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Fluorescence In Situ Hybridization Using an Old World Monkey Y Chromosome–specific Probe Combined With Immunofluorescence Staining on Rhesus Monkey Tissues

Xiujin Xia, Terri Rasmussen, Xavier Alvarez, Takahiro Taguchi, Marilyn Li and Vincent F. La Russa

Department of Pharmacology (XX,VFLR), Tulane Cancer Center (VFLR), and Hayward Genetics Center (ML), Tulane University Health Sciences Center, New Orleans, Louisiana; Division of Gene Therapy (XX,VFLR) and Division of Comparative Pathology (TR,XA), Tulane National Primate Research Center, Covington, Louisiana; and Division of Human Health and Medical Science, Graduate School of Kuroshio Science, Kochi University, Kochi, Japan (TT)

Correspondence to: Vincent F. La Russa, PhD, Tulane University Health Sciences Center, Cancer Center, 1415 Tulane Avenue, SL-34, New Orleans, LA. E-mail: vlaruss{at}tulane.edu

To date, there is no commercially available Y chromosome probe that can be used for fluorescence in situ hybridization (FISH) for the male rhesus monkey. We have recently generated a probe for FISH with high specificity to the short arm of the rhesus monkey Y chromosome. In this study, we further describe a method that keeps the integrity of tissue-specific antigenic structures for immunofluorescence staining subsequent to FISH on paraffin-embedded rhesus monkey tissues. We have examined this technique in combination with an epithelial cell–specific marker, cytokeratin 8/18 (CK8/18), on various tissues, including jejunum, liver, kidney, and pancreas. CK8/18 and Y chromosome signals were distinctly seen simultaneously on epithelial cells from the same tissue section from male but not female monkeys. These studies indicate that our FISH immunofluorescence technique can be reliably used to identify and phenotype male cells in paraffin-embedded rhesus monkey tissues. (J Histochem Cytochem 55:1115–1121, 2007)

Key Words: rhesus monkey • fluorescence in situ hybridization • Y chromosome immunofluorescence

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