Acetylcholine synthesis and release is enhanced by dibutyryl cyclic AMP in a neuronal cell line derived from mouse septum

Abstract

Cholinergic properties of the SN56.B5.G4 cell line derived from the fusion of neurons of the mouse postnatal day 21 septum and the murine neuroblastoma cell line N18TG2 were investigated and correlated with morphological differentiation. In basal serum-containing growth medium, few cells developed neurites. Neurite extension occurred in cells grown for 2 d with forskolin or dibutyryl cAMP (dbcAMP) but not with butyrate. In cells treated with these compounds, the activity of ChAT and ACh content were two- to threefold higher relative to controls. The cells synthesized ACh from choline taken up by the sodium-dependent high-affinity transport. Forskolin-, dbcAMP-, and butyrate-treated cells (but not the controls) were capable of spontaneous and depolarization-evoked ACh release. The results indicate that the morphological and the neurochemical aspects of SN56.B5.G4 cell differentiation are independently regulated.

Immortalized murine striatal neuronal cell lines expressing dopamine receptors and cholinergic properties

“…On the other hand, the fusion of N 18TG2 cells with primary septal cholinergic cells resulted in cell lines which do express ChAT activity (Hammond et al, 1986(Hammond et al, , 1990Lee et al, 1990a), suggesting that the expression of a cholinergic phenotype in cell lines produced by the somatic cell fusion method is most likely a function of the lineage of the primary cells participating in the fusion process. In addition, neither ChAT activity nor ACh synthesis was detected in N 18TG2 cells grown in basal medium or under differentiating conditions as reported by Blusztajn et al (1992). Thus, the expression of ChAT activity and ACh synthesis in the hybrid cells, together with K+-evoked ACh release from X52 cells (Wainwright et al, 1993) most likely reflects the expression of C57BY6J genes from cholinergic interneurons of the corpus striatum.…”

“…The neuroblastoma cell line was previously obtained through chemical mutagenesis of the N 18 neuroblastoma, a subclone of the C 1300 neuroblastoma isolated from an A/J mouse . The N 18TG2 cell line expresses negligible levels of choline acetyltransferase (ChAT) activity (Greene et al, 1975;Hammond et al, 1986Hammond et al, , 1990Blusztajn et al, 1992).…”

Role of Potassium Channels in Amyloid‐Induced Cell Death

“…When differentiated, SN56 cells show high levels of choline acetyltransferase activity (Lee et al, 1990;Blusztajn et al, 1992) and a well developed TEA-sensitive delayed rectifier K~current (IK). Here we present evidence that SN56 cells are susceptible to A/3 toxicity, that A/.3 induces increases in 'K density followed by apoptotic cell death in SN56 cells, and that A~3toxicity could be inhibited by TEA blockade of 'K.…”

“…Cell lines express phenotypes typical of differentiated septal neurons and high molecular weight neurofilament protein. Although several lines express high levels of choline acetyltransferase activity (e.g., SN56), others do not (e.g., SN48) (Lee et al, 1990;Blusztajn et al, 1992).…”

“…Six hours after plating, cells were differentiated by the addition of 1 mM dibutyryl-cyclic AMP. On differentiation, cells stop/slow dividing and were morphologically different, including extensive de novo neurite growth (Lee et al, 1990;Blusztajn et al, 1992).…”