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Pharmacologic characterization of opioid peptide release from chromaffin cell transplants using a brain slice superfusion method

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Abstract

A simple chamber and an inexpensive superfusion system for studying mammalian brain slices containing neural transplants is described. With this method, rat brain slices containing bovine chromaffin cell transplants can be maintained for several hours, allowing for the determination of neurochemical characteristics and pharmacologic responsiveness of the grafted cells. Using this technique, basal and nicotine-stimulated release of metenkephalin from rat periaqueductal gray slices containing bovine chromaffin cell transplants were measured. Results showed that met-enkephalin release can be increased by nicotinic stimulation in slices containing chromaffin cell, but not control implants, for at least 8 weeks postimplantation. Furthermore, this response was doserelated. These results are in good agreement with previous behavioral studies and provide corroborative evidence for the mechanism of pain reduction by the release of opioid peptides from chromaffin cell transplants in the periaqueductal gray. This study demonstrates that neurochemical and pharmacologic analyses of neural transplants using a superfused brain slice method can be a complementary approach in determining the underlying mechanisms of neural transplants in the central nervous system.

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References

  • Alger BE, Dhanjal S, Dingledine R, Garthweite J, Henderson F, King GL, Lipton P, North A, Schwartzkroin PA, Sears TA, Seagal M, Whittingham TS, Williams J (1984) Brain slice methods. In: Dingledine R (ed) Brain slices. Plenum, New York, pp 381–437

    Google Scholar 

  • Basbaum AI, Fields HL (1984) Endogenous pain control systems: brainstem spinal pathways and endorphin circuitry. Annu Rev Neurosci 7:309–338

    Article  CAS  PubMed  Google Scholar 

  • Becker JB, Freed WJ (1988) Adrenal medulla grafts enhance functional activity of the striatal dopamine system following substantia nigra lesions. Brain Res 462:401–406

    Google Scholar 

  • Black EW, Lakoski JM (1991) Maintenance of CNS tissues in vitro for subsequent pharmacologic evaluation: a simple and inexpensive superfusion chamber. J Pharmacol Methods 25:285–289

    Google Scholar 

  • Callewaert G, Johnson RG, Morad M (1991) Regulation of the secretory response in bovine chromaffin cells. Am J Physiol (Paris) [Suppl 29] 260: C851-C860

    Google Scholar 

  • Daszuta A, Kalen P, Strecker RE, Brundin P, Björklund A (1989) Serotonin neurons grafted to the adult rat hippocampus. II. 5-HT release as studied by intracerebral microdialysis. Brain Res 498:323–332

    Google Scholar 

  • Decombe R, Rivot JP, Aunis D, Abrous N, Peschanski M, Herman JP (1990) Importance of catecholamine release for the functional action of intrastriatal implants of adrenal medullary cells: pharmacological analysis and in vivo electrochemistry. Exp Neurol 107:143–153

    Google Scholar 

  • Delanoy RL, Hunter GD, Dunn GD (1982) Catecholamine metabolism in brain slices. Determination of relevant precursor pool and the effects of elevated K+. Biochem Pharmacol 31:3289–3296

    Google Scholar 

  • Dingledine R, Dodd J, Kelly JS (1980) The in vitro brain slice as a useful neurophysiological preparation for intracellular recording. J Neurosci Methods 2:323–362

    Google Scholar 

  • Doupe 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 

  • Dunwiddie T, Mueller A, Basile A (1983) The use of brain slices in central nervous pharmacology. Fed Proc 42:2891–2898

    Google Scholar 

  • Eiden LE, Giraud P, Dave JR, Hotchkiss AJ, Affolter H-U (1984) Nicotinic receptor stimulation activates enkephalin release and biosynthesis in adrenal chromaffin cells. Nature 312:661–663

    Google Scholar 

  • Haas HL, Schaerer B, Vosmansky M (1979) A simple perfusion chamber for the study of nervous tissue slices in vitro. J Neurosci Methods 1:323–325

    Google Scholar 

  • Hoffman AJ, Maxwell DM, Nichols DE (1986) A simple organ bath for electrical stimulation and superfusion of rat brain slices. J Pharmacol Methods 16:185–199

    Google Scholar 

  • Horellou P, Brundin P, Kalen P, Mallet J, Björklund A (1990) In vivo release of DOPA and dopamine from genetically engineered cells grafted to the denervated rat striatum. Neuron 5:393–402

    Google Scholar 

  • Kalen P, Cenci MA, Daszuta A, Lindvall O, Björklund A (1990a) In vivo microdialysis: a new approach for the study of functional activity of grafted monoaminergic neurons and their interaction with the host brain. Prog Brain Res 82:329–338

    Google Scholar 

  • Kalen P, Nilsson OG, Cenci MA, Rosengren E, Lindvall O, Björklund A (1990b) Intracerebral microdialysis as a tool to monitor transmitter release from grafted cholinergic and monoaminergic neurons. J Neurosci Methods 34:107–115

    Google Scholar 

  • Keppel G (1973) Design and analysis: a researcher's handbook. Prentice-Hall, New Jersey

    Google Scholar 

  • Kilpatrick DL, Howells RD, Fleminger G, Udenfriend S (1984) Denervation of rat adrenal glands markedly increases preproenkephalin mRNA. Proc Natl Acad Sci USA 81:7221–7223

    Google Scholar 

  • Leszczyszyn DJ, Jankowski JA, Viveros OH, Diliberto EJ Jr, Near JA, Wightman RM (1991) Secretion of catecholamines from individual adrenal medullary chromaffin cells. J Neurochem 56:1855–1863

    Google Scholar 

  • Lewis RV, Stern AS, Kilpatrick DL, Gerber LD, Rossier J, Stein S, Udenfriend S (1981) Marked increases in large enkephalin-containing polypeptides in the rat adrenal gland following denervation. J Neurosci 1:80–82

    Google Scholar 

  • Livett BG, Dean DM, Whelan LG, Udenfriend S, Rossier J (1981) Co-release of enkephalins and catecholamines from cultured adrenal chromaffin cells. Nature 28:317–319

    Google Scholar 

  • Nguyen TT, De Lean A (1987) Nonadrenergic modulation by clonidine of the cosecretion of catecholamines and enkephalins in adrenal chromaffin cells. Can J Physiol Pharmacol 65:823–827

    Google Scholar 

  • Nicoll RA, Alger BE (1981) A simple chamber for recording from submerged brain slices. J Neurosci Methods 4:153–156

    Google Scholar 

  • Olson L, Stromberg I, Herrera-Marschitz M, Ungerstedt U, Ebendal T (1985) Adrenal medullary tissue grafted to the dopaminedenervated rat striatum: histochemical and functional effects of additions of nerve growth factor. In: Björklund A, Stenevi U (eds) Neural grafting in the mammalian CNS. Elsevier, Amsterdam, pp 505–518

    Google Scholar 

  • Ortega JD, Sagen J, Pappas GD (1992a) Short-term immunosuppression enhances long-term survival of bovine chromaffin cell xenografts in rat CNS. Cell Transplant 1:33–41

    Google Scholar 

  • Ortega JD, Sagen J, Pappas GD (1992b) Survival and integration of bovine chromaffin cells transplanted into rat central nervous system without exogenous trophic factors. J Comp Neurol 323:13–24

    Google Scholar 

  • Pohorecky LA, Riezzi RS, Wurtman RJ (1970) Steroid induction of phenylethanolamine-N-methyl transferase in adrenomedullary explants: independence of adrenal innervation. Endocrinology 86:1466–1468

    Google Scholar 

  • Sagen J, Pappas GD (1988) Pharmacologic consequences of the vascular permeability of chromaffin cell transplants in CNS pain modulatory regions. Exp Neurol 102:290–297

    Google Scholar 

  • Sagen J, Kemmler JE (1989) Increased levels of met-enkephalin-like immunoreactivity in the spinal cord CSF of rats with adrenal medullary transplants. Brain Res 502:1–10

    Google Scholar 

  • Sagen J, Pappas GD, Perlow MJ (1987) Alterations in nociception following adrenal medullary transplants into the rat periaqueductal gray. Exp Brain Res 67:380–390

    Google Scholar 

  • Sagen J, Pappas GD, Ortega JD (1990) Host-graft relationships of isolated bovine chromaffin cells in rat periaqueductal grey. J Neurocytol 19:697–707

    Google Scholar 

  • Sagen J, Wang H, Tresco PA, Aebischer P (1993) Transplants of immunologically-isolated xenogeneic chromaffin cells provide a long-term source of pain reducing neuroactive substances. J Neurosci (in press)

  • Sagen J, Pappas GD, and Perlow MJ (1986a) Adrenal medullary tissue transplants in the rat spinal cord reduce pain sensitivity. Brain Res 384:188–194

    Google Scholar 

  • Sagen J, Pappas GD, and Pollard HB (1986b) Analgesia induced by isolated bovine chromaffin cells implanted in the rat spinal cord. Proc Natl Acad Sci USA 83:7522–7526

    Google Scholar 

  • Segal M (1981) The action of norepinephrine in the rat hippocampus: intracellular studies in the slice preparation. Brain Res 206:107–128

    Google Scholar 

  • Segal M (1987) Interactions between grafted serotonin neurons and adult host rat hippocampus. Ann NY Acad Sci 495:284–295

    Google Scholar 

  • Stine SM, Yang H-Y, Costa E (1980) Release of enkephalin-like immunoreactive material from isolated bovine chromaffin cells. Neuropharmacology 19:683–685

    Google Scholar 

  • Thiemann W, Malisch R, Reymann G (1986) A new microcirculation chamber for inexpensive long-term investigations of nervous tissue in vivo. Brain Res Bull 17:1–4

    Google Scholar 

  • Unsicker K, Tschenchne B, Tschechne D (1981) Differentiation and transdifferentiation of adrenal chromaffin cells of the guinea pig. I. Transplants to the anterior chamber of the eye. Cell Tissue Res 215:341–367

    Google Scholar 

  • Yaksh TL, Rudy TA (1978) Narcotic analgetics: CNS sites and mechanisms of action as revealed by intracerebral injection techniques. Pain 4:299–359

    Google Scholar 

  • Yamamoto C, McIlwain H (1966) Electrical activities in thin sections from the mammalian brain maintained in chemically defined media in vitro. J Neurochem 13:1333–1343

    Google Scholar 

  • Yoburn BC, Franklin SO, Calvano SE, Inturrisi CE (1987) Regulation of rat adrenal medullary enkephalins by glucocorticoids. Life Sci 40:2495–2503

    Google Scholar 

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Ortega, J.D., Sagen, J. Pharmacologic characterization of opioid peptide release from chromaffin cell transplants using a brain slice superfusion method. Exp Brain Res 95, 381–387 (1993). https://doi.org/10.1007/BF00227130

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  • DOI: https://doi.org/10.1007/BF00227130

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