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Expression of polycystin in mouse metanephros and extra-metanephric tissues
M D Griffin1, D A O'Sullivan, V E Torres
1Nephrology Research Unit, Mayo Clinic, Rochester, Minnesota, USA.
Abstract:
The presence of messenger RNA for the mouse homologue of the polycystic kidney disease 1 gene (PKD1) was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) methods in mouse embryo messenger RNA. A single locus for the PKD1 gene was detected on mouse chromosome 17 by fluorescent in situ hybridization. Immunoprecipitation of proteins from [35S] methionine-labeled mouse metanephric explants with an anti-polycystin antibody (Pc1) revealed high molecular weight bands, the highest being > 400 kDa. Immunoperoxidase staining of mouse embryos with Pc1 revealed expression of polycystin as early as day 8 gestation. The expression was seen in epithelial cells of the ureteric bud, in condensing blastemal cells of the developing metanephros and, subsequently, in cells of the nascent tubules. In addition, Pc1 immunoreactivity was seen in hepatocytes and biliary epithelium, cardiac and skeletal muscle, neural tissue, gut, and bronchial epithelium. In post-natal and adult mouse kidney and liver persistent slight to moderate immunoreactivity was observed. Immunofluorescent studies of cultured 13-day mouse metanephroi revealed polycystin expression in ureteric bud epithelium, early glomerular structures (that is, condensates, S-shaped and comma-shaped bodies) and in proximal and distal tubular epithelia. These data indicate that the mouse has a single gene homologous to human PKD1 on chromosome 17, and polycystin is expressed in a variety of tissues during embryonic development.
Insights
Researchers identified the mouse polycystic kidney disease 1 gene (PKD1) on chromosome 17. Polycystin protein is expressed in various embryonic mouse tissues, including the developing kidney, suggesting its crucial role in development.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- The polycystic kidney disease 1 (PKD1) gene is implicated in human autosomal dominant polycystic kidney disease.
- Understanding the homologous gene and protein expression in model organisms is crucial for disease research.
Purpose of the Study:
- To characterize the mouse homologue of the human PKD1 gene.
- To investigate the expression pattern of polycystin during mouse embryonic development.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) to detect PKD1 mRNA.
- Fluorescent in situ hybridization to determine gene locus.
- Immunoprecipitation and immunoperoxidase/immunofluorescent staining to detect polycystin protein expression.
Main Results:
- A single mouse PKD1 gene locus was mapped to chromosome 17.
- Polycystin protein, with high molecular weight bands (>400 kDa), was detected.
- Polycystin expression was observed as early as day 8 of gestation in various embryonic tissues, including the developing kidney, liver, muscle, and neural tissues.
Conclusions:
- The mouse possesses a single PKD1 gene homologous to the human gene, located on chromosome 17.
- Polycystin is widely expressed in diverse tissues during mouse embryonic development, particularly in the developing kidney.
- These findings provide a foundation for studying PKD1 function and related diseases in mice.