Neuroblastoma cells mouse

Sensitive to toxins, in this case means that the assay presents no false negative results. Primary hepatocytes can elucidate hepatotoxins, and mouse neuroblastoma cells can elucidate sodium channel-blocking neurotoxins therefore these assays can be used to screen for the appropriate toxins. 

Tissue Culture Assay. Kogure et al. (48) report a novel tissue culture assay for detecting several types of sodium channel blockers. The mouse neuroblastoma cell line ATCC CCL 131 is grown in RPMI 1640 supplemented with 13.5% fetal bovine serum and 100 pg/ml gentamycin, in an atmosphere of 5% C0 95% air at 37 C. Ninety-six well plates are seeded with 1 x 10 cells in 200 pi of medium containing 1 mM ouabain and 0.075 mM veratridine. Veratridine and ouabain cause neuroblastoma cells to round-up and die. In the presence of sodium channel blockers (e.g., TTXs or STXs), the lethal action of veratridine is obviated and cells retain normal morphology and viability. An important feature of this assay is that a positive test for sodium channel blockers results in normal cell viability. Since bacterial extracts can contain cytotoxic components, this assay offers an advantage over tests that use cell death as an endpoint. The minimum detectable level of TTX is approximately 3 nM, or approximately 1/1000 mouse unit. 

Tamplin et. al. (54) observed that V. cholerae and A. hydrophila cell extracts contained substances with TTX-like biological activity in tissue culture assay, counteracting the lethal effect of veratridine on ouabain-treated mouse neuroblastoma cells. Concentrations of TTX-like activity ranged from 5 to 100 ng/L of culture when compared to standard TTX. The same bacterial extracts also displaced radiolabelled STX from rat brain membrane sodium channel receptors and inhibited the compound action potential of frog sciatic nerve. However, the same extracts did not show TTX-like blocking events of sodium current when applied to rat sarcolemmal sodium channels in planar lipid bilayers. 

CTx that has been purified from muscles of Gymnothorax javanicus stimulates the release of neurotransmitters such as 7-aminobutyric acid and dopamine from rat brain nerve terminals. It causes a membrane depolarization of mouse neuroblastoma cells and, under appropriate conditions, it creates spontaneous oscillations of... 

Richelson, E. (1978). Histamine HI receptor-mediated guanosine 3, 5 -monophosphate formation by cultured mouse neuroblastoma cells. Science 201, 69-71. 

Platelet-activating factor is a transcriptional activator of cyclooxygenase-2. PAF induces mouse COX-2-promoter-driven luciferase activity transfected in neuroblastoma cells, such as NG108-15 or SH-SY5Y, and in NIH 3T3 cells (Fig. 33-5). The intracellular PAF antagonist BN 50730 inhibits PAF activation of this construct [41]. 

Holloway, S.F., V.L. Salgado, C.H. Wu, and T. Narahashi. 1989. Kinetic properties of single sodium channels modified by fenvalerate in mouse neuroblastoma cells. Pflugers Archiv. (European Jour. Physiol.) 414 613-621. 

Fredrickson, P. A., and Richelson, E. (1979) Hallucinogens antagonize histamine H, receptors of cultured mouse neuroblastoma cells. Eur. J. Pharmacol., 56 261-264. 

Narahashi T, Tsunoo A, Yoshii M (1987) Characterization of two types of calcium channels in mouse neuroblastoma cells. J Physiol (Lond) 383 231-249... 

The implementation of animal test protocols in the 1980s has been accompanied by the development of a host of alternative methods to study adverse effects of chemicals on reproductive and developmental parameters. For example, rat whole embryo culture stems from the seventies (16), as does the rat limb bud organ culture (17) and rat limb bud and brain micromass was developed in the eighties (18). An elegant nonvertebrate alternative model used regeneration of polyps of Hydra atUnuata from dissociated cells (19). Animal-free in vitro alternatives include those employing the proliferation of a human embryonic palatal mesenchymal cell line (20), the attachment of a mouse ovarian tumor cell line (21), and the differentiation of a neuroblastoma cell line (22) and a embryonal carcinoma cell line (23). Various overviews of methods have been published over the years (24). The predictability of... 

In contrast to this work on whole brain, Kemp and Stoolmiller138 used cultured mouse-neuroblastoma cells for their study of incorporation of [ lH]2-acetamido-2-deox> -D-mannose into the sialic acid moiety of gangliosides. They were able to demonstrate clearly the precursor-product relationship among CMS, GM2, and GM1. They found that cultured NB41A cells incorporated [3H]2-acetamido-2-deoxy-D-man-nose into the sialic acid moiety of GM3 in less than 10 minutes. Labeled GM2 was not detected in cells incubated for less than 30 minutes, and measurable activity did not appear in G I1 until alter 60 to 90 minutes. These results further supported the concept that the pathway of synthesis of ganglio-tvpe gangliosides proceeds by way of GM3 —> GM2 — GM1. 

The next channel to be dealt with is the Cx40 channel. The unique conductance and gating of gap junction channels formed by Cx40 has been investigated in Cx40-transfected mouse neuroblastoma cells (N2A cells). In... 

WHO (1993) Guidelines for Drinking Water Quality, 2nd Ed., Vol. 1, Recommendations, Geneva Yoda, K., Shimizu, M. Fujimura, S. (1982) Induction of morphological differentiation in cultured mouse neuroblastoma cells by alkylating agents. Carcinogenesis, 3, 1369-1371... 

Lang, D. G., Wang, C. M., Cooper, B. R. Lamotrigine, phenytoin and carbamezepine interactions on the sodium current present in N4TG1 mouse neuroblastoma cells, J. Pharmacol. Exp. Therap. 1993, 266, 829-835. 

The apparent Km values of the enzyme systems for CMP-NeuNAc assayed with 0.5 mg enzyme protein, was 0.13 mM (same with all four types of acceptors (15)). This value is comparable to that (0.15 mM) obtained (22) in cultured mouse neuroblastoma cells with lactosylceramide as the NeuNAc acceptor. The Km value reported previously (23) for the calf brain enzyme with desialy-lated ai-acid glycoprotein as the acceptor is 4-fold higher. 

For some time, the effects of and responses to vitamin E have been interpreted in terms of an antioxidant mechanism of action. However, several observations have raised the question as to whether other mechanisms could be involved. For example, the effects of selenium and vitamin E on growth and polyunsaturated fatty acid synthesis in cultured mouse fibroblasts could not be reproduced by artificial antioxidants [198, 199]. The specific requirement of (+ )-a-toco-pherol for the phenotypic differentiation of the rotifer [200] may not be through an antioxidant mechanism. The effects of vitamin E on differentiation of neuroblastoma cells [201] and metamorphosis of various species [202] are likely to be due to a growth-factor-like action. A study on the interaction... 

Ratio of nitrogen versus phosphate Cell line (Mouse neuroblastoma cells)... 

Before implantation several in vitro tests were performed. For evaluation of a possible toxic reaction, we investigated the material and the whole devices in vitro with cell culture methods. Direct contact and extraction tests with a mouse fibroblasts cell line (L 929) and a neuroblastoma cell line (neuro-2-a) were performed according to the international standard ISO 10993 ( Biological Evaluation of Medical Devices ). The materials and devices showed no toxicity, i.e. no significant differences in membrane integrity of the cell membranes, mitochondrial activity and DNA synthesis rate. The neuro-2-a cell line is so sensitive that even small changes in process technology are detectable. The flexible polyimide structures proved to be non toxic. 

Bisogno, T., Melck, D., De Petrocellis, L., and Di Marzo, V. (1999). Phosphatidic acid as the biosynthetic precursor of the endocannabinoid 2-arachidonoylglycerol in intact mouse neuroblastoma cells stimulated with ionomycin. J. Neurochem. 72, 2113-2119. 

Strikingly, tamoxifen, an anticancer agent in chnical use, significantly inhibited K+ currents and proliferation of mouse neuroblastoma cells NG108-15. The effect of tamoxifen on the cell prohferation was well correlated with the effect of tamoxifen on the resting K+ flux, indicating that the antitumor action of tamoxifen could be due to its interaction with K+ channels (Rouzaire-Dubois and Dubois 1990). 

FIGURE 12 Survival of NB-bearing nude mice after injection of oligonucleotides (free or encapsulated within liposomal formulations). Nude mice were injected intravenously with 3.5 x 106 HTLA-230 neuroblastoma cells. After 4h each mouse received 50 ig of oligonucleotides either free or encapsulated in targeted or nontargeted liposomes. Control mice received HEPES-buffered saline. (Reprinted from ref. 385 with permission of Elsevier.)... 

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