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Caspase-3 Cleaves Extracellular Vesicle Proteins During Auditory Brainstem Development.

Forrest Weghorst1, Yeva Mirzakhanyan2, Kian Samimi1

  • 1Department of Neurobiology and Behavior, University of California, Irvine, Irvine, CA, United States.

Frontiers in Cellular Neuroscience
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Caspase-3 protease activity in the auditory brainstem influences neural circuit development by cleaving proteins within extracellular vesicles (EVs), impacting sound localization.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Developmental Biology

Background:

  • Auditory brainstem circuit development is crucial for sound localization but its molecular mechanisms remain largely unknown.
  • The protease caspase-3 has a previously identified non-apoptotic role in chick auditory brainstem development.

Purpose of the Study:

  • To identify caspase-3 proteolytic substrates in the developing chick auditory brainstem.
  • To investigate the role of these substrates, particularly those within extracellular vesicles (EVs), in auditory circuit formation.

Main Methods:

  • Mass spectrometry was used to identify caspase-3 substrates.
  • Functional annotation analysis was performed on identified substrates.
  • Proteomic analysis of auditory brainstem extracellular vesicles (EVs) was conducted.
  • Expression analysis of specific substrates (NCAM, Ng-CAM) was performed.

Main Results:

  • Mass spectrometry identified numerous caspase-3 substrates, enriched for those cleaved in non-apoptotic contexts.
  • Substrates were significantly associated with extracellular vesicles (EVs).
  • Auditory brainstem EVs were highly enriched for caspase-3 substrates.
  • Two axon guidance molecules, NCAM and Ng-CAM, were identified in EVs and co-expressed with cleaved caspase-3 in the auditory pathway.

Conclusions:

  • Caspase-3 influences auditory brainstem circuit development through the non-apoptotic cleavage of proteins within extracellular vesicles (EVs).
  • This mechanism may involve EV-mediated intercellular communication and axon guidance processes impacting sound localization.