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Viability of adrenal chromaffin cells in the superior cervical ganglion of young adult and aged rats
Neuroscience Letters, 118 (1990)215-218
215
ElsevierScientificPublishers Ireland Ltd. NSL 07220
Viability of adrenal chromaffin cells in the superior cervical ganglion of young adult and aged rats J a a n a S u h o n e n , Jari K o i s t i n a h o a n d A n t t i H e r v o n e n Laboratory of Gerontology, Department of Public Health and Tampere Brain Research Center, University of Tampere, Tampere (Finland)
(Received5 April 1990;Revisedversionreceived25 June 1990;Accepted 26 June 1990) Key words: Rat; Aging; Adrenergicneuron; Transplantation; Formaldehyde-inducedfluorescence;Tyrosinehydroxylase
Adrenal medullarytissue was autotransplanted to the superior cervicalganglion (SCG) of aged (26 months old) and young adult (3 months old) rats. Four and 20 weeks after operation, the viability of the transplants was evaluated using the formaldehyde-inducedfluorescence(FIF) technique and tyrosine hydroxylase(TH) immunocytochemistry.Four weeks postgrafting, the transplant consisted of a densely-packedgroup of intensively fluorescentchromattin cells in both age groups. The cells showed strong TH immunoreactivityand some of them were elongated, but only a few displayed short processes.At 20 weeks,most of the cells were spindle shaped and sent out fluorescentprocessesand a few of them were transformed toward ganglion-like cells. The results suggest that both young and old adrenal chromaffincells are able to survive, produce neuronal processes and transform toward a neuronal phenotype in the rat superiorcervicalganglion.
Transplantation of adrenal chromaffin cells to the brain of laboratory animals has been used experimentally to replace lost aminergic neurons [1, 2] or to compensate 6-OH-dopamine- or MPTP-induced rotational behavior [1, 5, 6, 9, 14]. When fetal or adult chromaffin tissue is transplanted into the striatum of adult rat recipients whose nigrostriatal pathways have been lesioned, the surviving chromaffin cells grow out neuronal processes and continue to synthesize catecholamines [1, 2, 6, 9, 14]. For some time, Str6mberg and Ebendahl reported that aged adrenal chromaffin cells survive intraocular grafting, send out neuronal processes after sympathetic denervation, and respond to exogenous nerve growth factor (NGF) by transformation toward a neuronal phenotype [13]. Further, in vitro experiments with adrenal chromaffin cells show that these cells are capable of growing processes [10, 18-20] and show many neuronal features at electron microscopical level [18, 19]. The purpose of this study was to test the plasticity of young and old adrenal chromaffin cells in the peripheral neuronal tissue o f their own host animals. A total of 22 male Wistar rats were used. The animal colony was kept on - 2 0 % caloric restriction to postpone the age-related diseases, and maintain good health during the third year o f rat life span. Ninety % of the aniCorrespondence: J. Suhonen, Laboratory of Gerontology,Department of Public Health, University of Tampere, P.O. Box 607, SF-33101 Tampere, Finland.
0304-3940/90/$03.50 © 1990ElsevierScientificPublishers Ireland Ltd.
mals die before the age of 33 months. The rats were anesthetized with sodium pentobarbital (Mebunat, Orion, Finland; 30 mg/kg, i.p.) and given atropine (Atropine, Orion; 0.04 mg/kg, s.c.). The left adrenal gland was dissected out and placed in sterile minimal essential medium (Gibco, Grand Island, NY). Then the left superior cervical ganglion (SCG) was exposed and a small cavity was punctured through a capsule with a 22-gauge needle. A piece of adrenal medullary tissue was then placed into the cavity using microforceps. The right-side SCG served as control tissue. Four aged (26 months old) and 8 young (3 months old) rats were used at 4 weeks and 3 aged and 7 young rats at 20 weeks postoperation. The SCGs were dissected and immersed in liquid nitrogen. The specimens were processed for formaldehydeinduced fluorescence (FIF) microscopy as described earlier [4]. Briefly, the tissues were freeze-dried ( - 4 0 ° C ) under vacuum (1.3 x 10 -3 Pa) for 7 days, exposed to formaldehyde vapor (relative humidity 60%) at +60°C for 60 min and embedded in vacuo. The specimens were sectioned serially at 10/tm and then coverslipped using liquid paraffin. The sections were viewed and photographed using an Olympus Vanox T fluorescence microscope. The same sections used to demonstrate FIF were subsequently processed for tyrosine hydroxylase (TH) peroxidase--antiperoxidase (PAP) immunocytochemistry in order to correlate catecholamines with the enzyme expression. The sections were incubated with anti-TH (diluted 1:100, Eugene. Tech. Int., Allendale,
216
N J) for 12-24 h and processed according to Sternberger [12]. In the control sections the specific antiserum was omitted. Control SCG and adrenal medullary tissue from young adult and aged rats are shown in Figs. 1 and 2. Sympathetic ganglion cells showed FIF and TH immunoreactivity of varying intensity in both age groups and the amount of yellow fluorescent lipopigments was increased with age. There were also scattered groups of small intensely fluorescent (SIF) cells and a network of varicose fibers in the ganglia. Fluorescence and TH immunocytochemistry did not reveal clear age-related changes in the adrenal medulla, although there appeared to be a large volume of it in aged animals. Four weeks postgrafting, fluorescence microscopy revealed a group of intensively fluorescent chromaffin cells in the recipient SCG (Figs. 3 and 4) of both young and old rats. In some cases the ganglion cells around the transplant had disappeared due to the lesion caused by the operation procedure. The packing density and the intensity of TH immunoreactivity of the cells appeared to be unchanged compared with the control adrenal medullary cells. Some grafted cells, especially in the periphery of the transplant, were elongated and sent out short and thin processes. At 20 weeks following the operation, the packing density of the grafted chromaffin cells appeared to be decreased, most of the cells were elongated, and some cells in the periphery of the transplant were enlarged resembling small neurons (Fig. 5). Neuronal processes were seen more frequently than at four weeks, and when the transplant was found within or close to the ganglion capsule, a dense network of fluorescent nerve fibers was seen to project out from the grafted cells (Figs. 5 and 6). Most
of the grafted cells continued to show strong TH immunoreactivity (Fig. 7), whereas in some enlarged cells TH immunoreactivity was reduced. There was no difference in viability and morphology of the graft between young and aged animals. The characteristics of SIF cells in the adult SCG differ from the phenotype of adrenal chromaffin cells [3, 7, 11]. The catecholamine storing cells within the SCG show more neuronal features [3, 17]. The grafted adrenal chromaffin cells could be distinguished from the original 'SIF' cells by morphology and location of the graft. We have shown that young and old adrenal medullary cells survive and develop neuronal features when transplanted into the SCG. Transplantation of adult adrenal medullary tissue to the brain or anterior chamber of the eye causes extensive growth of nerve fibers by the graft, if the normal adrenergic innervation is destroyed before grafting [1, 5, 6] or if the transplant viability is supported with exogenous NGF [14-16]. In the present study, grafted adrenal medullary cells grew out neurites in the SCG 20 weeks after transplantation without NGF treatment or without previously destroyed pre- or postganglionic innervation. It is possible that the lesion in the ganglion caused by the transplantation procedure itself may lead to a release of NGF from Schwann cells [8]. At 20 weeks after transplantation, the cells in the periphery of the adrenal medullary tissue were more elongated and more frequently enlarged than the cells in the center, suggesting further that the transformation inducing effect originated from surrounding non-neuronal ganglion tissue rather than being mediated via blood vessels grown throughout the graft. Strrmberg and Ebendal [13] showed that both aged and young adult adrenal medullary tissue survive five
Fig. 1. Fluorescence micrographs of con)rol sympathetic ganglion cells from young (a) (3 months old) and old (b) (26 months old) rats. * Pigmented ganglion cells. 216 x . Fig. 2. Fluorescence (a) and TH-immunohistoehemical (b) micrographs of adrenal chromaffln cells from an old (26 months old) rat. Both catecholamine fluorescence and TH immunoreactivity are rather homogeneous. 436 x . Fig. 3. Four weeks postgrafting the transplant (arrows) contains a group of intensively fluorescent chromattin cells (a,b). TH immunoreactivity (c) of the transplanted cells (arrows) is unchanged compared with the control adrenal medullary cells. A young animal, *Ganglion cells, a,c, 108 x ; b, 436 ×. Fig. 4. Four weeks postgrafting the transplant (arrows) contains a group of intensively fluorescent chromaflin cells in an old animal. * Ganglion cells. 436 x Fig. 5 and 6. Twenty weeks after transplantation most of the grafted cells are elongated and some of the cells resemble small neurons (*). Fluorescent nerve fibers (arrow) are seen to project out from chromaffin ceils. An old animal (5); a young animal (6). 216 x . Fig. 7. TH immunoreactivity of young adrenal chromaflin cells 20 weeks postgrafting. Most of the cells are elongated and have an enlarged cytoplasmic area compared to controls. 216 ×.
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218 weeks in the d e n e r v a t e d a n t e r i o r c h a m b e r o f the y o u n g r a t eye, a n d f o r m nerve fibers in response to N G F . The present findings extend the viability o f aged a d r e n a l chromaffin cells in the b r a i n a n d eye to their survival in the p e r i p h e r a l ganglia, in which b o t h y o u n g a n d aged grafted chromaffin cells were able to p r o d u c e c a t e c h o l a mines a n d send o u t neurites even at 20 weeks after operation. T h e grafting was c a r r i e d o u t as an a u t o t r a n s p l a n tation. Since no evident difference in the survival o r t r a n s f o r m a t i o n between y o u n g a n d aged c h r o m a f l i n cell t r a n s p l a n t s was seen, it is i n d i c a t e d t h a t r a t a d r e n a l chromaffin cells m a i n t a i n their plasticity to the o l d age. A l t h o u g h several degenerative a n d c o m p e n s a t o r y changes t a k e place in the aging s u p e r i o r cervical ganglion, the aged s y m p a t h e t i c g a n g l i o n still p r o v i d e s the elements required for the viability o f t r a n s p l a n t e d old chromaffin cells. 1 Bj6rklund, A. and Stenevi, U., Reconstruction of the nigrostriatal dopamine pathway by intraeerebral nigral transplants, Brain Res., 177 (1979) 555-560. 2 Bohn, M.C., Cupit, L., Mareiano, F. and Gash, D.M., Adrenal medulla grafts enhance recovery of striatal dopaminerglc fibers, Science, 237 (1987) 913-915. 3 Elfvin, L.-G., H6gfelt, T. and Goldstein, M., Fluorescence microscopical, immunohistoehemical and ultrastructural studies on sympathetic ganglia of guinea-pig with special reference to the S.I.F. cells and their catecholamine content, J. Ultrastruct. Res., 51 (1975) 377-396. 4 Erfink6, O., The practical histochemical demonstration of catecholamines by formaldehyde-induced fluorescence, J.R. Microsc. Soc., 87 (1967) 259 276. 5 Freed, W.J., Morihisa, J.M., Spoor, E., Hoffer, B., Olson, L., Seiger, .~. and Wyatt, R., Transplanted adrenal chromattin cells in rat brain reduce lesion-induced rotational behavior, Nature, 292 (1981) 351 352. 6 Hansen, J., Bing, G., Notter, M. and Gash, D.M., Paraneuronal grafts in unilateral 6-hydroxydopamine lesioned rats: morphological aspects of adrenal chromattin and carotid body glomus cell implants, Prog. Brain Res., 78 (1988) 507-511. 7 Helen, P., Alho, H. and Hervonen, H., Ultrastructure and histochemistry of human SIF cells and paraganglia, Adv. Biochem. Psychopharmacol., 25 (1980) 145-152.
8 Heumann, R., Korsching, S., Bandtlow, C. and Thoenen, H., Changes of nerve growth factor synthesis in nonneuronal cells in response to sciatic nerve transsection, J. Cell. Biol., 104 (1987) 1623-1631. 9 Kamo, I., Kim, S., McGeer, P. and Shin, D.H., Transplantation of cultured fetal human adrenal chromafl]n cells to rat brain, Neurosci. Lett., 57 (1985) 43--48. I00lson, L., Seiger, .~., Ebendal, T. and Hoffer, B., Comparisons of nerve fiber growth from three major catecholamine producing cell systems; adrenal medulla, superior cervical ganglion and locus coerulens, Adv. Biochem. Psyehopharmacol., 25 (1980) 27-34. 11 Partanen, M., Hervonen, A. and Santer, R.M., Effect of aging on SIF cells of the rat. Adv. Biochem. Psychopharmacol., 25 (1980) 143-147. 12 Sternberger, L.A., Immunohistoehemistry, 2nd edn. Wiley, New York, 1979. 13 Str6mberg, I. and Ebendahl, T., Aged adrenal medullary tissue survives intraocular grafting, forms nerve fibers and responds to the nerve growth factor, J. Neurosei. Res., 23 (1989) 162-171. 14 Str6mberg, I., Herrera-Marsehitz, M., Ungerstedt, U., Ebendahl, T. and Olson, L., Chronic implants of ehromatfin tissue into the dopamine-denervated striatum. Effects of NGF on graft survival, fiber growth and rotational behavior, Exp. Brain Res., 60 (1985) 335-349. 15 Str6mberg, I., Hultgardh-Nilsson, A., Hedin, U. and Ebendal, T., Fate of intraocular chromaftin cell suspensions, role of initial nerve growth factor support, Cell Tissue Res., 254 (1988) 487-497. 16 Str6mberg, I. Ebendal, T., Seiger,/~,. and Olson, L., Nerve fiber production by intraocular adrenal medullary grafts: Stimulation by nerve growth factor or sympathetic denervation of the host iris, Cell Tissue Res., 241 (1985) 241-249. 17 Taxi, J., Derer, M. and Domich, A., Morphology and histophysiology of SIF cells in the autonomic ganglia. In L.-G. Elfvin (Ed.), Autonomic Ganglia, Wiley, New York, 1983, pp. 67 95. 18 Tischler, A.S., DeLellis, R.A., Biales, B., Nunnemacher, G., Carabba, V. and Wolfe, H.J., Nerve growth factor-induced neurite outgrowth from normal human chromaifin ceils, Lab. Invest., 43 (1980) 399-409. 19 Tischler, A.S. and Greene, L.A., Phenotypic plasticity of pheochromocytoma and normal adrenal medullary cells, Adv. Biochem. Psychopharmacol., 25 (1980) 61-68. 20 Unsicker, K., Cell and tissue culture studies on the sympathoadrenal system. In L.C. Elfvin (Ed.), Autonomic Ganglia, Wiley, New York, 1983, pp. 475-505.
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ElsevierScientificPublishers Ireland Ltd. NSL 07220
Viability of adrenal chromaffin cells in the superior cervical ganglion of young adult and aged rats J a a n a S u h o n e n , Jari K o i s t i n a h o a n d A n t t i H e r v o n e n Laboratory of Gerontology, Department of Public Health and Tampere Brain Research Center, University of Tampere, Tampere (Finland)
(Received5 April 1990;Revisedversionreceived25 June 1990;Accepted 26 June 1990) Key words: Rat; Aging; Adrenergicneuron; Transplantation; Formaldehyde-inducedfluorescence;Tyrosinehydroxylase
Adrenal medullarytissue was autotransplanted to the superior cervicalganglion (SCG) of aged (26 months old) and young adult (3 months old) rats. Four and 20 weeks after operation, the viability of the transplants was evaluated using the formaldehyde-inducedfluorescence(FIF) technique and tyrosine hydroxylase(TH) immunocytochemistry.Four weeks postgrafting, the transplant consisted of a densely-packedgroup of intensively fluorescentchromattin cells in both age groups. The cells showed strong TH immunoreactivityand some of them were elongated, but only a few displayed short processes.At 20 weeks,most of the cells were spindle shaped and sent out fluorescentprocessesand a few of them were transformed toward ganglion-like cells. The results suggest that both young and old adrenal chromaffincells are able to survive, produce neuronal processes and transform toward a neuronal phenotype in the rat superiorcervicalganglion.
Transplantation of adrenal chromaffin cells to the brain of laboratory animals has been used experimentally to replace lost aminergic neurons [1, 2] or to compensate 6-OH-dopamine- or MPTP-induced rotational behavior [1, 5, 6, 9, 14]. When fetal or adult chromaffin tissue is transplanted into the striatum of adult rat recipients whose nigrostriatal pathways have been lesioned, the surviving chromaffin cells grow out neuronal processes and continue to synthesize catecholamines [1, 2, 6, 9, 14]. For some time, Str6mberg and Ebendahl reported that aged adrenal chromaffin cells survive intraocular grafting, send out neuronal processes after sympathetic denervation, and respond to exogenous nerve growth factor (NGF) by transformation toward a neuronal phenotype [13]. Further, in vitro experiments with adrenal chromaffin cells show that these cells are capable of growing processes [10, 18-20] and show many neuronal features at electron microscopical level [18, 19]. The purpose of this study was to test the plasticity of young and old adrenal chromaffin cells in the peripheral neuronal tissue o f their own host animals. A total of 22 male Wistar rats were used. The animal colony was kept on - 2 0 % caloric restriction to postpone the age-related diseases, and maintain good health during the third year o f rat life span. Ninety % of the aniCorrespondence: J. Suhonen, Laboratory of Gerontology,Department of Public Health, University of Tampere, P.O. Box 607, SF-33101 Tampere, Finland.
0304-3940/90/$03.50 © 1990ElsevierScientificPublishers Ireland Ltd.
mals die before the age of 33 months. The rats were anesthetized with sodium pentobarbital (Mebunat, Orion, Finland; 30 mg/kg, i.p.) and given atropine (Atropine, Orion; 0.04 mg/kg, s.c.). The left adrenal gland was dissected out and placed in sterile minimal essential medium (Gibco, Grand Island, NY). Then the left superior cervical ganglion (SCG) was exposed and a small cavity was punctured through a capsule with a 22-gauge needle. A piece of adrenal medullary tissue was then placed into the cavity using microforceps. The right-side SCG served as control tissue. Four aged (26 months old) and 8 young (3 months old) rats were used at 4 weeks and 3 aged and 7 young rats at 20 weeks postoperation. The SCGs were dissected and immersed in liquid nitrogen. The specimens were processed for formaldehydeinduced fluorescence (FIF) microscopy as described earlier [4]. Briefly, the tissues were freeze-dried ( - 4 0 ° C ) under vacuum (1.3 x 10 -3 Pa) for 7 days, exposed to formaldehyde vapor (relative humidity 60%) at +60°C for 60 min and embedded in vacuo. The specimens were sectioned serially at 10/tm and then coverslipped using liquid paraffin. The sections were viewed and photographed using an Olympus Vanox T fluorescence microscope. The same sections used to demonstrate FIF were subsequently processed for tyrosine hydroxylase (TH) peroxidase--antiperoxidase (PAP) immunocytochemistry in order to correlate catecholamines with the enzyme expression. The sections were incubated with anti-TH (diluted 1:100, Eugene. Tech. Int., Allendale,
216
N J) for 12-24 h and processed according to Sternberger [12]. In the control sections the specific antiserum was omitted. Control SCG and adrenal medullary tissue from young adult and aged rats are shown in Figs. 1 and 2. Sympathetic ganglion cells showed FIF and TH immunoreactivity of varying intensity in both age groups and the amount of yellow fluorescent lipopigments was increased with age. There were also scattered groups of small intensely fluorescent (SIF) cells and a network of varicose fibers in the ganglia. Fluorescence and TH immunocytochemistry did not reveal clear age-related changes in the adrenal medulla, although there appeared to be a large volume of it in aged animals. Four weeks postgrafting, fluorescence microscopy revealed a group of intensively fluorescent chromaffin cells in the recipient SCG (Figs. 3 and 4) of both young and old rats. In some cases the ganglion cells around the transplant had disappeared due to the lesion caused by the operation procedure. The packing density and the intensity of TH immunoreactivity of the cells appeared to be unchanged compared with the control adrenal medullary cells. Some grafted cells, especially in the periphery of the transplant, were elongated and sent out short and thin processes. At 20 weeks following the operation, the packing density of the grafted chromaffin cells appeared to be decreased, most of the cells were elongated, and some cells in the periphery of the transplant were enlarged resembling small neurons (Fig. 5). Neuronal processes were seen more frequently than at four weeks, and when the transplant was found within or close to the ganglion capsule, a dense network of fluorescent nerve fibers was seen to project out from the grafted cells (Figs. 5 and 6). Most
of the grafted cells continued to show strong TH immunoreactivity (Fig. 7), whereas in some enlarged cells TH immunoreactivity was reduced. There was no difference in viability and morphology of the graft between young and aged animals. The characteristics of SIF cells in the adult SCG differ from the phenotype of adrenal chromaffin cells [3, 7, 11]. The catecholamine storing cells within the SCG show more neuronal features [3, 17]. The grafted adrenal chromaffin cells could be distinguished from the original 'SIF' cells by morphology and location of the graft. We have shown that young and old adrenal medullary cells survive and develop neuronal features when transplanted into the SCG. Transplantation of adult adrenal medullary tissue to the brain or anterior chamber of the eye causes extensive growth of nerve fibers by the graft, if the normal adrenergic innervation is destroyed before grafting [1, 5, 6] or if the transplant viability is supported with exogenous NGF [14-16]. In the present study, grafted adrenal medullary cells grew out neurites in the SCG 20 weeks after transplantation without NGF treatment or without previously destroyed pre- or postganglionic innervation. It is possible that the lesion in the ganglion caused by the transplantation procedure itself may lead to a release of NGF from Schwann cells [8]. At 20 weeks after transplantation, the cells in the periphery of the adrenal medullary tissue were more elongated and more frequently enlarged than the cells in the center, suggesting further that the transformation inducing effect originated from surrounding non-neuronal ganglion tissue rather than being mediated via blood vessels grown throughout the graft. Strrmberg and Ebendal [13] showed that both aged and young adult adrenal medullary tissue survive five
Fig. 1. Fluorescence micrographs of con)rol sympathetic ganglion cells from young (a) (3 months old) and old (b) (26 months old) rats. * Pigmented ganglion cells. 216 x . Fig. 2. Fluorescence (a) and TH-immunohistoehemical (b) micrographs of adrenal chromaffln cells from an old (26 months old) rat. Both catecholamine fluorescence and TH immunoreactivity are rather homogeneous. 436 x . Fig. 3. Four weeks postgrafting the transplant (arrows) contains a group of intensively fluorescent chromattin cells (a,b). TH immunoreactivity (c) of the transplanted cells (arrows) is unchanged compared with the control adrenal medullary cells. A young animal, *Ganglion cells, a,c, 108 x ; b, 436 ×. Fig. 4. Four weeks postgrafting the transplant (arrows) contains a group of intensively fluorescent chromaflin cells in an old animal. * Ganglion cells. 436 x Fig. 5 and 6. Twenty weeks after transplantation most of the grafted cells are elongated and some of the cells resemble small neurons (*). Fluorescent nerve fibers (arrow) are seen to project out from chromaffin ceils. An old animal (5); a young animal (6). 216 x . Fig. 7. TH immunoreactivity of young adrenal chromaflin cells 20 weeks postgrafting. Most of the cells are elongated and have an enlarged cytoplasmic area compared to controls. 216 ×.
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218 weeks in the d e n e r v a t e d a n t e r i o r c h a m b e r o f the y o u n g r a t eye, a n d f o r m nerve fibers in response to N G F . The present findings extend the viability o f aged a d r e n a l chromaffin cells in the b r a i n a n d eye to their survival in the p e r i p h e r a l ganglia, in which b o t h y o u n g a n d aged grafted chromaffin cells were able to p r o d u c e c a t e c h o l a mines a n d send o u t neurites even at 20 weeks after operation. T h e grafting was c a r r i e d o u t as an a u t o t r a n s p l a n tation. Since no evident difference in the survival o r t r a n s f o r m a t i o n between y o u n g a n d aged c h r o m a f l i n cell t r a n s p l a n t s was seen, it is i n d i c a t e d t h a t r a t a d r e n a l chromaffin cells m a i n t a i n their plasticity to the o l d age. A l t h o u g h several degenerative a n d c o m p e n s a t o r y changes t a k e place in the aging s u p e r i o r cervical ganglion, the aged s y m p a t h e t i c g a n g l i o n still p r o v i d e s the elements required for the viability o f t r a n s p l a n t e d old chromaffin cells. 1 Bj6rklund, A. and Stenevi, U., Reconstruction of the nigrostriatal dopamine pathway by intraeerebral nigral transplants, Brain Res., 177 (1979) 555-560. 2 Bohn, M.C., Cupit, L., Mareiano, F. and Gash, D.M., Adrenal medulla grafts enhance recovery of striatal dopaminerglc fibers, Science, 237 (1987) 913-915. 3 Elfvin, L.-G., H6gfelt, T. and Goldstein, M., Fluorescence microscopical, immunohistoehemical and ultrastructural studies on sympathetic ganglia of guinea-pig with special reference to the S.I.F. cells and their catecholamine content, J. Ultrastruct. Res., 51 (1975) 377-396. 4 Erfink6, O., The practical histochemical demonstration of catecholamines by formaldehyde-induced fluorescence, J.R. Microsc. Soc., 87 (1967) 259 276. 5 Freed, W.J., Morihisa, J.M., Spoor, E., Hoffer, B., Olson, L., Seiger, .~. and Wyatt, R., Transplanted adrenal chromattin cells in rat brain reduce lesion-induced rotational behavior, Nature, 292 (1981) 351 352. 6 Hansen, J., Bing, G., Notter, M. and Gash, D.M., Paraneuronal grafts in unilateral 6-hydroxydopamine lesioned rats: morphological aspects of adrenal chromattin and carotid body glomus cell implants, Prog. Brain Res., 78 (1988) 507-511. 7 Helen, P., Alho, H. and Hervonen, H., Ultrastructure and histochemistry of human SIF cells and paraganglia, Adv. Biochem. Psychopharmacol., 25 (1980) 145-152.
8 Heumann, R., Korsching, S., Bandtlow, C. and Thoenen, H., Changes of nerve growth factor synthesis in nonneuronal cells in response to sciatic nerve transsection, J. Cell. Biol., 104 (1987) 1623-1631. 9 Kamo, I., Kim, S., McGeer, P. and Shin, D.H., Transplantation of cultured fetal human adrenal chromafl]n cells to rat brain, Neurosci. Lett., 57 (1985) 43--48. I00lson, L., Seiger, .~., Ebendal, T. and Hoffer, B., Comparisons of nerve fiber growth from three major catecholamine producing cell systems; adrenal medulla, superior cervical ganglion and locus coerulens, Adv. Biochem. Psyehopharmacol., 25 (1980) 27-34. 11 Partanen, M., Hervonen, A. and Santer, R.M., Effect of aging on SIF cells of the rat. Adv. Biochem. Psychopharmacol., 25 (1980) 143-147. 12 Sternberger, L.A., Immunohistoehemistry, 2nd edn. Wiley, New York, 1979. 13 Str6mberg, I. and Ebendahl, T., Aged adrenal medullary tissue survives intraocular grafting, forms nerve fibers and responds to the nerve growth factor, J. Neurosei. Res., 23 (1989) 162-171. 14 Str6mberg, I., Herrera-Marsehitz, M., Ungerstedt, U., Ebendahl, T. and Olson, L., Chronic implants of ehromatfin tissue into the dopamine-denervated striatum. Effects of NGF on graft survival, fiber growth and rotational behavior, Exp. Brain Res., 60 (1985) 335-349. 15 Str6mberg, I., Hultgardh-Nilsson, A., Hedin, U. and Ebendal, T., Fate of intraocular chromaftin cell suspensions, role of initial nerve growth factor support, Cell Tissue Res., 254 (1988) 487-497. 16 Str6mberg, I. Ebendal, T., Seiger,/~,. and Olson, L., Nerve fiber production by intraocular adrenal medullary grafts: Stimulation by nerve growth factor or sympathetic denervation of the host iris, Cell Tissue Res., 241 (1985) 241-249. 17 Taxi, J., Derer, M. and Domich, A., Morphology and histophysiology of SIF cells in the autonomic ganglia. In L.-G. Elfvin (Ed.), Autonomic Ganglia, Wiley, New York, 1983, pp. 67 95. 18 Tischler, A.S., DeLellis, R.A., Biales, B., Nunnemacher, G., Carabba, V. and Wolfe, H.J., Nerve growth factor-induced neurite outgrowth from normal human chromaifin ceils, Lab. Invest., 43 (1980) 399-409. 19 Tischler, A.S. and Greene, L.A., Phenotypic plasticity of pheochromocytoma and normal adrenal medullary cells, Adv. Biochem. Psychopharmacol., 25 (1980) 61-68. 20 Unsicker, K., Cell and tissue culture studies on the sympathoadrenal system. In L.C. Elfvin (Ed.), Autonomic Ganglia, Wiley, New York, 1983, pp. 475-505.