Nephrotic syndrome is characterized by proteinuria and hypoalbuminemia, results from the dysregulation of glomerular podocytes and is a significant cause of chronic kidney disease. We previously reported that urinary protein excretion decreased by administration of yohimbine, α2-adrenoceptor antagonist, in 5/6 nephrectomy-induced chronic kidney disease rat model. Here, we examined the effect of yohimbine on puromycin aminonucleoside (PAN)-induced nephrosis rats. Male Sprague Dawley rats were randomly divided into the following groups: sham-operated, PAN (50 mg/kg, i.v.) and PAN + yohimbine (3 mg/L in drinking water). We found that proteinuria increased in the PAN group in a time-dependent manner. In immunostaining studies, PAN group showed decreased expressions of nephrin and podocin as well as increased desmin expression compared with sham group. Treatment with yohimbine improved proteinuria and increased expression of desmin in PAN group. Furthermore, yohimbine restored the expressions of nephrin and podocin. The α2C-adrenoceptors were partly distributed in some compartments of the podocytes. In contrast, α2A- and α2B-adrenoceptors were not detected in the glomeruli. These findings suggest that the yohimbine can regulate glomerular filtration and proteinuria through the induction of morphological changes in podocytes via α2C-adrenoceptors.