Expression of Gas1 in Mouse Brain: Release and Role in Neuronal Differentiation
Growth arrest-specific 1 (Gas1) is a pleiotropic protein that induces apoptosis of tumor cells and has important roles during development. Recently, the presence of two forms of Gas1 was reported: one attached to the cell membrane by a GPI anchor; and a soluble extracellular form shed by cells. Previously, we showed that Gas1 is expressed in different areas of the adult mouse CNS. Here, we report the levels of Gas1 mRNA protein in different regions and analyzed its expressions in glutamatergic, GABAergic, and dopaminergic neurons. We found that Gas1 is expressed in GABAergic and glutamatergic neurons in the Purkinje-molecular layer of the cerebellum, hippocampus, thalamus, and fastigial nucleus, as well as in dopaminergic neurons of the substantia nigra. In all cases, Gas1 was found in the cell bodies, but not in the neuropil. The Purkinje and the molecular layers show the highest levels of Gas1, whereas the granule cell layer has low levels. Moreover, we detected the expression and release of Gas1 from primary cultures of Purkinje cells and from hippocampal neurons as well as from neuronal cell lines, but not from cerebellar granular cells. In addition, using SH-SY5Y cells differentiated with retinoic acid as a neuronal model, we found that extracellular Gas1 promotes neurite outgrowth, increases the levels of tyrosine hydroxylase, and stimulates the inhibition of GSK3β. These findings demonstrate that Gas1 is expressed and released by neurons and promotes differentiation, suggesting an important role for Gas1 in cellular signaling in the CNS.
KeywordsGrowth arrest-specific 1 (Gas1) Glycogen synthase kinase 3β (GSK3β) Hippocampus Vesicular glutamate transporter 2 (VGLUT2) Glutamic acid decarboxylase of 67 kDa (GAD67) Retinoic acid (RA)
This study was partially supported by CONACyT (Mexico) Grants 239516 (JS) and PAPIIT/DGAPA IN205917 (R.A.-R.). The authors wish to thank Araceli Navarrete Alonso for their technical support and Rubén Sánchez for his laboratory assistance.
Designed research: EB, NZ, MLL, and JS; performed research: EB, MLL, NZ, NA-P, and PV; analyzed data: JAGB, EB, RAR, and JS; wrote the manuscript: EB and JS.
Compliance with Ethical Standards
Conflict of interest
The authors declare no competing financial interests nor other potential conflicts of interest.
Experiments were performed according to the current Mexican legislation NOM-062-ZOO-1999 (SAGARPA) and internal institutional guidelines, per authorization 0152-03, from Cinvestav (Comité Interno para el Cuidado y Uso de Animales de Laboratorio).
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