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Assay for Blood-brain Barrier Integrity in Drosophila melanogaster
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Elena Repnikova1, Kate Koles, Michiko Nakamura

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Area of Science:

  • Neuroscience
  • Glycobiology
  • Developmental Biology

Background:

  • Sialylated glycans are crucial for vertebrate nervous system development and function.
  • Investigating sialylation roles is challenging due to glycosylation complexity and enzyme redundancy in mammals.
  • Drosophila melanogaster offers a model system with a single vertebrate-type sialyltransferase, Drosophila sialyltransferase (DSiaT).

Purpose of the Study:

  • To investigate the biological function of sialylation in Drosophila.
  • To explore the role of DSiaT in the Drosophila nervous system.
  • To determine if DSiaT function is conserved between invertebrates and vertebrates.

Main Methods:

  • Gene targeting to inactivate DSiaT in Drosophila.
  • Analysis of DSiaT mutant phenotypes using behavioral, immunolabeling, electrophysiological, and pharmacological methods.
  • Assessment of sialyltransferase activity in vivo.

Main Results:

  • DSiaT is expressed in a subset of developing CNS neurons.
  • DSiaT mutations led to reduced lifespan, locomotor defects, temperature-sensitive paralysis, and neuromuscular junction abnormalities.
  • DSiaT regulates neuronal excitability and voltage-gated sodium channel function, indicating a defect in N-glycan sialylation.

Conclusions:

  • Sialylation has a critical biological function in protostomes, specifically in the nervous system.
  • DSiaT's role in neuronal function highlights an ancient, conserved function of sialic acids in metazoans.
  • This study provides evidence for conserved roles of sialylation in nervous system function between flies and mammals.