Advertisement

Cell and Tissue Research

, Volume 262, Issue 3, pp 431–443 | Cite as

Somatostatin-binding sites on rat telencephalic astrocytes

Light- and electron-microscopic studies in vitro and in vivo
  • Rolf Mentlein
  • Cornelia Buchholz
  • Brigitte Krisch
Article

Summary

Using a somatostatin-gold conjugate as ligand, high-affinity binding sites for this neuropeptide were demonstrated at three levels: (i) cultured astrocytes from rat cortex, (ii) hippocampal slice cultures, and (iii) frozen tissue sections of rat telencephalon. The conjugate proved as active as the native peptide in competing for the binding sites. Light-microscopic visualization of bound ligand was achieved by silver intensification of the colloidal gold. This method is faster and yields superior resolution compared with autoradiography. Cultured astrocytes from cortex and hippocampus could be labeled by the ligand. At the light- and electron-microscopic level, astrocytes could be double-labeled by the somatostatin-gold conjugate and immunostaining for glial fibrillary acidic protein (GFAP). In hippocampal slice cultures, the conjugate did not penetrate into the neuropil because of a covering glial layer. However, a portion of this completely GFAP-positive covering glia reacted with the somatostatin ligand. In frozen brain sections, apart from delicate punctate structures, two types of labeled glia cells were seen: single stellate astrocytes and perivascular glia cells.

Key words

Astrocytes Telencephalon Receptors, membrane Somatostatin (SRIF) Rat (Han: WIST) 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ackermann GA, Yang J, Wolken KW (1983) Differential surface labeling and internalization of glucagon by peripheral leukocytes. J Histochem Cytochem 31:433–440Google Scholar
  2. Balazs R, Patel AJ, Richter D (1972) Metabolic compartments in the brain: their properties and relation to morphological structures. In: Balazs R, Cremer JE (eds) Metabolic compartmentation in the brain. Macmillan, New York, pp 167–184Google Scholar
  3. Calker D van, Müller M, Hamprecht B (1980) Regulation by secretin, vasoactive intestinal peptide, and somatostatin of cyclic AMP accumulation in cultured brain cells. Proc Natl Acad Sci USA 77:6907–6911Google Scholar
  4. Cernik A, Petrack B (1983) Somatostatin receptor binding in rat cerebral cortex: characterization using a non reducible somatostatin analog. J Biol Chem 256:5525–5530Google Scholar
  5. Cholewinski AJ, Wilkin GP (1988) Astrocytes from forebrain, cerebellum, and spinal cord differ in their responses to vasoactive intestinal peptide. J Neurochem 51:1626–1633Google Scholar
  6. Enjalbert A, Rasolonjanahary R, Moyse E, Kordon C, Epelbaum J (1983) Guanine nucleotide sensitivity of (125I)-iodo-N-Tyr somatostatin binding in rat adenohypophysis and cerebral cortex. Endocrinology 113:822–824Google Scholar
  7. Gähwiler BH (1984) Slice cultures of cerebellar, hippocampal and hypothalamic tissue. Experientia 40:235–308Google Scholar
  8. Hansson E (1988) Astroglia from defined brain regions as studied with primary cultures. Prog Neurobiol 30:369–397Google Scholar
  9. Hazum E (1986) Preparation and use of biotinylated neuroendocrine peptides. In: Conn PM (ed) Methods in enzymology, vol 124. Academic Press, Orlando, pp 47–57Google Scholar
  10. Hazum E, Cuatrecas P, Marian J, Conn PM (1980) Receptor-mediated internalization of fluorescent gonadotropin-releasing hormone by pituitary gonadotropes. Proc Natl Acad Sci USA 77:6692–6695Google Scholar
  11. Hertz L, Schousboe A (1986) Role of astrocytes in compartmentation of amino acid and energy metabolism. In: Fedoroff S, Vernadakis A (eds) Astrocytes, vol 2. Academic Press, Orlando, pp 179–208Google Scholar
  12. Herzog V, Farquhar MG (1983) Use of electron-opaque tracers for studies on endocytosis and membrane recycling. In: Fleischer S, Fleischer B (eds) Methods in enzymology, vol 98. Academic Press, Orlando, pp 203–225Google Scholar
  13. Jennes L, Conn PM, Stumpf WE (1986) Synthesis and use of colloidal gold-coupled receptor ligands. In: Conn PM (ed) Methods in enzymology, vol 124. Academic Press, Orlando, pp 36–47Google Scholar
  14. Kimura N (1989) Developmental change and molecular properties of somatostatin receptors in the rat cerebral cortex. Biochem Biophys Res Commun 160:72–78Google Scholar
  15. Krisch B (1980) Immunocytochemistry of neuroendocrine systems. Prog Histochem Cytochem 13:1–163Google Scholar
  16. Krisch B, Buchholz C, Mentlein R (1990) Somatostatin-binding sites on rat diencephalic astrocytes. Light-microscopic study in vitro and in vivo. Cell Tissue Res (submitted)Google Scholar
  17. Lucius R, Mentlein R (1989) Inactivation of the neuropeptide somatostatin by brain cells and membranes. Biol Chem Hoppe-Seyler 370:1001–1002Google Scholar
  18. Markstein R, Stoeckli KA, Reubi JC (1989) Differential effects of somatostatin on adenylate cyclase as functional correlate for different brain somatostatin receptor subpopulations. Neurosci Lett 104:13–18Google Scholar
  19. Maurer R, Reubi JC (1985) Brain somatostatin receptor subpopulation visualized by autoradiography. Brain Res 333:178–181Google Scholar
  20. McCarthy KD, Vellis J de (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85:890–902Google Scholar
  21. Mentlein R, Buchholz C, Krisch B (1989) Binding and internalization of gold-conjugated somatostatin and growth hormone-releasing hormone in cultured rat somatotropes. Cell Tissue Res 258:309–317Google Scholar
  22. Milner TA, Bacon CE (1989) Ultrastructural localization of somatostatin-like immunoreactivity in the rat dentate gyrus. J Comp Neurol 290:544–560Google Scholar
  23. Palacios JM, Dietl MM (1989) Regulatory peptide receptors: visualization by autoradiography. In: Polak JM (ed) Regulatory peptides. Birkhäuser Verlag, Basel Boston Berlin, pp 70–97Google Scholar
  24. Palacios JM, Reubi JC, Maurer R (1986) Somatostatin receptors in rat hippocampus: localization to intrinsic neurons. Neurosci Lett 67:169–174Google Scholar
  25. Pelletier G, Leroux P, Morel G (1986) Localization of somatostatin receptors. In: Conn PM (ed) Methods in enzymology, vol 124. Academic Press, Orlando, pp 607–617Google Scholar
  26. Reubi JC, Maurer R (1986) Different ionic requirements for somatostatin receptor subpopulations in the brain. Regul Pept 14:301–311Google Scholar
  27. Roach PD, Zollinger M, Noel S-P (1987) Detection of the low density lipoprotein (LDL) receptor on nitrocellulose paper with colloidal gold-LDL conjugates. J Lipid Res 28:1515–1521Google Scholar
  28. Sakamoto C, Nagao M, Matozaki T, Nishizaki H, Konda Y, Baba S (1988) Somatostatin receptors on rat cerebrocortical membranes. Structural characterization of somatostatin-14 and somatostatin-28 receptors and comparison with pancreatic type receptors. J Biol Chem 263:14441–14445Google Scholar
  29. Slot JW, Geuze HJ (1985) A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol 38:87–93Google Scholar
  30. Smith RM, Jarett L (1985) The preparation of biologically active monomeric ferritin-insulin and its use as a high resolution electron-microscopic marker of occupied insulin receptors. In: Birnbauer L, O'Malley W (eds) Methods in enzymology, vol 109. Academic Press, Orlando, pp 187–203Google Scholar
  31. Srikant CB, Patel YC (1981) Somatostatin receptors: identification and characterization in rat brain membranes. Proc Natl Acad Sci USA 78:3930–3934Google Scholar
  32. Srikant CB, Dahan A, Craig C (1990) Receptor binding of somatostatin-14 and somatostatin-28 in rat brain: differential modulation by nucleotides and ions. Regul Pept 27:181–194Google Scholar
  33. Swanson LW, Köhler C Björklund A (1987) The limbic region. I: The septohippocampal system. In: Björklund A, Hökfelt T, Swanson LW (eds) Handbook of chemical neuroanatomy, vol 5: integrated systems of the CNS, part I. Elsevier, Amsterdam, pp 125–277Google Scholar
  34. Tran VT, Uhl GR, Perry D, Manning DC, Vale W, Perrin MH, Martin JB, Snyder SH (1984) Autoradiographic localization of somatostatin receptors in rat brain. Eur J Pharmacol 101:307–309Google Scholar
  35. Tran VT, Beal MF, Martin JB (1985) Two types of somatostatin receptors differentiated by cyclic somatostatin analogs. Science 228:492–495Google Scholar
  36. Walz W (1989) Role of glia cells in the regulation of the brain microenvironment. Prog Neurobiol 33:309–333Google Scholar
  37. Zoelen EJJ van (1989) Receptor-ligand interaction: a new method for determining parameters without a priori assumptions on non-specific binding. Biochem J 262:549–556Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Rolf Mentlein
    • 1
  • Cornelia Buchholz
    • 1
  • Brigitte Krisch
    • 1
  1. 1.Anatomisches Institut der Universität KielKielFederal Republic of Germany

Personalised recommendations