Fig. 2.

Over-expression of CaSR promotes neurite growth in the absence of NGF.

(A) Quantification of CaSR immunofluorescence in E18 SCG neurons 24 h after transfection with either pFLCaSR or pcDNA3.1, mean ± sem (****P < 0.0001, unpaired t-test with Welch’s correction, n = 24 cells per condition). Total length (B) and branch point number (C) of the neurite arbors of E18 SCG neurons transfected with either pcDNA3.1 or pFLCaSR and cultured without NGF for 24 h in medium containing 2.3 mM [Ca²⁺]_o. All cultures received 50 μM Boc-D-FMK. Mean ± sem of data from 592 to 620 neurons per condition. ***P < 0.001, two-tailed, unpaired t-test. (D) Quantification of CaSR immunofluorescence in non-transfected SCG neurons cultured for 24 h with and without 10 ng/ml NGF in media containing either 0.7 mM or 2.3 mM [Ca²⁺]_o. ***P < 0.001, ANOVA with Bonferroni’s post-hoc test (n = 24 per condition). (E) Representative images of SCG neurons double labelled for CaSR and βIII-tubulin after 24 h in media containing 10 ng/ml NGF with either 0.7 mM or 2.3 mM [Ca²⁺]_o. Scale bar = 20 μm.