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A microfluidic based in vitro model of synaptic competition.

Ainsley Coquinco1, Luba Kojic1, Wendy Wen1

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Cellular Biology

Background:

  • Synaptic competition is crucial for neuronal network formation and function.
  • The precise mechanisms governing synaptic competition remain largely undefined.
  • Understanding these mechanisms is key to deciphering neural development and plasticity.

Purpose of the Study:

  • To investigate the mechanisms of synaptic competition in vitro.
  • To develop and utilize a novel microfluidic device for studying neuronal competition.
  • To elucidate the factors influencing synapse formation and axon growth during competition.

Main Methods:

  • Developed a 3-compartment microfluidic device for in vitro neuronal culture.
  • Utilized a two-input pathway competition model with cultured rat cortical neurons.
  • Employed muscimol (GABAAR agonist) to inhibit one input's activity.
  • Conducted time-lapse imaging to observe axon growth and synapse formation.
  • Investigated the roles of NMDAR and CamKII in the observed plasticity.

Main Results:

  • Inhibition of one neuronal input led to increased synapse numbers and axon elongation in the opposing input.
  • Uninhibited inputs demonstrated superior outgrowth and connectivity compared to inhibited counterparts.
  • Synaptic competition occurred during a sensitive developmental period, concluding before 21 days in vitro (DIV).
  • The observed plasticity was dependent on N-methyl-D-aspartate receptors (NMDAR) and Calcium/calmodulin-dependent protein kinase II (CamKII).
  • Plasticity was influenced by the age of central compartment neurons but not the competing inputs.

Conclusions:

  • Activity-dependent synaptic competition actively prunes and strengthens neuronal connections.
  • The age of postsynaptic neurons is a critical factor in modulating competitive plasticity.
  • NMDAR and CamKII signaling pathways are essential mediators of this competitive process during a specific developmental window.