Skip to main content
SpringerLink
Log in

Immunocytochemical study of PC12 cells grafted to the brain of immature rats

  • Published:
Experimental Brain Research Aims and scope Submit manuscript

Summary

Aggregated PC12 cells were transplanted to the brain of infant Sprague Dawley (SD) rats and grown for up to two months. Histochemical and immunocytochemical methods served to identify and characterize the grafted PC12 cells. It was found that PC12 cells persist and continue to proliferate within the brain of SD rats whereas it has previously been reported that grafts placed into peripheral sites do not proliferate. Dispersed and aggregated PC12 cell grafts contained immunoreactivity to tyrosine hydroxylase and aromatic amino acid decarboxylase, but failed to express phenylethanolamine n-methyltransferase. Acetylcholinesterase activity was below histochemical detection levels in grafts of all ages. In aggregate grafts most PC12 cells were round but some small clusters of PC12 cells formed short processes. Process-forming PC12 cells were most prominent when positioned adjacent to deep layers of the cerebral cortex in one month old grafts. Hostderived capillaries and astroglia became incorporated into the PC12 aggregate grafts at successive stages. The blood brain barrier of the grafts was incomplete. This study indicates that grafted PC12 cells have some potential to restore function in brains that exhibit an endogenous dopamine deficiency, providing that mitosis in PC12 cells can be inhibited.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Algire GH, Chalkley HW (1945) Vascular reactions of normal and malignant tissues in vivo. I. Vascular reactions of mice to wounds and to normal and neoplastic transplants. J Natl Cancer Inst 6: 73–124

    Google Scholar 

  • Arendash GW, Gorski RA (1982) Enhancement of sexual behavior in female rats by neonatal transplantation of brain tissue from males. Science 217: 1276–1278

    Google Scholar 

  • Baetge EE, Kaplan BB, Reis DJ, Joh TH (1981) Translation of tyrosine hydroxylase from poly(A)-m RNA in pheochromocytoma cells is enhanced by dexamethasone. Proc Natl Acad Sci (USA) 78: 1269–1273

    Google Scholar 

  • Bernheimer H, Birkmayer W, Hornykiewicz O, Jellinger K, Seitelberger F (1973) Brain dopamine and the syndromes of Parkinson and Huntington: clinical, morphological and neurochemical correlations. J Neurol Sci 20: 415–455

    Article  CAS  PubMed  Google Scholar 

  • Bernd P, Greene LA (1983) Electron microscopic radioautographic localization of iodinated nerve growth factor bound to and internalized by PC12 cells. J Neurosci 3: 631–643

    Google Scholar 

  • Benowitz LI, Greene LA (1979) Nerve growth factor in the goldfish brain: biological assay studies using pheochromocytoma cells. Brain Res 162: 164–168

    Google Scholar 

  • Bignami A, Dahl D (1976) The astroglial response to stabbing. Immunofluorescence studies with antibodies to astrocyte-specific protein (GFA) in mammalian and submammalian vertebrates. Neuropath Appl Neurobiol 2: 99–110

    Google Scholar 

  • Björklund A, Stenevi U (1984) Intracerebral neural implants: neuronal replacement and reconstruction of damaged circuitries. Ann Rev Neurosci 7: 279–308

    Google Scholar 

  • Björklund A, Stenevi U, Dunnett SB, Iversen SD (1981) Functional reactivation of the deafferented neostriatum by nigral transplants. Nature 289: 497–499

    Google Scholar 

  • Björklund H, Dahl D (1982) Glial disturbances in isolated neocortex: evidence from immunohistochemistry of intraocular grafts. Dev Neurosci 5: 424–435

    Google Scholar 

  • Collins F (1984) An effect of nerve growth factor on the parasympathetic ciliary ganglion. J Neurosci 4: 1281–1288

    Google Scholar 

  • Cserr HF, Bundgaard M (1984) Blood-brain interfaces in vertebrates: a comparative approach. Am J Physiol 246: R277–288

    Google Scholar 

  • Dahl D, Bignami A (1976) Immunogenic properties of the glial fibrillary acidic protein. Brain Res 116: 150–157

    Google Scholar 

  • Dichter MA, Tischler AS, Greene LA (1977) Nerve growth factor induced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line. Nature 268: 501–504

    Google Scholar 

  • Dunnett SB, Björklund A, Stenevi U (1983) Dopamine-rich transplants in experimental Parkinsonism. Trends Neurosci 6: 266–270

    Google Scholar 

  • Dunnett SB, Björklund A, Stenevi U, Iversen SD (1981a) Behavioral recovery following transplantation of substantia nigra in rats subjected to 6-OHDA lesions of the nigrostriatal pathway. I. Unilateral lesions. Brain Res 215: 147–161

    Google Scholar 

  • Dunnett SB, Björklund A, Stenevi U, Iversen SD (1981b) Grafts of embryonic substantia nigra reinnervating the ventrolateral striatum ameliorate sensorimotor impairments and akinesia in rats with 6-OHDA lesions of the nigrostriatal pathway. Brain Res 229: 207–227

    Google Scholar 

  • Edgar DH, Thoenen H (1978) Selective enzyme induction in a nerve growth factor-responsive pheochromocytoma cell line (PC12). Brain Res 154: 186–190

    Google Scholar 

  • Edgar D, Barde Y-A, Thoenen H (1979) Induction of fiber outgrowth and choline acetyltransferase in PC12 pheochromocytoma cells by conditioned media from glial cells and organ extracts. Exp Cell Res 121: 353–361

    Google Scholar 

  • Edmonds-Smith M, Somera FP, Eng LF (1983) Immunocytochemical staining for glial fibrillary acidic protein and the metabolism of cytoskeletal proteins in experimental allergic encephalomyelitis. Brain Res 264: 241–253

    Google Scholar 

  • Eng LF (1980) The glial fibrillary acidic protein. In: Bradshaw RA, Schneider DM (eds) Proteins of the nervous system. Raven Press, New York, pp 85–117

    Google Scholar 

  • Folkman J, Cotran RS (1976) Relation of vascular proliferation to tumor growth. Int Rev Exp Path 16: 207–248

    Google Scholar 

  • Fontana A, Fierz W, Wekerle H (1984) Astrocytes present myelin basic protein to encephalitogenic T-cell lines. Nature 307: 273–276

    Google Scholar 

  • Fray PJ, Dunnett SB, Iversen SD, Björklund A, Stenevi U (1983) Nigral transplants reinnervating the dopamine-depleted neostriatum can sustain intracranial self-stimulation. Science 219: 416–419

    Google Scholar 

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

    Google Scholar 

  • Gash D, Sladek JR, Sladek CD (1980) Functional development of grafted vasopressin neurons. Science 210: 1367–1369

    Google Scholar 

  • Gash DM, Notter MFD, Kraus AL, Okawara SH, Lopez-Lozano JH, Wechkin SW, Joynt RJ (1984) Cholinergic neurons transplanted into septo-hippocampal lesioned and intact african green monkey's (cercopithecus aethiops). In: Björklund A (ed) Transplantation in the mammalian CNS. Proc Fernström Symposium, Elsevier/North Holland Biomed Press (in press)

  • Geneser-Jensen FA, Blackstad TW (1971) Distribution of acetylcholinesterase in the hippocampal region of the guinea pig. I. Entorhinal area, parasubiculum and presubiculum. Z Zellforsch Mikrosk Anat 114: 460–481

    Google Scholar 

  • Greene LA, Rein G (1977) Release, storage and uptake of catecholamines by a clonal cell line of nerve growth factor (NGF) responsive pheochromocytoma cells. Brain Res 129: 247–263

    Google Scholar 

  • Greene LA, Rukenstein A (1981) Regulation of acetylcholinesterase activity by nerve growth factor. J Biol Chem 256: 6363–6367

    Google Scholar 

  • Greene LA, Shooter EM (1980) The nerve growth factor: biochemistry, synthesis and mechanism of action. Ann Rev Neurosci 3: 353–402

    Google Scholar 

  • Greene LA, Tischler AS (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73: 1414–1418

    Google Scholar 

  • Greene LA, Tischler AS (1982) PC12 pheocromocytoma cultures in neurobiological research. In: Fedoroff S, Hertz L (eds) Cellular neurobiology. Academic Press, New York, pp 373–414

    Google Scholar 

  • Hatten ME, Liem RKH (1981) Astroglial cells provide a template for the positioning of developing cerebellar neurons in vitro. J Cell Biol 90: 622–630

    Google Scholar 

  • Heinicke EA, Kiernan JA (1978) Vascular permeability and axonal regeneration in skin autotransplanted into the brain. J Anat 125: 409–420

    Google Scholar 

  • Herrup K, Thoenen H (1979) Properties of the nerve growth factor receptor of a clonal line of rat pheochromocytoma (PC12) cells. Exp Cell Res 121: 71–78

    Google Scholar 

  • Jaeger CB, Lund RD (1980) Transplantation of embryonic occipital cortex to the brain of newborn rats. An autoradiographic study of transplant histogenesis. Exp Brain Res 40: 265–272

    Google Scholar 

  • Jaeger CB, Lund RD (1982) Influence of grafted glia cells and host mossy fibers on anomalously migrated host granule cells surviving in cortical transplants. Neuroscience 7: 3069–3076

    Google Scholar 

  • Jaeger CB (1985) Cytoarchitectonics of substantia nigra grafts. A light and electronmicroscopic study of immunocytochemically identified dopaminergic neurons and fibrous astrocytes. J Comp Neurol 213: 121–135

    Google Scholar 

  • Joh TH, Ross ME (1983) Preparation of catecholamine-synthesizing enzymes as immunogens for immunohistochemistry. In: Cuello AC (ed) Immunohistochemistry. IBRO Handbook Series. Methods in neuroscience, Vol 3. John Wiley & Sons, Chichester, pp 121–138

    Google Scholar 

  • Karnovsky MJ, Roots L (1964) A “direct-coloring” thiocholine method for cholinesterases. J Histochem Cytochem 12: 219–221

    Google Scholar 

  • Kordower JH, Notter MFD, Gash DM (1984) Survival of intrahippocampal transplants of cholinergic neuroblastoma into septal lesioned rats. Soc Neurosci Abst 10: 981

    Google Scholar 

  • Krieger DT, Perlow MJ, Gibson MJ, Davies TF, Zimmerman EA, Ferin M, Charlton HM (1982) Brain grafts reverse hypogonadism of gonadotropin releasing hormone deficiency. Nature 298: 468–471

    Google Scholar 

  • Levi A, Shechter Y, Neufeld EJ, Schlessinger J (1980) Mobility, clustering and transport of nerve growth factor in embryonal sensory cells and in sympathetic neuronal cell line. Proc Natl Acad Sci USA 77: 3469–3473

    Google Scholar 

  • Levitt M, Spector S, Sjoerdsma A, Udenfriend S (1965) Elucidation of the rate limiting step in norepinephrine biosynthesis in the perfused guinea pig heart. J Pharmacol Exp Ther 148: 1–8

    Google Scholar 

  • Liem R (1982) Simultaneous separation and purification of neurofilament and glial filament proteins from brain. J Neurochem 38: 142–150

    Google Scholar 

  • Lucas CA, Edgar D, Thoenen H (1979) Regulation of tyrosine hydroxylase and choline acetyltransferase activities by cell density in the PC12 rat pheochromocytoma clonal cell line. Exp Cell Res 121: 79–86

    Google Scholar 

  • Manger WM, Gifford RW Jr (1977) Pheochromocytoma. Springer, New York

    Google Scholar 

  • Mesulam MM (1976) Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents. J Histochem Cytochem 26: 106–117

    Google Scholar 

  • Olson L, Stromberg I, Herrera-Marchitz H (1984) Adrenal medullary tissue grafted to the dopamine-denervated rat striatum. In: Björklund A (ed) Transplantation in the mammalian CNS. Proc. Fernström Symposium; Elsevier/North-Holland Biomed Press (in press)

  • Perlow MJ, Freed WJ, Hoffer BJ, Seiger A, Olson L, Wyatt RJ (1979) Brain grafts reduce motor abnormalities produced by destruction of nigrostriatal dopamine system. Science 204: 643–647

    Google Scholar 

  • Pickel VM (1981) Immunocytochemical methods. Heimer L, Robards MJ (eds) Neuroanatomical tract-tracing methods. Plenum Publ Corp, New York, pp 483–509

    Google Scholar 

  • Reese TS, Karnovsky MJ (1967) Fine structural localization of a blood-barin barrier to exogenous peroxidase. J Cell Biol 34: 207–217

    Article  CAS  PubMed  Google Scholar 

  • Rosenstein JM, Brightman NW (1983) Circumventing the bloodbrain barrier with autonomic ganglion transplants. Science 221: 879–881

    Google Scholar 

  • Schubert D, Heinemann S, Kidokoro Y (1977) Cholinergic metabolism and synapse formation by a rat nerve cell line. Proc Natl Acad Sci (Wash) 74: 2579–2583

    Google Scholar 

  • Schwab ME, Otten U, Agid Y, Thoenen H (1979) Nerve growth factor (NGF) in the rat CNS: absence of specific retrograde axonal transport and tyrosine hydroxylase induction in locus coeruleus and substantia nigra. Brain Res 168: 473–483

    Google Scholar 

  • Stallcup WB (1979) Sodium and calcium fluxes in a clonal nerve cell line. J Physiol (Lond) 286: 525–540

    Google Scholar 

  • Sternberger LA (1979) Immunocytochemistry, 2nd edn. John Wiley and Sons, New York

    Google Scholar 

  • Stöckel K, Schwab M, Thoenen H (1975) Comparison between the retrograde axonal transport of nerve growth factor and tetanus toxin in motor, sensory and adrenergic neurons. Brain Res 99: 1–16

    Google Scholar 

  • Thoenen H, Barde YA (1980) Physiology of nerve growth factor. Physiol Rev 60: 1284–1335

    Google Scholar 

  • Tischler AS, Greene LA (1978) Morphologic and cytochemical properties of a clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Lab Invest 39: 77–89

    Google Scholar 

  • Tischler AS, Perlman RL, Nunnemacher GM, Morse RA, 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 Tiss Res 225: 525–542

    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 

  • Warren S, Chute R (1972) Pheochromocytoma. Cancer 29: 327–331

    Google Scholar 

  • White FP, Dutton GR, Norenberg MD (1981) Microvessels isolated from rat brain: localization of astrocyte processes by immunohistochemical techniques. J Neurochem 36: 328–332

    Google Scholar 

  • Yanker BA, Shooter EM (1979) Nerve growth factor in the nucleus: interaction with receptors on the nuclear membrane. Proc Natl Acad Sci USA 76: 1269–1273

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiology and Biophysics, New York University Medical Center, 550 First Avenue, 10016, New York, NY, USA

    C. B. Jaeger

Authors
  1. C. B. Jaeger
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by United States Public Health Service grant NS-19699

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jaeger, C.B. Immunocytochemical study of PC12 cells grafted to the brain of immature rats. Exp Brain Res 59, 615–624 (1985). https://doi.org/10.1007/BF00261353

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00261353

Key words

Search

Navigation