LncRNA Gm16638-201 Is Involved In Depressive-Like Behaviors By Inhibiting The 14-3-3ε Pathway In Murine Prefrontal Cortex

Objective: To explore the molecular mechanism of lncRNA Gm16638-201 in the prefrontal cortex of depressive mouse.Methods: 1.The depressive mouse model was constructed by chronic unpredictable mild stress(CUMS).The body weight and behaviors were measured by sucrose preference test(SPT),tail suspension test(TST),forced swimming test(FST)and bead embedding.2.The expression of Gm16638-201 in prefrontal cortex was detected by qRT-PCR.3.The subcellular localization of Gm16638-201,cis or trans regulatory genes and possible pathways of were predicted and analyzed by bioinformatics methods.4.The primary mouse cortical neurons were cultured in vitro,neurofilament protein(NF-L)was detected by immunofluorescence.Gm16638-201 was overexpressed by adenoviral vector mediated infection.5.RT-PCR was used to detect the expression of Gm16638-201 and its cis /trans regulatory genes,including Elmo2,Satb1,Hnrnpul1,Sipa1l3,Mapt,Tada3,Sgip1 and Star D5.6.ELISA assays was used to determine interleukin-16(IL-16).7.Western Blot was used to detected 14-3-3ε,MAPT and BAD in the prefrontal cortex of CUMS mice and the primary cells of mouse cortical neurons overexpressing Gm16638-201.Results: 1.Compared with normal BALB/c male mice,the mice in CUMS group showed depressive behavioral manifestations including body weight loss,sucrose intake decrease in SPT,struggle time reduction in TST and FST and decreased number of buried beads.2.The qRT-PCR showed that the expression of Gm16638-201 was up-regulated in the prefrontal cortex of CUMS mice(P<0.01).3.Bioinformatics analysis showed that Gm16638-201 is mainly expressed in the cytoplasm and nine neural related genes(Elmo2,Satb1,Hnrnpul1,Sipa1l3,Mapt,Tada3,Sgip1,Star D5 and IL-16)may be regulated by Gm16638-201 in cis or trans.These nine genes may intact with 14-3-3ε pathway.4.Immunofluorescence staining showed that neuronal specifically protein NF-L was expressed in the primary cortical neurons.5.The expression of Elmo2,Satb1,Hnrnpul1,Sipa1l3,Mapt,Tada3 and Star D5 were down-regulated at the transcriptional level in Gm16638-201 overexpressed primary mouse cortical neurons(P<0.01)While the expression of Sgip1 was no significantly(P>0.05).6.Compared with the NOR group,the expression of IL-16 was up-regulated in CUMS group(P<0.01).Compared with the PrimeN and NC group,the expression of IL-16 was also up-regulated in Gm16638-201-Exp group(P<0.01).7.Compared with the NOR group,the expression of 14-3-3ε,MAPT and BAD was down-regulated in in the prefrontal cortex of CUMS mice(P<0.01).Compared with the Prime-N and NC group,the expression of IL-16 was also up-regulated in Gm16638-201-Exp group(P<0.01).Conclusion: LncRNA Gm16638-201 may be involved in mouse depressivelike behavior by regulating the expression of Elmo2,Satb1,Hnrnpul1,Sipa1l3,Mapt,Tada3,Star D5 and IL-16 and inhibiting the 14-3-3ε pathway.