Alteration of selective neurotransmitters in fetal brains of prenatally alcohol‐treated C57BL/6 mice: quantitative analysis using liquid chromatography/tandem mass spectrometry

Abstract

We previously demonstrated that prenatal alcohol exposure results in brain defects at different embryonic stages. This study is aimed at characterizing the influence of prenatal alcohol exposure on the levels of several neurotransmitters at early embryonic stage 13 (E13). Pregnant C57BL/6 mice were exposed to either a 25% ethanol derived calorie diet (ALC) or pair-fed (PF) liquid diet from E7 to E13. At E13, fetal brains were collected from dams of the ALC and PF groups. Liquid chromatography/tandem mass spectrometry (LC-MS) was then used to evaluate neurotransmitter levels. This approach involved the use of an LC column in conjunction with multiple-reaction monitoring mass spectrometry. Quantitative analyses of catecholamines, idolamine, and amino acid neurotransmitters revealed significant reductions in the levels of dopamine (p=0.004), norepinephrine (p=0.0009), epinephrine (p=0.0002), serotonin (p=0.004), and GABA (p=0.002) in the ALC group compared to the PF group. However, there was no significant change in the levels of glutamate in E13 fetal brains. These findings demonstrate that prenatal alcohol exposure reduces the concentrations of some catecholamines, idolamine, and amino acid neurotransmitters in E13 fetal brains. This study suggests that alterations of selective neurotransmitters may be the cause of abnormalities in brain function and behavior found in fetal alcohol spectrum disorders.

Bioconcentration, metabolism and neurotoxicity of the organophorous flame retardant 1,3-dichloro 2-propyl phosphate (TDCPP) to zebrafish

“…Monoamine extraction from both larvae (50, n=3 replicates) and from adult fish brains was performed as previously described (Wang et al, 2012;Sari et al, 2010).…”

“…Before extraction, 1 ng of DHB was added as internal standard (Sari et al, 2010). After weighing and homogenizing in 400 μL of 1% formic acid dissolved in pre-cooled acetonitrile, the samples were incubated for 15 min at -20°C, before centrifuging at 12000×g for 20 min at 4°C.…”

Transgenerational endocrine disruption and neurotoxicity in zebrafish larvae after parental exposure to binary mixtures of decabromodiphenyl ether (BDE-209) and lead

“…The extraction and measurement of selective neurotransmitters in 5 dpf larval offspring were performed as described previously (Sari et al, 2010). Briefly, the samples were homogenized in 400 mL of acetonitrile (1% formic acid) and incubated for 15 min at À20 C. After centrifugation at 12,000 Â g for 20 min at 4 C, the supernatant was freeze-dried and resuspended in 50 mL of 50% acetonitrile/water (v/v) for analysis.…”

Metabolomics and fetal alcohol spectrum disorder

“…In the study by Sari et al (2010), pregnant C57BL/6 mice were exposed to either a diet where 25% of the calories came from EtOH (ALC) or a pair-fed (PF) liquid diet from embryonic stages 7 to 13. Significant reductions in the levels of dopamine (p = 0.004), norepinephrine (p = 0.0009), epinephrine (p = 0.0002), serotonin (p = 0.004), and GABA (p = 0.002) were found in the ALC-fed group; however, no significant changes in Glu were found in the E13 fetal brains.…”