Molecular fingerprinting of the podocyte reveals novel gene and protein regulatory networks.
A thorough characterization of the transcriptome and proteome of endogenous podocytes has been hampered by low cell yields during isolation. Here we describe a double fluorescent reporter mouse model combined with an optimized bead perfusion protocol and efficient single cell dissociation to yield more than 500,000 podocytes per mouse allowing for global, unbiased downstream applications. Combining mRNA and miRNA transcriptional profiling with quantitative proteomic analyses revealed programs of highly specific gene regulation tightly controlling cytoskeleton, cell differentiation, endosomal transport, and peroxisome function in podocytes. Strikingly, the analyses further predict that these podocyte-specific gene regulatory networks are accompanied by alternative splicing of respective genes. Thus, our 'omics' approach will facilitate the discovery and integration of novel gene, protein, and organelle regulatory networks that deepen our systematic understanding of podocyte biology.
Freiburg Institute for Advanced Studies-LifeNet, Albert-Ludwigs-University Freiburg, Freiburg, Germany.
Melanie
M
Boerries
Florian
F
Grahammer
Sven
S
Eiselein
Moritz
M
Buck
Charlotte
C
Meyer
Markus
M
Goedel
Wibke
W
Bechtel
Stefan
S
Zschiedrich
Dietmar
D
Pfeifer
Denis
D
Laloë
Christelle
C
Arrondel
Sara
S
Gonçalves
Marcus
M
Krüger
Scott J
SJ
Harvey
Hauke
H
Busch
Joern
J
Dengjel
Tobias B
TB
Huber
D017398:Alternative Splicing; D000818:Animals; D002469:Cell Separation; D019295:Computational Biology; D020869:Gene Expression Profiling / Q000379:methods; D053263:Gene Regulatory Networks; D017930:Genes, Reporter; D005838:Genotype; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D050199:Podocytes / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction / Q000235:genetics
D008164:Luminescent Proteins; D035683:MicroRNAs