SNAP25 protects primary cortical neurons from hypoxic‐ischemic injury associated with CREB signal

Abstract

Background: Hypoxic-ischemic encephalopathy (HIE) could induce exacerbated changes and unpredictable efects in brain cells, and the mechanism remains unclear.Methods: HIE model was established in neonatal rats, Zea-Longa score and TTC staining were used to observe the neurobehavior and brain infarct volume in rats subjected to cerebral hypoxia-ischemia (HI). Primary cortical neurons were then cultured in vitro to establish an oxygen and glucose deprivation model. To determine the role of synaptosomal-associated protein-25 (SNAP25) in HIE, PC12 cells were cultured and efective siRNA fragments were screened, and SNAP25 was transfected into primary neurons. Then, quantitative real-time polymerase chain reaction was used to detect the mRNA expression level and immunofluorescence staining was used to observe the morphological changes of neurons before and after the injury. Finally, the abundance values of SNAP25 and its associated genes were iltered using the NCBI and GeneMANIA, respectively. Results: HI leads to a decrease in neuronal number and an increase in SNAP25 expression. Whereas, the interference of SNAP25 caused marked decrease in neuronal number and the length of neurite. Moreover, the expression levels of CREB and SYP were signiicantly decreased after interference of SNAP25. Conclusion:SNAP25 exhibited several neuroprotective efects to neuronal protection in neonatal cerebral HI by regulating CREB and SYP. others promote neuronal death (Papadopoulos MC, et al., 2000). In addition, HIE is often difficult to diagnose in real time because of convulsions and other non-speciic encephalopathy, and is the most common reason of perinatal asphyxia (Zhang Y, et al., 2016). HIE can also lead to severe brain damage and is a common cause of neurological handicaps in adulthood (Kojima T, et al., 2013). Currently, there is no efective clinical treatments to mitigate HI-induced brain injury, due to the lack of understanding of the neural networks associated with HIinduced neurodegeneration and its mechanisms (Liu S, Article history:

ER Stress Impedes Regulated Secretion by Governing Key Exocytotic and Granulogenic Molecular Switches

“…SNAP25 is one important membrane-associated t-SNARE required for the fusion of specialised secretory vesicles with the plasma membrane (Arora et al, 2017;Tsuboi and Fukuda, 2005;Walter et al, 2010). Such molecular switches are further regulated by master genes such as CREB (cAMP Response Element Binding protein), which is wellknown for regulating SNAP25 expression (Jin et al, 2021;Liew et al, 2010;Wang et al, 2019).…”

“…8C). It is noteworthy that CREB is an established positive regulator of SNAP25 expression (Jin et al, 2021;Wang et al, 2019). This unexpected finding led us to test the expression profile of Atf4, a competitive inhibitor of CREB (also known as CREB 2) and a downstream effector molecule of the PERK-EIF2α axis (Adjibade et al, 2017;Dey et al, 2012;Liew et al, 2010;Smith et al, 2020).…”

Downregulating EVA1C exerts the potential to promote neuron growth after neonatal hypoxic‐ischemic encephalopathy injury associated with alternative splicing

“…Then the brain damage was detected by behavior and morphology test. Zea‐longa score is usually applied to assess the degree of neurological impairment post constructing a brain injury model 23 . In addition, TTC staining after modeling can intuitively observe cerebral infarction, so as to confirm the success of the model 24 .…”

“…Zea-longa score is usually applied to assess the degree of neurological impairment post constructing a brain injury model. 23 In addition, TTC staining after modeling can intuitively observe cerebral infarction, so as to confirm the success of the model. 24 Moreover, HE and Nissl staining were performed to detect the neuronal damage in the brain, which can display the neuron morphology with normal and injury.…”

Differences between cultured cortical neurons by trypsin and papain digestion

“…After the volume was adjusted to 3 ml, the cells were counted, inoculated in a six‐well plate with 1 × 10 6 /well, and cultured in an incubator filled with 5% CO 2 at 37°C. After culturing for 4 h, the medium was replaced with neuron medium (Neurobasal + 2% B27 + 1% glutamine + 0.5% penicillin–streptomycin solution) 15–17 . Photographs were taken using an inverted phase‐contrast microscope.…”