Journal: The Journal of Cell Biology
Article Title: Centrosome amplification disrupts renal development and causes cystogenesis
Figure Lengend Snippet: CA in adult mice sensitizes kidneys to renal injury. (A) H E staining of control and Ksp-Plk4 kidneys isolated at P7 and P30. No prominent cysts were evident at these stages. (B) Kidney samples from P30 Ksp-Plk4 mice immunostained with anti-mCherry to identify cells expressing mChPlk4. There was robust expression of Plk4 in DBA-positive collecting ducts and CLC-K–positive distal tubules, but absent in LTL-positive proximal tubules, consistent with Ksp-Cre–mediated expression. Nuclei were stained with DAPI. (C) Immunostaining of P30 control and Ksp-Plk4 kidneys to mark centrosomes (γ-tubulin), cilia (acetylated α-tubulin), collecting ducts (DBA), and DNA (DAPI). In contrast to the normal centrosome number in control kidneys, CA is evident in Ksp-Plk4. White box denotes regions shown in insets. Arrowheads point to cells with multiple centrosomes and cilia. Bars (insets), 2 µm. (D) Quantification of CA in P30 WT and Ksp-Plk4 mice. n = 747 cells (WT) and 562 cells (Ksp-Plk4). (E) Quantification of the percentage of cells expressing zero, one, or more than one primary cilium in P30 Ksp-Plk4 mice containing excess centrosomes compared with cells with normal centrosome number. CA caused an increase in the fraction of superciliated cells. n = 274 cells (WT) and 172 cells (Ksp-Plk4). Statistical significance was determined using a two-tailed unpaired t test (*, P
Article Snippet: For postnatal kidneys, samples were costained with nephron segment markers including LTL (marker of proximal tubules; Vector Laboratories), antibodies against the chloride channel CLC-K (marker of distal tubules; Alomone Labs; gift of F. Chen, Washington University, St. Louis, MO), and DBA (marker of collecting ducts; Vector Laboratories).
Techniques: Mouse Assay, Staining, Isolation, Expressing, Immunostaining, Two Tailed Test