Histological analysis of kidneys and placentas
Kidneys and placentas of rats were harvested from the dams on GD 20.5.
The tissues were then fixed in 10% buffered formalin overnight at 4°C.
Processed tissues were embedded in paraffin. Serial
sections (5 µm) were cut from each
uteroplacental unit and serial sections (3 µm) were cut from each
kidney. The sections of kidney were deparaffinized, rehydrated and
stained with Hematoxylin and Eosin and periodic acid-Schiff for
histopathological assessment of renal alterations. The degree of
glomerular pathology was assessed by a blinded observer based on
criteria and scoring methods adapted from prior studies
(Li et al., 2007;
Strevens et al., 2003) and after
consultation with an experienced renal pathologist at Xiangya Hospital.
For each kidney, 20 glomeruli were individually scored and the severity
of glomerular pathology was determined from the mean score for each
kidney. The sections of placentas were de-waxed, hydrated, and stained
with Hematoxylin and Eosin, and the morphology was examined by analyzing
10 fields in 5 sections from 3 placentas in each rat under light
microscopy.