Skip to main content
SpringerLink
Log in

Expression of mRNAs of multiple growth factors and receptors by astrocytes and glioma cells: Detection with reverse transcription-polymerase chain reaction

  • Review Article
  • Published:
Cellular and Molecular Neurobiology Aims and scope Submit manuscript

Summary

1. Although glial cells in culture are known to secrete growth factors and are also known to be responsive to some of them, detailed comparisons are difficult because the bulk of information was based on various animals of origin, developmental stages, growth properties, culture age, and culture conditions.

2. To present a unified picture of the growth factors and their receptors found in glial cells, we surveyed the expression of messenger RNAs of a panel of growth factors and receptors, using reverse transcription-polymerase chain reaction (RT-PCR), in three common glial cell types: rat astrocytes in primary culture, rat glioma line C6, and human glioma line A172.

3. We observed that normal and neoplastic glial cells in culture express multiple growth factors and also possess most of the receptors to these factors, suggesting multiple autocrine functions. In addition, glia produce growth factors known to be capable of acting on neurons, implicating paracrine function involving glia-neuron interaction. Glial cells also produce growth factors and receptors that are capable of communicating with hematopoietic cells, suggesting neuroimmunologic interaction. What is most interesting is that glial cells express receptors for growth factors previously thought to be acting on neurons only.

4. The current study demonstrates the feasibility of screening from a small sample a large number of growth factors and receptors. The method portends future clinical application to biopsy or necropsy samples from brain tumors or pathologic brains suffering from degenerative diseases such as Alzheimer's or Parkinson's disease.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Abraham, J. A., Whang, J. L., Tumolo, A., Mergia, A., Friedman, J., Gospodarowicz, D., and Fiddes, J. C. (1986). Human basic fibroblast growth factor: Nucleotide sequence and genomic organization.EMBO J. 52523–2528.

    PubMed  Google Scholar 

  • Bell, G. I., Fong, N. M., Stempien, M. M., Wormsted, M. A., Caput, D., Ku, L., Urdea, M. S., Rall, L. B., and Sanchez-Pescador, R. (1986). Human epidermal growth factor: cDNA sequence, expressionin vitro and gene organization.Nucleic Acids Res. 148427–8446.

    PubMed  Google Scholar 

  • Betsholtz, C., Johnsson, A., Helding, C.-H., Westermark, B., Lind, P., Urdea, M. S., Eddy, R., Shows, T. B., Philpott, K., Mellor, A. L., Knott, T. J., and Scott, J. (1986). cDNA sequence and chromosomal localization of human platelet-derived growth factor A-chain and its expression in tumor cell lines.Nature 320695–699.

    PubMed  Google Scholar 

  • Birren, S. J., and Anderson, D. J. (1990). A v-myc-immortalized sympathoadrenal progenitor cell line in which neuronal differentiation is initiated by FGF but not NGF.Neuron 4189–201.

    PubMed  Google Scholar 

  • Brenner, C. A., Tam, A. W., Nelson, P. A., Engleman, E. G., Suzuki, N., Fry, K. E., and Larrick, J. W. (1989). Message amplification phenotyping (MAPPing): A technique to simultaneously measure multiple mRNAs from small number of cells.Biotechniques 71096–1103.

    PubMed  Google Scholar 

  • Burt, D. W., and Paton, I. R. (1992). Evolutionary origins of the transforming growth factor-beta gene family.DNA Cell. Biol. 11497–510.

    PubMed  Google Scholar 

  • Chomczynski, P., and Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.Anal. Biochem. 162156–159.

    PubMed  Google Scholar 

  • Claesson-Welsh, L., Ericksson, A., Moren, A., Severinsson, L., Ek, B., Ostman, A., Betsholtz, C., and Heldin, C.-H. (1988). cDNA cloning and expression of a human platelet-derived growth factor (PDGF) receptor specific for B-chain-containing PDGF molecules.Mol. Cell Biol. 83476–3486.

    PubMed  Google Scholar 

  • Claesson-Welsh, L., Ericksson, A., Westermark, B., and Heldin, C.-H. (1989). cDNA cloning and expression of the human A-type platelet-derived growth factor (PDGF) receptor establishes structural similarity to the B-type PDGF receptor.Proc. Natl. Acad. Sci. USA 864917–4921.

    PubMed  Google Scholar 

  • Constam, D. B., Philipp, J., Malipiero, U. V., Ten Dijke, P., Schachner, M., and Fontana, A. (1992). Differential expression of transforming growth factor-beta 1, -beta 2, and -beta 3 by glioblastoma cells, astrocytes, and microglia.J. Immunol. 1481404–1410.

    PubMed  Google Scholar 

  • Cordell, B., Bell, G., Tischer, E., DeNoto, F. M., Ullrich, A., Pictet, R., Rutter, W. J., and Goodman, H. M. (1979). Isolation and characterization of a cloned rat insulin gene.Cell 18533–543.

    PubMed  Google Scholar 

  • Davis, J. B., and Stroobant, P. (1990). Platelet-derived growth factors and fibroblast growth factors are mitogens for rat Schwann cells.J. Cell Biol. 1101353–1360.

    PubMed  Google Scholar 

  • Davis, S., Aldrich, T. H., Valenzuela, D. M., Wong, V., Furth, M. E., Squinto, S. P., and Yancopoulos, G. D. (1991). The receptor for ciliary neurotrophic factor.Science 25359–63.

    PubMed  Google Scholar 

  • Gearing, D. P., Thut, C. J., VandenBos, T., Gimpel, S. D., Delaney, P. B., King, J., Price, V., Cosman, D., and Beckmann, M. P. (1991). Leukemia inhibitory factor receptor is structurally related to the IL-6 signal transducer, gp130.EMBO J. 102839–2848.

    PubMed  Google Scholar 

  • Giulian, D., and Lachman, L. B. (1985). Interleukin-1 stimulation of astroglial proliferation after brain injury.Science 228497–499.

    PubMed  Google Scholar 

  • Goldstein, B. J., and Dudley, A. L. (1990). The rat insulin receptor: primary structure and conservation of tissue-specific alternative messenger RNA splicing.Mol. Endocrinol. 4235–244.

    PubMed  Google Scholar 

  • Hermanson, M., Funa, K., Hartman, M., Claesson-Welsh, L., Heldin, C. H., Westermark, B., and Nister, M. (1992). Platelet-derived growth factor and its receptors in human glioma tissue: expression of messenger RNA and protein suggests the presence of autocrine and paracrine loops.Cancer Res. 523213–3219.

    PubMed  Google Scholar 

  • Hibi, M., Murakami, M., Saito, M., Hirano, T., Taga, T., and Kishimoto, T. (1990). Molecular cloning and expression of an Il-6 signal transducer, gp130.Cell 631149–1157.

    PubMed  Google Scholar 

  • Hirota, S., Ito, A., Morii, E., Wanaka, A., Tohyama, M., Kitamura, Y., and Nomura, S. (1992). Localization of mRNA for c-kit receptor and its ligand in the brain of adult rats.Mol. Brain Res. 1547–54.

    PubMed  Google Scholar 

  • Hughes, S. M., Lillien, L. E., Raff, M. C., Rohrer, H., and Sandtner, M. (1988). Ciliary neurotrophic factor induces type-2 astrocyte differentiation in culture.Nature 33570–73.

    PubMed  Google Scholar 

  • Hutton, L. A., de Vellis, J., and Perez-Polo, J. R. (1992). Expression of p75, Trk A and Trk B mRNA in rat C6 glioma and type I astrocyte cultures.J. Neurosci. Res. 32375–383.

    PubMed  Google Scholar 

  • Jaye, M., Howk, R., Burgess, W., Ricca, G. A., Chiu, I.-M., Ravera, M. W., O'Brien, S. J., Modi, W. S., Maciag, T., and Drohan, W. N. (1986). Human endothelial cell growth factor: Cloning, nucleotide sequence, and chromosome localization.Science 233541–545.

    PubMed  Google Scholar 

  • Keshet, E., Lyman, S. D., Williams, D. E., Anderson, D. M., Jenkins, N. A., Copeland, N. G., and Parada, L. F. (1991). Embryonic RNA expression patterns of the c-kit receptor and its cognate ligand suggest multiple functional role in mouse development.EMBO J. 102425–2435.

    PubMed  Google Scholar 

  • Klein, R., Parada, L. F., Coulier, F., and Barbacid, M. (1989). TrkB, a novel tyrosine protein kinase receptor expressed during mouse neural development.EMBO J. 83701–3709.

    PubMed  Google Scholar 

  • Korsching, S. (1993). The neurotrophic factor concept: A reexamination.J. Neurosci. 132739–2748.

    PubMed  Google Scholar 

  • Lamballe, F., Klein, R., and Barbacid, M. (1991). Trk C, a new member of the trk family of tyrosine protein kinases, is a receptor for neurotrophin-3.Cell 66967–979.

    PubMed  Google Scholar 

  • LeRoith, D., Roberts, C. T., Jr., Werner, H., Bondy, C., Raizada, M., and Adamo, M. L. (1993). Insulin-like growth factors in the brain. InNeurotrophic Factors (S. E. Longhlin and J. H. Fallon, Eds.), Academic Press, New York, pp. 339–357.

    Google Scholar 

  • Lim, R., and Bosch, E. P. (1990). Preparation of astrocytes and Schwann cells.Methods Neurosci. 247–56.

    Google Scholar 

  • Lim, R., Zhong, W., and Zaheer, A. (1990). Antiproliferative function of glia maturation factor beta.Cell Regul. 1741–746.

    PubMed  Google Scholar 

  • Lindholm, D., Castrin, E., Kiefer, R., Zafra, F., and Thoenen, H. (1992). Transforming growth factor-beta 1 in the rat brain: Increase after injury and inhibition of astrocyte proliferation.J. Cell Biol. 117395–400.

    PubMed  Google Scholar 

  • Maisonpierre, P. C., Belluscio, L., Squinto, S., Ip, N. Y., Furth, M. E., Lindsay, R. M., and Yancopoulos, G. D. (1990). Neurotrophin-3: A neurotrophic factor related to NGF and BDNF.Science 2471446–1451.

    PubMed  Google Scholar 

  • Manova, K., Bachvarova, R. F., Huang, E. J., Sanchez, S., Pronovost, S. M., Velazquez, E., McGuire, B., and Besmer, P. (1992). c-kit receptor and ligand expression in postnatal development of the mouse cerebellum suggests a function for c-kit in inhibitory neurons.J. Neurosci. 124663–4676.

    PubMed  Google Scholar 

  • Martin, F. H., Suggs, S. V., Langley, K. E., Lu, H. S., Ting, J., Okino, K. H., Morris, C. F., Mcniece, I. K., Jacobson, F. W., Mendiaz, E. A., Birkett, N. C., Smith, K. A., Johnson, M. J., Parker, V. P., Flores, J. C., Patel, A. C., Fisher, E. F., Erjavec, H. O., Herrera, C. J., Wypych, J., Sachdev, R. K., Pope, J. A., Leslie, I., Wen, D., Lin, C.-H., Cupples, R. L., and Zsebo, K. M. (1990). Primary structure and functional expression of rat and human stem cell factor DNAs.Cell 63203–211.

    PubMed  Google Scholar 

  • Martin-Zanca, D., Oskam, R., Mitra, G., Copeland, T., and Barbacid, M. (1989). Molecular and biochemical characterization of the human trk proto-oncogene.Mol. Cell. Biol. 924–33.

    PubMed  Google Scholar 

  • Masiakowski, P., Liu, H., Radziejewski, C., Lottspeich, F., Oberthuer, W., Wong, V., Lindsay, R. M., Furth, M. E., and Panayotatos, M. (1991). Recombinant human and rat ciliary neurotrophic factors.J. Neurochem. 571003–1012.

    PubMed  Google Scholar 

  • Matsui, Y., Zsebo, K., and Hogan, B. L. (1990). Embryonic expression of a haematopoietic growth factor encoded by the S1 locus and the ligand for c-kit.Nature 347667–669.

    PubMed  Google Scholar 

  • Moretto, G., Xu, R. Y., Walker, D. G., and Kim, S. U. (1994). Co-expression of mRNA for neurotrophic factors in human neurons and glial cells in culture.J. Neuropathol. Exp. Neurol. 5378–85.

    PubMed  Google Scholar 

  • Morgan, D. O., Edman, J. C., Standring, D. N., Fried, V. A., Smith, M. C., Roth, R. A., and Rutter, W. J. (1987). Insulin-like growth factor II receptor as a multifunctional binding protein.Nature 329301–307.

    PubMed  Google Scholar 

  • Morrison, R. S. (1991). Suppression of basic fibroblast growth factor expression by antisense oligodeoxynucleotides inhibits the growth of transformed human astrocytes.J. Biol. Chem. 266728–734.

    PubMed  Google Scholar 

  • Morrison, R. S. (1993). Epidermal growth factors: Structure, expression, and functions in the central nervous system. InNeurotrophic Factors (S. E. Loughlin and J. H. Fallon, Eds.), Academic Press, New York, pp. 339–357.

    Google Scholar 

  • Natali, P. G., Nicotra, M. R., Sures, I., Santoro, E., Bigotti, A., and Ullrich, A. (1992). Expression of c-kit receptor in normal and transformed human nonlymphoid tissues.Cancer Res. 526139–6143.

    PubMed  Google Scholar 

  • Nichols, N. R., Osterburg, H. H., Masters, J. N., Miller, S. L., and Finch, C. E. (1990). Messenger RNA for glial fibrillary acidic protein is decreased in rat brain following corticosterone treatment.Mol. Brain. Res. 71–7.

    PubMed  Google Scholar 

  • Nishida, T., Nishino, N., Takano, M., Sekiguchi, Y., Kawai, K., Mizuno, K. U., Nakai, S., Masui, Y., and Hirai, Y. (1989). Molecular cloning and expression of rat interleukin-1 alpha cDNA.J. Biochem. 105351–357.

    PubMed  Google Scholar 

  • Noble, M., Murray, K., Stroobant, P., Waterfield, M. D., and Riddle, P. (1988). Platelet-derived growth factor promotes division and motility and inhibits premature differentiation of the oligodendrocyte/type-2 astrocyte progenitor cell.Nature 333560–562.

    PubMed  Google Scholar 

  • Patterson, P. H. (1992). The emerging neuropoietic cytokine family.Curr. Opin. Neurobiol. 294–97.

    PubMed  Google Scholar 

  • Pizzuti, A., Borsani, G., Falini, A., Rugarli, E. I., Sidoli, A., Baralle, F. E., Scarlato, G., and Silani, V. (1990). Detection of β-nerve growth factor mRNA in the human fetal brain.Brain Res. 518337–341.

    PubMed  Google Scholar 

  • Radeke, M. J., Misko, T. P., Hsu, C., Herzenberg, L. A., and Shooter, E. M. (1987). Gene transfer and molecular cloning of the rat nerve growth factor receptor.Nature 325593–597.

    PubMed  Google Scholar 

  • Rall, L. B., Scott, J., and Bell, G. I. (1987). Human insulin-like growth factor I and II messenger RNA: isolation of complementary DNA and analysis of expression.Methods Enzymol. 146239–248.

    PubMed  Google Scholar 

  • Rosenthal, A., Goeddel, D. V., Nguyen, T., Martin, E., Burton, L. E., Shih, A., Laramee, G. R., Wurm, F., Mason, A., Nikolics, K., and Winslow, J. W. (1991). Primary structure and biological activity of human brain-derived neurotrophic factor.Endocrinology 1291289–1294.

    PubMed  Google Scholar 

  • Saad, B., Constam, D. B., Ortmann, R., Moos, M., Fontana, A., and Schachner, M. (1991). Astrocyte-derived TGF-beta 2 and NGF differentially regulate neural recognition molecule expression by cultured astrocytes.J. Cell Biol. 115473–484.

    PubMed  Google Scholar 

  • Scooltink, H., Stoyan, T., Lenz, D., Schmitz, H., Hirano, T., Kishimoto, T., Heinrich, P. C., and Rose-Jhon, S. (1991). Structural and functional studies on the human hepatic interleukin-6 receptor. Molecular cloning and over expression in Hep G2 cells.Biochem. J. 277659–664.

    PubMed  Google Scholar 

  • Sims, J. E., March, C. J., Cosman, D., Widmer, M. B., Robson MacDonald, H., McMahan, C. J., Grubin, C. E., Wignal, J. M., Jackson, J. L., Call, S. M., Friend, D., Alpert, A. R., Gillis, S., Urdal, D. L., and Dower, S. K. (1988). cDNA expression cloning of the IL-1 receptor, a member of the immunoglobulin superfamily.Science 241585–589.

    PubMed  Google Scholar 

  • Smits, A., Kato, M., Westermark, B., Nister, M., Heldin, C. H., and Funa, K. (1991). Neurotrophic activity of platelet-derived growth factor (PDGF): rat neuronal cells possess functional PDGF beta-type receptors and respond to PDGF.Proc. Natl. Acad. Sci. USA 888159–8163.

    PubMed  Google Scholar 

  • Sporn, M. B., and Roberts, A. B. (1992). Transforming growth factor-beta: Recent progress and new challenges.J. Cell Biol. 1191017–1021.

    PubMed  Google Scholar 

  • Stockli, K. A., Lillien, L. E., Näher-Noe, M., Bratfeld, G., Hughes, R. A., Raff, M. C., Thoenen, H., and Sendtner, M. (1991). Regional distribution, developmental changes, and cellular localization of CNTF-mRNA and protein in the rat brain.J. Cell Biol. 115447–459.

    PubMed  Google Scholar 

  • Taga, T., and Kishimoto, T. (1992). Cytokine receptors and signal transduction.FASEB J. 63387–3396.

    PubMed  Google Scholar 

  • Ullrich, A. (1991). Insulin-like growth factor I receptor cDNA cloning.Methods Enzymol. 19817–26.

    PubMed  Google Scholar 

  • Ullrich, A., Coussens, L., Hayflick, J. S., Dull, T. J., Gray, A., Tam, A. W., Lee, J., Yarden, Y., Libermann, T. A., Schlessinger, J., Downward, J., Mayes, E. L. V., Whittle, N., Waterfield, M. D., and Seeburg, P. H. (1984). Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells.Nature 309418–425.

    PubMed  Google Scholar 

  • Unsicker, K., Grothe, C., Otto, D., and Westermann, R. (1991). Basic fibroblast growth factor in neurons and its putative functions.Ann. N.Y. Acad. Sci. 638300–305.

    PubMed  Google Scholar 

  • Valenzuela, D. M., Maisonpierre, P. C., Glass, D. J., Rojas, E., Nuñez, L., Kong, Y., Gies, D. R., Stitt, T. N., Ip, N. Y., and Yancopoulos, G. D. (1993). Alternative forms of rat Trk C with different functional capabilities.Neuron. 10963–974.

    PubMed  Google Scholar 

  • Wang, X.-F., Lin, H. Y., Ng-Eaton, E., Downward, J., Lodish, H. F., and Weinberg, R. A. (1991). Expression, cloning and characterization of the TGF-beta Type III receptor.Cell 67797–805.

    PubMed  Google Scholar 

  • Wennstrom, S., Sandstrom, C., and Claesson-Welsh, L. (1991). cDNA cloning and expression of a human FGF receptor which binds acidic and basic FGF.Growth Factors 4197–208.

    PubMed  Google Scholar 

  • Wesselingh, S. L., Gough, N. M., Finlay-Jones, J. J., and McDonald, P. J. (1991). Detection of cytokine mRNA in astrocyte cultures using the polymerase chain reaction.Lymphokine Res. 9177–185.

    Google Scholar 

  • Westermark, R., Grothe, C., and Unsicker, K. (1990). Basic fibroblast growth factor (bFGF), a multifunctional growth factor for neuroectodermal cells.J. Cell Sci. Suppl. 1397–117.

    PubMed  Google Scholar 

  • Witte, O. N. (1990). Steel locus defines new multipotent growth factor.Cell 635–6.

    PubMed  Google Scholar 

  • Yamamori, T. (1991). Localization of cholinergic differentiation factor/leukemia inhibitory factor mRNA in the rat brain and peripheral tissues.Proc. Natl. Acad. Sci. USA 887298–7302.

    PubMed  Google Scholar 

  • Yarden, Y., Kuang, W.-J., Yang-Feng, T., Coussens, L., Munemitsu, S., Dull, T. J., Chen, E., Schlessinger, J., Francke, U., and Ullrich, A. (1987). Human proto-oncogene c-kit: A new cell surface receptor tyrosine kinase for an unidentified ligand.EMBO J. 63341–3351.

    PubMed  Google Scholar 

  • Zaheer, A., Fink, B., and Lim, R. (1993). Expression of glia maturation factorβ mRNA and protein in rat organs and cells.J. Neurochem. 60914–920.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurology (Division of Neurochemistry and Neurobiology), The University of Iowa College of Medicine and Veterans Affairs Medical Center, 52242, Iowa City, Iowa

    Asgar Zaheer, Weixiong Zhong, Ergun Y. Uc & Ramon Lim

  2. The University of Iowa DNA Facility, 52242, Iowa city, Iowa

    David R. Moser

Authors
  1. Asgar Zaheer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Weixiong Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ergun Y. Uc
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. David R. Moser
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ramon Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zaheer, A., Zhong, W., Uc, E.Y. et al. Expression of mRNAs of multiple growth factors and receptors by astrocytes and glioma cells: Detection with reverse transcription-polymerase chain reaction. Cell Mol Neurobiol 15, 221–237 (1995). https://doi.org/10.1007/BF02073330

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02073330

Key words

Search

Navigation