Administration of Histidine to Female Rats Induces Changes in Oxidative Status in Cortex and Hippocampus of the Offspring

Abstract

Histidinemia is an inherited metabolic disorder biochemically characterized by high concentrations of histidine in biological fluids. Usually affected patients are asymptomatic although some individuals have mental retardation and speech disorders. Considering the high prevalence of histidinemia and the scarce information on the effects of maternal histidinemia on their progeny, we investigated various parameters of oxidative stress in brain cortex and hippocampus of the offspring from female rats that received histidine (0.5 mg/g of body weight) in the course of pregnancy and lactation. At 21 days of age we found a significant increase of thiobarbituric acid reactive substances (TBARS), 2',7'-dihydrodichlorofluorescein oxidation, superoxide dismutase (SOD) activity, catalase (CAT) activity, total sulfhydryls and glutathione (GSH) content in cerebral cortex and hippocampus. We also verified that at 60 days of age, GSH, SOD and total sulfhydryls returned to normal levels in brain cortex, while the other parameters decreased in the same structure. In the hippocampus, at 60 days of age GSH returned to normal levels, CAT persisted elevated and the other parameters decreased. These results indicate that histidine administration to female rats can induce oxidative stress in the brain from the offspring, which partially recovers 40 days after breastfeeding stopped.

The effect of imatinib administered in the prenatal period on testis development in rats

“…Briefly, 1 ml of this buffer was added to 100 mg tissue from each specimen and was centrifuged for 10 min at 800 g at 4°C. 19…”

“…Standard 20, 10, 5, 2.5, 1.25, and 0.625 nmol/mL solutions were prepared from this main stock solution. 19…”

“…Tissue homogenization: A 7.4 pH homogenization buffer was prepared using 20 mM 1 L sodium phosphate and 140 mM potassium chloride pH for use in tissue homogenization. Briefly, 1 ml of this buffer was added to 100 mg tissue from each specimen and was centrifuged for 10 min at 800 g at 4 C. 19 TBARS assay: The TBARS assay was performed as described by Ohkawa et al For each specimen used, 200 ml testis tissue supernatant, 50 ml 8.1% sodium dodecyl sulfate (SDS), 375 ml 20% acetic acid (v/v), and 375 ml 0.8 pH 3.5 thiobarbituric acid (TBA) were placed into Eppendorf tubes. 20 A mixture involving the specified amounts was prepared from all solutions and vortexed.…”

“…TBARS concentrations were calculated as nmol/wet tissue. 19 Standard solution preparation: 1,1,3,3-tetra methoxy propane was used for a standard solution for standard curve production. A solution was then obtained by mixing 82.5 mL 1,1,3,3-tetra methoxy propane and 0.01 M 50 mL HCl, and was left to incubate for 1 h at 50 C. Standard 20, 10, 5, 2.5, 1.25, and 0.625 nmol/mL solutions were prepared from this main stock solution.…”

“…A solution was then obtained by mixing 82.5 mL 1,1,3,3-tetra methoxy propane and 0.01 M 50 mL HCl, and was left to incubate for 1 h at 50 C. Standard 20, 10, 5, 2.5, 1.25, and 0.625 nmol/mL solutions were prepared from this main stock solution. 19 Thiol analysis protocol: The Ellman method was employed for the thiol assay in testis tissue. 21 The sulfhydryl groups were determined in tissue using an Ellman separator: 4 mg dithiobis 2 nitrobenzoic acids (DTNB) was dissolved in 1% sodium citrate solution for this analysis.…”