Fig. 5.

Possible mechanism of action of choline a mRNA expression of selected neurotrophic factors in shCtrl and shMeCP2 neuronal cultures with or without choline supplementation. Expression levels were normalized to the housekeeping gene Gapdh, and expressed relative to shCtrl levels. n = 5 cultures. Bdnf, Brain-derived neurotrophic factor; Igf1, Insulin-like growth factor 1; Ngf, Nerve growth factor; Nt-3, Neurotrophin 3. b–e Quantification of total dendritic length of shCtrl and shMeCP2 neurons with (+) or without (−) choline supplementation, expressed as a percentage of shCtrl levels. Neurons were cultured in the presence of specific inhibitors: 2 μM HC-3 to block acetylcholine synthesis b, 1.7 mM Dimbunol to block DNA methylation c, 5 μM GG to inhibit phosphatidylcholine synthesis d, and double inhibition with both Dimbunol and HC-3 e. n = 3 cultures. All values are presented as mean ± s.e.m. * p < 0.05 and ** p < 0.01. One-way ANOVA with Bonferroni post-hoc. f Illustration of the possible mechanism of choline in ameliorating RTT-like neuronal deficits and behaviour through its role in the synthesis of phosphatidylcholine