National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
F31NS132517
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R35GM151999
United States
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
NS062829
United States
National Institutes of Health/National Institute of Mental Health (NIH/NIMH)
MH137505
United States
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
R01NS101686
United States
Citation
Journal: Mol Brain / Year: 2026 Title: Primary cortical neurons precipitate and extrude large mitochondria-associated calcium-phosphate sheets with a bone-precursor-like ultrastructure. Authors: Erik D Anderson / Christopher A Cronkite / Philip R Baldwin / Carlota P Abella / Joseph G Duman / Ashleigh N Simmonds / M Neal Waxham / Kimberley F Tolias / Steven J Ludtke / Abstract: Calcium-phosphate (CaP) is a ubiquitous inorganic compound that plays an important structural role in healthy bone and teeth formation, but its pathologic buildup can occur in dyshomeostatic calcium ...Calcium-phosphate (CaP) is a ubiquitous inorganic compound that plays an important structural role in healthy bone and teeth formation, but its pathologic buildup can occur in dyshomeostatic calcium disorders like Alzheimer's disease and Leigh syndrome. The nexus of pathologic extracellular CaP in the nervous system is not well understood, but prior evidence suggests mitochondria could be a source. We have observed mitochondria-sized sheet-like CaP aggregates within functional wild type cortical neuron cultures at 1 and 20 days in vitro. Neurons were extracted from embryonic day 18 (E18) rat embryos following standard protocols to study neuronal structure and function. We have used a combination of cryo-ET, cryo-CLEM, and LDSAED to demonstrate that these aggregates are octacalcium phosphate-like, are associated with mitochondria, and that at least a portion are extruded via migrasomes. Visually similar aggregates were previously observed in Huntington's disease model neurons, but in that study they were not observed in WT controls. These findings show that this CaP aggregation process occurs routinely in WT neurons and may reveal an important link for how mitochondria may participate in calcification, highlighting them as potential therapeutic targets in neurological disorders characterized by pathological calcification, such as Alzheimer's disease.
Tomogram was reconstructed with EMAN2 as described previously: Chen, M., J.M. Bell, X. Shi, S.Y. Sun, Z. Wang, and S.J. Ludtke. 2019. A complete data processing workflow for cryo-ET and subtomogram averaging. Nat Methods. 16:1161?1168. doi:10.1038/s41592-019-0591-8.
Final reconstruction
Software - Name: EMAN2 / Number images used: 45
CTF correction
Software - Name: EMAN2 / Details: Tomogram was CTF corrected with EMAN2 / Type: PHASE FLIPPING ONLY
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