High Content Analysis of Hippocampal Neuron-Astrocyte Co-cultures Shows a Positive Effect of Fortasyn Connect on Neuronal Survival and Postsynaptic Maturation.

Anne-Lieke F Van Deijk,J Martin Verkuyl,August B Smit,Mark H G Verheijen,Laus M Broersen

doi:10.3389/fnins.2017.00440

Anne-Lieke F Van Deijk, J Martin Verkuyl + Show 3 more

Open Access

https://doi.org/10.3389/fnins.2017.00440

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Abstract

Neuronal and synaptic membranes are composed of a phospholipid bilayer. Supplementation with dietary precursors for phospholipid synthesis –docosahexaenoic acid (DHA), uridine and choline– has been shown to increase neurite outgrowth and synaptogenesis both in vivo and in vitro. A role for multi-nutrient intervention with specific precursors and cofactors has recently emerged in early Alzheimer's disease, which is characterized by decreased synapse numbers in the hippocampus. Moreover, the medical food Souvenaid, containing the specific nutrient combination Fortasyn Connect (FC), improves memory performance in early Alzheimer's disease patients, possibly via maintaining brain connectivity. This suggests an effect of FC on synapses, but the underlying cellular mechanism is not fully understood. Therefore, we investigated the effect of FC (consisting of DHA, eicosapentaenoic acid (EPA), uridine, choline, phospholipids, folic acid, vitamins B12, B6, C and E, and selenium), on synaptogenesis by supplementing it to primary neuron-astrocyte co-cultures, a cellular model that mimics metabolic dependencies in the brain. We measured neuronal developmental processes using high content screening in an automated manner, including neuronal survival, neurite morphology, as well as the formation and maturation of synapses. Here, we show that FC supplementation resulted in increased numbers of neurons without affecting astrocyte number. Furthermore, FC increased postsynaptic PSD95 levels in both immature and mature synapses. These findings suggest that supplementation with FC to neuron-astrocyte co-cultures increased both neuronal survival and the maturation of postsynaptic terminals, which might aid the functional interpretation of FC-based intervention strategies in neurological diseases characterized by neuronal loss and impaired synaptic functioning.

Highlights

During neuronal development, neurites sprout from the cell body and elongate, followed by the formation of dendrites and axons and the formation of synapses, allowing synaptic transmission and plasticity (Govek et al, 2005; Wurtman et al, 2009b)
Fortasyn Connect (FC) increased postsynaptic PSD95 levels in both immature and mature synapses. These findings suggest that supplementation with FC to neuron-astrocyte co-cultures increased both neuronal survival and the maturation of postsynaptic terminals, which might aid the functional interpretation of FC-based intervention strategies in neurological diseases characterized by neuronal loss and impaired synaptic functioning
We used a new method of analysis to examine the effect of nutritional phospholipid precursors and cofactors, as provided by Fortasyn Connect (FC) supplementation, on synapse formation and maturation of primary hippocampal neurons co-cultured with astrocytes

Summary

Introduction

Neurites sprout from the cell body and elongate, followed by the formation of dendrites and axons and the formation of synapses, allowing synaptic transmission and plasticity (Govek et al, 2005; Wurtman et al, 2009b). Synapses contribute in important ways to learning and memory processes. Several studies have shown that supplementation with these phospholipid precursors enhances neurite outgrowth and synaptogenesis both in vivo and in vitro. It was shown that oral administration of a combination of uridine-5′-monophosphate (UMP; a source of uridine) and DHA in rats and gerbils increased hippocampal spine density and synaptic protein levels (Sakamoto et al, 2007; Cansev et al, 2009). Dietary supplementation with a combination of uridine, DHA and choline improved spatial learning and memory in healthy gerbils (Holguin et al, 2008). It has been demonstrated that DHA supplementation enhanced neurite outgrowth in cultured rat hippocampal neurons (Calderon and Kim, 2004)

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