Unspecific labelling of oligodendrocytes by sulforhodamine 101 depends on astrocytic uptake via the thyroid hormone transporter OATP1C1 (SLCO1C1)

Neurosci Lett. 2016 Sep 19:631:13-18. doi: 10.1016/j.neulet.2016.08.010. Epub 2016 Aug 9.

Abstract

The red fluorescent dye Sulforhodamine 101 (SR101) is often used as a marker for astrocytes, although variations of the staining protocol have been shown to influence the preferentially labeled cell type. Here we analyzed SR101-labeling of oligodendrocytes in the hippocampal slices preparation of PLP-EGFP mice. Using different staining protocols, we found robust SR101-labeled oligodendrocytes in the CA1 stratum radiatum of the hippocampus. Application of L-thyroxin, which is known to block SR101 transport into astrocytes via competitive inhibition of the multi-specific OATP1C1 (SLCO1C1) transporter, significantly reduced oligodendrocyte labeling. Since OATP1C1 is not expressed in oligodendrocytes, we conclude that oligodendrocyte labeling with SR101 requires SR101-uptake by astrocytes, which then diffuses to oligodendrocytes via heterotypic gap junctions of the pan-glial network. In summary, unequivocal identification of a particular cell type is not possible by SR101 only, hence caution is recommended when using SR101 in future studies.

Keywords: Astrocyte; Brain imaging; Cells specific dye; Gap-junctions; Thyroid hormone transporter.

MeSH terms

  • Animals
  • Astrocytes / cytology
  • Astrocytes / metabolism*
  • CA1 Region, Hippocampal / cytology
  • CA1 Region, Hippocampal / metabolism
  • Fluorescent Dyes / administration & dosage*
  • Fluorescent Dyes / pharmacology
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence / methods*
  • Oligodendroglia / cytology
  • Oligodendroglia / metabolism*
  • Organic Cation Transport Proteins / metabolism*
  • Rhodamines / administration & dosage*
  • Rhodamines / pharmacology

Substances

  • Fluorescent Dyes
  • Oatp2 protein, mouse
  • Organic Cation Transport Proteins
  • Rhodamines
  • sulforhodamine 101