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Fetal Brain Injury Models of Fetal Alcohol Syndrome: Examination of Neuronal Morphologic Condition Using Sholl Assay

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Part of the Methods in Molecular Biology book series (MIMB,volume 2311)

Abstract

The lack of a convenient in vitro human neuronal model to study alcohol-induced neurodegenerative diseases, such as fetal alcohol syndrome (FAS), prompted us to develop human neuronal culture and in vitro human FAS model by incubating cells with physiologically relevant EtOH concentration (50 mM). Here, we describe the detailed method of isolation of human neuronal culture, and ability to analyze neurites extension using Sholl assay. We utilized highly efficient transfection method of neuronal cells to study morphology of neurons with or without EtOH treatment.

Key words

  • Human neuronal cells
  • Alcohol
  • FAS
  • Sholl assay

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  • DOI: 10.1007/978-1-0716-1437-2_16
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References

  1. Eysseric H, Gonthier B, Soubeyran A, Bessard G, Saxod R, Barret L (1997) There is no simple method to maintain a constant ethanol concentration in long-term cell culture: keys to a solution applied to the survey of astrocytic ethanol absorption. Alcohol 14(2):111–115

    CAS  CrossRef  Google Scholar 

  2. Rodriguez FS, Siminsson P, Alling C (1992) A method for maintaining constant ethanol concentrations in cell culture media. Alcohol Alcohol 27:309–313

    CAS  PubMed  Google Scholar 

  3. De Simone FI, Darbinian N, Amini S, Muniswamy M, White MK, Elrod JW, Datta PK, Langford D, Khalili K (2016) HIV-1 Tat and cocaine impair survival of cultured primary neuronal cells via a mitochondrial pathway. J Neuroimmune Pharmacol 11(2):358–368. https://doi.org/10.1007/s11481-016-9669-6. Epub 2016 Mar 31

    CrossRef  PubMed  PubMed Central  Google Scholar 

  4. Darbinyan A, Pozniak P, Darbinian N, White MK, Khalili K (2013a) Compartmentalized neuronal cultures. Methods Mol Biol 1078:147–152. https://doi.org/10.1007/978-1-62703-640-5_13

    CrossRef  PubMed  PubMed Central  Google Scholar 

  5. Darbinyan A, Kaminski R, White MK, Darbinian N, Khalili K (2013b) Isolation and propagation of primary human and rodent embryonic neural progenitor cells and cortical neurons. Methods Mol Biol 1078:45–54. https://doi.org/10.1007/978-1-62703-640-5_5

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  6. Darbinian N, Khalili K, Amini S (2014) Neuroprotective activity of pDING in response to HIV-1 Tat. J Cell Physiol 229(2):153–161. https://doi.org/10.1002/jcp.24392

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  7. Dell’Ovo V, Rosenzweig J, Burd I, Merabova N, Darbinian N, Goetzl L (2015) An animal model for chorioamnionitis at term. Am J Obstet Gynecol 213(3):387.e1–387.10. https://doi.org/10.1016/j.ajog.2015.05.007. Epub 2015 May 12

    CAS  CrossRef  Google Scholar 

  8. Braunewell KH, Dwary AD, Richter F, Trappe K, Zhao C, Giegling I, Schönrath K, Rujescu D (2011) Association of VSNL1 with schizophrenia, frontal cortical function, and biological significance for its gene product as a modulator of cAMP levels and neuronal morphology. Transl Psychiatry 1:e22

    CAS  CrossRef  Google Scholar 

  9. Yanni PA, Lindsley TA (2000) Ethanol inhibits development of dendrites and synapses in rat hippocampal pyramidal neuron cultures. Dev Brain Res 120:233–243

    CAS  CrossRef  Google Scholar 

  10. Wang JY, Darbinyan A, White MK, Darbinian N, Reiss K, Amini S (2014) Involvement of IRS-1 interaction with ADAM10 in the regulation of neurite extension. J Cell Physiol 229(8):1039–1046. https://doi.org/10.1002/jcp.24528

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  11. Li H, Xue Q, Xu X (2020) Involvement of the nervous system in SARS-CoV-2 infection. Neurotox Res 38(1):1–7

    CrossRef  Google Scholar 

  12. Zhou Z, Kang H, Li S, Zhao X (2020) Understanding the neurotropic characteristics of SARS-CoV-2: from neurological manifestations of COVID-19 to potential neurotropic mechanisms. J Neurol 267(8):2179–2184

    CAS  CrossRef  Google Scholar 

  13. Natoli S, Oliveira V, Calabresi P, Maia LF, Pisani A (2020) Does SARS-Cov-2 invade the brain? Translational lessons from animal models. Eur J Neurol 27(9):1764–1773. https://doi.org/10.1111/ene.14277

    CAS  CrossRef  PubMed  Google Scholar 

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Correspondence to Nune Darbinian .

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Darbinian, N., Darbinyan, A., Khalili, K., Amini, S. (2021). Fetal Brain Injury Models of Fetal Alcohol Syndrome: Examination of Neuronal Morphologic Condition Using Sholl Assay. In: Amini, S., White, M.K. (eds) Neuronal Cell Culture. Methods in Molecular Biology, vol 2311. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1437-2_16

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  • DOI: https://doi.org/10.1007/978-1-0716-1437-2_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1436-5

  • Online ISBN: 978-1-0716-1437-2

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