The rat adrenal cortex has the uncommon ability to demonstrate morphological and functional regeneration after injury-induced loss of cortical tissue. Peripheral nerves are involved in tissue regeneration and healing after injury, implying that nerves may also be involved in modulating the regeneration of the adrenal cortex. Studies were initiated to assess changes in adrenal innervation during cortical tissue regeneration subsequent to adrenal enucleation. Innervation of regenerating adrenals was assessed from 3 to 62 days postenucleation by immunohistofluorescent detection of neuronal markers for primary afferent, preganglionic sympathetic, and postganglionic sympathetic fibers. The regenerating adrenal contained few nerves at 3 days postenucleation, but became differentially innervated, with extensive innervation by nerve fibers positive for calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH), neuropeptide Y (NPY), and neuronal nitric oxide synthase (nNOS). In contrast, there was only minimal innervation by nerve fibers positive for vasoactive intestinal peptide. By 14 days postenucleation, the CGRP-, TH-, and NPY-positive innervation included areas of hyperinnervation in the capsule, cortex, and central inflammatory site of the regenerating gland. In addition, many chromaffin cells were present at all time points postenucleation. Quantification of the regenerating gland content of CGRP, norepinephrine, epinephrine, and nNOS verified the immunohistofluorescent observations. The period of extensive innervation correlated temporally with the time (3–30 days) during which the regenerating glands recovered steroidogenic function. Moreover, splanchnic nerve transection at the time of adrenal enucleation decreased the innervation by CGRP-positive and vesicular acetylcholine transporter-positive fibers and delayed regeneration. These results support the hypothesis that adrenal innervation modulates tissue regeneration and functional recovery of the enucleated adrenal gland.

Keywords:
Adrenal, Regeneration, Adrenal medulla, Rat, Hyperinnervation, Corticotropin, Catecholamines, Tyrosine hydroxylase, Calcitonin gene-related peptides, Neuropeptide Y
1.
Jennings RW, Hunt TK: Overview of postnatal wound healing; in Adzick NA, Longaker MT (eds): Fetal Wound Healing. New York, Elsevier, 1992, pp 25–52.
2.
Ingle DJ, Higgins GM: Regeneration of the adrenal gland following enucleation. Am J Med Sci 1935;196:232–239.
3.
Greep RO, Deane HW: Histological, cytochemical and physiological observations on the regeneration of the rat’s adrenal gland following enucleation. Endocrinology 1949;45:42–56.
4.
Brogi MP, Pellegrino C: The secretion of corticosterone and aldosterone by the rat adrenal cortex regenerating after enucleation. J Physiol (Lond) 1959;146:165–178.
5.
Skelton FR: Adrenal regeneration and adrenal-regeneration hypertension. Physiol Rev 1959;39:163–182.
6.
Kjartansson J, Dalsgaard CJ, Jonsson CE: Decreased survival of experimental critical flaps in rats after sensory denervation with capsaicin. Plast Reconstr Surg 1987;79:218–221.
7.
Lusthaus S, Shoshan S, Benmeir P, Livoff A, Ashur H, Vardy D: Effect of denervation on incision wound scars in rabbits. J Geriatr Dermatol 1993;1:11–14.
8.
Westerman RA, Carr RW, Delaney CA, Morris MJ, Roberts RGD: The role of skin nociceptive afferent nerves in blister healing. Clin Exp Neurol 1993;30:39–60.
9.
Khalil Z, Helme R: Sensory peptides as neuromodulators of wound healing in aged rats. J Gerontol 1996;51:B354–B361.
10.
Mikulec AA, Tanelian DL: CGRP increases the rate of corneal re-epithelialization in an in vitro whole-mount preparation. J Ocul Pharmacol Ther 1996;12:417–423.
11.
Jones MA, Marfurt CF: Sympathetic stimulation of corneal epithelial proliferation in wounded and nonwounded rat eyes. Invest Ophthalmol Vis Sci 1996;37:2535–2547.
12.
Goss RJ: Regeneration versus repair; in Cohen K, Diegelmann RF, Landbland WJ (eds): Wound Healing: Biochemical and Clinical Aspects. Philadelphia, Saunders, 1992, pp 20–39.
13.
Stocum DL: Wound Repair, Regeneration and Artificial Tissues. Austin, Landes, 1995.
14.
Kuramoto H, Kondo H, Fujita T: Calcitonin gene-related peptide (CGRP)-like immunoreactivity in scattered chromaffin cells and nerve fibers in the adrenal gland of rats. Cell Tissue Res 1987;247:309–315.
15.
Pelto-Huikko M: Immunocytochemical localization of neuropeptides in the adrenal medulla. J Electron Microsc Tech 1989;12:364–379.
16.
Zhou XF, Oldfield BJ, Livett BG: Substance P-containing sensory neurons in the rat dorsal root ganglia innervate the adrenal medulla. J Auton Nerv Syst 1991;33:247–254.
17.
Holgert H, Aman K, Cozzari C, Hartman BK, Brimijoin S, Emson P, Goldstein M, Hokfelt T: The cholinergic innervation of the adrenal gland and its relation to enkephalin and nitric oxide synthase. Neuroreport 1995;6:2576–2580.
18.
Kondo H: Immunohistochemical analysis of the localization of neuropeptides in the adrenal gland. Arch Histol Jpn 1985;48:453–481.
19.
Holgert H, Dagerlind A, Hokfelt T: Immunohistochemical characterization of the peptidergic innervation of the rat adrenal gland. Horm Metab Res 1998;30:315–322.
20.
Kleitman N, Holzwarth MA: Catecholaminergic innervation of the rat adrenal cortex. Cell Tissue Res 1985;241:139–147.
21.
Engeland WC, Dallman MF: Compensatory adrenal growth is neurally mediated. Neuroendocrinology 1975;19:352–362.
22.
Holzwarth MA, Shinsako J, Dallman MF: Adrenal regeneration: Time course, effect of hypothalamic hemi-islands and response to unilateral adrenalectomy. Neuroendocrinology 1980;31:168–176.
23.
Jasper MS, Engeland WC: Splanchnic neural activity modulates ultradian and circadian rhythms in adrenocortical secretion in awake rats. Neuroendocrinology 1994;59:97–109.
24.
Jasper MS, Engeland WC: Synchronous ultradian rhythms in adrenocortical secretion detected by microdialysis in awake rats. Am J Physiol 1991;261:R1257–R1268.
25.
Kennedy WR, Wendelschafer-Crabb G, Johnson T: Quantitation of epidermal nerves in diabetic neuropathy. Neurology 1996;47:1042–1048.
26.
Grutzner EH, Garry MG, Hargreaves KM: Effect of injury on pulpal levels of immunoreactive substance P and immunoreactive calcitonin gene-related peptide. J Endod 1992;18:553–557.
27.
Engeland WC, Dempsher DP, Byrnes GJ, Presnell K, Gann DS: The adrenal medullary response to graded hemorrhage in awake dogs. Endocrinology 1981;101:1539–1544.
28.
Klatt P, Schmidt K, Lehner D, Glatter O, Bachinger HP, Mayer B: Structural analysis of porcine brain nitric oxide synthase reveals a role of tetrahydrobiopterin and L-arginine in the formation of an SDS-resistant dimer. EMBO J 1995;14:1687–1695.
29.
Lara HE, Dees WL, Hiney JK, Dissen GA, Rivier C, Ojeda SR: Functional recovery of the developing rat ovary after transplantation: Contribution of the extrinsic innervation. Endocrinology 1991;129:1849–1860.
30.
Engeland WC: Functional innervation of the adrenal cortex by the splanchnic nerve. Horm Metab Res 1998;30:311–314.
31.
Hokfelt T, Lundberg JM, Schultzberg M, Fahrenkrug J: Immunohistochemical evidence for a local VIP-ergic neuron system in the adrenal gland of the rat. Acta Physiol Scand 1981;113:575–576.
32.
Holzwarth MA: The distribution of vasoactive intestinal peptide in the rat adrenal cortex and medulla. J Auton Nerv Syst 1984;11:269–283.
33.
Varndell IM, Polak JM, Allen JM, Terenghi G, Bloom SR: Neuropeptide tyrosine (NPY) immunoreactivity in norepinephrine-containing cells and nerves of the mammalian adrenal gland. Endocrinology 1984;114:1460–1462.
34.
Afework M, Tomlinson A, Burnstock G: Distribution and colocalization of nitric oxide synthase and NADPH-diaphorase in adrenal gland of developing, adult and aging Sprague-Dawley rats. Cell Tissue Res 1994;276:133–141.
35.
Dijkstra CD, Dopp EA, Joling P, Kraal G: The heterogeneity of mononuclear phagocytes in lymphoid organs: Distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3. Immunology 1985;54:589–599.
36.
Gallo-Payet N, Pothier P, Isler H: On the presence of chromaffin cells in the adrenal cortex: Their possible role in adrenocortical function. Biochem Cell Biol 1987;65:588–592.
37.
Inglis GC, Kenyon CJ, Hannah JA, Connell JM, Ball SG: Does dopamine regulate aldosterone secretion in the rat? Clin Sci 1987;73:93–97.
38.
Dagerlind A, Goldstein M, Hokfelt T: Most ganglion cells in the adrenal medulla are noradrenergic. Neuroreport 1990;1:137–140.
39.
Oomori Y, Okuno S, Fujisawa H, Iuchi H, Ishikawa K, Satoh Y, Ono K: Ganglion cells immunoreactive for catecholamine-synthesizing enzymes, neuropeptide Y and vasoactive intestinal peptide in the rat adrenal gland. Cell Tissue Res 1994;275:201–213.
40.
Lundberg JM, Franco-Cereceda A, Lacroix JS, Pernow J: Neuropeptide Y and sympathetic neurotransmission. Ann NY Acad Sci 1990;611:166–174.
41.
Unsicker K, Krisch B, Otten U, Thoenen H: Nerve growth factor-induced fiber outgrowth from isolated rat adrenal chromaffin cells: Impairment by glucocorticoids. Proc Natl Acad Sci USA 1978;75:3498–3502.
42.
Doupe AJ, Landis SC, Patterson PH: Environmental influences in the development of neural crest derivatives: Glucocorticoids, growth factors and chromaffin cell plasticity. J Neurosci 1985;5:2119–2142.
43.
Woolf CJ, Safieh-Garabedian B, Ma QP, Crilly P, Winters J: Nerve growth factor contributes to the generation of inflammatory sensory hypersensitivity. Neuroscience 1994;62:327–331.
44.
Klimaschewski L, Tran T, Nobiling R, Heym C: Plasticity of postganglionic sympathetic neurons in the rat superior cervical ganglion after axotomy. Microsc Res Tech 1994;29:120–130.
45.
Fristad I, Heyeraas KJ, Kvinnsland IH: Neuropeptide Y expression in the trigeminal ganglion and mandibular division of the trigeminal nerve after inferior alveolar nerve axotomy in young rats. Exp Neurol 1996;142:276–286.
46.
Raivich G, Kreutzberg G: Nerve growth factor and regeneration of peripheral nervous system. Clin Neurol Neurosurg 1993;95:S84–S88.
47.
Hoyle GW, Graham RM, Finkelstein JB, Nguyen KPT, Gozal D, Friedman M: Hyperinnervation of the airways in transgenic mice overexpressing nerve growth factor. Am J Respir Cell Mol Biol 1998;18:149–157.
48.
Isaacson LG, Saffran BN, Crutcher KA: Intracerebral NGF infusion induces hyperinnervation of cerebral blood vessels. Neurobiol Aging 1990;11:51–55.
49.
Jackson S, Hodgkinson S, Estivariz FE, Lowry PJ: IGF1 and 2 in two models of adrenal growth. J Steroid Biochem 1991;40:399–404.
50.
Raivich G, Kreutzberg G: Peripheral nerve regeneration: Role of growth factors and their receptors. Int J Dev Neurosci 1993;11:311–324.
51.
Strand F, Zuccarelli L, Williams K, Lee S, Lee T, Autonawich F, Alves S: Melanotropins as growth factors. Ann NY Acad Sci 1993;680:29–50.
52.
Wilkinson CW, Shinsako J, Dallman MF: Return of pituitary-adrenal function after adrenal enucleation or transplantation: Diurnal rhythms and responses to ether. Endocrinology 1981;109:162–169.
53.
Pratt JH, Turner D, McAteer J, Henry D: β-Adrenergic stimulation of aldosterone production by rat adrenal capsular explants. Endocrinology 1985;117:1189–1194.
54.
Cunningham LA, Holzwarth MA: Vasoactive intestinal peptide stimulates adrenal aldosterone and corticosterone secretion. Endocrinology 1988;122:2090–2097.
55.
Murakami M, Suzuki H, Nakajima S, Nakamoto H, Kageyama Y, Saruta T: Calcitonin gene-related peptide is an inhibitor of aldosterone secretion. Endocrinology 1989;125:2227–2229.
56.
Hinson JP, Vinson GP: Calcitonin gene-related peptide stimulates adrenocortical function in the isolated perfused rat adrenal gland in situ. Neuropeptides 1990;16:129–133.
57.
Bernet F, Bernard J, Laborie C, Montel V, Maubert E, Dupouy JP: Neuropeptide Y (NPY)- and vasoactive intestinal peptide (VIP)-induced aldosterone secretion by rat capsule/glomerular zone could be mediated by catecholamines via β1-adrenergic receptors. Neurosci Lett 1994;166:109–112.
58.
Kimberly C, Byers M: Inflammation of rat molar pulp and periodontium causes increased calcitonin gene-related peptide and axonal sprouting. Anat Rec 1988;222:289–300.
59.
Taylor P, Byers M, Redd P: Sprouting of CGRP nerve fibers in response to dentin injury in rat molar. Brain Res 1990;461:371–376.
60.
Mihara M: Regenerated cutaneous nerves in human epidermal and subepidermal regions. An electron microscopy study. Arch Dermatol Res 1984;276:115–122.
61.
Richter J, Andersson R, Edvinsson L, Gullberg U: Calcitonin gene-related peptide (CGRP) activates human neutrophils – Inhibition by chemotactic peptide antagonist BOC-MLP. Immunology 1992;77:416–421.
62.
Reynier-Rebuffel AM, Mathiau P, Callebert J, Dimitriadou V, Farjaudon N, Kacem K, Launey JM, Seylaz J, Aubineau P: Substance P, calcitonin gene-related peptide, and capsaicin release serotonin from cerebrovascular mast cells. Am J Physiol 1994;267:R1421–R1429.
63.
Wang X, Fiscus RR, Yang L, Mathews HL: Suppression of the functional activity of IL-2-activated lymphocytes by CGRP. Cell Immunol 1995;162:105–113.
64.
Ichinose M, Sawada M: Enhancement of phagocytosis by calcitonin gene-related peptide (CGRP) in cultured mouse peritoneal macrophages. Peptides 1996;17:1405–1414.
65.
Mazzocchi G, Macchi C, Malendowicz LK, Nussdorfer GG: Evidence that endogenous substance-P (SP) is involved in the maintenance of the growth and steroidogenic capacity of rat adrenal zona glomerulosa. Neuropeptides 1995;29:53–58.
66.
Mazzocchi G, Malendowicz LK, Macchi C, Gottardo G, Nussdorfer GG: Further investigations of the effects of neuropeptide Y on the secretion and growth of rat adrenal zona glomerulosa. Neuropeptides 1996;30:19–27.
67.
Haegerstrand A, Dalsgaard CJ, Jonzon B, Larsson O, Nilsson J: Calcitonin gene-related peptide stimulates proliferation of human endothelial cells. Proc Natl Acad Sci USA 1990;87:3299–3303.
68.
Ziche M, Morbidelli L, Pacini M, Geppetti P, Alessandri G, Maggi C: Substance P stimulates neovascularization in vivo and proliferation of cultured endothelial cells. Microvasc Res 1990;40:264–278.
69.
Fan TPD, Hu DE, Guard S, Gresham GA, Watling KJ: Stimulation of angiogenesis by substance P and interleukin-1 in the rat and its inhibition by NK1 or interleukin-1 receptor antagonists. Br J Pharmacol 1993;110:43–49.
70.
Brain SD, Williams TJ, Tippins JR, Morris HR, MacIntyre I: Calcitonin gene-related peptide is a potent vasodilator. Nature 1985;313:54–56.
© 2000 S. Karger AG, Basel
2000
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.