Regulation of acetylcholine release from neuroblastoma x glioma hybrid cells.

Abstract

Neuroblastoma X glioma NGIOS-15 hybrid cells exposed to N6,02'-dibutyryladenosine 3':5'-cyclic mono- (1), and the fetal bovine serum concentration was decreased from 10% to 5%. Cells to be used for experiments were dissociated and transferred to 200-Ml disposable glass capillary pipettes (total volume, 300 gl) bent in the form of a "U" and connected in series (1.5-2.0 X 105 cells per capillary) and incubated for 3 hr at 370 to promote cell attachment to the glass. The tubes then were perfused with 2 ml of medium per hr for 2 days unless indicated otherwise. and other 3H-labeled metabolites of choline were to be determined, the tubes were perfused for 1.5 min (0.7 ml/min) with medium A, a small air bubble was introduced into the line, and the capillary was filled with acetone/1 M formic acid, 85:15 (vol/vol) The 3H-labeled compounds extracted from cells were fractionated by high-voltage paper electrophoresis with 1.5 M acetic Abbreviations: Bt2cAMP, N6,02'-dibutyryladenosine 3':5'-cyclic monophosphate; ACh, acetylcholine; PG, prostaglandin; DMEM, Dulbecco's modification of Eagle's minimum essential medium; 5-HT, 5-hydroxytrypamine (serotonin).

Neurotransmitter release: vacuolar ATPase V0 sector c-subunits in possible gene or cell therapies for Parkinson’s, Alzheimer’s, and psychiatric diseases

“…N18TG-2 neuroblastoma cells were typical such cell type. Contrarily, other culture cells were very competent for ACh release provided they were loaded with the transmitter: the C6BU-1 glioma cell line followed by the fibroblastic L cell line, PC12, Neuro 2A and NG108-15 cells [13][14][15][21][22][23][24][25]. Interestingly, the ACh release mechanism that is expressed by the latter cells was calcium dependent, when release was elicited by an influx of calcium with the calcium ionophore A23187 [11,13,16].…”

“…Of particular interest is the series of work of Higashida and his colleagues who studied in co-culture with myocytes and neuroblastoma (Neuro 2A) or hybrid cell lines (NG108-15, N18TG-2 x C6Bu-1; NBr-10A, N18TG-2 x BRL-30E liver) [19][20][21][22][23], in which the detected release was measured as endplate potential of the ACh receptor origin [24]. The N18TG2 cells that were transfected with ChAT was insufficient to induce transmission at the synapse in co-cultures, and were unable to elicit miniature endplate potentials, as similarly as those experiments after loading them with ACh using the chemiluminescent ACh detection [16,21,22,27].…”

“…The N18TG2 cells that were transfected with ChAT was insufficient to induce transmission at the synapse in co-cultures, and were unable to elicit miniature endplate potentials, as similarly as those experiments after loading them with ACh using the chemiluminescent ACh detection [16,21,22,27]. In Table 1 opposite cell lines are listed: One has a very efficient ACh release mechanism (the C6BU-1 glioma cells);…”

A factor from neurons increases the number of acetylcholine receptor aggregates on cultured muscle cells.

“…In the three experiments described in Table 1, A-C, mouse myotube cultures treated with control medium had an average A/M of 0.335 (SEM = 0.032, n = 6), whereas cultures treated with NG108-15 cell-conditioned medium had an average A/M of 1.17 (SEM = 0.037, n = 6). (17). However, the addition of AcCho (1 mM) to muscle cultures in control medium did not increase the A/M, but decreased it to one-third of the control A/M.…”

Neuropeptide Y in neuroblastoma × glioma hybrid cells

“…The NG108-15 cell line possesses many neuronal properties including eIectricaI excitability [lt2] mediated by activation of specific sodium charmels f3] and the ability to form synapses in vitro [ 1, 2,4]. These cells synthesize the neurotransmitter acetylcholine which is rekeased in response to stimulation [5]. The cells were derived by hybridization of the N18TG2 mouse neuroblastoma with the C6BU-1 rat gIioma clone.…”