Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1987 Oct 1; 105(4): 1917–1923.
PMCID: PMC2114645

Characterization of concentration gradients of a morphogenetically active retinoid in the chick limb bud


It has long been suggested that the generation of biological patterns depends in part on gradients of diffusible substances. In an attempt to bridge the gap between this largely theoretical concept and experimental embryology, we have examined the physiology of diffusion gradients in an actual embryonic field. In particular, we have generated in the chick wing bud concentration gradients of the morphogenetically active retinoid TTNPB, (E)-4-[2-(5,6,7,8-tetrahydro- 5,5,8,8-tetramethyl-2-naphthalenyl)-1-prope nyl] benzoic acid, a synthetic vitamin A compound. Upon local application of TTNPB the normal 234 digit pattern is duplicated in a way that correlates with the geometry of the underlying TTNPB gradient; low doses of TTNPB lead to a shallow gradient and an additional digit 2, whereas higher doses result in a steep, far-reaching gradient and patterns with additional digits 3 and 4. The experimentally measured TTNPB distribution along the anteroposterior axis, can be modeled by a local source and a dispersed sink. This model correctly predicts the site of specification of digit 2, and provides an empirical estimate of the diffusion coefficient (D) of retinoids in embryonic limb tissue. The numerical value of approximately 10(-7) cm2s-1 for D suggests that retinoids are not freely diffusible in the limb rudiment, but interact with the previously identified cellular retinoic acid binding protein. In addition, D affords an estimate of the time required to establish a diffusion gradient as 3 to 4 h. This time span is in a range compatible with the time scale of pattern specification in developing vertebrate limbs. Our studies support the view that diffusion of morphogenetic substances is a plausible mechanism of pattern formation in secondary embryonic fields.

Full Text

The Full Text of this article is available as a PDF (742K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Crick F. Diffusion in embryogenesis. Nature. 1970 Jan 31;225(5231):420–422. [PubMed]
  • Eichele G, Tickle C, Alberts BM. Microcontrolled release of biologically active compounds in chick embryos: beads of 200-microns diameter for the local release of retinoids. Anal Biochem. 1984 Nov 1;142(2):542–555. [PubMed]
  • Eichele G, Tickle C, Alberts BM. Studies on the mechanism of retinoid-induced pattern duplications in the early chick limb bud: temporal and spatial aspects. J Cell Biol. 1985 Nov;101(5 Pt 1):1913–1920. [PMC free article] [PubMed]
  • Hinchliffe JR, Garcia-Porrero JA, Gumpel-Pinot M. The role of the zone of polarizing activity in controlling the differentiation of the apical mesenchyme of the chick wing-bud: histochemical techniques in the analysis of a developmental problem. Histochem J. 1981 Jul;13(4):643–658. [PubMed]
  • Honig LS. Positional signal transmission in the developing chick limb. Nature. 1981 May 7;291(5810):72–73. [PubMed]
  • Jargiello DM, Caplan AI. The establishment of vascular-derived microenvironments in the developing chick wing. Dev Biol. 1983 Jun;97(2):364–374. [PubMed]
  • Kreis TE, Geiger B, Schlessinger J. Mobility of microinjected rhodamine actin within living chicken gizzard cells determined by fluorescence photobleaching recovery. Cell. 1982 Jul;29(3):835–845. [PubMed]
  • Kwarta RF, Jr, Kimmel CA, Kimmel GL, Slikker W., Jr Identification of the cellular retinoic acid binding protein (cRABP) within the embryonic mouse (CD-1) limb bud. Teratology. 1985 Aug;32(1):103–111. [PubMed]
  • Labarca C, Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. [PubMed]
  • Mastro AM, Babich MA, Taylor WD, Keith AD. Diffusion of a small molecule in the cytoplasm of mammalian cells. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3414–3418. [PMC free article] [PubMed]
  • Maden M, Summerbell D. Retinoic acid-binding protein in the chick limb bud: identification at developmental stages and binding affinities of various retinoids. J Embryol Exp Morphol. 1986 Sep;97:239–250. [PubMed]
  • Paine PL, Moore LC, Horowitz SB. Nuclear envelope permeability. Nature. 1975 Mar 13;254(5496):109–114. [PubMed]
  • Strickland S, Breitman TR, Frickel F, Nürrenbach A, Hädicke E, Sporn MB. Structure-activity relationships of a new series of retinoidal benzoic acid derivatives as measured by induction of differentiation of murine F9 teratocarcinoma cells and human HL-60 promyelocytic leukemia cells. Cancer Res. 1983 Nov;43(11):5268–5272. [PubMed]
  • Thaller C, Eichele G. Identification and spatial distribution of retinoids in the developing chick limb bud. Nature. 1987 Jun 18;327(6123):625–628. [PubMed]
  • Tickle C, Alberts B, Wolpert L, Lee J. Local application of retinoic acid to the limb bond mimics the action of the polarizing region. Nature. 1982 Apr 8;296(5857):564–566. [PubMed]
  • Tickle C, Lee J, Eichele G. A quantitative analysis of the effect of all-trans-retinoic acid on the pattern of chick wing development. Dev Biol. 1985 May;109(1):82–95. [PubMed]
  • Wolpert L. Positional information and the spatial pattern of cellular differentiation. J Theor Biol. 1969 Oct;25(1):1–47. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...