A mouse neuroblastoma x rat glioma hybrid cell produces immunoreactive substance P-like material

A new hybrid cell line, designated NG115-401, produced a significant amount of immunoreactive (IR) substance P-like material. The production was measured by radioimmunoassay using specific antisera raised to substance P. The production rate was dependent upon cell growth, and maximum production occurred at the same stage (late logarithmic stage) as the time of appearance of choline acetyltransferase activity, one of the characteristic neuronal properties of the cells. On the other hand, no IR-substance P was found in hybrid cells other than NG115-401, i.e. NG115-301 and NG115-303 cells, which were derived from the same parent mouse neuroblastoma N115TG-2 cells. Furthermore, in an attempt to survey IR-substance P production in several clonal cells, consisting of some mouse neuroblastoma cells and a rat glioma cell, and several of their hybrid cells, we could find no cell line producing a significant amount of IR-substance P except for NG115-401 hybrid cells. The cellular production of IR-substance P in NG115-401 hybrid cells was also confirmed by detecting the material in all 14 subpopulations of this hybrid cells. Partial characterization of IR-substance P-like material produced by NG115-401 hybrid cells was carried out by its application to gel-filtration column chromatography. Only high molecular weight material (approximately 26,000) appeared in the column eluates.