Two neuronal cell lines expressing the myelin basic protein gene display differences in their in vitro survival and in their response to glia

Abstract

We have generated two conditionally immortalized neuronal cell lines from primary cultures of embryonic day 13 (E13) and postmitotic (postnatal day 0; P0) cortical neurons transformed with the temperature-sensitive SV-40 large-T antigen. Two clonal cell lines (CN1.4 from E13 cultures and SJ3.6 from P0 cultures) were isolated and stable maintained in vitro. Both cell lines expressed a number of neuronal markers such as the neurofilaments, glutamic acid decarboxylase 67, neuron-specific enolase, and the BG21 isoform of the myelin basic protein gene. At 34 degrees C, the CN1.4 cell line had elaborated short processes, whereas the SJ3.6 cell line produced long processes that formed a delicate network. When these cell lines were cultured at 39 degrees C, some of the cellular processes grew longer, adopting a more mature neuronal morphology. Interestingly, at 39 degrees C, the in vitro survival of these cell lines differed significantly. Whereas the survival of CN1.4 cell line was greatly unaffected, SJ3.6 cells died soon after they were cultured at 39 degrees C. The cell death of SJ3.6 cells was accompanied by fragmentation and condensation of DNA in their nuclei, indicative of an apoptotic event. Under these conditions, SJ3.6 showed an upregulation of the p75 receptor. When this cell line was cocultured with oligodendrocytes, astrocytes, or glial conditioned media (GCM), there was a marked increase in survival. In contrast, little effect of glial cells or GCM was observed on the CN1.4 cell line. These lines appear to be useful models to study neuronal-glial interactions in addition to neuronal cell death and the effects of glial factors that promote the survival of neurons.

Immortalized cortical neurons expressing caspase-cleaved tau are sensitized to endoplasmic reticulum stress induced cell death

“…Immortalized cortical neurons (Bongarzone et al, 1998) expressing inducible full-length (T4) or Asp421 truncated (T4C3) tau were prepared as described previously (Krishnamurthy and Johnson, 2004). Cells were cultured in DMEM with High Glucose (Mediatech) and supplemented with 5% FBS, 0.1% gentamicin, and 4 mM glutamine, penicillin, and streptomycin at 33 °C.…”

Identification of a New Exon in the Myelin Proteolipid Protein Gene Encoding Novel Protein Isoforms That Are Restricted to the Somata of Oligodendrocytes and Neurons

“…Cultures of cerebellar granular neurons were prepared from postnatal day 7 (P7) cerebellar cortices as described elsewhere (Gao et al, 1991). The L BRM T-cell line (a gift of Dr. J. Merrill) and the C N1.4 neuronal line (Bongarzone et al, 1998b) were grown in 10% FC S-DM EM for 4 d before collection.…”

Cytoarchitecture of the mouse neocortex revealed by the low-molecular-weight neurofilament protein subunit

“…This commercial antiserum has been used in fluorescence immunocytochemical studies on rodent brainderived cell cultures (Bongarzone et al 1998;Evans et al 2002) and to characterize the similarities of NFPs in different species (Shaw et al 1984(Shaw et al , 1986. To assess its specificity in mouse, Western blotting was performed on different tissue samples (Fig.…”