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Measurement of glomerular diameter and glomerular counting will be performed in paraffin slides stained with H&E using BIOQUANT True Color Image Analysis System, as outlined on the respective pages. In addition glomerular counting will be performed on freshly isolated adult mouse kidneys following acid digestion, as described (1). See protocol for details.

Glomerular counting > Protocol Section

 Glomerular quantification (based on technique described in Godley et al. G&D 1996) (1)

  1. Remove kidney and remove capsule. Cut kidney into 2 mm3 pieces. Place in a 50 ml tube.
  2. Incubate in 5 ml of 6 N HCl at 37¢ªC for 90 min.
  3. To disrupt kidney fragments, aspirate up and down quickly using a Pipette-Aid. If necessary, press kidney fragments against the side. Add 25 ml of distilled water.
  4. Incubate at 4¢ªC overnight. 
  5. Mix by inversion and count 1 ml in a counting chamber (scored 35 mm culture dish). It may be necessary to count 1:10 dilution of this sample. Calculate total glomerular number.

Publications for Glomerular counting (1)

Godley LA, Kopp JB, Eckhaus M, Paglino JJ, Owens J, Varmus HE. Wild-type p53 transgenic mice exhibit altered differentiation of the ureteric bud and possess small kidneys. Genes Dev (1996) 10:836-50
View abstract View in PubMed

Transgenic mice expressing wild-type murine p53 under the control of the mouse mammary tumor virus long terminal repeat (MMTV LTR) undergo progressive renal failure due to abnormal kidney development. Similar phenotypes are observed in two transgenic lines that express wild-type p53 within the ureteric bud but not in transgenic animals expressing a dominant-negative p53 mutant allele. Defective differentiation of the ureteric bud, as evidenced by altered marker expression during development, accompanies expression of the p53 transgene. At E17.5-18.5, metanephric mesenchymal cells undergo high rates of apoptosis, and fewer cells than normal are converted to tubular epithelium. As a result, p53 transgenic kidneys grow to only half of their expected size and contain about half of the normal number of nephrons, with compensatory hypertrophy of the glomeruli. In this setting, rather than arrest the cell cycle or induce apoptosis directly, abnormally high levels of wild-type p53 appear to alter cellular differentiation in embryonic ureteric buds and cause secondary effects (apoptosis and inefficient conversion to epithelium) in the adjacent undifferentiated mesenchyme.

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Last updated on 2013-11-06 Moderated by Ming-Zhi Zhang