Abstract
The CNS-derived cell line, CAD cell line, when grown in a protein free medium (PFM), differentiates to neuron-like cells with very long processes. It was previously studied biochemically and found to express TH activity, some neurospecific proteins, but no glial proteins. We have now further studied the CAD cells and focused on the expression of various neuropeptides, GAP-43 and GFAP. All peptides studied were present, including TH, but also GFAP, in contrast to earlier studies. A different kind of processes, short, slender and distributed like a “fringe” around cell body and along processes was observed, NESP55 but not other chromogranins was present in these “fringes”, GAP43 showed some degree of overlapping with NESP55. The results show that even after differentiation in PFM, the CAD cells express a palette of neuropeptides and chromogranins, catecholaminergic markers as well as the glia-specific GFAP. Our efforts to induce exocytosis/endocytosis from the peptide granules by high K+ were, however, unsuccessful. Due to long processes, the CAD cells may represent a good model for studying intracellular transport, and, since the cells express both neuronal and glial characteristics, it may be useful for investigating the influence of different trophic/growth factors on the expression of various neuronal characteristics.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Allen RD, Weiss DG, Hayden JH, Brown DT, Fujiwake H, Simpson M (1985) Gliding movement of and bidirectional transport along single native microtubules from squid axoplasm: evidence for an active role of microtubules in cytoplasmic transport. J Cell Biol 100:1736–1752
Bradford MM (1976) A rapid and sensitive method for the quntitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Calakos N, Scheller RH (1996) Synaptic vesicle biogenesis, docking, and fusion; a molecular description. Physiol Rev 76:1–29
Castaneda-Castellanos DR, Cano M, Wang JK, Corbett A, Benson D, Blanck TJ, Thornhill WB, Recio-Pinto E (2000) CNS voltage-dependent Na channel expression and distribution in an undifferentiated and differentiated CNS cell line. Brain Res 866:281–285
Chanat E, Pimplikar SW, Stinchcombe JC, Huttner WB (1991) What the granins tell us about the formation of secretory granules in neuroendocrine cells. Cell Biophys 19:85–91
Dahlstrom A, Li JY (1994) Fast and slow axonal transport—different methodological approaches give complementary information: contributions of the stop-flow/crush approach. Neurochem Res 19:1413–1419
Dahlstrom A, Czernik AJ, Li JY (1992) Organelles in fast axonal transport—what molecules do they carry in anterograde versus retrograde directions, as observed in mammalian systems? Mol Neurobiol 6:157–177
De Potter WP, Kurzawa R, Miserez B, Coen EP (1995) Evidence against diferential release of noradrenaline, neuropeptide Y, and dopamine-beta-hydroxylase from adrenergic nerves in the isolated perfused sheep spleen. Synapse 19:67–76
Ferro-Novick S, Jahn R (1994) Vesicle fusion from yeast to man. Nature 370:191–193
Hashemi SH, Li JY, Faigle R, Dahlstrom A (2003) Adrengergic differentiation and SSR receptor expression in CAD cells cultured in serum-free medium. Neurochem Int 42:9–17
Jakobsen AM, Ahlman H, Kolby L, Abrahamsson J, Fischer-Colbrie R, Nilsson O (2003) NESP55, a novel chromograinin-like peptide, is expressed in endocrine tumours of the pancreas and adrenal medulla but not in ileal carcinoids. Br J Cancer 88:1746–1754
Li JY, Kling-Petersen A, Dahlstrom A (1992) Influence of spinal cord transaction on the presence and axonal transport of CGRP-, chromogran A-, VIP-, synapsin- and synaptophysin-like immunoreactivities in rat motor nerve. J Neurobiol 23:1094–1110
Li JY, Leitner B, Lovisetti-Scamihorn P, Winkler H, Dahlstrom A (1999) Proteolytic processing, axonal transport and differential distribution of chromogranins A and B, and secretogranin II (secretoneurin) in rat sciatic nerve and spinal cord. Eur J Neurosci 11:528–544
Natori S, Huttner WB (1994) Peptides derived form the granins (chromogranins/secretogranins). Biochem 76:277–282
Ozawa H, Takata K (1995) The granin family—its role in sorting and secretory granule formation. Cell Struct Funct 20:415–420
Qi Y, Wang JK, McMillian M, Chikaraishi DM (1997) Characterization of a CNS cell line, in which morphological differentiation is initiated by serum deprivation. J Neurosci 17:1217–1225
Rothman JE, Warren G (1994) Implications of the SNARE hypothesis for intracellular membrane topology and dynamics. Curr Biol 4:220–223
Schubert D, Heinemann S, Carlisle W, Tarikas H, Kimes B, Patrick J, Steinbach JH, Culp W, Brandt BL (1974) Clonal cell lines from the rat central nervous system. Nature 249:224–227
Suri C, Fung BP, Tischler AS, Chikaraishi DM (1993) Catecholaminergic cell lines from the brain and adrenal glands of tyrosine hydroxylase-SV40 T antigen transgenic mice. J Neurosci 13:1280–1291
Thureson-Klein AK, Klein RL (1990) Exocytosis from neuronal large dense-cored vesicles. Int Rev Cytol 121:67–126
Vale RD, Reese TS, Sheetz MP (1985) Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell 42:39–50
Wang H, Oxford GS (2000) Voltage-dependent ion channels in CAD cells: A catecholaminergic neuronal line that exhibits inducible differentiation. J Neurophysiol 84:2888–2895
Acknowledgements
This study was supported by the Göteborg Medical Society, the Swedish Medical Research Council (14X-2207), the Medical Faculty, Göteborg University and the Royal Society of Arts and Science in Göteborg. We are grateful to Drs. H. Winkler and R. Fischer-Colbrie (Department of Pharmacology, University of Innsbruck, Innsbruck, Austria) for a generous supply of antisera against Chromogranins and to Ellinor Andersson (Lundberg Laboratory for Cancer Research, Sahlgrenska University Hospital, Gothenburg, Sweden) for technical help. We also thank Ms. Eva Lyche for secretarial help.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, Y., Hou, L.XE., Aktiv, A. et al. Immunohistochemical characterisation of differentiated CAD cells: expression of peptides and chromogranins. Histochem Cell Biol 124, 25–33 (2005). https://doi.org/10.1007/s00418-005-0017-9
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-005-0017-9