Establishment and characterization of immortalized neuronal cell lines derived from the spinal cord of normal and trisomy 16 fetal mice, an animal model of Down syndrome

Cárdenas AM; Allen, DD; Arriagada C.; Olivares, A.; Bennett, LB; Caviedes R.; Dagnino-Subiabre, A; Mendoza, IE; Segura-Aguilar, J; Rapoport, SI; Caviedes P.

Abstract

We report the establishment of continuously growing cell lines from spinal cords of normal and trisomy 16 fetal mice. We show that both cell lines, named M4b (derived from a normal animal) and MTh (trisomic) possess neurological markers by immunohistochemistry (neuron specific enolase, synaptophysin, microtubule associated protein-2 [MAP-2], and choline acetyltransferase) and lack glial traits (glial fibrillary acidic protein and S100). MTh cells were shown to overexpress mRNA of Cu/Zn superoxide dismutase, whose gene is present in autosome 16. We also studied intracellular Ca2+ signals ([Ca2+]i) induced by different agonists in Indo-1 loaded cells. Basal [Ca2+]i was significantly higher in MTh cells compared to M4b cells. Glutamate (200 ?M) and (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACDP) (100 ?M) induced rapid, transient increases in [Ca2+]i in M4b and MTh cells, indicating the presence of glutamatergic metabotropic receptors. N-methyl-D-aspartate (NMDA) and kainate, but not alphaamino-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), produced [Ca2+]i rises in both cell types. MTh cells exhibited faster time-dependent decay phase kinetics in glutamate-induced responses compared to M4b cells. Nicotine induced a transient increase in [Ca2+]i in M4b and MTh cells, with significantly greater amplitudes in the latter compared to the former. Further, both cell types responded to noradrenaline. Finally, we examined cholinergic function in both cell lines and found no significant differences in the [3H]-choline uptake, but fractional acetylcholine release induced by either K+, glutamate or nicotine was significantly higher in MTh cells. These results show that M4b and MTh cells have neuronal characteristics and the MTh line shows differences which could be related to neuronal pathophysiology in Down's syndrome. © 2002 Wiley-Liss, Inc.

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Título según WOS: Establishment and characterization of immortalized neuronal cell lines derived from the spinal cord of normal and trisomy 16 fetal mice, an animal model of Down syndrome
Título según SCOPUS: Establishment and characterization of immortalized neuronal cell lines derived from the spinal cord of normal and trisomy 16 fetal mice, an animal model of down syndrome
Título de la Revista: JOURNAL OF NEUROSCIENCE RESEARCH
Volumen: 68
Número: 1
Editorial: Wiley
Fecha de publicación: 2002
Página de inicio: 46
Página final: 58
Idioma: English
URL: http://doi.wiley.com/10.1002/jnr.10205
DOI:

10.1002/jnr.10205

Notas: ISI, SCOPUS