Skip to main content
Log in

Heterogeneity of smooth muscle-associated proteins in mammalian brain microvasculature

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

In the brain, the microvascular system is composed of endothelial cells surrounded by a layer of pericytes. The lack of smooth muscle cells in this tissue suggests that any contractile function must be performed by one or both of these cell types. The present study was undertaken in order to identify cells in terminal blood vessels that contain smooth muscle-like contractile machinery. Endothelial cells were reactive with antibodies against smooth muscle myosin but showed no other smooth muscle-related features. In contrast, pericytes of intact microvessels showed a pattern of protein expression similar to that of smooth muscle cells. Pericytes also behaved in tissue culture like cultured smooth muscle cells, with regard to the changes in expression of smooth muscle-related proteins. These data confirm the close relationship between smooth muscle cells and pericytes, and point to their contractile function in the brain microvessels.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Abd-El-Basset EM, Fedoroff S (1991) Immunolocalization of the α isoform of smooth muscle actin in mouse astroglia in cultures. Neuroscience Lett 125:117–120

    Google Scholar 

  • Amberger A, Bauer H, Tontsch U, Gabbiani G, Kocher O, Bauer HC (1991) Reversible expression of sm α-actin protein and sm α-actin mRNA in cloned cerebral endothelial cells. FEBS Lett 287:223–225

    Google Scholar 

  • Bär T (1980) The vascular system of the cerebral cortex. Advances in anatomy, embryology and cell biology, vol 59. Springer, Berlin New York, pp 1–62

    Google Scholar 

  • Borrione AC, Zanellato AMC, Giurato L, Scannapieco G, Pauletto P, Sartore S (1990) Nonmuscle and smooth muscle myosin isoforms in bovine endothelial cells. Exp Cell Res 190:1–10

    Google Scholar 

  • Boswell CA, Majno G, Joris I, Ostrom KA (1992) Acute endothelial cell contraction in vitro: a comparison with vascular smooth muscle cells and fibroblasts. Microvasc Res 43: 178–191

    Google Scholar 

  • Campbell JH, Kocher O, Skalli O, Gabbiani G, Campbell GR (1989) Cytodifferentiation and expression of α-smooth muscle actin mRNA and protein during primary culture of aortic smooth muscle cells. Arteriosclerosis 9:633–643

    Google Scholar 

  • Coffin JD, Harrison J, Schwartz S, Heimark R (1991) Angioblast differentiation and morphogenesis of the vascular endothelium in the mouse embryo. Dev Biol 148:51–62

    Google Scholar 

  • Darby I, Skalli O, Gabbiani G (1990) α-Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29

    Google Scholar 

  • Debus E, Weber K, Osborn M (1983) Monoclonal antibodies to desmin, the muscle-specific intermediate filament protein. EMBO J 2:2305–2312

    Google Scholar 

  • DeNofrio D, Hoock TC, Herman IM (1989) Functional sorting of actin isoforms in microvascular pericytes. J Cell Biol 109: 191–202

    Google Scholar 

  • Desmouliere A, Rubbia-Brandt L, Gabbiani G (1991) Modulation of actin isoform expression in cultured arterial smooth muscle cells by heparin and culture conditions. Arteriosclerosis Thromb 11:244–253

    Google Scholar 

  • Desmouliere A, Rubbia-Brandt L, Abdiu A, Walz T, Macieira-Coelho A, Gabbiani G (1992) α-Smooth muscle actin is expressed in a subpopulation of cultured and cloned fibroblasts and is modulated by γ-interferon. Exp Cell Res 201:64–73

    Google Scholar 

  • Desmouliere A, Geinoz A, Gabbiani F, Gabbiani G (1993) Transforming growth factor-β1 induces α-Smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts. J Cell Biol 122:103–111

    Google Scholar 

  • Diaz-Flores L, Gutierrez R, Varela H, Rancel N, Valladares F (1991) Microvascular pericytes: a review of their morphological and functional characteristics. Histol Histopathol 6: 269–286

    Google Scholar 

  • Dodge AB, Hechtman HB, Shepro D (1991) Microvascular endothelial-derived autacoids regulate pericyte contractility. Cell Motil Cytoskel 18:180–188

    Google Scholar 

  • Draeger A, Gimona M, Stuckert A, Small JV (1991) Calponin. Developmental isoforms and a low molecular weight variant. FEBS Lett 291:24–28

    Google Scholar 

  • Eddinger TJ, Murphy RA (1991) Developmental changes in actin and myosin heavy chain isoform expression in smooth muscle. Arch Biochem Biophys 284:232–237

    Google Scholar 

  • Farrell CR, Stewart PA, Farrell CL, Del Maestro RF (1987) Pericytes in human microvasculature. Anat Rec 218:466–469

    Google Scholar 

  • Frid MG, Shekhonin BV, Koteliansky VE, Glukhova MA (1992) Phenotypic changes of human smooth muscle cells during development: late expression of heavy caldesmon and calponin. Dev Biol 153:185–193

    Google Scholar 

  • Fürst DO, Cross RA, De Mey J, Small JV (1986) Caldesmon is an elongated, flexible molecule localized in the actomyosin domains of smooth muscle. EMBO J 5:251–257

    Google Scholar 

  • Fujimoto T, Singer SJ (1986) Immunocytochemical studies of endothelial cells in vivo. I. The presence of desmin only, or of desmin plus vimentin, or vimentin only, in the endothelial cells of different capillaries of the adult chicken. J Cell Biol 103:2775–2786

    Google Scholar 

  • Fujimoto T, Singer SJ (1987) Immunocytochemical studies of desmin and vimentin in pericapillary cells of chicken. J Histochem Cytochem 35:1105–1115

    Google Scholar 

  • Galmiche MC, Koteliansky VE, Briere J, Herve P, Charbord P (1993) Stromal cells from human long-term marrow cultures are mesenchymal cells that differentiate following a vascular smooth muscle differentiation pathway. Blood 82:66–76

    Google Scholar 

  • Gimona M, Herzog M, Vandekerckhove J, Small JV (1990) Smooth muscle specific expression of calponin. FEBS Lett 274:159–162

    Google Scholar 

  • Gimona M, Vandekerckhove J, Goethals M, Herzog M, Lando Z, Small JV (1994) β-Actin specific monoclonal antibody. Cell Motil Cytoskel 27:108–116

    Google Scholar 

  • Glukhova MA, Kabakov AE, Belkin AM, Frid MG, Ornatsky OI, Zhidkova NI, Koteliansky VE (1986) Mela-vinculin distribution in adult human tissues and cultured cells. FEBS Lett 207:139–141

    Google Scholar 

  • Goncharova EJ, Kam Z, Geiger B (1992) The involvement of adherens junction components in myofibrillogenesis in cultured cardiac myocytes. Development 114:173–183

    Google Scholar 

  • Gown AM, Vogel AM, Gordon D, Lu PL (1985) A smooth muscle-specific monoclonal antibody recognizes smooth muscle actin isozymes. J Cell Biol 100:807–813

    Google Scholar 

  • Haeberle JR, Hathaway DR, Smith CL (1992) Caldesmon content of mammalian smooth muscles. J Muscle Res Cell Motil 13:81–89

    Google Scholar 

  • Herman IM, D'Amore PA (1985) Microvascular pericytes contain muscle and nonmuscle actins. J Cell Biol 101:43–52

    Google Scholar 

  • Hoock TC, Newcomb PM, Herman IM (1991) β Actin and its mRNA are localized at the plasma membrane and the regions of moving cytoplasm during the cellular response to injury. J Cell Biol 112:653–664

    Google Scholar 

  • Hormia M (1982) Expression of factor VIII-related antigen and Ulex lectin binding sites in endothelial cells during long term culture. Cell Biol Int Rep 6:1123–1134

    Google Scholar 

  • Johnson GD, Davidson RS, McNamee KC, Russell G, Goodwin D, Holborow EJ (1982) Fading of immunofluorescence during microscopy: a study of the phenomena and its remedy. J Immunol Methods 55:231–242

    Google Scholar 

  • Joyce NC, Haire MF, Palade GE (1985a) Contractile proteins in pericytes. I. Immunoperoxidase localization of tropomyosin. J Cell Biol 100:1379–1386

    Google Scholar 

  • Joyce, NC, Haire, MF, Palade GE (1985a) Contractile proteins in pericytes. II. Immunocytochemical evidence for the presence of two isomyosins in graded concentrations. J Cell Biol 100:1387–1395

    Google Scholar 

  • Kelley C, D'Amore P, Hechtman HB, Shepro D (1987) Microvascular pericyte contractility in vitro: comparison with other cells of the vascular wall. J Cell Biol 104:483–490

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the heads of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lazard D, Sastre X, Frid MG, Glukhova MA, Thiery JP, Koteliansky VE (1993) Expression of smooth muscle-specific proteins in myoepithelium and stromal fibroblasts of normal and malignant human breast tissue. Proc Natl Acad Sci USA 90:999–1003

    Google Scholar 

  • Lecain E, Alliot F, Laine MC, Calas B, Pessac B (1991) α Isoform of smooth muscle actin is expressed in astrocytes in vitro and in vivo. J Neurosci Res 28:601–606

    Google Scholar 

  • Lin JJ-C, Lin JL-C, Davis-Nanthakumar EJ, Lourim D (1988) Monoclonal antibodies against caldesmon, a Ca++/calmodulinand actin-binding protein of smooth muscle and nonmuscle cells. Hybridoma 7:273–288

    Google Scholar 

  • Matsudaira P, Burgess DR (1978) SDS microslab linear gradient polyacrylamide gel electrophoresis. Anal Biochem 87:386–396

    Google Scholar 

  • Moeremans M, Daneels G, Van Dijck A, Langanger G, De Mey J (1984) Sensitive visualization of antigen-antibody reactions in dot and blot immune overlay assays with immunogold and immunogold/silver staining. J Immunol Methods 74:353–360

    Google Scholar 

  • Nehls V, Drenckhahn D (1991) Heterogeneity of microvascular pericytes for smooth muscle type alpha-actin. J Cell Biol 113:147–154

    Google Scholar 

  • Nehls V, Drenckhahn D (1993) The versatility of microvascular pericytes: from mesenchyme to smooth muscle?. Histochemistry 99:1–12

    Google Scholar 

  • Nehls V, Denzer K, Drenckhahn D (1992) Pericyte involvement in capillary sprouting during angiogenesis in situ. Cell Tissue Res 270:469–474

    Google Scholar 

  • North AJ, Galazkiewicz B, Byers TJ, Glenney JR, Small JV (1993) Complementary distributions of vinculin and dystrophin define two distinct sarcolemma domains in smooth muscle. J Cell Biol 120:1159–1167

    Google Scholar 

  • Osborn M, Caselitz J, Weber K (1981) Heterogeneity of intermediate filament expression in vascular smooth muscle: a gradient in desmin positive cells from the rat aortic arch to the level of the arteria iliaca communis. Differentiation 20:196–202

    Google Scholar 

  • Osborn M, Debus E, Weber K (1984) Monoclonal antibodies specific for vimentin. Eur J Cell Biol 34:137–143

    Google Scholar 

  • Pardridge WM, Boado RJ, Farrel CR (1990) Brain-type glucose transporter (GLUT-1) is selectively localized to the bloodbrain barrier. J Biol Chem 265:18035–18040

    Google Scholar 

  • Paul RJ, Hewett TE, Martin AF (1991) Myosin heavy chain isoforms and smooth muscle function. In: Moreland RS (ed) Regulation of Smooth Muscle Contraction. Plenum Press, New York, pp 139–145

    Google Scholar 

  • Pilch PF (1990) Glucose transporters: What's in a name? Endocrinology 126:3–5

    Google Scholar 

  • Ragan DMS, Schmidt EE, MacDonald IC, Groom AC (1988) Spontaneous cyclic contractions of the capillary wall in vivo, impeding red cell flow: A quantitative analysis-evidence for endothelial contractility. Microvasc Res 36:13–30

    Google Scholar 

  • Rinnerthaler G, Geiger B, Small JV (1988) Contact formation during fibroblast locomotion: Involvement of membrane ruffles and microtubules. J Cell Biol 106:747–760

    Google Scholar 

  • Robinson RA, ten Eyck CJ, Hart MN (1986) Establishment and preliminary growth characteristics of a transformed mouse cerbral microvessel endothelial cell line. Lab Invest 54:579–588

    Google Scholar 

  • Ronnov-Jessen L, Petersen OW (1993) Induction of α-smooth muscle actin by transforming growth factor-β1 in quiescent human breast fibroblasts. Implications for myofibroblast generation in breast neoplasia. Lab Invest 68:696–707

    Google Scholar 

  • Rovner AS, Murphy RA, Owens GK (1986) Expression of smooth muscle and non-muscle myosin heavy chains in cultured vascular smooth muscle cells. J Biol Chem 261:14740–14745

    Google Scholar 

  • Sartore S, De Marzo N, Borrione AC, Zanellato AMC, Saggin L, Fabbri L, Schiaffino S (1989) Myosin heavy-chain isoforms in human smooth muscle. Eur J Biochem 179:79–85

    Google Scholar 

  • Schor AM, Schor SL (1986) The isolation and culture of endothelial cells and pericytes from the bovine retinal microvasculature: a comparative study with large vessel vascular cells. Microvasc Res 32:21–38

    Google Scholar 

  • Schor AM, Canfield AE, Sloan P, Schor SL (1991) Differentiation of pericytes in culture is accompanied by changes in the extracellular matrix. In Vitro Cell Dev Biol 27A:651–659

    Google Scholar 

  • Scott PAE, Bicknell R (1993) The isolation and culture of microvascular endothelium. J Cell Sci 105:269–273

    Google Scholar 

  • Shirinsky VP, Birukov KG, Koteliansky VE, Glukhova MA, Spanidis E, Rogers JD, Campbell JH, Campbell GR (1991) Density-related expression of caldesmon and vinculin in cultured rabbit aortic smooth muscle cells. Exp Cell Res 194:186–189

    Google Scholar 

  • Sims DE (1986) The pericyte—a review. Tissue Cell 18:153–174

    Google Scholar 

  • Sims DE (1991) Recent advances in pericyte biology—Implications for health and disease. Can J Cardiol 7:431–443

    Google Scholar 

  • Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillesen D, Gabbiani G (1986) A monoclonal antibody against α-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103:2787–2796

    Google Scholar 

  • Skalli O, Pelte MF, Peclet MC, Gabbiani G, Gugliotta P, Bussolati G, Ravazzola M, Orci L (1989) α-Smooth muscle actin, a differentiation marker of smooth muscle cells, is present in microfilamentous bundles of pericytes. J Histochem Cytochem 37:315–321

    Google Scholar 

  • Small JV, Fürst DO, De Mey J (1986) Localization of filamin in smooth muscle. J Cell Biol 102:210–220

    Google Scholar 

  • Small JV, Herzog M, Barth M, Draeger A (1990) Supercontracted state of vertebrate smooth muscle cell fragments reveals myofilament lengths. J Cell Biol 111:2451–2461

    Google Scholar 

  • Takeuchi K, Takahashi K, Abe M, Nishida W, Hiwada K, Nabeya T, Maruyama K (1991) Co-localization of immunoreactive forms of calponin with actin cytoskeleton in platelets, fibroblasts and vascular smooth muscle. J Biochem 109:311–316

    Google Scholar 

  • Tokuyasu KT (1980) Immunochemistry on ultrathin frozen sections. Histochem J 12:381–403

    Google Scholar 

  • Tokuyasu KT (1990) Use of poly(vinylpyrrolidone) and poly(vinylalcohol) for cryoultramicrotomy. Histochem J 21:163–171

    Google Scholar 

  • Tokuyasu KT, Singer SJ (1976) Improved procedure for immunoferritin labeling of ultrathin frozen sections. J Cell Biol 71:891–906

    Google Scholar 

  • Tontsch U, Bauer HC (1989) Isolation, characterization, and longterm cultivation of porcine and murine cerebral capillary endothelial cells. Microvasc Res 37:148–161

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Turner CE, Burridge K (1989) Detection of metavinculin in human platelets using a modified talin overlay assay. Eur J Cell Biol 49:202–206

    Google Scholar 

  • Voyta JC, Via DP, Butterfield CE, Zetter BR (1984) Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J Cell Biol 99:2034–2040

    Google Scholar 

  • Woodcock-Mitchell J, Mitchell JJ, Low RB, Kieny M, Sengel P, Rubbia L, Skalli O, Jackson B, Gabbiani G (1988) α-Smooth muscle actin is transiently expressed in embryonic rat cardiac and skeletal muscles. Differentiation 9:161–166

    Google Scholar 

  • Zannellato AMC, Borrione AC, Tonello M, Scannapieco G, Pauletto P, Sartore S (1990) Myosin isoform expression and smooth muscle cell heterogeneity in normal and atherosclerotic rabbit aorta. Arteriosclerosis 10:996–1009

    Google Scholar 

  • Zimmermann KW (1923) Der feinere Bau der Blutcapillaren. Z Anat Entw Gesch 68:3–109

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ehler, E., Karlhuber, G., Bauer, HC. et al. Heterogeneity of smooth muscle-associated proteins in mammalian brain microvasculature. Cell Tissue Res 279, 393–403 (1995). https://doi.org/10.1007/BF00318497

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00318497

Key words

Navigation