Coexistence of cholinergic, catecholaminergic, serotonergic, and glutamatergic neurotransmitter markers in mouse clonal hybrid neurons derived from the septal region

Abstract

Two clonal immortalized neurons designated SN6.1b and SN6.2a were isolated by limiting dilution from a mouse embryonic septal cholinergic neuronal hybrid cell line SN6 (Hammond et al., 1986). In the serum-containing medium without extra differentiating agents, one-third of SN6.1b cells stably exhibited a morphology of differentiated neurons with extensive elaborate neurites, while a majority of SN6.2a cells, along with the parent cell line SN6, were round in shape with poorly branched short processes. Neurochemical studies showed that both clones synthesized choline acetyltransferase (ChAT), dopamine, norepinephrine, serotonin, and glutamate. Immunocytochemically, they expressed a number of neuronal antigens, such as 200-kDa neurofilament protein, neuron-specific enolase, microtubule-associated protein 2, tau protein, tubulin, neural cell adhesion molecule, Thy-1.2, saxitoxin-binding sodium channel protein, ChAT, tyrosine hydroxylase, serotonin, and glutamate. The coexistence of cholinergic, catecholaminergic, serotonergic, and glutamatergic neurotransmitter markers in the clonal hybrid septal neurons that express a variety of immunocytochemical properties of differentiated neurons suggests that embryonic septal cholinergic neurons are potentially multiphenotypic with respect to neurotransmitter synthesis.

HSP72 induction by heat stress is not universal in mammalian neural cell lines

“…SN6. l b is a subclone isolated from the SN6 cell line by limiting dilution in our laboratory (Satoh et al, 1992b). Both CL8c4.7 and CL8a5.2 clones were generated by somatic cell fusion between N 18TG2 and mouse cerebelladbrain stem neurons (Satoh et al, 1992~).…”

“…l b cells consist of two morphologically distinct cell types: small polygonal cells with short processes and large multinucleated cells with long elaborate processes. This heterogeneity may be attributable to the cells in different stages of the cell cycle (Satoh et al, 1992b). The level of HSP72 expression was not different between both cell types.…”

“…C92F3B-1 specifically recognizes stress-inducible 72-kDa protein (HSP72) but does not react with constitutive 73-kDa protein (HSC73) (Welch and Suhan, 1986;Vass et al, 1988;Gonzalez et al, 1989;Marini et al, 1990), while N27F3-4 recognizes both HSP72 and HSC73 (Kost et al, 1989). Anti-tubulin antibody that labels microtubules distributed in the cytoplasm in both N 18TG2 and SN6.1b cells (Satoh et al, 1992b) was used to identify the whole cell contour. The cells were washed and incubated with a mixture of rhodamine-conjugated antimouse IgG (1:80; Organon Teknika, West Chester, PA) and fluorescein isothiocyanate (F1TC)-conjugated antirabbit immunoglobulin (1:XO; Organon Teknika).…”

Neurotransmitter synthesis by SN6 cell lines, a family of hybrid cell lines of embryonic septal origin

“…Further detailed characteristics of the SN6 cell line remain unclear, although it was found to respond to trophic factors (Hammond et al, 1990) including the hemopoietic factors erythropoietin and erythroid differentiation factor Masuda et al, 1993), interleukin-3, and granulocytemacrophage colony-stimulating factor (Kamegai et al, 1990a,b). We found the presence of multiple neurotransmitters in the two subclones (Satoh et a]., 1992). To Transmitter Synthesis by SN6 Cell Lines 785 min at 2°C.…”

“…In a previous study (Satoh et al, 1992), we described the isolation of two different subclones, designated as SN6.1b and SN6.2a, from a cholinergic hybrid cell line, SN6.10.2.2. The mother cell line SN6 (Hammond et al, 1986) was established by somatic cell fusion between the mouse neuroblastoma N18TG2 (Amano et al, 1972) and embryonic septal cholinergic neurons.…”