Detection of mRNA of Kinesin Superfamily 3A in the Cerebrum and Cerebellum : Biotin-Tyramine-Catalyzed Signal Amplification for In Situ Hybridization

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To improve the sensitivity of in situ hybridization (ISH) carried out using oligonucleotide probes without radionuclides, we employed a new method entailing amplification of ISH signals by catalyzed reporter deposition. This method is based on biotin-tyraminecatalyzed signal amplification (BT-CSA) and was used to detect expression of a relatively scarce polynucleotid, kinesin 3A mRNA, in the entral nervous system. Strong singals were detected in the hippocampus and in the granular cell layer of the cerebellum, but expression of kinesin 3A mRNA was not detected in Purkinje cells. ISH performed without the use of radionuclides has the advantage of being comparatively easy to perform, but has the disadvantage of low sensitivity. We showed that the low sensitivity can be overcome by using BT-CSA. Our method also had the advantage of enabling visualization of ISH signals via enzymatic reaction with horseradish peroxidase or by fluorescent staining.
日本組織細胞化学会の論文