Calcium signals in cell lines derived from the cerebral cortex of normal and trisomy 16 mice

Abstract

We established two immortalized cell lines from cerebral cortex of normal (CNh) and trisomy 16 (CTb) mouse fetuses, an animal model of human trisomy 21. Those cells loaded with the fluorescent Ca2+ dyes, Indo-1 and Fluo-3, exhibited increments of intracellular Ca2+ ([Ca2+]i) in response to external glutamate, NMDA, AMPA and kainate. CTb cells exhibited higher basal Ca2+ concentrations and had higher amplitude and slower time-dependent kinetics in the decay than CNh cells, suggesting an impaired Ca2+ buffering capacity in the trisomy 16-derived cell line. Nicotine also induced increments of [Ca2+]i. The CTb cell line could represent a model for studying cellular alterations related to Down syndrome.

Corticosterone down-regulates dopamine D4 receptor in a mouse cerebral cortex neuronal cell line

“…However, D 4 receptor protein expression was observed to be mainly associated to caudate putamen and hippocampus, suggesting that the D 4 receptor protein found in these areas probably represents presynaptic receptors of input neurons whose somas are located in areas rich in D 4 mRNA, like the frontal cortex, amygdala or thalamus . Cárdenas et al (1999) established continuously growing cell lines from the cerebral cortex of a normal fetal mouse using an original method (Liberona et al, 1997). The cell line retains differentiated traits, expressed in the presence of neuronal markers and lack of glial traits by immunohistochemistry after almost 6 years in continuous culture (Cárdenas et al, 1999).…”

“…Cárdenas et al (1999) established continuously growing cell lines from the cerebral cortex of a normal fetal mouse using an original method (Liberona et al, 1997). The cell line retains differentiated traits, expressed in the presence of neuronal markers and lack of glial traits by immunohistochemistry after almost 6 years in continuous culture (Cárdenas et al, 1999). In the present work, we have employed this cerebral cortex cell line to investigate the effect of different concentrations of corticosterone on the expression of D 4 DA receptors.…”

“…The CNh cell line was originally prepared from cerebral cortex dissected from mouse fetuses (Cárdenas et al, 1999). The cells are immunohistochemically positive to several neuronal markers such as MAP-2, synaptophysin, neuron specific enolase (NSE), choline acetyl transferase, and lack glial traits such as glial fibrillary acid protein (GFAP), S-100 and galactocerebroside, after almost 6 years in continuous culture (Cárdenas et al, 1999).…”

“…cm 2 culture flasks to a density of 75% confluence. The culture medium consisted of DME/F-12 (1:1) modified to contain 6 g/l glucose, 10% adult bovine serum, 1 g/l sodium bicarbonate, 2.5% fetal bovine serum, 1000U/ml penicillin, and 1000 µg/ml streptomycin (Cárdenas et al, 1999). Cells were detached by trypsinization with 0.1% trypsin (w/v) and aliquots of this suspension were seeded onto polylysinated round glass coverslips placed in multiwell plates (24 wells, 48,000 cells/well) for corticosterone incubation and later immunohistochemical analysis.…”

“…2). f Immortalized cell lines from the cerebral cortex of normal mouse fetuses has been successfully established and characterized by Cárdenas et al (1999) and Allen et al (2000). This CNh cell line was reported to express several functional neuronal properties like the expression of NMDA and AMPA/kainate receptors, evidenced by elevations of the levels of intracellular Ca 2+ in response to glutamate, AMPA, kainate, nicotine, muscarine and GABA (Cárdenas et al, 1999).…”