Trace elements during primordial plexiform network formation in human cerebral organoids

Abstract

Systematic studies of micronutrients during brain formation are hindered by restrictions to animal models and adult post-mortem tissues. Recently, advances in stem cell biology have enabled recapitulation of the early stages of human telencephalon development. In the present work, we exposed cerebral organoids derived from human pluripotent stem cells to synchrotron radiation in order to measure how biologically valuable micronutrients are incorporated and distributed in the exogenously developing brain. Our findings indicate that elemental inclusion in organoids is consistent with human brain tissue and involves calcium, iron, phosphorus, potassium, sulfur, and zinc. Local trends in concentrations suggest a switch from passive to actively mediated transport across cell membranes. Finally, correlational analysis for pairs of elements shows spatially conserved patterns, suggesting they may physically associate, be stored in similar compartments or used in related biological processes. These findings might reflect which trace elements are important during human brain development and will support studies aimed to unravel the consequences of disrupted metal homeostasis for neurodevelopmental diseases, including those manifested in adulthood.PeerJ Preprints | https://doi.org/10.7287/peerj.preprints.2126v1 | CC BY 4.0 Open Access |

Short term changes in the proteome of human cerebral organoids induced by 5-MeO-DMT

“…In 45 day-old cerebral organoids, basal immunostaining was observed in AMPA (selective glutamate receptor 2) and NMDA (ionotropic glutamate receptor) – both characteristic of glutamatergic synapses – along with the neuronal marker MAP2 (Fig. 2A–D ), as previously described 29 . Glial cells (GFAP+) are also present in organoids, as shown in Fig.…”

“…45-day-old cerebral organoids were treated with either 5-MeO-DMT, vehicle, or left untreated for 24 h. Samples were analyzed using label-free state-of-the-art quantitative proteomics, using two-dimensional fractionation and high-resolution mass spectrometry. Workflow art was modified from 29 . (B) Venn diagram comparing the number of proteins identified by shotgun mass spectrometry in control human cerebral organoids, those treated with vehicle (EtOH), and 5-MeO-DMT.…”

“…Differentiation of hESCs into cerebral organoids was based on a previously described protocol 26 , 29 . Briefly, hESC cells were inoculated into a spinner flask, and to enable embryoid body formation, after six days the medium was changed to neural induction media (DMEM/F12, 1X N2 supplement (Gibco), 2 mM Glutamax (Invitrogen), 1% MEM-NEAA, and 1 μg/mL heparin (Sigma)) and the aggregates were cultured for five more days.…”

Short term changes in the proteome of human cerebral organoids induced by 5-methoxy-N,N-dimethyltryptamine

“…Basal immunostaining of ionotropic receptors AMPA and NMDA, characteristic of glutamatergic synapses, along with the neuronal marker MAP2 was observed in 45-days-old cerebral organoids ( Fig. 2A-D), as previously described (Sartore et al 2017). Glial cells (GFAP+) are also present in organoids, as shown in Fig.…”

“…Differentiation of hESCs into cerebral organoids was based on previously described protocol (Lancaster et al 2013;Sartore et al 2017). Briefly, hESC were inoculated into a spinner flask and after six days to enable embryoid bodies formation, media was changed to neural induction media (DMEM/F12, 1x N2 supplement (Gibco), 2 mM Glutamax (Invitrogen), 1% MEM-NEAA and 1 µg/mL heparin (Sigma) and the aggregates were cultured for more five days.…”