(P09-018-25) Glycerophosphocholine and Betaine Improve Markers of Neuronal Development

Objectives: The goal of this study was to assess the effects of glycerophosphocholine (GPC) and betaine, alone and in tandem, upon markers of neuronal development in an in vitro N2A cell culture model. We hypothesized GPC and betaine together would exert a synergistic effect upon the selected outcomes.

Methods: N2A cells were cultured for 24 hours (h) in Minimum Essential Medium – Alpha (MEM-α) supplemented with 10% of fetal bovine serum (FBS). Cells were split into four treatment groups: control, GPC (25 µM), betaine (50 µM), or GPC+betaine (25 and 50 µM, respectively) and treated for 48 h in MEM-α supplemented with 1% FBS, after which they underwent analysis for neurite outgrowth and harvest to assess protein markers of neuronal development. Neurite outgrowth was assessed using ImageJ. Western blots were used to assess markers of neuronal development including glucose transporters 1 and 3 (GLUT1 and GLUT3), aquaporin 4 (AQP4), microtubule-associated protein 2 (MAP2), and βIII-tubulin. Statistics were performed with GraphPad Prism version 10.2.3 for Windows.

Results: GPC+betaine treatment increased neurite outgrowth compared to control by 47% (p < 0.0001). GPC and betaine alone respectively improved neurite outgrowth by 17% (p=0.0293) and 14% (p=0.0117). Expression of GLUT1 (42%; p=0.0199), MAP2 (34%; p=0.0017), and AQP4 (74%; p=0.0415), was increased by GPC+betaine compared to control or GPC and betaine individually. βIII-tubulin and GLUT3 expression did not differ between treatment groups.

Conclusions: Both GPC and betaine alone improved neurite outgrowth. However, the combination of GPC+betaine substantially improved neurite outgrowth and markers for metabolism and cytoskeletal growth compared to control, GPC, or betaine treatments. These findings indicate treatment with GPC and betaine together may convey benefits for neurodevelopment and neuroplasticity.

Funding Sources: This research was funded by Abbott Laboratories.