TRPC3 is required for the survival, pluripotency and neural differentiation of mouse embryonic stem cells (mESCs)

Science China Life Sciences, Jan 2018

Transient receptor potential canonical subfamily member 3 (TRPC3) is known to be important for neural development and the formation of neuronal networks. Here, we investigated the role of TRPC3 in undifferentiated mouse embryonic stem cells (mESCs) and during the differentiation of mESCs into neurons. CRISPR/Cas9-mediated knockout (KO) of TRPC3 induced apoptosis and the disruption of mitochondrial membrane potential both in undifferentiated mESCs and in those undergoing neural differentiation. In addition, TRPC3 KO impaired the pluripotency of mESCs. TRPC3 KO also dramatically repressed the neural differentiation of mESCs by inhibiting the expression of markers for neural progenitors, neurons, astrocytes and oligodendrocytes. Taken together, our new data demonstrate an important function of TRPC3 with regards to the survival, pluripotency and neural differentiation of mESCs.

A PDF file should load here. If you do not see its contents the file may be temporarily unavailable at the journal website or you do not have a PDF plug-in installed and enabled in your browser.

Alternatively, you can download the file locally and open with any standalone PDF reader:

https://link.springer.com/content/pdf/10.1007%2Fs11427-017-9222-9.pdf

TRPC3 is required for the survival, pluripotency and neural differentiation of mouse embryonic stem cells (mESCs)

TRPC3 is required for the survival, pluripotency and neural differentiation of mouse embryonic stem cells (mESCs) Helen Baixia Hao 2 Sarah E. Webb 2 Jianbo Yue 1 Marc Moreau 0 Catherine Leclerc 0 Andrew L. Miller 2 0 Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université de Toulouse , CNRS, UPS, Toulouse F-31062 , France 1 Department of Biomedical Sciences, City University of Hong Kong , Hong Kong , China 2 Division of Life Science and State Key Laboratory of Molecular Neuroscience, HKUST, Clear Water Bay , Hong Kong , China •COVER ARTICLE• . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .March 2018 Vol.61 No.3: 253-265 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .https://doi.org/10.1007/s11427-017-9222-9 Transient receptor potential canonical subfamily member 3 (TRPC3) is known to be important for neural development and the formation of neuronal networks. Here, we investigated the role of TRPC3 in undifferentiated mouse embryonic stem cells (mESCs) and during the differentiation of mESCs into neurons. CRISPR/Cas9-mediated knockout (KO) of TRPC3 induced apoptosis and the disruption of mitochondrial membrane potential both in undifferentiated mESCs and in those undergoing neural differentiation. In addition, TRPC3 KO impaired the pluripotency of mESCs. TRPC3 KO also dramatically repressed the neural differentiation of mESCs by inhibiting the expression of markers for neural progenitors, neurons, astrocytes and oligodendrocytes. Taken together, our new data demonstrate an important function of TRPC3 with regards to the survival, pluripotency and neural differentiation of mESCs. - Citation: INTRODUCTION Embryonic stem cells (ESCs) have two distinguishing properties, they can proliferate in an unlimited manner (i.e., have the ability to self-renew) and they can differentiate into any type of somatic cell (i.e. they are pluripotent) (Bradley et al., 1984; Beddington and Robertson, 1989) . It is because of these properties that ESCs have the potential for use in cellbased therapies to treat a range of congenital, developmental, and degenerative conditions, including neurodegenerative diseases (McNeish, 2004; Hipp and Atala, 2008) . Thus, elucidating the cellular and molecular mechanisms that regulate neural fate determination from stem cells is a crucial first step before they can be successfully applied to regenerative medicine (Liu et al., 2013; Engel et al., 2016) . The neural differentiation of ESCs is a dynamic and complex process, which is regulated by combinatorial signaling pathways, and involves intrinsic transcription factors, cytokines, and growth factors, as well as epigenetic modifications (Dhara and Stice, 2008; Hayashi et al., 2008; Chuang et al., 2015) . In addition, Ca2+ is a versatile and universal intra- and inter-cellular second messenger, which is known to be vital for embryonic neurogenesis (Berridge et al., 2003; Leclerc et al., 2012) . Ca2+ signaling is also reported to be key for regulating the differentiation of ESCs into neuronal progenitor cells. For example, antagonizing the © Science China Press and Springer-Verlag GmbH Germany 2018 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . life.scichina.com link.springer.com activity of RyR2 (an ER-located Ca2+ release channel), CD38 (an enzyme that catalyzes the synthesis of cADPR and NAADP), neuronatin (a SERCA2 inhibitor), or STIM1 (an ER Ca2+ sensor), resulted in an inhibition of the neural differentiation of mouse ESCs (mESCs) (Yu et al., 2008; Lin et al., 2010; Hao et al., 2014; Wei et al., 2015; Hao et al., 2016) . The role of several of the transient receptor potential canonical (TRPC) channels in neural differentiation has also been investigated. For example, the inhibition of TRPC channel members 1 and 4 (TRPC1, TRPC4; both Ca2+ influx channels), has been shown to impair the proliferation of neuroepithelial cells and block neurite extension in post-mitotic neurons derived from human ESCs (hESCs) (Weick et al., 2009) . The contribution, however, of many other Ca2+ channels or pumps to the regulation of neural differentiation of ESCs is still largely unknown. However, numerous studies have shown that TRPC member 3 (TRPC3) is abundantly expressed in the brain, and various attributed functions in the central nervous system (CNS) have been described (Fusco et al., 2004; Amaral and Pozzo-Miller, 2007) . For example, using a TRPC3 knockout (KO) mouse model, it was proposed that TRPC3 mediates mGluR-dependent synaptic transmission in cerebellar Purkinje cells, and is crucial for motor coordination (Hartmann et al., 2008) . TRPC3 is a nonselective cation channel, which is permeable to both Na+ and Ca2+ (Lichtenegger and Groschner, 2014) . It is proposed to contribute to downstrea (...truncated)


This is a preview of a remote PDF: https://link.springer.com/content/pdf/10.1007%2Fs11427-017-9222-9.pdf

Helen Baixia Hao, Sarah E. Webb, Jianbo Yue, Marc Moreau, Catherine Leclerc, Andrew L. Miller. TRPC3 is required for the survival, pluripotency and neural differentiation of mouse embryonic stem cells (mESCs), Science China Life Sciences, 2018, pp. 253-265, Volume 61, Issue 3, DOI: 10.1007/s11427-017-9222-9