Use of dual section mRNA in situ hybridisation/immunohistochemistry to clarify gene expression patterns during the early stages of nephron development in the embryo and in the mature nephron of the adult mouse kidney
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The kidney is the most complex organ within the urogenital system. The adult mouse kidney contains in excess of 8,000 mature nephrons, each of which can be subdivided into a renal corpuscle and 14 distinct tubular segments. The histological complexity of this organ can make the clarification of the site of gene expression by in situ hybridisation difficult. We have defined a panel of seven antibodies capable of identifying the six stages of early nephron development, the tubular nephron segments and the components of the renal corpuscle within the embryonic and adult mouse kidney. We have analysed in detail the protein expression of Wt1, Calb1 Aqp1, Aqp2 and Umod using these antibodies. We have then coupled immunohistochemistry with RNA in situ hybridisation in order to precisely identify the expression pattern of different genes, including Wnt4, Umod and Spp1. This technique will be invaluable for examining at high resolution, the structure of both the developing and mature nephron where standard in situ hybridisation and histological techniques are insufficient. The use of this technique will enhance the expression analyses of genes which may be involved in nephron formation and the function of the mature nephron in the mouse.
KeywordsImmunohistochemistry mRNA in situ hybridisation Gene expression Nephron Kidney
We would like to thank Darrin Taylor and Sean Grimmond for technical advice and assistance in designing the primers for the mRNA riboprobes. Images and annotated protein expression patterns using these antibodies are available on the GUDMAP gene expression database (http://www.gudmap.org). We would like to thank the Edinburgh GUDMAP Editorial Office. This work was supported by the National Institute of Diabetes Digestion and Kidney Diseases, National Institutes of Health, USA (DK070136-02).
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