Hostname: page-component-586b7cd67f-rdxmf Total loading time: 0 Render date: 2024-11-26T22:32:34.498Z Has data issue: false hasContentIssue false
  • English
  • Français

Morphological Alterations of CAD Cells Overexpressing AKT1

Published online by Cambridge University Press:  30 July 2020

James Wachira
James Wachira*
Affiliation:
Morgan State University, Baltimore, Maryland, United States

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

CAD cells are neuronal cells used in studies of cell differentiation and in cellular models of neuropathology. When cultured in differentiation medium, CAD cells exhibit characteristics of mature neurons including the generation of action potential. In addition to being a central signaling kinase in cell survival, AKT1 plays important roles in the nervous system including neuroplasticity and this study examined the localization of exogenous AKT1 in CAD cells. Neuropeptides modulate many signal transduction pathways and melacortins are implicated in regulating growth factor signal transduction pathways, including the PI3K/AKT pathway. AKT1-DsReD was transfected into CAD cells that were stably expressing melanocortin 3-receptor-GFP (MC3R-GFP), a G-protein coupled receptor. The cells were imaged with confocal microscopy to determine the fluorescent protein localization patterns. AKT1-DsRed was predominantly localized in the cytoplasm and the nucleus. Further, expression of exogenous AKT1 in these cell lines led to morphological changes reminiscent of apoptosis. As expected, MC3R-GFP localized to the plasma membrane but it internalized upon cell stimulation with the cognate ligand. In limited areas of the plasma membrane, AKT1-DsRed and MC3R-GFP were colocalized. In conclusion, quantitative studies to understand the role of relative levels of AKT1 in determining cell survival are needed.

Type
Biomedical and Pharmaceutical Research on the Development, Diagnosis, Prevention, and Treatment of Diseases
Information
Microscopy and Microanalysis , Volume 26 , Supplement S2 , August 2020 , pp. 1354 - 1358
Check for updates
Copyright
Copyright © Microscopy Society of America 2020

References

Breit, A., Wolff, K., Kalwa, H., Jarry, H., Büch, T. & Gudermann, T. (2006). The natural inverse agonist agouti-related protein induces arrestin-mediated endocytosis of melanocortin-3 and -4 receptors. The Journal of Biological Chemistry 281, 37447–37456.10.1074/jbc.M605982200CrossRefGoogle ScholarPubMed
Fremuntova, Z., Mosko, T., Soukup, J., Kucerova, J., Kostelanska, M., Hanusova, Z. B., Filipova, M., Cervenakova, L. & Holada, K. (2020). Changes in cellular prion protein expression, processing and localisation during differentiation of the neuronal cell line CAD 5. Biology of the Cell 112, 1–21.10.1111/boc.201900045CrossRefGoogle ScholarPubMed
Fu, F., Li, L.-S., Li, R., Deng, Q., Yu, Q.-X., Yang, X., Pan, M., Han, J., Zhen, L., Zhang, L.-N., Lei, T.-Y., Li, D.-Z. & Liao, C. (2020). All-trans-retinoid acid induces the differentiation of P19 cells into neurons involved in the PI3K/Akt/GSK3β signaling pathway. Journal of Cellular Biochemistry.10.1002/jcb.29659CrossRefGoogle ScholarPubMed
Hers, I., Vincent, E. E. & Tavaré, J. M. (2011). Akt signalling in health and disease. Cellular Signalling 23, 1515–1527.10.1016/j.cellsig.2011.05.004CrossRefGoogle ScholarPubMed
Jean-Charles, P.-Y., Kaur, S. & Shenoy, S. K. (2017). GPCR signaling via β-arrestin-dependent mechanisms. Journal of cardiovascular pharmacology 70, 142–158.10.1097/FJC.0000000000000482CrossRefGoogle Scholar
Li, Y., Hou, L. X.-E., Aktiv, A. & Dahlström, A. (2005). Immunohistochemical characterisation of differentiated CAD cells: expression of peptides and chromogranins. Histochemistry and Cell Biology 124, 25–33.10.1007/s00418-005-0017-9CrossRefGoogle ScholarPubMed
Nyan, D. C., Anbazhagan, R., Hughes-Darden, C. A. & Wachira, S. J. M. (2008). Endosomal colocalization of melanocortin-3 receptor and beta-arrestins in CAD cells with altered modification of AKT/PKB. Neuropeptides 42, 355–366.10.1016/j.npep.2007.12.007CrossRefGoogle ScholarPubMed
Yang, L.-K. & Tao, Y.-X. (2017). Biased Signaling at Neural Melanocortin Receptors in Regulation of Energy Homeostasis. Biochimica et biophysica acta 1863, 2486–2495.10.1016/j.bbadis.2017.04.010CrossRefGoogle ScholarPubMed