The International Journal of Developmental Biology

Int. J. Dev. Biol. 58: 35 - 43 (2014)

https://doi.org/10.1387/ijdb.130305md

Vol 58, Issue 1

Focal adhesion kinase as a mechanotransducer during rapid brain growth of the chick embryo

Original Article | Published: 21 March 2014

Mary E. Desmond1, Janice E. Knepper1, Angela J. DiBenedetto1, Elizabeth Malaugh1, Sagrario Callejo2, Raquel Carretero2, Maria-Isabel Alonso2 and Angel Gato2

1Department of Biology, Villanova University, Villanova, PA, USA and 2Departamento de Anatomia y Radiologia, Facultad de Medicina, Universidad de Valladolid, Valladolid, Spain

Abstract

Expansion of the hollow fluid-filled embryonic brain occurs by an increase in intraluminal pressure created by accumulation of cerebrospinal fluid (CSF). Experiments have shown a direct correlation between cavity pressure and cell proliferation within the neuroepithelium. These findings lead us to ask how mechanistically this might come about. Are there perhaps molecules on the luminal surface of the embryonic neuroepithelium, such as focal adhesion kinases (FAKs) known to respond to tension in other epithelial cells? Immunodetection using antibodies to total FAK and p-FAK was performed with subsequent confocal analysis of the pattern of their activation under normal intraluminal pressure and induced chronic pressure. Western analysis was also done to look at the amount of FAK expression, as well as its activation under these same conditions. Using immunolocalization, we have shown that FAK is present and activated on both apical and basolateral surfaces and within the cytoplasm of the neuroepithelial cells. This pattern changed profoundly when the neuroepithelium was under pressure. By Western blot, we have shown that FAK was upregulated and activated in the neuroepithelium of the embryos just after the neural tube becomes a closed pressurized system, with phosphorylation detected on the luminal instead of the basal surface, along with an increase in cell proliferation. Chronic hyper-pressure does not induce an increase in phosphorylation of FAK. In conclusion, here we show that neuroepithelial cells respond to intraluminal pressure via FAK phosphorylation on the luminal surface.

Keywords

cerebrospinal fluid, neuroepithelium, mitosis, hydrostatic pressure, FAK, cell proliferation

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