Longitudinal investigation of the metabolome of 3D aggregating brain cell cultures at different maturation stages by 1H HR-MAS NMR

Abstract

The aim of the present study was to establish the developmental profile of metabolic changes of 3D aggregating brain cell cultures by H high-resolution magic angle spinning (HR-MAS) NMR spectroscopy. The histotypic 3D brain aggregate, containing all brain cell types, is an excellent model for mechanistic studies including OMICS analysis; however, their metabolic profile has not been yet fully investigated. Chemometric analysis revealed a clear separation of samples from the different maturation time points. Metabolite concentration evolutions could be followed and revealed strong and various metabolic alterations. The strong metabolite evolution emphasizes the brain modeling complexity during maturation, possibly reflecting physiological processes of brain tissue development. The small observed intra- and inter-experimental variabilities show the robustness of the combination ofH-HR-MAS NMR and 3D brain aggregates, making it useful to investigate mechanisms of toxicity that will ultimately contribute to improve predictive neurotoxicology. Graphical Abstract ᅟ.

Organophosphorus flame retardants are developmental neurotoxicants in a rat primary brainsphere in vitro model

“…material 1). Creatine is a metabolite that has shown to be a marker of gliosis when increased (Konaka et al 2003) as well as a marker of neurotoxicity when decreased (Diserens et al 2018;van Vliet et al 2008). Exposure to IPP, IDDP and TMPP significantly decreased the levels of creatine ( Fig.…”

High‐resolution magic angle spinning NMR for intact biological specimen analysis: Initial discovery, recent developments, and future directions

“…A recent HRMAS NMR study examined 3D brain cell aggregates at maturation times ranging from 7 to 35 days, and presented a longitudinal metabolomic follow-up of these aggregates. 154 The metabolomic alterations thus measured showed large changes across time, underscoring the complexity of brain modeling during maturation, perhaps reflecting physiological processes of brain tissue development. The study's observation of small intraexperimental and interexperimental variabilities supported the usefulness of HRMAS NMR in metabolomics studies of 3D brain aggregates, and supported these studies' potential contributions within future physiological and pathological investigations.…”

“…A recent HRMAS NMR study examined 3D brain cell aggregates at maturation times ranging from 7 to 35 days, and presented a longitudinal metabolomic follow‐up of these aggregates 154 . The metabolomic alterations thus measured showed large changes across time, underscoring the complexity of brain modeling during maturation, perhaps reflecting physiological processes of brain tissue development.…”

Sampling Method Affects HR-MAS NMR Spectra of Healthy Caprine Brain Biopsies

“…All experiments were performed in a 500.13 MHz Bruker Avance II spectrometer (Bruker Biospin AG, Fällanden, Switzerland) equipped with a 4 mm HR-MAS dual inverse ( 1 H, 13 C) probe. Spectra were acquired in a randomized order using a 1D 1 H sequence with water presaturation and with a T2-filter eliminating J-modulation ("PROJECT" Periodic Refocusing of J Evolution by Coherence Transfer [26]) at a 3 kHz MAS rate and nominal 278 K. Each 1D 1 H NMR spectrum was acquired by applying 512 transients, a spectral width of 6010 Hz, a data size of 32 K points, an acquisition time of 2.73 s, a relaxation delay of 4 s, an echo time (TE) of 400 ms, and a rotor-synchronized interpulse delay of 1.33 ms [27]. After testing several TEs, a TE of 400ms was chosen to achieve an optimal balance between a sufficient signal of the small metabolite peaks and the suppression of broad components from macromolecules and lipids, which otherwise may have introduced bias into the integration of the small metabolite peaks.…”