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Ultrastructural immunocytochemical co-localization of serotonin and PNMT in adrenal medullary vesicles

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Summary

Previous immunocytochemical studies at the light microscopic level have demonstrated serotonin immunoreactivity in rat adrenal epinephrine-containing cells. In this study we have used electron microscopic immunocytochemical methods to study the subcellular distribution of serotonin and the enzyme responsible for epinephrine biosynthesis, phenylethanolamine-N-methyltransferase (PNMT). The distribution of the immunostaining was compared in adjacent serial thin sections using a post-embedding method in conjunction with peroxidase-antiperoxidase (PAP) immunocytochemistry. Serotonin immunoreactivity was associated with the limiting membrane as well as with the core of the chromaffin vesicles. In adjacent sections PNMT immunoreactivity was also seen in the serotonin-containing vesicles. However, its intravesicular distribution was different from that of serotonin; PNMT occupied the eccentric zone of the vesicles between the serotonin immunoreactive sites.

These results are interpreted to be in support of biochemical studies claiming a serotonin uptake and storage capacity of adrenal chromaffin vesicle fractions as well as those which suggest serotonin is synthesized by chromaffin cells. The relative contribution of uptake and synthesis to the pool of serotonin that is stored in the vesicles is an open question. The co-localization of serotonin and PNMT in the same vesicle is suggestive of a capacity for co-release of serotonin and epinephrine by the adrenal medulla.

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References

  • Brusco A, Peressini S, Saavedra JP (1983) Serotonin-like immunoreactivity and anti-5-hydroxytryptamine (5HT) antibodies. J Histochem Cytochem 31:524–530

    Google Scholar 

  • Carlsson A, Hillarp N-A, Waldeck P (1963) Analysis of the Mg-ATP dependent storage mechanism in the amine granules of the adrenal medulla. Acta Physiol Scand 59:(Suppl 215):1–38

    Google Scholar 

  • Coupland RE, Hopwood D (1966) The mechanism of the differential staining reaction for adrenaline and noradrenaline-storing granules in tissues fixed in glutaraldehyde. J Anat 100:227–243

    Google Scholar 

  • De May J (1983) Immunogold probes in immunocytochemistry. In: Polak JM, Van Noorden S (eds) Immunocytochemistry, Wright, Boston, pp 82–112

    Google Scholar 

  • Farnum CE, Wilsman N (1984) Lectin-binding histochemistry of non decalicified growth plate cartilage: a post-embedding method for light microscopy of epon-embedded tissue. J Histochem Cytochem 32:593–607

    Google Scholar 

  • Gershon MD, Ross LL (1966a) Radioisotope studies of the binding exchange, and distribution of 5-hydroxytryptamine synthesized form its radioactive precursor. J Physiol 186:451–476

    Google Scholar 

  • Gershon MD, Ross LL (1966b) Location of sites of 5-hydroxytryptamine storage and metabolism by radioautography. J Physiol 186:477–492

    Google Scholar 

  • Holzwarth MA, Brownfield MS (1985) Serotonin coexists with epinephrine in rat adrenal medulla. Neuroendocrinology (in press)

  • Holzwarth MA, Sawetawan C, Brownfield MS (1984) Serotonin-immunoreactivity in the adrenal medulla: Distribution and response to pharmacological manipulation. Brain Res Bull 13:299–308

    Google Scholar 

  • Huang WM, Gibson SJ, Facer P, Gu J, Polak JM (1983) Improved section adhesion for immunocytochemistry using high molecular weight polymers of l-lysine as a slide coating. Histochemistry 77:275–279

    Google Scholar 

  • Kent C Coupland RE (1984) On the uptake and storage of 5-hydroxy-tryptamine, 5-hydroxytryptophan and catecholamines by adrenal chromaffin cells and nerve endings. Cell Tissue Res 236:189–195

    Google Scholar 

  • Nagatsu I, Karasawa N, Kondo Y, Inagaki S (1979) Immunocytochemical localization of tyrosine hydroxylase, dopamine-beta-hydroxylase and phenylethanolamine-N-methyltransferase in the adrenal glands of frog and rat by a peroxidase-antiperoxidase method. Histochemistry 64:131–144

    Google Scholar 

  • Ritzen M, Hammarstrom L, Ullbery S (1965) Autoradiographic distribution of 5-hydroxytryptamine and 5-hydroxytryptophan in the mouse. Biochem Pharmacol 14:313–332

    Google Scholar 

  • Schipper J, Tilders FJH (1983) A new technique for studying specificity of immunocytochemical procedures. J Histochem Cytochem 31:12–18

    Google Scholar 

  • Slotkin TA, Kirschner N (1971) Uptake, storage and distribution distribution of amines in bovine adrenal medullary vesicles. Mol Pharmacol 7:581–592

    Google Scholar 

  • Snyder SH, Axelrod J, Zweig M (1965) A sensitive and specific fluorescence assay for tissue preparation. Biochem Pharmacol 14:831–835

    Google Scholar 

  • Steinbusch HWM, Verhofstad AAJ, Joosten HWJ (1978) Localization of serotonin in the central nervous system by immunohistochemistry: description of a specific and sensitive technique and some applications. Neuroscience 3:811–819

    Google Scholar 

  • Sternberger LA (1979) Immunocytochemistry. 2 Edn. John Wiley & Sons, New York

    Google Scholar 

  • Ungar A, Phillips JH (1983) Regulation of the adrenal medulla. Physiol Rev 63:787–843

    Google Scholar 

  • Van de Kar LD, Wilkinson CW, Ganong WF (1981) Pharmacological evidence for a role of brain serotonin in the maintenance of plasma renin activity unanaesthetized rats. J Pharmacol Exp Ther 219:85–90

    Google Scholar 

  • Van Orden LS, III, Burke JP, Redick JA, Rybarczyk KE, Van Orden DE, Baker HA, Hartman BK (1977) Immunocytochemical evidence for particulate localization of phenylethanolamine-N-methyltransferase in adrenal medulla. Neuropharmacology 16:129–133

    Google Scholar 

  • Verhofstad AAJ, Jonsson G (1983) Immunohistochemical and neurochemical evidence for the presence of serotonin in the adrenal medulla of the rat. Neuroscience 10:1443–1453

    Google Scholar 

  • Winkler H (1977) The biogenesis of adrenal chromaffin granules. Neuroscience 2:657–683

    Google Scholar 

  • Winkler H, Westhead E (1980) The molecular organization of adrenal chromaffin granules. Neuroscience 5:1803–1823

    Google Scholar 

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Brownfield, M.S., Poff, B.C. & Holzwarth, M.A. Ultrastructural immunocytochemical co-localization of serotonin and PNMT in adrenal medullary vesicles. Histochemistry 83, 41–46 (1985). https://doi.org/10.1007/BF00495298

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