Summary
In order to study the cellular distribution of larval and adult hemoglobins during larval development ofPleurodeles waltlii a double specific immunofluorescent labelling technique was developed.
Rabbit antibodies specific for larval and adult hemoglobin components were prepared and conjugated with tetramethyl-rhodamine isothiocyanate for the anti-larval antibodies and fluorescein isothiocyanate for the anti-adult hemoglobin antibodies.
Both simultaneous and sequential staining with the two types of fluorescent antibodies indicated that larval and adult hemoglobins were never observed within the same erythrocyte during development. The results provide evidence that two distinct cell populations exist, one synthesizing exclusively larval hemoglobins which is progressively replaced by the other one synthesizing exclusively adult hemoglobins. It remains to be determined if these two populations arise from two distinct types of stem cells (adult and larval) or from the same stem cell type.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altèr BP, Goff SC, Efremov GD, Gravely ME, Huisman TH (1980) Globin chain electrophoresis: a new approach to the determination of the Gγ/Aγ ratio in fetal hemoglobin and to studies of globin synthesis. Br J Haematol, 44:527–534
Barker JE, Pierce JE, Kefauver BC, Nienhuis AW (1977) Hemoglobin switching in sheep and goats: Induction of hemoglobin C synthesis in cultures of sheep fetal erythroid cells. Proc Natl Acad Sci USA 74:5078–5082
Benbassat J (1974) The transition from tadpole to frog haemoglobin during natural amphibian metamorphosis. II. Immunofluorescence studies. J Cell Sci 16:143–156
Brotherton TW, Chui DHK, Gauldie J, Patterson M (1979) Hemoglobin ontogeny during normal mouse fetal development. Proc Natl Acad Sci USA, 76:2853–2857
Chapman BS, Tobin AJ (1979) Distribution of developmentally regulated hemoglobins in embryonic erythroid populations. Dev Biol 69:375–387
Cirotto C, Panara F, Geraci G (1977) Two different populations of primitive erythroid cells in the chick embryo. Dev Biol 61:384–387
Clegg JB, Naughton MA, Weatherall DJ (1966) Abnormal human haemoglobin. Separation and characterization of the α and β chains by chromatography and the determination of two new variants, Hb Chesepeak and Hb J (Bangkok). J Mol Biol 19:91–018
Dan M, Hagiwara A (1967) Detection of two types of hemoglobin (HbA and HbF) in single erythrocytes by fluorescent antibody technique. Exp Cell Res 46:596–598
Deparis P (1968) Recherches expérimentales sur l'origine des érythrocytes de l'individu adulte par la méthode des greffes embryonnaires d'ilots sanguins deiploïdes et triploïdes chezPleurodeles waltlii (Amphibiens Urodèles). Ann Embryol Morphol 1:351–359
Deparis P, Jaylet A (1975) Recherches sur l'origine des différentes lignées de cellules sanguines chez l'Amphibien UrodèlePleurodeles waltlii. J Embryol Exp Morphol 33:665–683
De Witt W (1968) Microcytic response to thyroxine administration. J Mol Biol 32:502–504
Flavin M, Blouquit Y, Rosa J (1978a) Biochemical studies of the hemoglobin switch during metamorphosis in the SalamanderPleurodeles waltlii. I. Partial characterization of the adult hemoglobin. Comp Biochem Physiol 61 B:533–537
Flavin M, Blouquit Y, Duprat AM, Rosa J (1978b) Biochemical studies of the hemoglobin switch during metamorphosis in the salamanderPleurodeles waltlii. II. Comparative studies of larval and adult hemoglobins. Comp Biochem Physiol 61 B:539–544
Flavin M, Duprat AM, Rosa J (1979) Ontogenic changes in the haemoglobins of the salamanderPleurodeles waltlii. Cell Differentiation 8:405–410
Flavin M, Duprat AM, Rosa J (1981) Effect of thyroid hormones on the switch from larval to adult hemoglobin synthesis in the SalamanderPleurodeles waltlii. Cell Different
Gallien L et Durocher M (1957) Table chronologique du développement chezPleurodeles waltlii Michah. Bull Biol Fr Belg 91:97–114
Garrick MD, Reichlin M, Mattioli M, Manning R (1973) The anemia-induced reversible switch from hemoglobin A to hemoglobin C in caprine ruminants: immunochemical evidence that both hemoglobins are found in the same cell. Dev Biol 30:1–12
Goding JW (1976) Conjugation of antibodies with fluorochromes: Modification of the standard methods. J Immunol Methods 13:215–226
Hall JG, Motulsky AG (1968) Production of foctal hemoglobin in marrow culture of human adults. Nature (Lond) 217:569–571
Ingram VM (1972) Embryonic red blood cell formation. Nature (Lond) 235:338–339
Ingram VM, Keane RW, Lindblad PC (1979) In: Stamatoyannopoulos G, Nienhuis AW (eds) “Cellular and molecular regulation of hemoglobin switching”. Grune and Stratton, New York, pp 193–204
Jurd RD, Maclean N (1970) An immunofluorescent study of the haemoglobins in metamorphosingXenopus laevis. J Embryol Exp Morphol 23:299–309
Just JJ, Schwager J, Weber R, Fey H, Pfister H (1980) Immunological analysis of hemoglobin transition during metamorphosis of normal and isogenicXenopus. Wilhelm Roux's Arch 188:75–80
Maniatis GM, Ingram VM (1971) Erythropoiesis during amphibian metamorphosis. III. Immunochemical detection of tadpole and frog hemoglobins (Rana catesbeiana) in single erythrocytes. J Cell Biol 19:290–401
Moss B, Ingram VM (1968) Hemoglin synthesis during amphibian metamorphosis. II. Synthesis of adult hemoglobin following thyroxine administration. J Mol Biol 32:493–504
Papayannopoulou Th, Brice M, Stamatoyannopoulos G (1977) Hemoglobin F synthesis in vitro: evidence for control at the level of primitive erythroid stem cells. Proc Natl Acad Sci USA 74:2923–2927
Shimizu K (1976) Identification of hemoglobin types contained in single chicken erythrocytes by fluorescent antibody technique. Dev Biol 48:317–326
Zanjani ED, McGlave PB, Bhakthavathsalam A, Stamatoyannopoulos G (1979) Sheep fetal haemopoietic cells produce adult haemoglobin when transplanted in the adult animal. Nature 280:495–496
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Flavin, M., Ton That, H., Deparis, P. et al. Hemoglobin switching in the salamanderPleurodeles waltlii . Wilhelm Roux' Archiv 191, 185–190 (1982). https://doi.org/10.1007/BF00848334
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00848334