Modeling Fetal Alcohol Spectrum Disorder: validating an ex vivo primary hippocampal cell culture system
| Autor: | Elif Tunc-Ozcan, Adriana Ferreira, Eva E. Redei |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male medicine.medical_specialty GRB10 Adaptor Protein Primary Cell Culture Medicine (miscellaneous) Gene Expression Cell Cycle Proteins Hippocampal formation Toxicology Hippocampus Models Biological Article 03 medical and health sciences 0302 clinical medicine Insulin-Like Growth Factor II Pregnancy Internal medicine Gene expression medicine Hippocampus (mythology) Animals Genes Tumor Suppressor reproductive and urinary physiology Fetus Sex Characteristics biology Ethanol ras-GRF1 Gene Expression Profiling Receptors Somatomedin Receptor Insulin Rats Psychiatry and Mental health Insulin receptor 030104 developmental biology Endocrinology Real-time polymerase chain reaction Cell culture Fetal Alcohol Spectrum Disorders Prenatal Exposure Delayed Effects biology.protein Female 030217 neurology & neurosurgery Ex vivo Signal Transduction Transcription Factors |
| Popis: | Background Fetal alcohol spectrum disorder (FASD) is the leading nongenetic cause of mental retardation. There are no treatments for FASD to date. Preclinical in vivo and in vitro studies could help in identifying novel drug targets as for other diseases. Here, we describe an ex vivo model that combines the physiological advantages of prenatal ethanol (EtOH) exposure in vivo with the uniformity of primary fetal hippocampal culture to characterize the effects of prenatal EtOH. The insulin signaling pathways are known to be involved in hippocampal functions. Therefore, we compared the expression of insulin signaling pathway genes between fetal hippocampi (in vivo) and primary hippocampal culture (ex vivo). The similarity of prenatal EtOH effects in these 2 paradigms would deem the ex vivo culture acceptable to screen possible treatments for FASD. Methods Pregnant Sprague–Dawley rats received 1 of 3 diets: ad libitum standard laboratory chow (control-C), isocaloric pair-fed (nutritional control), and EtOH containing liquid diets from gestational day (GD) 8. Fetal male and female hippocampi were collected either on GD21 (in vivo) or on GD18 for primary culture (ex vivo). Transcript levels of Igf2, Igf2r, Insr, Grb10, Rasgrf1, and Zac1 were measured by reverse transcription quantitative polymerase chain reaction. Results Hippocampal transcript levels differed by prenatal treatment in both males and females with sex differences observed in the expression of Igf2 and Insr. The effect of prenatal EtOH on the hippocampal expression of the insulin pathway genes was parallel in the in vivo and the ex vivo conditions. Conclusions The similarity of gene expression changes in response to prenatal EtOH between the in vivo and the ex vivo conditions ascertains that these effects are already set in the fetal hippocampus at GD18. This strengthens the feasibility of the ex vivo primary hippocampal culture as a tool to test and screen candidate drug targets for FASD. |
| Databáze: | OpenAIRE |
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