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Development, Vol 117, Issue 3 937-945, Copyright © 1993 by Company of Biologists

JOURNAL ARTICLES

Isoform-specific immunological detection of newt retinoic acid receptor delta 1 in normal and regenerating limbs

DS Hill, CW Ragsdale and JP Brockes
Ludwig Institute for Cancer Research, Middlesex Hospital/University College London Branch, UK.

Retinoic acid (RA) exerts a variety of effects on the regenerating urodele limb including positional respecification of the blastema. The major RA receptor expressed in the newt limb and blastema is the delta 1 isoform and, in order to detect delta 1 in this context, we have made five affinity-purified antibodies against fusion proteins and peptides from non-overlapping regions of the molecule. These antibodies have been evaluated by reaction with transfected COS-7 cells, newt limb cells in culture and newt limb tissue sections. The most informative antibodies were RP6, directed against N-terminal region A sequence, and RP8, directed against C-terminal sequence. In western blots of blastemal extracts, delta 1 protein was detected as two major bands of immunoreactivity at positions consistent with the employment of two candidate methionine initiators identified by cDNA sequencing. Staining of adult limb sections with RP6 and RP8 showed reactivity in half of the nuclei in epidermal and mesenchymal tissues, a heterogeneity that was observed with adjacent nuclei in muscle fibres. In the regenerating limb, nuclei in the blastemal mesenchyme and wound epidermis were strongly reactive, although no axial variation in expression was detected.


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J. White, M. Boffa, B Jones, and M Petkovich
A zebrafish retinoic acid receptor expressed in the regenerating caudal fin
Development, January 7, 1994; 120(7): 1861 - 1872.
[Abstract] [PDF]

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 DevelopmentHome page
L. Pecorino, D. Lo, and J. Brockes
Isoform-specific induction of a retinoid-responsive antigen after biolistic transfection of chimaeric retinoic acid/thyroid hormone receptors into a regenerating limb
Development, January 2, 1994; 120(2): 325 - 333.
[Abstract] [PDF]


© The Company of Biologists Ltd 1993