forms in the anterior half, while the posterior half forms an inverted partial embryo. These results can be explained in terms of gradients at the poles of the embryo that turn on one set of structures and repress the formation of others. (After Sander, 1960, and French, 1988.)

31. The first maternal morphogen found--Bicoid • Drosophila—A maternal morphogen called Bicoid (a transcription factor) is concentrated in what will become the anterior end. It diffuses out and forms a gradient and dependent upon its concentration activates different subsets of other transcription factors in different cells that subdivides the body axis.

32. _ inhibits Inhibits --

33. Double gradient of 4 maternal RNAs for transcription factors sets up the Ant-post axis in Drosophila Anterior end: Bicoid and Hunchback Caudal and Nanos- Posterior end From Sanes,DH, Et al, 2000

34. Homeobox genes (HOX genes) have a homeodomain that codes for transcription factors

36. Human From Purves, et al, 2004

37. HOX genes have sequential overlapping expression domains From Squire, et al, 2003