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An immunohistochemical study of endothelial cell heterogeneity in the rat: observations in “en face” Häutchen preparations

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Summary

“En face” sheets of endothelium, in which cellular spatial relationships were maintained, were prepared from proximal pulmonary and femoral arteries and aortae of the Wistar rat, in order to visualise patterns of heterogeneity in populations of endothelial cells. These preparations, termed Häutchen, were immunolabelled with antibodies to angiotensin II, endothelin and Factor-VIII-related antigen, and visualised by an avidin/biotin peroxidase complex. Clusters of cells, which accounted for approximately 30% of the total endothelial cell population, and which were positively immunostained for angiotensin II, were found perpendicular to the longitudinal axis of the aorta and femoral artery. Cells in the pulmonary artery were immunonegative for angiotensin II. The majority of cells in all three vessels were immunopositive for endothelin; groups of intensely stained cells were present in both the femoral artery and aorta, but not in the pulmonary artery. Immunoreactivity to Factor-VIII-related antigen was heterogeneous, with intensely stained amorphous patches of endothelial cells present in the femoral artery and aorta. Häutchen preparations present an opportunity for the investigation of endothelial cell heterogeneity, both within and between vessels; this may provide a basis for the interpretation of the heterogeneity of endothelium-dependent responses in vessels of differing origin.

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References

  • Boulanger C, Lüscher TF (1990) Release of endothelin from the porcine aorta. J Clin Invest 85:587–590

    Google Scholar 

  • Burnstock G (1988) Regulation of local blood flow by neurohumoral substances released from perivascular nerves and endothelial cells. Acta Physiol Scand Vol 133 [Suppl] 571: 53–59

    Google Scholar 

  • Burnstock G, Lincoln J, Fehér E, Hopwood AM, Kirkpatrick K, Milner P, Ralevic V (1988) Serotonin is localised in endothelial cells of coronary arteries and released during hypoxia: a possible new mechanism for hypoxia-induced vasodilatation of the rat heart. Experientia 40:705–707

    Google Scholar 

  • Catravas JD, Bassingthwaighte JB, Sparks HV Jr (1988) Adenosine transport and uptake by cardiac and pulmonary endothelial cells. In: Ryan US (ed) Endothelial cells, vol 1.CRC Press Inc, Boca Raton, Fla, pp 65–82

    Google Scholar 

  • Cocks TM, Angus JA (1983) Endothelium-dependent relaxation of coronary arteries by noradrenaline and serotonin. Nature 305:627–630

    Google Scholar 

  • De Bault LE, Esmond NL, Smith GL, Esmon CT (1986) Localization of thrombomodulin antigen in rabbit endothelial cells in culture: an immunofluorescence and immunoelectron microscope study. Lab Invest 54:179–187

    Google Scholar 

  • De Mey JG, Claeys M, Vanhoutte PM (1982) Endothelium-dependent inhibitory effects of acetylcholine, adenosine triphosphate, thrombin and arachidonic acid in the canine femoral artery. J Pharmacol Exp Ther 222:166–173

    Google Scholar 

  • De Nucci G, Thomas R, D'Orleans-Juste P, Antunes E, Walder C, Warner TD, Vane JR (1988) Pressor effects of circulating endothelin are limited by its removal in the pulmonary circulation and by the release of prostacyclin and endothelium-derived relaxing factor. Proc Natl Acad Sci USA 85:9797–9800

    Google Scholar 

  • Dzau VJ (1984) Vascular wall renin-angiotensin pathway in control of the circulation. Am J Med [Suppl 4A] 77:31–36

    Google Scholar 

  • Furchgott RF (1981) The requirement for endothelial cells in the relaxation of arteries by acetylcholine and some other vasodilators. Trends Pharmacol Sci 2:173–176

    Google Scholar 

  • Furchgott RF, Zawadzki JV (1980) The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature 288:373–376

    Google Scholar 

  • Hamel E, Assumel-Lurdin C, Edvinsson L, Fage D, Mackenzie ET (1987) Neuronal versus endothelial origin of vasoactive acetylcholine in pial vessels. Brain Res 420:391–396

    Google Scholar 

  • Hodge RL, Ng KKF, Vane JR (1967) Disappearance of angiotensin from the circulation of the dog. Nature 215:138–141

    Google Scholar 

  • Jaffe EA, Nachman RL, Becker CG, Mimick CR (1973) Culture of human endothelial cells derived from umbilical veins. J Clin Invest 52:2745–2756

    Google Scholar 

  • Johnson AR (1984) The metabolism of vasoactive peptides by human endothelial cells. In: Jaffe EA (ed) Biology of endothelial cells. Martinus Nijhof, Boston The Hague Dordrecht Lancaster, pp 302–316

    Google Scholar 

  • Junod AF, Ody C (1977) Amine uptake and metabolism by endothelium of pig pulmonary artery and aorta. Am J Physiol 232:C88-C94

    Google Scholar 

  • Katusic Z, Shepherd JT, Vanhoutte PM (1984a) Arginie vasopressin induces endothelium-dependent relaxations of canine basilar and coronary arteries. Fed Proc 43:1084

    Google Scholar 

  • Katusic ZS, Shepherd JT, Vanhoutte PM (1984b) Vasopressin causes endothelium-dependent relaxations of the canine basilar artery. Circ Res 55:575–579

    Google Scholar 

  • Kifor I, Dzau VJ (1987) Endothelial renin-angiotensin pathway: evidence for intracellular synthesis and secretion of angiotensins. Circ Res 60:422–428

    Google Scholar 

  • Lincoln J, Burnstock G (1990) Neural-endothelial interactions in control of local blood flow. In: Warren J (ed) The endothelium: an introduction to current research. Liss, New York, (in press)

    Google Scholar 

  • Lincoln J, Loesch A, Burnstock G (1990) Localization of vasopressin, serotonin and angiotensin II in endothelial cells of the renal and mesenteric arteries of the rat. Cell Tissue Res 259:341–344

    Google Scholar 

  • Linnik MD, Moskowitz MA (1989) Identification of immunoreactive substance P in human and other mammalian endothelial cells. Peptides 10:957–962

    Google Scholar 

  • Loesch A, Burnstock (1988) Ultrastructural localisation of serotonin and substance P in vascular endothelial cells of rat femoral and mesenteric arteries. Anat Embryol 178:137–142

    Google Scholar 

  • Milner P, Ralevic V, Hopwood AM, Fehér E, Lincoln J, Kirkpatrick KA, Burnstock G (1989) Ultrastructural localisation of substance P and choline acetyltransferase in endothelial cells of rat coronary artery and release of substance P and acetylcholine during hypoxia. Experientia 45:121–125

    Google Scholar 

  • Nakatsu K, Karwamoto JH, Brien JF, Marks GS (1988) A facile reliable method for staining blood vessel endothelium. J Pharmacol Methods 19:149–154

    Google Scholar 

  • Ng KKF, Vane JR (1967) Conversion of angiotensin I to angiotensin II. Nature 216:762–766

    Google Scholar 

  • Parnavelas JG, Kelly W, Burnstock G (1985) Ultrastructural localization of choline acetyltransferase in vascular endothelial cells in rat brain. nature 316:724–725

    Google Scholar 

  • Pearson JD, Gordon JL (1985) Nucleotide metabolism by endothelium. Ann Rev Physiol 47:617–627

    Google Scholar 

  • Pearson JD, Carleton JS, Hutchings A, Gordon JL (1978) Uptake and metabolism of adenosine by pig aortic endothelial and smooth muscle cells in culture. Biochem J 170:265–271

    Google Scholar 

  • Poole JCF, Sanders AG, Florey HW (1958) The regeneration of aortic endothelium. J Pathol Bacteriol 75:133–143

    Google Scholar 

  • Ralevic V, Milner P, Hudlická O, Kristek F, Burnstock G (1990) Substance P is released from the endothelium of normal and capsaicin-treated rat hindlimb vasculature, in vivo, by increased flow. Circ Res 66:1178–1183

    Google Scholar 

  • Robinson-White A, Peterson S, Hechtman HB, Shepro D (1981) Serotonin uptake by isolated adipose capillary endothelium. J Pharmacol Exp Ther 216:125–128

    Google Scholar 

  • Rone JD, Goodman AL (1987) Heterogeneity of rabbit aortic endothelial cells in primary culture. Proc Soc Exp Biol Med 184:495–503

    Google Scholar 

  • Ryan JW, Stewart JM, Leary WP, Ledingham JG (1970) Metabolism of angiotensin I in the pulmonary circulation. Biochem J 120:221–223

    Google Scholar 

  • Ryan JW, Niemeyer RS, Goodwin DW, Smith V, Stewart JM (1971) Metabolism of (8-L[14C]phenylalanine)-angiotensin I in the pulmonary circulation. Biochem J 125:921–923

    Google Scholar 

  • Ryan JW, Rvan US, Schultz DR, Whitaker C, Chung A, Dorer FE (1975) Subcellular localization of pulmonary angiotensin converting enzyme (kininase II). Biochem J 146:497–499

    Google Scholar 

  • Toda N (1984) Endothelium-dependent relaxation induced by angiotensin II and histamine in isolated arteries of dog. Br J Pharmacol 81:301–307

    Google Scholar 

  • Toda N, Miyazaki M (1981) Angiotensin-induced relaxation in isolated dog renal and cerebral arteries. Am J Physiol 240:H247-H254

    Google Scholar 

  • Tomobe Y, Miyauchi T, Saito A, Yanagisawa N, Kimura S, Goto K, Masaki T (1988) Effects of endothelin on the renal artery from spontaneously hypertensive and Wistar Kyoto rats. Eur J Pharmacol 152:373–376

    Google Scholar 

  • Vane JR (1969) The release and fate of vaso-active hormones in the circulation. Br J Pharmacol 35:209–242

    Google Scholar 

  • Vanhoutte PM, Miller VM (1985) Heterogeneity of endothelium-dependent responses in mammalian blood vessels. J Cardiovasc Pharmacol 3:S12-S23

    Google Scholar 

  • Wagner DD, Olmsted JB, Marder VJ (1982) Immunolocalization of von-Willebrand protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol 95:355–360

    Google Scholar 

  • Warner TD, Mitchell JA, De Nucci G, Vane JR (1989) Endothelin-1 and endothelin-3 release EDRF from isolated perfused arterial vessels of the rat and rabbit. J Cardiovasc Pharmacol 13:S85-S88

    Google Scholar 

  • Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T (1988) A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 332:411–415

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy and Developmental Biology, and Centre for Neuroscience, University College London, Gower Street, London, UK

    A. Tomlinson, H. Van Vlijmen, A. Loesch & G. Burnstock

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  1. A. Tomlinson
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  2. H. Van Vlijmen
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  3. A. Loesch
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  4. G. Burnstock
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Tomlinson, A., Van Vlijmen, H., Loesch, A. et al. An immunohistochemical study of endothelial cell heterogeneity in the rat: observations in “en face” Häutchen preparations. Cell Tissue Res 263, 173–181 (1991). https://doi.org/10.1007/BF00318413

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