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Functional characterization of an aquaporin from a microsporidium, Nosema bombycis

Gong Chen1, Wei Wang1, Hongli Chen1

  • 1Jiangsu University of Science and Technology, Zhenjiang, Jiangsu Province, China.

Plos One
|July 28, 2017
PubMed

Insights

Researchers identified a functional water channel protein, Nosema bombycis aquaporin (NbAQP), in the microsporidian parasite Nosema bombycis. This discovery offers potential targets for preventing microsporidiosis.

Area of Science:

  • Microbiology
  • Parasitology
  • Molecular Biology

Background:

  • Microsporidia are eukaryotic parasites causing infections in various hosts.
  • Spore germination involves osmotic pressure changes, suggesting a role for water channels.
  • The specific water transport mechanisms in Nosema bombycis remain largely uncharacterized.

Purpose of the Study:

  • To clone and characterize a water channel protein (aquaporin) in Nosema bombycis.
  • To investigate the localization, expression, and function of the identified aquaporin.
  • To explore the potential of this aquaporin as a target for anti-microsporidial strategies.

Main Methods:

  • Cloning of the Nosema bombycis aquaporin (NbAQP) gene.
  • Sequence analysis and homology comparisons with other aquaporins.
  • Indirect immunofluorescence (IFA) and immunogold electron microscopy for protein localization.
  • Quantitative reverse transcription PCR (qRT-PCR) for gene expression analysis.
  • Xenopus laevis oocyte expression system to assess water permeability.
  • Antibody blocking experiments to evaluate the effect on spore germination.

Main Results:

  • The NbAQP gene encodes a 249-amino acid polypeptide with homology to other microsporidian aquaporins.
  • NbAQP is predominantly localized in the spore wall of N. bombycis spores.
  • NbAQP expression levels fluctuate post-inoculation, peaking at 6 days.
  • Expressed NbAQP significantly increased water permeability in Xenopus oocytes.
  • Antibody blocking of NbAQP inhibited spore germination by approximately 28% (P < 0.05).

Conclusions:

  • Nosema bombycis possesses functional water channel proteins (NbAQP).
  • NbAQP plays a role in the osmotic regulation during spore germination.
  • NbAQP represents a potential therapeutic target for controlling microsporidiosis.

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