Summary
Distribution of prolactin has been examined in regenerating forelimbs from the newt Notophthalmus viridescens. Specific prolactin binding was demonstrated in homogenates of unamputated tissue, and of regenerating limbs at from 3 to 21 days postamputation. Labeled prolactin that was injected intraperitoneally into animals with one regenerating limb accumulated in the most distal portion of the regenerate at 7 and 14 days postamputation. Light microscopic autoradiography demonstrated that labeled prolactin was localized most heavily in the apical, outer layer of the wound epithelium. Scanning electron microscopy demonstrated that, in addition to changes in prolactin affinity following amputation, morphological changes occurred in the apical wound epithelium as well. Cell surfaces of the stump epidermis were characterized by periodic dispersion of papillae among a network of interconnecting structures 1–2 μm across. By contrast, the surfaces of cells from the area in which labeled prolactin was found to localize most intensely were characterized by lack of papillae and, depending on the stage of regeneration, a pattern of microvilli and microplicae. These morphological alterations appear to reflect functional and biochemical differences between stump epidermis and wound epithelium.
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Barkey RJ, Shani J, Amit T, Barzilai D (1979) Characterization of the specific binding of prolactin to binding sites in the seminal vesicle of the rat. J Endocrinol 80:181–189
Borgens RB (1982) What is the role of naturally produced electric current in vertebrate regeneration and healing? Int Rev Cytol 76:245–301
Borgens RB, Vanable JW, Jaffe LF (1977) Bioelectricity and regeneration: Large currents leave the stumps of regenerating newt limbs. Proc Natl Acad Sci USA 74:4528–4532
Bromley SC (1977) Accumulation of glucocorticoids in regenerating areas of limbs of the newt, Notophthalmus viridescens. J Exp Zool 201:101–108
Bromley SC, Thornton CS (1974) Effect of a highly purified growth hormone on limb regeneration in the hypophysectomized newt, Notophthalmus viridescens. J Exp Zool 190:143–154
Brown PS, Brown SC (1973) Prolactin and thyroid hormone interactions in salt and water balance in the newt Notophthalmus viridescens. Gen Comp Endocrinol 20:456–466
Bryant SV, French V, Bryant PJ (1982) Distal regeneration and symmetry. Science 212:993–1002
Carlone RL, Gonagarajoh M, Rathbone MP (1981) Bovine pituitary fibroblast growth factor has neurotrophic activity on newt limb regenerates and skeletal muscle in vitro. Exp Cell Res 132:15–21
Carr FE, Jaffe RC (1981) Solubilization and molecular weight estimation of prolactin receptors from Rana catesbeiana tadpole liver and tail fin. Endocrinology 109:943–949
Carr FE, Jacobs PJ, Jaffe RC (1981) Changes in specific prolactin binding in Rana catesbeiana tadpole tissues during metamorphosis and following prolactin and thyroid hormone treatment. Mol Cell Endocrinol 23:65–76
Catt KJ, Baukal AJ, Davies TF, Dufou ML (1979) Luteinizing hormone-releasing hormone-induced regulation of gonadotropin and prolactin receptors in rat testis. J Endocrinol 104:17–25
Connelly TG, Tassava RA, Thornton CS (1968) Limb regeneration and survival of prolactin treated hypophysectomized adult newts. J Morphol 126:365–372
Davis WL, Goodman DB, Martin JH, Matthews JL, Rasmussen H (1974) Vasopressin-induced changes in the toad urinary bladder epithelial surface. J Cell Biol 61:544–547
Dent JN (1975) Integumentary effects of prolactin in the lower vertebrates. Am Zool 15:923–935
Dent JN, Eng LA, Forbes MS (1973) Relations of prolactin and thyroid hormone to molting, skin texture and cutaneous secretion in the red-spotted newt. J Exp Zool 184:369–382
French V, Bryant PJ, Bryant SV (1976) Pattern regulation in epimorphic fields. Science 193:969–981
Furlong ST (1982) An examination of the interaction of prolactin with the regenerating limb of the newt Notophthalmus viridescens. Doctoral dissertation. Michigan State University
Furlong ST, Heidemann MH, Bromley SC (1985) Fine structure of the forelimb regenerate of the African Clawed Toad, Xenopus laevis. Anat Rec 211:444–449
Geraudie J, Singer M (1981) Scanning electron microscopy of the normal and denervated limb regenerate in the newt, Notophthalmus, including observations on embryonic limb-bud mesenchyme and blastema of fish-fin regenerates. Am J Anat 162:73–87
Greenwood FC, Hunter WM, Glover JS (1963) The preparation of I131 labeled human growth hormone of high specific activity. Biochem J 89:114
Hessler AC, Landesman R (1981 a) An investigation of the prolactin-thyroxine synergism in newt limb regeneration. J Morphol 167:103–108
Hessler AC, Landesman R (1981 b) Hormone dependent changes in the apical connective tissue during early stages of forelimb regeneration in the hypophysectomized newt Notophthalmus viridescens. J Morphol 168:297–308
Hoffman CW, Dent JN (1977a) Hormonal effects on mitotic rhythmn in the epidermis of the red-spotted newt. Gen Comp Endocrinol 32:512–521
Hoffman CW, Dent JN (1977b) Hormonal regulation of cellular proliferation in the epidermis of the red-spotted newt. Gen Comp Endocrinol 32:522–530
Humason GL (1979) Animal tissue techniques. W.H. Freeman and Co., New York
Humphrey CD, Pittman FE (1979) A simple methylene blue/azure II/basic fuchsin stain for epoxy embedded tissue sections. Stain Technol 49:9–14
Jasch LG (1980) Fine structure of the surfaces of the epithelium and mesenchyme of the newt limb regenerate separated in salt solution: 14 to 25 days. Am J Anat 158:171–191
Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 27:137A
LaSalle B (1980) Are surface potentials necessary for amphibian limb regeneration? Dev Biol 75:460–466
Leaf A (1967) Membrane effects of antidiuretic hormones. Am J Med 42:745
Liversage RA, Banks BJ (1980) Insulin levels in the serum of intact and regenerating adult newts (Notophthalmus viridescens). J Exp Zool 211:259–265
Liversage RA, Korneluk RG (1978) Serum levels of thyroid hormone during forelimb regeneration in the adult newt, Notophthalmus viridescens. J Exp Zool 206:223–227
Liversage RA, Stewart WE, McLaughlin DS (1984) In vitro studies of the influence of prolactin on tail regeneration in the adult newt Notophthalmus viridescens. Roux's Arch Dev Biol 193:379–387
Lodi G, Biciotti M, Sacerdote M (1978) Osmoregulatory activity of prolactin in the skin of the crested newt. Gen Comp Endocrinol 36:7–15
Maden M (1977) The regeneration of positional information in the amphibian limb. J Theor Biol 69:735–753
Meinhardt H (1983a) A boundary model for pattern formation in vertebrate limbs. J Embryol Exp Morphol 76:115–137
Meinhardt H (1983b) A bootstrap model for proximodistal pattern formation in vertebrate limbs. J Embryol Exp Morphol 76:139–146
Mescher AL, Loh JJ (1981) Newt forelimb regeneration blastemas in vitro: cellular response to explantation and effects of various growth-promoting substances. J Exp Zool 216:235–245
Nardi JB, Stocum DL (1983) Surface properties of regenerating limb cells: Evidence for gradation along the proximodistal axis. Differentiation 25:27–31
Norman WP, Schmidt AJ (1967) The fine structure of tissues in the amputated-regenerating limb of the adult newt, Diemictylus viridescens. J Morphol 123:271–312
Read SM, Northcote DH (1981) Minimization of variation in the response to different proteins of the Coomassie blue G-dyebinding assay for protein. Anal Biochem 116:53–64
Sakai S, Banerjee MR (1979) Glucocorticoid modulation of prolactin receptors on mammary cells of lactating mice. Biochem Biophys Acta 582:79–88
Schauble MK, Nentwig MR (1974) Temperature and prolactin as control factors in newt forelimb regeneration. J Exp Zool 187:335–344
Schotté OE (1926) Hypophysectomie et régénération chez les batraciens urodèles. C R Soc Phys Hist Nat 43:67–72
Schotté OE, Hall AB (1952) Effects of hypophysectomy upon phases of regeneration in progress (Triturus viridescens). J Exp Zool 121:521–559
Singer M (1974) Neurotrophic control of limb regeneration in the newt. Ann NY Acad Sci 228:308–322
Singer M, Saltpeter MM (1961) Regeneration in vertebrates: the role of the wound epithelium. In: Zarrow MX (ed) Growth in living systems. Basic Books, Inc., New York
Slack JMW (1982) Protein synthesis during limb regeneration in the axolotl. J Embryol Exp Morphol 70:241–260
Stocum DL (1975) Outgrowth and pattern formation during limb ontogeny and regeneration. Differentiation 3:167–182
Tassava RA (1969) Hormonal and nutritional requirements for limb regeneration and survival of adult newts. J Exp Zool 170:33–54
Tassava RA (1983) Limb regeneration to digit stages occurs in well-fed adult newts after hypophysectomy. J Exp Zool 225:433–441
Thornton CS (1965) Influence of the wound skin on blastemal cell aggregation. In: Kiortsis V (ed) Regeneration in animals and related problems. North Holland Publishers, Amsterdam
Vethamany-Globus S, Liversage RA (1973) Effects of insulin insufficiency on forelimb and tail regeneration in adult Diemictylus viridescens. J Embryol Exp Morphol 30:427–447
Vethamany-Globus S, Globus M, Tomlinson B (1978) Neural and hormonal stimulation of DNA and protein synthesis in cultured regeneration blastemata in the newt Notophthalmus viridescens. Dev Biol 65:183–192
Vethamany-Globus S, Globus M, Darch A, Milton G, Tomlinson BL (1984) In vitro effects of insulin on macromolecular events in newt limb regeneration blastemata. J Exp Zool 231:63–74
White BA (1981) Occurrence and binding affinity of prolactin receptors in amphibian tissues. Gen Comp Endocrinol 45:153–161
White BA, Nicoll CS (1979) Prolactin receptors in Rana catesheiana during development and metamorphosis. Science 204:851–853
White BA, Lebovic GS, Nicoll CS (1981) Prolactin inhibits the induction of its own renal receptors in Rana catesbeiana tadspoles. Gen Comp Endocrinol 43:30–38
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Furlong, S.T., Chancy, W.G., Heideman, M.K. et al. Increased prolactin binding and morphological changes in the wound epithelium of regenerating limbs of Notophthalmus viridescens . Cell Tissue Res. 249, 411–419 (1987). https://doi.org/10.1007/BF00215525
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DOI: https://doi.org/10.1007/BF00215525