Histochemical fluorescent staining of Sendai virus-infected cells with a novel sialidase substrate

Virology. 2014 Sep:464-465:206-212. doi: 10.1016/j.virol.2014.04.005. Epub 2014 Aug 2.

Abstract

Sialidases, enzymes that remove terminal sialic acid residues, are pivotal in various biological processes such as malignancy and infection with pathogens. For histochemical staining of sialidase activity, we have developed a new synthetic sialidase substrate, sialic acid-conjugated fluorescent benzothiazolylphenol derivative (BTP3-Neu5Ac), for rapid, sensitive, and specific fluorescent staining of sialidase activity. Here, we showed the usefulness of BTP3-Neu5Ac for histochemical fluorescent staining of cells infected with Sendai virus (SV), which possesses sialidase activity. BTP3-Neu5Ac also visualised SV-infected regions of lung sections from SV-infected mice. We succeeded in histochemical fluorescent staining of SV both in vitro and in vivo. SV has been utilised in many virological and biotechnological studies such as developments of an oncolytic virus, a gene therapy vector, and a vaccine candidate. BTP3-Neu5Ac should contribute to rapid progress of such studies and researches on viral sialidase.

Keywords: Easy protocol; Fluorescent staining; Hemagglutinin-neuraminidase; Histochemical staining; Infection detection; Rapid protocol; Sendai virus; Sensitive detection; Sialidase activity; Synthetic substrate.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzothiazoles / chemistry
  • Benzothiazoles / metabolism
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Humans
  • N-Acetylneuraminic Acid / chemistry
  • N-Acetylneuraminic Acid / metabolism
  • Neuraminidase / chemistry*
  • Neuraminidase / metabolism
  • Respirovirus Infections / diagnosis
  • Respirovirus Infections / virology*
  • Sendai virus / chemistry
  • Sendai virus / enzymology*
  • Staining and Labeling / instrumentation
  • Staining and Labeling / methods*
  • Substrate Specificity
  • Viral Proteins / chemistry*
  • Viral Proteins / metabolism

Substances

  • Benzothiazoles
  • Fluorescent Dyes
  • Viral Proteins
  • Neuraminidase
  • N-Acetylneuraminic Acid