ABSTRACT
@#AIM:To evaluate the effects of Beta-B2 crystallin(CRYBB2)knockout on autophagy of mouse lens.<p>METHODS:Six-month-old WT and <i>Crybb2</i>KO mice were selected respectively. The morphological changes of autophagy of lens were observed by transmission electron microscopy. The expression of autophagy related proteins in the two groups were detected by Western blot method. <p>RESULTS:Compared with the control group, transmission electron microscopy revealed that mitochondria was accumulated and the number of autophagosomes in lens were higher in <i>Crybb2</i>KO mice. The relative expression of LC3B in <i>Crybb2</i>KO group was lower(0.09±0.01 <i>vs</i> 0.26±0.05). The P62 protein and p-mTOR(0.64±0.09 and 0.41±0.03)was higher than WT group(0.43±0.07 and 0.27±0.02).<p>CONCLUSION:The deletion of CRYBB2 may affect the process of lens autophagy by mTOR pathway and lead to cataract formation.
ABSTRACT
Objective To explore the expression profile of lncRNAs in the testis tissue of CRYBB2 gene knockout (KO) mice and its possible role in the testis development. Methods Testis tissues(n=3)from wild-type (WT) and CRYBB2 KO mice were subjected to lncRNA and mRNA microarray profiling. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to group the differentially expressed lncRNAs into regulated gene pathways and functions. The correlation matrix method was used to establish a network of lncRNA and mRNA co-expression. Quantitative (q)RT-PCR was used to verify expression of some differentially expressed lncRNAs and mRNAs. Results There were 140 differentially expressed lncRNAs and 477 differentially expressed mRNAs between testis tissues from WT and KO mice. There were 12 differentially expressed lncRNAs through the analyses of the GO, with 7 up-regulated and 5 down-regulated. The KEGG analysis showed that these differentially expressed mRNAs played important roles in Ca2+ signaling, ligand and receptor interactions, and so on. The correlation matrix method established an lncRNA and mRNA co-expression network, consisting of 9 lncRNAs and 8 mRNAs, with 17 nodes and 12 connections. Furthermore, expression of gene Rsl1 was regulated by three lncRNAs, expression of gene Lpo and gene Mpo was regulated by two lncRNAs, and expression of gene Hdac1 and gene Ephb4 was regulated by one lncRNA. qRT-PCR confirmed the significant down-regulation of lncRNA A-30-P01019163 expression, which significantly down-regulated its downstream gene P2rx7 in testis tissues of CRYBB2 KO mice(P<0.05). Conclusion LncRNA is closely related to the non-crystalline lens function of CRYBB2. LncRNA A-30-P01019163 may affect testicular cell cycle and signaling pathway by regulating P2rx7 expression in the testis tissues.