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Fate of intraocular chromaffin cell suspensions: role of initial nerve growth factor support

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Adrenal medullary tissue from adult rats was dissociated into cell suspensions and injected into the anterior chamber of the eye, where the cells were made to attach to the previously sympathectomized irides with the use of fibronectin. Short- and long-term survival of the chromaffin cells was examined in whole mounts of irides using Falck-Hillarp fluorescence histochemistry or indirect immunohistochemistry with antibodies against adrenaline and dopamine-β-hydroxylase (DBH). After 6 days in oculo all cells were immunoreactive for adrenaline; almost none displayed processes even if β-nerve growth factor (NGF) was given at grafting. One month after weekly intraocular injections of NGF, many cells were surrounded by nerve fiber net-works and all cells were DBH-immunoreactive. Eight months postgrafting and 7 months after the last injection of NGF almost the entire iris was reinnervated and resembled a normal, sympathetically innervated iris. Both at 1 and 8 months, chromaffin cells, ganglion cells and transitional cell forms (chromaffin cells transforming towards ganglion-like cells) were found in irides from the NGF-treated eyes. The number of ganglion cells was remarkably increased with time by NGF, while the number of chromaffin cells decreased compared to controls. A single treatment with NGF at grafting had no marked effects as examined up to 3 months; at this time there was a certain outgrowth of nerve terminals, which, however, was not as pronounced as 1 month after repeated NGF injections. In conclusion, it is shown that some cells in a chromaffin cell suspension attach to the iris, transform to ganglion cells after an induction with exogenous NGF, and reinnervate the sympathically denervated iris. Such cells remain ganglion-like in character and continue to form processes even after cessation of exogenous NGF treatment.

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References

  • Akers RM, Mosher DF, Lilien JE (1981) Promotion of neurite outgrowth by substratum-bound fibronectin. Dev Biol 86:179–188

    Google Scholar 

  • Aloe L, Levi-Montalcini R (1979) Nerve growth factor induced transformation of immature chromaffin cells in vivo into sympathetic neurons: Effect of antiserum to nerve growth factor. Proc Natl Acad Sci USA 76:1246–1250

    Google Scholar 

  • Backlund E-O, Granberg PO, Hamberger B, Knutsson E, Mårtenson A, Sedvall G, Seiger Å, Olson L (1985) Transplantation of adrenal medullary tissue to striatum in parkinsonism. First clinical trials. J Neurosurg 62:169–173

    Google Scholar 

  • Coons AH (1958) Fluroescent antibody methods. In: Danielli JF (ed) General cytochemical methods, Vol 1. Academic Press, New York, pp 399–422

    Google Scholar 

  • Costa M, Buffa R, Furness JB, Solcia E (1980) Immunohistochemical localization of polypeptides in peripheral autonomic nerves using whole mount preparations. Histochemistry 65:157–165

    Google Scholar 

  • Coupland RE (1965) The Natural History of the Chromaffin Cell. Langmans, London, pp 47–87

    Google Scholar 

  • Coupland RE (1972) The chromaffin system. In: Blaschko H, Muscholl E (eds) Handbook of Experimental Pharmacology Vol. XXXIII, Catecholamines, pp 16–45, Springer, Berlin Heidelberg New York

    Google Scholar 

  • Doupé AJ, Landis SC, Patterson PH (1985) Environmental influences in the development of neural crest derivatives: glucocorticoids, growth factors, and chromaffin cell plasticity. J Neurosci 5:2119–2142

    Google Scholar 

  • Ebendal T, Olson L, Seiger Å, Hedlund K-O (1980) Nerve growth factors in the rat iris. Nature 286:25–28

    Google Scholar 

  • Ebendal T, Olson L, Seiger Å (1983) The level of nerve growth factor as a function of innervation: A correlative radioimmunoassay and bioassay study of the rat iris. Exp Cell Res 148:311–317

    Google Scholar 

  • Ebendal T, Olson L, Seiger Å, Belew M (1984) Nerve growth factors in chick and rat tissues. In: Black IB (ed) Cellular and molecular biology of neuronal development. Plenum Press, New York, pp 231–242

    Google Scholar 

  • Falck B, Hillarp N-Å, Thieme G, Torp A (1962) Fluorescence of catecholamines and related compounds condensed with formaldehyde. J Histochem Cytochem 10:348–354

    CAS  Google Scholar 

  • Freed WJ, Morihisa JM, Spoor E, Hoffer B, Olson L, Seiger Å, Wyatt R (1981) Transplanted adrenal chromaffin cells in rat brain reduce lesion-induced rotational behaviour. Nature 292:351–352

    Google Scholar 

  • Freed WJ, Cannon-Spoor HE, Krauthamer E (1986) Intrastriatal adrenal medulla grafts in rats: Long-term survival and behavioral effects. J Neurosurg 65:664–670

    Google Scholar 

  • Greene LA, McGuire JC (1978) Induction of ornithine decarboxylase by nerve growth factor dissociated from effects on survival and neurite outgrowth. Nature 276:191–193

    Google Scholar 

  • Hedin U, Thyberg J (1987) Plasma fibronectin promotes modulation of arterial smooth muscle cells from contractile to synthetic phenotype. Differentiation 33:239–246

    Google Scholar 

  • Herrera-Marschitz M, Strömberg I, Olsson D, Olson L, Ungerstedt U (1984) Adrenal medullary implants in the dopamine-denervated rat striatum. II. Rotational behavior during the first seven hours as a function of graft amount and location and its modulation by neuroleptics. Brain Res 297:53–61

    Google Scholar 

  • Hillarp N-Å, Hökfelt B (1953) Evidence of adrenaline and noradrenaline in separate adrenal medullary cells. Acta Physiol Scand 30:55–68

    Google Scholar 

  • Johnson GD, De C Nogueira-Araujo GM (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349–350

    Article  CAS  PubMed  Google Scholar 

  • Kohn A (1902) Das chromaffine Gewebe. Z ges Anat 3, Ergebn Anat Entwickl-Gesch 12:253–348

    Google Scholar 

  • Kohn A (1903) Die Paraganglien. Arch Mikrosk Anat 62:263–365

    Google Scholar 

  • Korsching S, Auburger G, Heumann R, Scott J, Thoenen H (1985) Levels of nerve growth factor and its mRNA in the central nervous system of the rat correlate with cholinergic innervation. EMBO J 4:1389–1393

    Google Scholar 

  • Levi-Montalcini R, Aloe L (1980) Tropic, trophic, and transforming effects of nerve growth factor. Adv Biochem Psychopharmacol 25:3–15

    Google Scholar 

  • Levi-Montalcini R, Angeletti PU (1968) Nerve growth factor. Physiol Rev 48:534–569

    Google Scholar 

  • Lindvall O, Backlund E-O, Farde L, Sedvall G, Freedman R, Hoffer B, Nobin A, Seiger Å, Olson L (1987) Transplantation in Parkinson's disease: two cases of adrenal medullary grafts to the putamen. Ann Neurol 22:457–468

    Google Scholar 

  • Madrazo I, Drucker-Colin R, Diaz V, Martinez-Mata J, Torres C, Becarril JJ (1987) Open microimplantation of graft of adreneral medulla to the right caudate nucleus in two patients with intractable Parkinson's disease. N Engl J Med 316:831–834

    Google Scholar 

  • Müller TH, Unsicker K (1981) High-performance liquid chromatography with electrochemical detection as a highly efficient tool for studying catecholaminergic systems. I. Quantification of noradrenaline, adrenaline and dopamine in cultured adrenal medullary cells. J Neurosci Methods 4:39–52

    Google Scholar 

  • Olson L (1970) Fluorescence histochemical evidence for axonal growth and secretion from transplanted adrenal medullary tissue. Histochemie 22:1–7

    Google Scholar 

  • Olson L, Hamberger B, Hoffer B, Miller R, Seiger Å (1981) Nerve fiber formation by grafted adult adrenal medullary cells. In: Stjärne L, Hedqvist P, Lagercrantz H, Wennmalm Å (eds) Chemical neurotransmission 75 years. Second Nobel Conference. Academic Press, London, pp 35–48

    Google Scholar 

  • Rogers SL, Letourneau PC, Palm SL, McCarthy J, Furcht LT (1983) Neurite extension by peripheral and central nervous system neurons in response to substratum-bound fibronectin and laminin. Dev Biol 98:212–220

    Google Scholar 

  • Rovasio RA, Delouvee A, Yamada KM, Timpl R, Thiery JP (1983) Neural crest cell migration: Requirements for exogenous fibronectin and high cell density. J Cell Biol 96:462–473

    Google Scholar 

  • Ruoslahti E, Hayman EG, Piersbacher M, Engvall E (1982) Fibronectin: Purification, immunochemical properties, and biological activities. Methods Enzymol 82:803–831

    Google Scholar 

  • Seiber-Blum M, Sieber G, Yamada KM (1981) Cellular fibronectin promotes adrenergic differentiation of quail neural crest cells in vitro. Exp Cell Res 133:285–295

    Google Scholar 

  • Seiger Å, Olson L (1977) Quantitation of fiber growth in transplanted central monoamine neurons. Cell Tissue Res 179:285–316

    Google Scholar 

  • Strömberg I, Herrera-Marschitz M, Hultgren L, Ungerstedt U, Olson L (1984) Adrenal medullary implants in the dopaminedenervated rat striatum. I. Acute catecholamine levels in grafts and host caudate as determined by HPLC-electrochemistry and fluorescence histochemical image analysis. Brain Res 297:41–51

    Google Scholar 

  • Strömberg I, Ebendal T, Seiger Å, Olson L (1985a) Nerve fiber production by intraocular adrenal medullary grafts: Stimulation by nerve growth factor or sympathetic denervation of the host iris. Cell Tissue Res 241:241–249

    Google Scholar 

  • Stromberg I, Herrera-Marschitz M, Ungerstedt U, Ebendal T, Olson L (1985b) Chronic implants of chromaffin tissue into the dopamine-denervated striatum. Effects of NGF on graft survival, fiber growth and rotational behavior. Exp Brain Res 60:335–349

    Google Scholar 

  • Thoenen H, Barde Y-A (1980) Physiology of nerve growth factor. Physiol Rev 60:1284–1323

    Google Scholar 

  • Tischler AS, Greene LA (1980) Phenotypic plasticity of pheochromocytoma and normal adrenal medullary cells. In: Eränkö O et al. (eds) Histochemistry and cell biology of autonomic neurons, SIF cells, and paraneurons. Raven Press, New York, pp 61–68

    Google Scholar 

  • Tischler AS, DeLellis RA, Biales B, Nunnemacher G, Carabba V, Wolfe HJ (1980) Nerve growth factor-induced neurite outgrowth from normal human chromaffin cells. Lab Invest 43:399–409

    Google Scholar 

  • Tischler AS, Perlman RL, Nunnemacher G, Morse GM, DeLellis RA, Wolfe HJ, Sheard BE (1982) Long-term effects of dexamethasone and nerve growth factor on adrenal medullary cells cultured from young adult rats. Cell Tissue Res 225:525–542

    Google Scholar 

  • Unsicker K (1967) Über die Ganglienzellen im Nebennierenmark des Goldhamsters. Z Zellforsch 76:187–219

    Google Scholar 

  • Unsicker K, Chamley J (1977) Growth characteristics of postnatal rat adrenal medulla in culture. Cell Tissue Res 177:247–268

    Google Scholar 

  • Unsicker K, Krisch B, Otten U, Thoenen H (1978) Nerve growth factor induced fiber outgrowth from isolated rat adrenal chromaffin cells: Impairment by glucocorticoids. Proc Natl Acad Sci USA 75:3498–3502

    Google Scholar 

  • Unsicker K, Rieffert B, Ziegler W (1980) Effects of cell culture conditions, nerve growth factor, dexamethasone, and cyclic AMP on adrenal chromaffin cells in vitro. In: Eränkö O et al. (eds) Histochemical and cell biology of autonomic neurons, SIF cells, and paraneurons. Raven Press, New York, pp 51–59

    Google Scholar 

  • Unsicker K, Millar TJ, Hofmann HD (1982) Nerve growth factor requirement of postnatal rat adrenal medullary cells in vitro for survival, aggregate formation and maintenance of extended neurites. Dev Neurosci 5:412–417

    Google Scholar 

  • Unsicker K, Skaper SD, Varon S (1985) Developmental changes in the responses of rat chromaffin cells to neuronotrophic and neurite-promoting factors. Dev Biol 111:425–433

    Google Scholar 

  • Verhofstad AAJ, Coupland RE, Parker TR, Goldstein M (1985) Immunohistochemical and biochemical study on the development of the noradrenaline-and adrenaline-storing cells of the adrenal medulla of the rat. Cell Tissue Res 242:233–243

    Google Scholar 

  • Whittemore SR, Ebendal T, Lärkfors L, Olson L, Seiger Å, Strömberg I, Persson H (1986) Developmental and regional expression of β-nerve growth factor messenger RNA and protein in the rat central nervous system. Proc Natl Acad Sci USA 83:817–821

    Google Scholar 

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Authors and Affiliations

  1. Department of Histology and Neurobiology, Karolinska Institute, Stockholm, Sweden

    Ingrid Strömberg & Anna Hultgårdh-Nilsson

  2. Department of Medical Cell Genetics, Karolinska Institute, Stockholm, Sweden

    Ulf Hedin

  3. Department of Developmental Biology, Biomedical Center, Uppsala University, Uppsala, Sweden

    Ted Ebendal

Authors
  1. Ingrid Strömberg
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  2. Anna Hultgårdh-Nilsson
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  3. Ulf Hedin
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  4. Ted Ebendal
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Strömberg, I., Hultgårdh-Nilsson, A., Hedin, U. et al. Fate of intraocular chromaffin cell suspensions: role of initial nerve growth factor support. Cell Tissue Res. 254, 487–497 (1988). https://doi.org/10.1007/BF00226498

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