Abstract
The purpose of the current study was to investigate whether peptidyl-prolyl cis/trans isomerase NIMA-interacting 1 (Pin1) can interact with calpastatin (CAST) and regulate CAST/calpain2, under excessive glutamate conditions, and subsequently regulate necrosis in rat retinal neurons. Glutamate triggered CAST/calpain2-mediated necrosis regulation in primary cultured retinal neurons, as demonstrated by propidium iodide-staining and lactate dehydrogenase assay. Co-IP results and a computer simulation suggested that Pin1 could bind to CAST. Western blot, real-time quantitative polymerase chain reaction, immunofluorescence, and phosphorylation analysis results demonstrated that CAST was regulated by Pin1, as proven by the application of juglone (i.e., a Pin1 specific inhibitor). The retinal ganglion cell 5 cell line, combined with siRNA approach and flow cytometry, was then used to verify the regulatory pathway of Pin1 in CAST/calpain2-modulated neuronal necrosis that was induced by glutamate. Finally, in vivo studies further confirmed the role of Pin1 in CAST/calpain2-modulated necrosis following glutamate excitation, in the rat retinal ganglion cell and inner nuclear layers. In addition, a flash electroretinogram study provided evidence for the recovery of impaired visual function, which was induced by glutamate, with juglone treatment. Our work aims to investigate the involvement of the Pin1-CAST/calpain2 pathway in glutamate-mediated excitotoxicity.
Highlights
Glutamate, which is one of the twenty essential amino acids, is the main physiological excitatory neurotransmitters in the central nervous system (CNS) (Veyrat-Durebex et al, 2016; Yan et al, 2016)
We investigated the role of prolyl cis/trans isomerase NIMA-interacting 1 (Pin1) in CAST/calpain2 pathway-modulated regulated necrosis, which was induced by glutamate, in rat retinal neurons, RGC-5 cell line, and in vivo
We demonstrated that the overload of glutamate may lead to regulated necrosis in retinal injury
Summary
Glutamate, which is one of the twenty essential amino acids, is the main physiological excitatory neurotransmitters in the central nervous system (CNS) (Veyrat-Durebex et al, 2016; Yan et al, 2016). Role of Pin1-CAST/Calpain in Regulated Necrosis “reversed uptake” (Rossi et al, 2000; Grewer et al, 2008). All of these processes maintain the glutamate concentration in the synaptic cleft, regulate activation of high-affinity extrasynaptic glutamate receptors (Diamond and Jahr, 1997; Fang et al, 2010). It should be noting that, glial cells play critical roles in neurons survival and function in numerous ways, such as maintenance of synaptic glutamate homeostasis and modulation of neuronal susceptibility to glutamate excitotoxicity, or lactatemediated metabolic coupling (Veyrat-Durebex et al, 2016; Madji Hounoum et al, 2017)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
