Characterization of a CNS cell line, CAD, in which morphological differentiation is initiated by serum deprivation.

Published

Journal Article

A CNS catecholaminergic cell line, Cath.a, was established by targeted oncogenesis in transgenic mice. Cath.a cells express neuronal properties but lack neuronal morphology. Here, we describe a variant of Cath.a, called CAD (Cath.a-differentiated), in which reversible morphological differentiation can be initiated by removal of serum or exogenously added protein from the medium. In serum- or protein-free media, CAD cells stop proliferating and extend long processes. Differentiated CAD cells can be maintained without serum or protein for at least 6 weeks. CAD cells are distinct from Cath.a cells; most significant, the original immortalizing oncogene, SV40 T antigen, was spontaneously lost. By immunostaining or immunoblotting, we show that CAD cells express neuron-specific proteins, such as class III beta-tubulin, GAP-43, SNAP-25, and synaptotagmin, but not GFAP. Ultrastructurally, processes from differentiated CAD cells have abundant parallel microtubules and intermediate filaments, and bear varicosities that contain both large dense-core vesicles/granules (120-160 nm) and smaller clear vesicles (60-80 nm). Additionally, CAD cells express enzymatically active tyrosine hydroxylase and accumulate L-DOPA. CAD cells exhibit biochemical and morphological characteristics of primary neurons and provide an unique tool for studying neuronal differentiation.

Full Text

Duke Authors

Cited Authors

  • Qi, Y; Wang, JK; McMillian, M; Chikaraishi, DM

Published Date

  • February 15, 1997

Published In

Volume / Issue

  • 17 / 4

Start / End Page

  • 1217 - 1225

PubMed ID

  • 9006967

Pubmed Central ID

  • 9006967

International Standard Serial Number (ISSN)

  • 0270-6474

Language

  • eng

Conference Location

  • United States