Jan 13, 2001

Detection of peroxisomal proteins and their mRNAs in serial sections of fetal and newborn mouse organs

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
Markus GrabenbauerEveline Baumgart


We present a protocol for detection of peroxisomal proteins and their corresponding mRNAs on consecutive serial sections of fetal and newborn mouse tissues by immunohistochemistry (IHC) and nonradioactive in situ hybridization (ISH). The use of perfusion-fixation with depolymerized paraformaldehyde combined with paraffin embedding and digoxigenin-labeled cRNA probes provided a highly sensitive ISH protocol, which also permitted immunodetection with high optical resolution by light and/or fluorescence microscopy. Signal enhancement was achieved by the addition of polyvinyl alcohol (PVA) for ISH color development. For IHC, signal amplification was obtained by antigen retrieval combined with biotin-avidin-HRP and Nova Red as substrate or by the catalyzed reporter deposition of fluorescent tyramide. Using this protocol, we studied the developmental changes in localization of the peroxisomal marker enzymes catalase (CAT) and acyl-CoA oxidase 1 (AOX), the key regulatory enzyme of peroxisomal beta-oxidation, at the protein and mRNA levels in mice from embryonic Day 14.5 to birth (P0.5). The mRNA signals for CAT and AOX were detected in sections of complete fetuses, revealing organ- and cell-specific variations. Here we focus on the lo...Continue Reading

  • References42
  • Citations19


  • References42
  • Citations19


Mentioned in this Paper

Establishment and Maintenance of Localization
Acyl-CoA oxidase
Enzymes, antithrombotic
ACOX1 gene
Sample Fixation

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