Cell and Bioscience
·
2025
·Vol. 15
·期 1
·1-23
·
IF 6.2
Ji Hao-Nan,Zhou Hai-Qian,Qie Jing-Bo,Lu Wen-Mei,Gao Hai-Tao,Wu Dan-Hong
Background The identification and intervention of Alzheimer’s Disease (AD) in its early-stage allows for the timely implementation of lifestyle modifications and therapeutic strategies. Although dysregulation of protein expression has been reported in the brain from AD patients and AD animal models, the underlying mechanisms remain poorly understood. N4-acetylcytidine (ac4C), the only known form of RNA acetylation in eukaryotes, has recently been shown to regulate mRNA stability and translation efficiency. However, the dysregulation of ac4C associated with abnormal protein expression levels in the brain of early-stage mouse models of AD remains to be elucidated. Methods This study investigated ac4C modifications, mRNA and protein expression in the hippocampus of 3 and 6-month-old 5×FAD mice, a mouse model of AD, and wild-type (WT) littermates. The multi-omics analysis was performed: acetylated RNA immunoprecipitation followed by next-generation sequencing (acRIP-seq) to identify ac4C mRNAs, deep RNA sequencing (RNA-seq) to quantify mRNA abundance, and label-free quantitative proteomics to assess protein expression levels. In addition, we used acRIP-qPCR, regular qPCR and western blots to verify the ac4C, mRNA and protein levels of some key genes that were identified by the high-throughput assays. Results Proteomic analysis revealed significant change of protein expression in the hippocampus of 3-months-old 5×FAD mice, compared with WT littermates. In contrast, RNA-seq analysis indicated that there were no substantial alterations in mRNA expression levels in the hippocampus of 3-months-old 5×FAD mice, compared to WT littermates. Strikingly, acRIP-seq revealed notable variations in ac4C modification on mRNAs, particularly those associated with synaptic structure and function, in the hippocampus of 3-months-old 5×FAD mice, compared with WT littermates. The ac4C modifications were found to be correlated with protein expression changes. Genes that are essential for synaptic function and cognition, including GRIN1, MAP2, and DNAJC6, exhibited reduced ac4C and protein levels in 3-months-old 5×FAD mice, without any corresponding changes in the mRNA levels, compared with WT littermates. Moreover, only a small part of dysregulated ac4C mRNAs identified in the 3-month-old 5×FAD mice were found in the 6-month-old 5×FAD mice. Conclusions Altogether these results identified abnormal ac4C modification of mRNAs that may contribute to the dysregulation of protein synthesis in the hippocampus from an early-stage mouse model of AD.
研究领域:
神经生物学
·
分子生物学
·
表观遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Advanced Science
·
2025
·
IF 14.1
Yue Sun,Dan Liu,Yu Liang,Xue Yang,Xinxin Liu,Baofeng Zhao,Zhen Liang,Yukui Zhang,Lihua Zhang
Spatially resolved characterization of proteoforms has substantial potential to significantly advance the understanding of physiological and disease mechanisms. However, challenges remain regarding throughput and coverage. A robust method is developed for high-throughput proteoform imaging (HTPi) by combining matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI) and region-specific top-down proteomic analysis. MALDI MSI enables the imaging of proteoforms on tissue sections at a rate of 7 h cm −2 (100-µm spatial resolution), and the identification sensitivity of the proteoforms is improved by narrow-bore monolithic columns with low adsorption, yielding 366 annotated proteoform images from the mouse brain. The obtained proteoform images reveals differential expression of individual proteoforms across the brain regions, and distinct spatial distribution patterns of various proteoforms generated from a single gene. Given its ability to visualize proteoform, HTPi is further applied to explore spatial pathological changes associated with Alzheimer's disease (AD) in 5 × FAD mice. 158 annotated proteoform images are obtained in hippocampal regions at 50-µm spatial resolution, illuminating 14 differential proteoforms in the subiculum region and highlighting their significant associations with amyloid-β pathology in AD. The results highlight the power of HTPi in unraveling the intricate molecular landscape of brain tissues and its potential in elucidating disease mechanisms.
研究领域:
生物化学
·
神经科学
·
病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NEUROPATHOLOGY AND APPLIED NEUROBIOLOGY
·
2022
·Vol. 48
·期 7
·e12839
·
IF 6.25
Xiaolian Ye,Lu Chen,He Wang,Shixiao Peng,Mengjia Liu,Liyang Yao,Yizhi Zhang,Yun Stone Shi,Ying Cao,Jian-Jun Yang,Guiquan Chen
Aims Abundant recent evidence has shown that 3-phosphoinositide-dependent protein kinase 1 (PDK1) is activated in Alzheimer's disease (AD). However, it remains unknown whether inhibition of PDK1 in neurons may affect AD-like pathology in animal models of AD. Here, we aim to examine the effects of specific inactivation of neuronal PDK1 on pathology and behaviour in 5×FAD mice and to identify the underlying molecular mechanisms. Methods The Cre-loxP system was employed to generate Pdk1 cKO/5×FAD mice, in which PDK1 is inactivated in excitatory neurons in the adult forebrain. Cellular and behavioural techniques were used to examine plaque burden, inflammatory responses and spatial working memory in mice. Biochemical and molecular analyses were conducted to investigate relevant mechanisms. Results First, Aβ deposition was massively decreased and gliosis was highly attenuated in Pdk1 cKO/5×FAD mice compared with 5×FAD mice. Second, memory deficits were significantly improved in Pdk1 cKO/5×FAD mice. Third, APP levels were notably decreased in Pdk1 cKO/5×FAD mice. Fourth, mammalian target of rapamycin (mTOR) signalling and ribosome biogenesis were reduced in Pdk1 cKO/5×FAD mice. Conclusions Neuron-specific deletion of PDK1 robustly ameliorates AD-like pathology and improves spatial working memory in 5×FAD mice. We propose that genetic approach to inhibit PDK1 may be an effective strategy to slow AD.
研究领域:
神经科学
·
分子生物学
·
药理学
·
遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）