NMDA and NRG1 do not modify basal migration of wild-type ST14A neural progenitors. A: Agarose gel electrophoresis of RT-PCR amplification products. Wt ST14A cells were cultured in 2% FBS DMEM for 18 hrs before total RNA isolation. Splicing-specific primer pairs were employed to recognize N-terminal (1.a/1.b) and C-terminal (1.1/1.2; 1.3/1.4) splicing sites of the NR1 subunit. Wt ST14A express NR1.4a (combination of 1.a N-term and 1.4 C-term splicing forms, see arrows), NR2C and NR2D subunits. No PCR product was observed in the absence of reverse transcriptase (RT-) or cDNA (C-, water). "C-" reactions contained all primer pairs. C+ = various brain regions from adult rats. B: NMDA dose-response assay on wt ST14A cells. Cells were incubated in increasing concentrations of NMDA (1, 2, 4, 10 μM) in 2% FBS DMEM for 18 hrs. Data are expressed as mean + standard deviation (SD, n = 3). C: Effect of NMDA on wt ST14A cell migration measured by transwell assays performed in 2% FBS DMEM for 18 hrs with different concentrations of NMDA. D: wt ST14A migration was measured by transwell assay with addition of NMDA (8 μM) and NRG1 (5 nM) either alone or in combination. Representative images of migrated cells are shown above each bar/treatment. Migration was calculated as percentage of total migrated cells (see Material and Methods). Biological triplicate experiments were carried out in technical triplicate. Data are presented as means + SD. No statistically significant difference was found among different treatments.
Pregno et al. BMC Neuroscience 2011 12:103 doi:10.1186/1471-2202-12-103