Secondary Ion Mass Spectrometry Imaging Reveals Changes in the Lipid Structure of the Plasma Membranes of Hippocampal Neurons following Drugs Affecting Neuronal Activity.

Paola Agüi-Gonzalez,Maria A Gomes De Castro,Bao Guobin,Nhu T N Phan,Silvio O Rizzoli

doi:10.1021/acschemneuro.1c00031

Paola Agüi-Gonzalez, Maria A Gomes De Castro + Show 3 more

Open Access

https://doi.org/10.1021/acschemneuro.1c00031

Copy DOI

Abstract

The cellular functions of lipids in the neuronal plasma membranes have been increasingly acknowledged, particularly their association to neuronal processes and synaptic plasticity. However, the knowledge of their regulatory mechanisms in neuronal cells remains sparse. To address this, we investigated the lipid organization of the plasma membranes of hippocampal neurons in relation to neuronal activity using secondary ion mass spectrometry imaging. The neurons were treated with drugs, particularly tetrodotoxin (TTX) and bicuculline (BIC), to induce chronic activation and silencing. Distinct lipid organization was found in the plasma membrane of the cell body and the neurites. Moreover, significant alterations of the levels of the membrane lipids, especially ceramides, phosphatidylserines, phosphatidic acids, and triacylglycerols, were observed under the TTX and BIC treatments. We suggest that many types of membrane lipids are affected by, and may be involved in, the regulation of neuronal function.

Highlights

In the past decades, numerous studies have shown that lipids are the structural components and highly dynamic biomolecules involved in neuronal processes, in ion-channel regulation and synaptic plasticity.[1,2] The cellular functions of lipids at neuronal plasma membranes have been increasingly interested in neuroscience and cell biology due to their association with physiological and pathological processes
We investigated the lipid organization of the plasma membranes of hippocampal neurons and how it relates to the neuronal activity using ToF-secondary ion mass spectrometry (SIMS) imaging
Following the protocol of Kaech and Banker,[24] rat hippocampal neurons were cultured on indium tin oxide (ITO) coated glass slides to keep a physical distance with the astrocytic monolayer in a sandwich fashion, the socalled Banker cell culture

Summary

■ INTRODUCTION

Numerous studies have shown that lipids are the structural components and highly dynamic biomolecules involved in neuronal processes, in ion-channel regulation and synaptic plasticity.[1,2] The cellular functions of lipids at neuronal plasma membranes have been increasingly interested in neuroscience and cell biology due to their association with physiological and pathological processes. Significant alterations of the lipid levels, ceramides, PSs, PAs, and TGs, were observed under the TTX and BIC treatments This correlation suggests that membrane lipids change their organization to fit to, or even to mediate, neuronal and synaptic activity. The cells were washed with PBS for 5 min and brane of cell body and neurites obtained by ICA, assigned peaks in positive and negative SIMS mode, relative change of membrane lipid compositions in hippocampal neurons following drug treatments, and figures for comparison of CCD values between control and TTX treatment and between control and BIC treatment in the cell body and the neurites in positive and negative ion modes (PDF). ■ AUTHOR INFORMATION quickly rinsed four times with ammonium formate 150 mM at pH 7.4

Corresponding Author

■ ACKNOWLEDGMENTS

■ REFERENCES