ADNP Differential Nucleus/Cytoplasm Localization in Neurons Suggests Multiple Roles in Neuronal Differentiation and Maintenance
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Complete deficiency in activity-dependent neuroprotective protein (ADNP) results in neural tube closure defects and death at gestation day 9 in mice. ADNP-deficient embryos exhibit dramatic increases in gene transcripts associated with lipid metabolism coupled to reduction in organogenesis/neurogenesis-related transcripts. In the pluripotent teratocarcinoma cell line P19, ADNP was shown to interact with specific chromatin regions in the neurodifferentiated state, which was associated with binding to the heterochromatin protein 1 alpha. In this study, using P19 cells as a differentiation model, we showed that ADNP expression and cytoplasm/nucleus distribution is unique in neuronal-differentiated cells compared to cardiovascular and nondifferentiated pluripotent cells. ADNP-like immunohistochemical localization to the neuronal cytoplasm and neurites was shown in this study not only in the cellular model but also in the brain cerebral cortex and olfactory bulb. Small hairpin RNA ADNP downregulation was used to further investigate ADNP involvement in p19 neurodifferentiation. An approximately 80% robust reduction in ADNP led to a substantial reduction in embryoid body formation and a significant reduction (approximately 50%) in neurite numbers. These results position ADNP in direct association with neuronal cell differentiation and maturation.
KeywordsADNP Differential nucleus/cytoplasm localization Neurons Neuronal differentiation and maintenance
This work is in partial fulfillment of the Ph.D. thesis requirements of Mr. Shmuel Mandel. We thank Dr. Joanna M. Hill for her initial help with techniques in immunohistochemistry and Professor Colin Barnstable for the monoclonal anti-tubulin antibodies. Support was provided by the BSF, ISF and Allon Therapeutics Inc. Professor Gozes is the incumbent of the Lily and Avraham Gildor Chair for the Investigation of Growth Factors, and the Director of the Adams Super Center for Brain Studies, the Levie-Edersheim-Gitter Institute for Functional Brain Imaging and the Dr. Diana and Zelman Elton (Elbaum) Laboratory for Molecular Neuroendocrinology and Allon Therapeutics. Professor Illana Gozes serves as the Chief Scientific Officer of Allon Therapeutics. NAP is in phase II clinical trials for neuroprotection.
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