Over-expression of adenosine deaminase in mouse podocytes does not reverse puromycin aminonucleoside resistance

<p>Abstract</p> <p>Background</p> <p>Edema in nephrotic syndrome results from renal retention of sodium and alteration of the permeability properties of capillaries. Nephrotic syndrome induced by puromycin aminonucleoside (PAN) in rats reproduces the biological and clinical signs of the human disease, and has been widely used to identify the cellular mechanisms of sodium retention. Unfortunately, mice do not develop nephrotic syndrome in response to PAN, and we still lack a good mouse model of the disease in which the genetic tools necessary for further characterizing the pathophysiological pathway could be used. Mouse resistance to PAN has been attributed to a defect in glomerular adenosine deaminase (ADA), which metabolizes PAN. We therefore attempted to develop a mouse line sensitive to PAN through induction of normal adenosine metabolism in their podocytes.</p> <p>Methods</p> <p>A mouse line expressing functional ADA under the control of the podocyte-specific podocin promoter was generated by transgenesis. The effect of PAN on urinary excretion of sodium and proteins was compared in rats and in mice over-expressing ADA and in littermates.</p> <p>Results</p> <p>We confirmed that expression of ADA mRNAs was much lower in wild type mouse than in rat glomerulus. Transgenic mice expressed ADA specifically in the glomerulus, and their ADA activity was of the same order of magnitude as in rats. Nonetheless, ADA transgenic mice remained insensitive to PAN treatment in terms of both proteinuria and sodium retention.</p> <p>Conclusions</p> <p>Along with previous results, this study shows that adenosine deaminase is necessary but not sufficient to confer PAN sensitivity to podocytes. ADA transgenic mice could be used as a background strain for further transgenesis.</p

Ultraviolet resonance Raman study of DNA and of its interaction with actinomycin D.

The DNA-Actinomycin D interaction has been studied by resonance Raman effect using DNA as chromophore. First, the resonance Raman spectra of DNA obtained with a U.V. excitation at wavelengths of 300 nm and 280 nm are presented. The main Raman hands are assigned to the convenient nucleic bases by comparison with the spectra of mononucleotides obtained under the same experimental conditions. In particular, with a 300 nm excitation, the 1582 cm-1 line is provided by adenine, while the 1492 cm-1 one is almost exclusively due to guanine. Then, the DNA-Actinomycin D complex has been studied: the line enhancements and the specificity of the resonance permits the displaying of the DNA spectrum free of any contribution of Actinomycin. The interaction provides a large intensity decrease of the 1492 cm-1 guanine line: this is a direct consequence of the orbital overlapping of the guanine 2-aminogroup with the ring nitrogen of Actinomycin in the DNA-Actinomycin pi complex