The GIPC1-Akt1 Pathway Is Required for the Specification of the Eye Field in Mouse Embryonic Stem Cells

Abstract

During early patterning of the neural plate, a single region of the embryonic forebrain, the eye field, becomes competent for eye development. The hallmark of eye field specification is the expression of the eye field transcription factors (EFTFs). Experiments in fish, amphibians, birds and mammals have demonstrated largely conserved roles for the EFTFs. Although some of the key signaling events that direct the synchronized expression of these factors to the eye field have been elucidated in fish and frogs, it has been more difficult to study these mechanisms in mammalian embryos. In this study, we have used two different methods for directed differentiation of mouse embryonic stem cells (mESCs) to generate eye field cells and retina in vitro to test for a role of the PDZ domain-containing protein GIPC1 in the specification of the mammalian eye primordia. We find that the overexpression of a dominant-negative form of GIPC1 (dnGIPC1), as well as the downregulation of endogenous GIPC1, is sufficient to inhibit the development of eye field cells from mESCs. GIPC1 interacts directly with IGFR and participates in Akt1 activation, and pharmacological inhibition of Akt1 phosphorylation mimics the dnGIPC1 phenotype. Our data, together with previous studies in Xenopus, support the hypothesis that the GIPC1-PI3K-Akt1 pathway plays a key role in eye field specification in vertebrates.

Retinal Organoids from Pluripotent Stem Cells Efficiently Recapitulate Retinogenesis

“…Pluripotent embryonic stem cells (PSCs) facilitate research on mammalian neuronal development, neurodegenerative disorders, and regenerative therapies. It has been shown in the retina that developmental processes such as optic-vesicle (OV) and optic-cup (OC) morphogenesis and signaling cascades can be reproduced using mouse and human embryonic stem cells (mESCs and hESCs) ( Eiraku et al., 2011 , Nakano et al., 2012 , Hiler et al., 2015 , La Torre et al., 2015 ). Retinal organoid ( Boucherie et al., 2013 , Decembrini et al., 2014 , Gonzalez-Cordero et al., 2013 ) and 2D culture approaches ( Lamba et al., 2006 , Osakada et al., 2008 ) have been used for cell replacement therapy studies because efficient derivation of sufficient numbers of integration-competent cells remains a major limitation for regenerative medicine.…”

Effects of Adult Müller Cells and Their Conditioned Media on the Survival of Stem Cell-Derived Retinal Ganglion Cells

“…Due to this potential, pluripotent cells offer an unlimited source of donor cells to replace damaged cells in retinal degenerations. In the last decade, the ability to generate retinal neurons from pluripotent stem cells using three-dimensional (3D) organoid cultures has become well established [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. This method is based on plating dissociated pluripotent cells onto low-adhesion plates.…”

Synchrony and asynchrony between an epigenetic clock and developmental timing

“…The use of iPSCs in age-related disease modeling, show biomarkers of age-acceleration before the onset of age-related manifestations of the disease 12–17 . Evidence from many labs for both mouse and human indicates that developmental timing is conserved in tissues derived in vitro from pluripotent stem cells, such as cerebral and retinal organoids 18–22 , but it is not known whether tissues derived from iPSCs follow the same epigenetic clock as their corresponding fetal tissues.…”