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Abstract Abstract：【OBJECTIVE】To establish the method of in situ hybridization (ISH) with digoxin-labeled probe and detect the expression of FcRn in the small intestine of piglets .【METHOD】A digoxin-labeled single-strand cDNA probe for ISH detection of FcRn mRNA was plotted according to the conserved region of the gene for α chain of porcine FcRn. The ISH method was optimized through a series of trials such as the elimination of endogenous HRP, fixing agent, cellular membrane proteopepsis agent , hybridization solution, time and temperature of hybridization treatments. 【RESULTS】①The optimal thickness of the porcine small intestine tissue slice was 14 μm; ② Endogenous HRP was present in the intestine tissues and could be eliminated by the solution of 3％ H2O2 , whereas this step of treatment was proved unnecessary in this research because the enzyme did not apparently interfere with the ISH detection effect; ③The limpid hybridization signals of bright brown particles were detected in the intestinal glands and submucosa of duodenum, jejunum and ileum.【CONCLUSION】The established ISH method was proved to be effective. FcRn mRNA was expressed in the tissues of duodenum, jejunum and ileum of piglets, which indicated the correlation of the presence of FcRn and IgG transportation in the intestine.
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Received: 04 January 2008
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Corresponding Authors:
Li-Ying CHEN
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