Summary
-
1.
The developmental potencies of limb regeneration blastemas of the axolotl (Ambystoma mexicanum) were tested by transplanting them to the flank or to the orbit under various experimental conditions.
-
2.
Early upper-arm blastemas transplanted singly to the damaged flank musculature or to the orbit can form forearm skeletal elements in addition to hand cartilages (digital and carpal elements).
-
3.
Fusion of the mesenchyme of several early upper-arm blastemas into one transplant leads to the formation of a single regenerate. In this case an upper-arm element may differentiate in addition to the more distal limb regions.
-
4.
In both transplantation sites single hand plates of paddle-shaped upper-arm blastemas form exclusively hand structures. Combination of mesenchyme of several hand plates, however, can result in the differentiation of forearm elements in addition to hand structures. In one case even an upper-arm element was formed.
-
5.
The single transplantation of carpal blastemas of various stages leads to the development of hand structures only. Combination of the mesenchyme of several carpal blastemas on the flank shows no improvement of regional differentiation. When, however, the same combination experiment is performed in the orbit, the carpal blastemas of the oldest stage used are able fo form forearm elements as well.
-
6.
The first conclusion that can be drawn from these results is that the early upperarm blastema possesses the information for the development of all the structures lost by amputation. Morphogenetic stump influences are not necessary for the formation of these structures.
-
7.
The second conclusion is that the limb pattern in the transplants is established autonomously, because mesenchyme of several blastemas combined in random orientation gives rise to a single, normally oriented regenerate.
-
8.
The third conclusion is that mesenchyme which is already differentiating into hand structures at the time of transplantation (hand plate of paddle-shaped upper-arm blastema, and oldest carpal blastema) upon dedifferentiation can be induced to form more proximal structures as well. Therefore this mesenchyme still possesses the information for the development of structures which it would never have formedin situ.
-
9.
In the transplants that do not regress completely digital elements are always formed. According as regression is less extensive, or the mass of transplanted mesenchyme is enlarged experimentally, carpal elements are realized next, then forearm elements, and finally also the upper-arm element. This suggests that in the normal blastema distal differentiation tendencies appear first — under the influence of the epidermis — while successively more proximal differentiation tendencies arise as the blastema increases in mass. The disto-proximal succession of differentiation tendencies would continue until definitive differentiation begins in the most proximal mesenchyme, whence it proceeds in a proximo-distal direction.
-
10.
Since transplanted mesenchyme can form more structures than are lost by amputation if its mass is increased, it is likely that during normal regeneration the level of amputation determines what will be formed by regulating the mass of the blastema, while inhibitory stump influences may prevent the development of supernumerary structures from the blastema.
Similar content being viewed by others
References
Abeloos, M., Lecamp, M.: Sur la production de formations anormales et multiples dans les membres du Triton par transplantation de régénérats. C. R. Acad. Sci. (Paris)192, 639–643 (1931).
Amprino, R.: Aspects of limb morphogenesis in the chicken. In: Organogenesis, ed. by R. L. DeHaan and H. Ursprung, p. 256–282. New York: Holt, Rinehart & Winston 1965.
Balinsky, B. I.: Das Extremitätenseitenfeld, seine Ausdehnung und Beschaffenheit. Wilhelm Roux' Arch. Entwickl.-Mech. Org.130, 704–746 (1933).
—: Über die zeitliche Verhältnisse bei der Extremitäteninduktion. Wilhelm Roux' Arch. Entwickl.-Mech. Org.136, 250–285 (1937).
Bell, E., Saunders, J. W., Zwilling, E.: Limb development in the absence of the ectodermal ridge. Nature (Lond.)184, 1736–1737 (1959).
Bischler, V.: L'influence du squelette dans la régénération et les potentialités des divers territoires du membre duTriton cristatus. Rev. suisse Zool.33, 431–560 (1926).
Bodemer, C. W.: The development of nerve-induced supernumerary limbs in the adult newt,Triturus viridescens. J. Morph.102, 555–582 (1958).
Bohn, H.: Analyse der Regenerationsfähigkeit der Insektenextremität durch Amputations- und Transplantationsversuche an Larven der afrikanischen Schabe (Blattaria). I. Mitteilung: Regenerationspotenzen. Wilhelm Roux' Arch. Entwickl.-Mech. Org.156, 49–74 (1965a).
Bohn, H.: Analyse der Regenerationsfähigkeit der Insektenextremität durch Amputations- und Transplantationsversuche an Larven der afrikanische Schabe (Blattaria). II. Mitteilung: Achsendetermination. Wilhelm Roux' Arch. Entwickl.-Mech. Org.156, 449–503 (1965b).
—: Transplantationsexperimente mit interkalarer Regeneration zum Nachweis eines sich segmental wiederholenden Gradienten im Bein vonLeucophaea (Blattaria). Verh. dtsch. zool. Ges., Suppl.30, 499–508 (1966).
Both, N. J. de: Enhancement of the self-differentiation capacity of the early limb blastema by various experimental procedures. In: Regeneration in animals and related problems, ed. by V. Kiortsis and H. A. L. Trampusch, p. 420–426. Amsterdam: North-Holland Publ. Co. 1965.
—: Transplantation immunity in the axolotl (Ambystoma mexicanum) studies by blastemal grafts. J. exp. Zool.173, 147–158 (1970).
Bovet, D.: Les territoires de régénération; leurs propriétés étudiées par la méthode de déviation du nerf. Rev. suisse Zool.37, 83–145 (1930).
Breedis, C.: Induction of accessory limbs and of sarcoma in the newt (Triturus viridescens) with carcinogenic substances. Cancer Res.12, 861–866 (1952).
Burgess, A. M. C.: The developmental potentialities of regeneration blastema cell nuclei as determined by nuclear transplantation. J. Embryol. exp. Morph.18, 27–41 (1967).
Butler, E. G., Blum, H. F.: Supernumerary limbs of urodele larvae resulting from localized ultraviolet light. Develop. Biol.7, 218–233 (1963).
Carlson, B. M.: Studies on the mechanism of implant-induced supernumerary limb formation inUrodeles. I. The histology of supernumerary limb formation in the adult newt,Triturus viridescens. J. exp. Zool.164, 227–241 (1967).
Church, R. B., McCarthy, B. J.: Ribonucleic acid synthesis in regenerating and embryonic liver. I. The synthesis of new species of RNA during regeneration of mouse liver after partial heptatectomy. J. molec. Biol.23, 459–475 (1967a).
— —: Ribonucleic acid synthesis in regenerating and embryonic liver. II. The synthesis of RNA during embryonic liver development and its relationship to regenerating liver. J. molec. Biol.23, 477–486 (1967b).
Coward, S. J.: Effects of actinomycin D on regeneration give evidence of sequential gene activation. Nature (Lond.)219, 1257–1258 (1968).
David, L.: Das Verhalten von Extremitätenregeneraten des weißen und pigmentierten Axolotl bei heteroplastischer, heterotoper und orthotoper Transplantation und sukzessiver Regeneration. Wilhelm Roux' Arch. Entwickl.-Mech. Org.126, 457–511 (1932).
Efimov, M. I.: Über den Mechanismus des Regenerationsprozesses. II. Mitteilung — Die Rolle der Haut im Prozeß der Regeneration eines Organs beim Axolotl. Z. Biol. U.R.S.S.2 214–219 (1933).
—: Kann der Entwicklungsgang junger Blastemzellen durch Übertragung auf die Amputations-Wundfläche eines anderen Organs verändert werden ? Bull. Biol. Méd. exp. U.R.S.S.6, 75–77 (1938).
Faber, J.: An experimental analysis of regional organization in the regenerating fore limb of the axolotl (Ambystoma mexicanum). Arch. Biol. (Liege)71, 1–72 (1960).
—: Autonomous morphogenetic activities of the amphibian regeneration blastema. In: Regeneration in animals and related problems, ed. by V. Kiortsis and H. A. L. Trampusch, p. 404–419. Amsterdam: North-Holland Publ. Co.1965.
Giorgi, P. de: Les potentialités des régénérats chezSalamandra maculosa. Rev. suisse Zool.31, 1–52 (1924).
—, Guyénot, E.: Les potentialités des régénérats; croissance et différenciation. C. R. Soc. Biol. (Paris)89, 488–491 (1923).
Goetinck, P. F.: Genetic aspects of skin and limb development. Curr. Top. develop. Biol.1, 253–283 (1966).
Goss, R. J.: The unification of regenerates from symmetrically duplicated fore limbs. J. exp. Zool.133, 191–210 (1956a).
—: The relation of bone to the histogenesis of cartilage in regenerating fore limbs and tails of adultTriturus viridescens. J. Morph.98, 89–116 (1956b).
Gräper, L.: Extremitätentransplantationen an Anuren. I. Mitteilung. Wilhelm Roux' Arch. Entwickl.-Mech. Org.51, 284–309 (1922a).
—: Extremitätentransplantationen an Anuren. II. Mitteilung: Reverse Transplantationen. Wilhelm Roux' Arch. Entwickl.-Mech. Org.51, 587–609 (1922b).
Gurdon, J. B.: The developmental capacity of nuclei taken from intestinal epithelium cells of feeding tadpoles. J. Embryol. exp. Morph.10, 622–640 (1962).
—: Nuclear transplantation in Amphibia and the importance of stable nuclear changes in promoting cellular differentiation. Quart. Rev. Biol.38, 54–78 (1963).
—, Graham, C. F.: Nuclear changes during cell differentiation. Sci. Progr. Oxford55, 259–277 (1967).
Gurwitsch, A.: Über den Begriff des embryonalen Feldes. Wilhelm Roux' Arch. Entwickl.-Mech. Org.51, 383–415 (1922).
Guyénot, E.: Le problème morphogénétique dans la régénération des Urodèles; détermination et potentialités des régénérats. Rev. suisse Zool.34, 127–154 (1927).
—, Dinichert-Favarger, J., Galland, M.: L'exploration du territoire de la patte anterieure du Triton (asymétrie, duplicature, orientation des régénérats). Rev. suisse Zool.55, Suppl. 2, 1–120 (1948).
—, Schotté, O. E.: Démonstration de l'existence de territoires spécifiques de régénération par la méthode de la déviation des troncs nerveux. C. R. Soc. Biol. (Paris)94, 1050–1052 (1926).
— —: Greffe de régénérat et différenciation induite. C. R. Soc. Phys. Hist. nat. Genève44, 21–23 (1927).
Harrison, R. G.: On the reversal of laterality in the limbs ofAmblystoma embryos. Anat. Rec.10, 197 (1916).
—: On relations of symmetry in transplanted limbs. J. exp. Zool.32, 1–136 (1921).
—: The effect of reversing the medio-lateral or transverse axis of the fore limb bud in the salamander embryo (Amblystoma punctatum Linn.) Wilhelm Roux' Arch. Entwickl.-Mech. Org.106, 469–502 (1925).
Hearson, L. E.: An analysis of apical proliferation in the fore limb regeneration blastema of the axolotlAmbystoma mexicanum. Ph. D. thesis: Michigan State University. East Lansing, Mich. 1966.
Holtzer, H., Avery, G., Holtzer, S.: Some properties of the regenerating limb blastema cells of salamanders. Biol. Bull.107, 313 (1954).
Jordan, M.: Development of regeneration blastemas implanted into the brain. Folia biol. (Kraków)8, 41–53 (1960).
—: Investigations of the differentiation of blastemas implanted into the brain of Amphibia. Folia biol. (Krakow)13, 205–255 (1965).
Kieny, M.: Role inducteur du mésoderme dans la différenciation précoce du bourgeon de membre chez l'embryon de poulet. J. Embryol. exp. Morph.8, 457–467 (1960).
Kiortsis, V.: Potentialités du territoire patte chez le Triton (adultes, larves, embryons). Rev. suisse Zool.60, 301–110 (1953).
—, Droin, A.: La régénération caudale des Urodèles (Induction et réactivité du territoire). J. Embryol. exp. Morph.9, 77–96 (1961).
Lender, Th.: Recherches expérimentales sur la nature et les propriétés de l'inducteur de la régénération des yeux de la planairePolycelis nigra. J. Embryol. exp. Morph.4, 196–216 (1956).
—: L'inhibition spécifique de la différenciation du cerveau des planaires d'eau douce en régénération. J. Embryol. exp. Morph.8, 291–301 (1960).
Liosner, L. D.: Untersuchungen über die Eigenschaften der Regenerationsknospe des Amphibienschwanzes. Bull. Biol. Méd. exp. U.R.S.S.6, 262–265 (1938).
—, Woronzowa, M. A., Kusmina, N. A.: Regenerationspotenz der knochenlosen Extremität. Wilhelm Roux' Arch. Entwickl.-Mech. Org.134, 738–750 (1936).
Locatelli, P.: Der Einfluß des Nervensystems auf die Regeneration. Wilhelm Roux' Arch. Entwickl.-Mech. Org.114, 686–770 (1929).
Lodyzhenskaya, V.: La transplantation des bourgeons de régénération des extrémités de l'axolotl. C. R. Acad. Sci. U.R.S.S.15, 99–101 (1928).
]Mettetal, C.: La régénération des membres chez la salamandre et le Triton. Arch. Anat. (Strasbourg)28, 1–214 (1939).
—: Influence du niveau d'amputation sur les potentialités des régénérats chez les amphibiens urodèles. C. R. Acad. Sci. (Paris)234, 1808–1810 (1952).
Michael, M. I., Faber, J.: The self-differentiation of the paddle-shaped limb regenerate, transplanted with normal and reversed proximo-distal orientation after removal of the digital plate (Ambystoma mexicanum). Arch. Biol. (Liege)72, 301–330 (1961).
Milojević, B. D.: Beiträge zur Frage über die Determination der Regenerate (Vorläufige Mitteilung). Arch. mikr. Anat.130, 80–94 (1924).
Monroy, A.: Richerche sulla rigenerazione degli arti negli Anfibi Urodeli. Nota III. Osservazioni su rigenerati formatisi su doppie superfici di sezione e considerazioni sui processi determinativi della rigenerazione. Arch. Zool. ital.31, 151–172 (1946).
Nassonov, N. V.: Morphogenesis following the insertion of parts of various organs under the skin of the axolotl. C. R. Acad. Sci. U.R.S.S.19, 127–144 (1938).
Needham, J.: Biochemistry and morphogenesis. Cambridge: University Press 1942.
Oberheim, K. W., Luther, W.: Versuche über die Extremitätenregeneration von Salamanderlarven bei umgekehrter Polarität des Amputationsstumpfes. Wilhelm Roux' Arch. Entwickl.-Mech. Org.150, 373–382 (1958).
Pietsch, P.: The effects of heterotopic musculature on myogenesis during limb regeneration inAmblystoma larvae. Anat. Rec.141, 295–304 (1961 a).
—: Differentiation in regeneration. 1. The development of muscle and cartilage following deplantation of regenerating limb blastemata ofAmblystoma larvae. Develop. Biol.3, 255–264 (1961 b).
Polezhaev, L. V.: La valeur de la structure de l'organe et les capacités du blastème régénératif dans le processus de la détermination du régénérat. Bull. biol. Fr. Belg.70, 54–85 (1936).
—: Über die Determination des Regenerats. Izv. Akad. Nauk U.S.S.R.2, 151–247 (1937).
—: The loss and restoration of regenerative capacity in the limbs of tailless Amphibia. Biol. Rev.21, 141–147 (1946).
—, Teplits, N. A., Tuchkova, S. Y.: Factors governing recovery of the ability of axolotls to regenerate limbs after its suppression by X-rays. Dokl.(Proc.)Acad.Sci.U.S.S.R., Biological Science Section150, 690–693 (1963).
Rose, S. M.: Methods of initiating limb regeneration in adult Anura. J. exp. Zool.95, 149–170 (1944).
—: Specific inhibition during differentiation. Ann. N. Y. Acad. Sci.60, 1136–1153 (1955).
—: Polarized inhibitory effects during regeneration inTubularia. J. Morph.100, 187–206 (1957).
—: Polarized control of regional structure inTabularia. Develop. Biol.7, 488–501 (1963).
Ruben, L. N.: The effects of implanting anuran cancer into non-regenerating and regenerating larval urodele limbs. J. exp. Zool.128, 29–47 (1955).
—, Frothingham, M. L.: The importance of innervation and superficial wounding in urodele accessory limb formation. J. Morph.102, 91–118 (1958).
Saunders, J. W.: The proximo-distal sequence of origin of the parts of the chick wing and the role of the ectoderm. J. exp. Zool.108, 363–403 (1948).
Schaxel, J.: Zur Determination der Regeneration der Axolotl-extremität. C. R. Acad. Sci. U.R.S.S.4, 246–248 (1934).
Schneider, G.: Der Einfluß des Nervensystems auf die Regeneration der Extremitäten der Axolotl. Zool. Jb. Abt. allg. Zool. Physiol. Tiere60, 73–102 (1942).
Schotté, O. E., Hummel, K. P.: Lens induction at the expense of regenerating tissues of amphibians. J. exp. Zool.80, 131–165 (1939).
Schwidefsky, G.: Entwicklung und Determination der Extremitätenregenerate bei den Molchen. Wilhelm Roux' Arch. Entwickl.-Mech. Org.132, 57–114 (1935).
Searls, R. L.: Development of the embryonic chick limb bud in avascular culture. Develop. Biol.17, 382–399 (1968).
Singer, M., Ray, E. K., Peadon, A. M.: Regional growth differences in the early regenerate of the adult newt,Triturus viridescens, correlated with the position of the larger nerves. Folia biol. (Kraków)12, 347–362 (1964).
Skowron, S., Walknowska, J.: The fate of regeneration blastemas implanted into the body cavity. Folia biol. (Kraków)7, 113–126 (1959).
Stocum, D. L.: The urodele limb regeneration blastema: a self-organizing system. I. Differentiationin vitro. Develop. Biol.18, 441–456 (1968a).
—: The urodele limb regeneration blastema: a self-organizing system. II. Morphogenesis and differentiation of autografted whole and fractional blastemas. Develop. Biol.18, 457–480 (1968b).
Stone, L. S.: Regeneration of the retina, iris and lens. In: Regeneration in vertebrates, ed. by C. S. Thornton, p. 3–14. Chicago: University of Chicago Press 1959.
—: Experiments dealing with the inhibition and release of lens regeneration in eyes of adult newts. J. exp. Zool.161, 83–94 (1966).
Swett, F. H.: Experiments in splitting the regenerating limb bud (Diemictylus viridescens). Anat. Rec.40, 297–309 (1928).
Tardent, P.: Developmental aspects of regeneration in coelenterates. In: Regeneration in animals and related problems, ed. by V. Kiortsis and H. A. L. Trampusch, p. 71–88. Amsterdam: North-Holland Publ. Co. 1965.
Thornton, C. S.: Influence of head skin on limb regeneration in urodele amphibians. J. exp. Zool.150, 5–15 (1962).
—, Thornton, M. T.: The regeneration of accessory limb parts following epidermal cap transplantation in Urodeles. Experientia (Basel)21, 146 (1965).
Tschumi, P. A.: The growth of the hind limb bud ofXenopus laevis and its dependence upon the epidermis. J. Anat. (Lond.)91, 149–173 (1957).
Verwoerd, C. D. A.: Het herstel van het regeneratievermogen van bestraalde extremiteiten bij axolotl-larven. M. D. Thesis: Amsterdam 1968.
Waddington, C. H.: Organisers and genes. Cambridge: University Press 1947.
Weiss, P.: Regeneration aus doppeltem Extremitätenquerschnitt (amTriton cristatus). Anz. Akad. Wiss. Wien61, 45–46 (1924).
—: Unabhängigkeit der Extremitätenregeneration vom Skelett (beiTriton cristatus). Arch. mikrosk. Anat. Entwickl.-Mech.104, 359–394 (1925).
—: Ganzregenerate aus halbem Extremitätenquerschnitt. Wilhelm Roux' Arch. Entwickl.- Mech. Org.107, 1–53 (1926).
—: Potenzprüfung am Regenerationsblastem. I. Extremitätenbildung aus Schwanzblastem im Extremitätenfeld bei Triton. Wilhelm Roux' Arch. Entwickl.-Mech. Org.111, 317–341 (1927).
—: Principles of development. New York: Holt 1939.
Wolff, E., Hampé, A.: Sur la régulation de la patte de poulet après résection d'un segment intermédiaire du bourgeon de membre. C. R. Soc. Biol. (Paris)148, 154–156 (1954).
Ziller-Sengel, C.: Recherches sur la inhibition de la régénération du pharynx chez les planaires. I. Mise en évidence d'un facteur auto-inhibiteur de la régénération du pharynx. J. Embryol. exp. Morph.18, 91–105 (1967).
Zwilling, E.: Interaction between limb bud ectoderm and mesoderm in the chick embryo. II. Experimental limb duplication. J. exp. Zool.132, 173–187 (1956).
—: Limb morphogenesis. Advanc. Morphogenes.1, 301–330 (1961).
—: Development of fragmented and of dissociated limb bud mesoderm. Develop. Biol.9, 20–37 (1964).
Author information
Authors and Affiliations
Additional information
Aided by a grant from the Netherlands Organization for the Advancement of Pure Research (Z.W.O.).
I wish to thank Prof. P. D. Nieuwkoop for the hospitality extended to me as a guest worker at the Hubrecht Laboratory. I am very grateful to Dr. J. Faber for stimulating advice and for critical reading the manuscript. Mr. P. C. Sanders, M. Sc., kindly helped with the statistical treatment of the data. Finally I thank Mrs. M. van Deventer for technical assistance, and Mr. W. Veringa for animal care.
Rights and permissions
About this article
Cite this article
de Both, N.J. The developmental potencies of the regeneration blastema of the axolotl limb. W. Roux' Archiv f. Entwicklungsmechanik 165, 242–276 (1970). https://doi.org/10.1007/BF01380787
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01380787