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Basic Science and Pathogenesis.

Maria Clara Zanellati1

  • 1University of North Carolina, Chapel Hill, NC, USA.

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This study reveals a new role for VAPB-mediated ER-peroxisome contacts in neuronal differentiation. Multispectral imaging identified key organelle changes during neuron development, offering insights into neurodegenerative diseases.

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Area of Science:

  • Cell Biology
  • Neuroscience
  • Biochemistry

Background:

  • Dysfunctional organelle communication is central to neurodegenerative diseases like Alzheimer's disease (AD).
  • Studying organelle interactions in induced pluripotent stem cell (iPSC)-derived neurons with disease mutations aids understanding neurodegeneration mechanisms.

Purpose of the Study:

  • To develop and apply a multispectral imaging method for simultaneous visualization of eight organelles in live cells.
  • To analyze organelle morphology and dynamics (morphodynamics) during neuronal differentiation and maturation.

Main Methods:

  • Developed a multispectral imaging technique for live-cell analysis of eight organelles.
  • Utilized genetically encoded markers in iPSCs and iNeurons, collecting images throughout differentiation (5 time points).
  • Applied linear unmixing and a custom image analysis pipeline (Napari-InferSubC) to quantify ~1400 morphometrics and organelle contacts.

Main Results:

  • Observed significant organelle remodeling during iPSC to iNeuron differentiation, with increased endoplasmic reticulum (ER) and mitochondria volumes.
  • Documented an increase in organelle contacts, including higher-order contacts (3- and 4-way), during iNeuron maturation.
  • Identified increased ER-mitochondria, mitochondria-lysosome, and ER-peroxisome contacts; found VAPB expression increases with differentiation, mediating ER-peroxisome contacts crucial for plasmalogen production and synapse formation.

Conclusions:

  • Discovered a novel function for VAPB-mediated ER-peroxisome contacts in supporting neuronal differentiation.
  • Demonstrated the utility of multispectral imaging for studying organelle dynamics in neuronal development and disease.
  • Proposed future application of this method to investigate organelle communication defects in AD-related neurons.