spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

First published online June 28, 2004
doi: 10.1242/10.1242/dev.01222

Development 131, 3391-3399 (2004)
Published by The Company of Biologists 2004
This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Aerne, B.
Right arrow Articles by Ish-Horowicz, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Aerne, B.
Right arrow Articles by Ish-Horowicz, D.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

receptor tyrosine phosphatase {psi} is required for Delta/Notch signalling and cyclic gene expression in the presomitic mesoderm

Birgit Aerne and David Ish-Horowicz

Developmental Genetics Laboratory, Cancer Research UK, 44 Lincoln's Inn Fields, London WC2A 3PX, UK



View larger version (69K):

[in a new window]
  		Fig. 1. Domain structure of zebrafish RPTP{psi} and its expression pattern during early zebrafish development. (A) RPTP{psi} protein domains. MAM, meprin/A5/PTPµ domain; FN, fibronectin type III-like domain; Ig, immunoglobulin-like domain; PTPase, phosphatase domains. (B) RPTP{psi} expression pattern in 15h and 26h zebrafish embryos. BA, branchial arches; MHB, midbrain hindbrain boundary; OV, otic vesicle; PD, pronephric duct; R, retina; S, somites.

 


View larger version (46K):

[in a new window]
  		Fig. 2. Morpholinos RPTPmo1 and RPTPmo2 specifically block RPTP{psi} translation. (A) Morpholino target sequences. Part of the 5' UTR of the RPTP{psi} mRNA is shown illustrating the binding sites of the two morpholinos RPTPmo1 (spanning the ATG) and RPTPmo2 (binding 5' to ATG). (B) In vitro assay showing specific inhibition of RPTP{psi} translation in the presence of RPTPmo1 and RPTPmo2 morpholinos. RPTP{psi} cDNA (0.5 µg) was transcribed and translated in vitro in the presence of 35S-labeled methionine in the absence of morpholino (lane 2) or in the presence of 5-base mismatch control morpholino (5m, lane 3), unrelated morpholinos, e.g. against deltaC (DlC, lane 4) and various concentrations of RPTP{psi} morpholinos (RPTPmo1, lanes 5 and 6; RPTPmo2, lanes 7 and 8). Lane 1, in vitro transcription translation in the absence of RPTP{psi} cDNA. 35S-RPTP{psi} (arrow) was analysed by SDS-PAGE followed by autoradiography.

 


View larger version (95K):

[in a new window]
  		Fig. 3. (A) RPTPmo embryos show defects in somitogenesis and a shortening of the body axis. (B) The effect of increasing RPTPmo dose on myod expression at 26 hpf. Uninjected control, 4 ng, 6 ng and 8 ng RPTPmo/embryo.

 


View larger version (119K):

[in a new window]
  		Fig. 4. Paraxial mesoderm specification and maturation is unaffected in RPTPmo embryos. In RPTPmo embryos, the levels and pattern of expression of markers for paraxial mesoderm formation (spadetail; spt) and maturation (fgf8) appear normal.

 


View larger version (100K):

[in a new window]
  		Fig. 5. RPTPmo embryos show a defect in segment polarity. The expression patterns of markers of rostral (papC, A,D,G; mespa, B,E,H; mespb, C,F,I) and caudal (myod, deltaC; see Fig. 3B and Fig. 8) half-segment identity reveal a requirement for RPTP{psi} for the specification of anteroposterior polarity within somites. (A-C) Wild-type control, (D-F) RPTPmo embryos, (E-I) aei mutant embryos.

 


View larger version (126K):

[in a new window]
  		Fig. 8. Reduction of RPTP{psi} function affects convergent extension. RPTPmo embryos show shortening and broadening of the body axis (yellow arrows in B,E), a characteristic of convergent extension mutants. This effect is confirmed by using markers for the nascent notochord (A,D; ntl, arrowheads), the boundaries of the neuroectoderm (B,E; dlx3), the anteriormost prechordal plate hgg1 (hatching gland gene 1; B,E) and somites (C,F; deltaC). (A-C) Wild-type control embryos, (D-F) RPTPmo embryos. (G) Cycling deltaC expression in the psm of wild-type and knypek (kny) mutant embryos.

 


View larger version (87K):

[in a new window]
  		Fig. 6. Cyclic expression of deltaC, her1 and her7 in the presomitic mesoderm is disrupted in RPTPmo-injected embryos. Histogram shows number of embryos affected dependent on concentration of injected morpholino (uninjected; control, 10 ng/embryo; RPTPmo, 1-8 ng/embryo (from right to left)]. Phenotypes are classified into four groups: wild-type expression pattern (blue), partial clock arrest phenotype (red), complete clock arrest phenotype (yellow) and gastrulation defect (sky blue).

 


View larger version (68K):

[in a new window]
  		Fig. 7. RPTP{psi} acts upstream or in parallel to Delta/Notch signalling and is required for transcriptional activation of both her1 and her7. (A) Comparison of deltaC, her1 and her7 expression in RPTPmo embryos, her1 morphants, aei mutants and embryos co-injected with RPTPmo and her1mo. All embryos are stained equivalently. (B) Reduction of neither RPTP{psi} nor her1 in a aei mutant background results in an enhancement of the aei mutant phenotype, indicating a role for RPTP{psi} either upstream or in parallel to Delta/Notch signalling.

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?



© The Company of Biologists Ltd 2004