Subcellular localization of alpha-tubulin and opsin mRNA in the goldfish retina using digoxigenin-labeled cRNA probes detected by alkaline phosphatase and HRP histochemistry

J Neurosci Methods. 1993 Nov;50(2):145-52. doi: 10.1016/0165-0270(93)90002-9.


This paper describes a method for non-radioactive in situ hybridization providing subcellular localization of mRNA in 3 microns cryosections. We used two alternative colorimetric reactions to detect digoxigenin-labeled cRNA probes: alkaline phosphatase and HRP (horseradish peroxidase). With some probes the signal with the alkaline phosphatase reaction was intense, and diffusion of the reaction product was noticeable. Using HRP-conjugated antibodies improved the resolution but decreased the sensitivity of the signal. Photoamplification of the HRP reaction product increased the contrast and improved the sensitivity of the technique.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alkaline Phosphatase
  • Animals
  • Digoxigenin
  • Goldfish
  • Horseradish Peroxidase
  • In Situ Hybridization / methods
  • RNA Probes
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism*
  • Restriction Mapping
  • Retina / cytology
  • Retina / metabolism*
  • Rod Opsins / biosynthesis*
  • Transcription, Genetic
  • Tubulin / biosynthesis*


  • RNA Probes
  • RNA, Messenger
  • Rod Opsins
  • Tubulin
  • Horseradish Peroxidase
  • Alkaline Phosphatase
  • Digoxigenin