Animal Cells and Systems最新文献

Parkinson's disease (PD) is a progressive neurological disorder characterized by the degeneration of midbrain dopaminergic neurons and disabling motor impairments. Heat shock protein family A member 9 (HSPA9) play a crucial role in neuronal homeostasis by regulating the import of various mitochondrial proteins. HSPA9 is down-regulated in neurodegenerative diseases such as Alzheimer's disease and PD, and its loss leads to excessive mitochondrial fragmentation with oxidative stress, which subsequently causes damage to dopaminergic neurons. Moreover, HSPA9 interacts with multiple PD-associated proteins, including Pink1, DJ-1, and α-synuclein, however precise roles of HSPA9 in PD pathophysiology remain unclear. To further explore the contributions of HSPA9 in PD pathogenesis, we developed an HSPA9 knockout mouse. Haploinsufficiency of Hspa9 (Hspa9 ^+/-) was associated with the loss of tyrosine hydroxylase-positive neurons in the striatum and substantia nigra. Furthermore, Hspa9 haploinsufficiency induced excessive mitochondrial fission, enhanced apoptotic signaling, and resulted in diminished motor performance during the rotarod test. Administration of the mitochondrial neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in Hspa9 ^+/- mice further exacerbated the loss of dopaminergic neurons, aggravated motor impairments, and enhanced activation of apoptosis effector caspase-3. These results suggest that down-regulation of HSPA9 may contribute to the development and progression of PD, potentially offering a new therapeutic strategy for PD treatment.

Bisphenol A (BPA) is a widely used xenoestrogen that can disrupt neuroendocrine and immune regulation through multiple hormone receptors. This study investigated BPA-induced long non-coding RNA (lncRNA)-mRNA interactions in the cerebral cortex and hypothalamic-pituitary-thyroid (HPT) axis of adult male mice. Transcriptome sequencing and comprehensive lncRNA annotation identified 14,858 novel lncRNA transcripts. Integrated network analysis using weighted gene co-expression network analysis (WGCNA) revealed four distinct tissue-specific modules: neuronal signaling alterations (Tac1, Htr1b, Npy), RNA splicing modifications (Srsf5), PI3K/Akt-mediated cellular dysfunction (Creb5, Cdkn1a), and immune receptor signaling disruptions (Trbv15, Fcrla). These findings suggest that BPA reprograms transcriptional networks in a tissue-specific manner, potentially disrupting hormone-related neurotransmission, metabolic regulation, and immune signaling via lncRNA-mediated mechanisms. Such systems-level reprogramming of the immune-neuroendocrine network (INEN) provides novel mechanistic insights and biomarker candidates for assessing and mitigating the health impacts of environmental endocrine disruptors.

Pro-opiomelanocortin (POMC) neurons in the hypothalamic arcuate nucleus (ARC) play a pivotal role in regulating brown adipose tissue (BAT) thermogenesis via the sympathetic nervous system. The activation of transient receptor potential ankyrin 1 (TRPA1) has been demonstrated to enhance heat production, particularly in BAT. However, no direct evidence has been reported regarding BAT thermogenesis mediated by TRPA1-regulated ARC POMC neurons. This study aimed to investigate the role of TRPA1-expressing hypothalamic POMC neurons in BAT thermogenesis. To confirm TRPA1 expression in ARC POMC neurons, we employed single-cell reverse transcriptase polymerase chain reaction and immunolabeling techniques. Selective TRPA1 agonists, including capsiate and ASP7663, induced depolarization of ARC POMC neurons, an effect that was inhibited by A967079, a TRPA1-selective antagonist. Furthermore, intracerebroventricular (i.c.v.) administration of ASP7663 increased BAT and core body temperature. The thermogenic effect of ASP7663 in BAT was abolished by co-administration of A967079. Among the BAT thermogenic markers, peroxisome proliferator-activated receptor gamma coactivator 1-alpha and PR domain containing 16 (PRDM16) expressions were considerably upregulated following i.c.v. administration of ASP7663. However, this increase was reversed by A967079, except for PRDM16. These findings indicate that TRPA1-mediated activation of hypothalamic POMC neurons is critical in regulating BAT thermogenesis and promoting energy expenditure.

Nesfatin-1, initially identified as an appetite-regulating hormone, has also been detected in various cancer tissues and implicated in tumorigenesis. However, its role in the proliferation and migration of lung cancer cells remains unclear. This study aims to investigate the effects of nesfatin-1 on the proliferation and migration of human lung cancer cells and elucidate the underlying molecular mechanisms. The expression of nesfatin-1 protein and NUCB2 mRNA was detected in the immortalized normal human bronchial cell line BEAS-2B and the non-small-cell lung cancer cell line H1299. Immunohistochemical staining revealed the localization of nesfatin-1 binding sites in both cell lines. Nesfatin-1 treatment significantly increased the proliferation and migration of BEAS-2B cells but not of H1299 cells. The expression levels of cell proliferation-related genes, such as TGFα, PXN, MTOR, and CCND1, were upregulated in BEAS-2B cells, with no significant changes observed in H1299 cells. In addition, phosphorylation of FAK, PI3 K, and AKT was increased in BEAS-2B cells, whereas only FAK phosphorylation was increased in H1299 cells. To further assess the role of endogenous nesfatin-1, NUCB2 expression was silenced using small interfering RNA. Knockdown of NUCB2 suppressed proliferation and migration of BEAS-2B cells, as well as their expression of TGFα, PXN, MTOR, and CCND1; however, it had no significant effect on H1299 cells. These results suggest that nesfatin-1 promotes proliferation and migration in normal lung epithelial cells but not in lung cancer cells. Further research is needed to elucidate the molecular mechanisms underlying the differential effects of nesfatin-1 on normal and cancerous lung cells.

Mammalian spermatozoa acquire fertilizing ability in response to environmental factors enriched in the female reproductive tract, a process called capacitation. During capacitation, sperm undergo physiological changes that are accompanied by functional regulation of sperm proteins. However, the mechanism by which capacitation orchestrates sperm protein functions to modulate physiological characteristics remains unclear. Here, I analyzed capacitation-mediated global proteomic changes in mouse spermatozoa to unravel the underlying molecular association with the biological processes in sperm capacitation. I quantitatively compared 4,587 proteins identified by liquid chromatography-tandem mass spectrometry. Among them, the amounts of 47 and 180 proteins were reduced to over 1.5-fold (p < 0.05) and 1.3-fold (p < 0.1), respectively, and those of 11 and 60 proteins were increased over 1.5-fold (p < 0.05) and 1.3-fold (p < 0.1), respectively, in capacitated mouse sperm. Functional annotation of proteins reduced in capacitated sperm revealed that these proteins could be associated with lipid metabolism, RNA processing, and remodeling of the nuclear envelope structure. This result suggests that reactive oxygen species might be more generated for cholesterol efflux and the nucleus might undergo decondensation to form pronucleus in sperm during capacitation. In addition, functional annotation of proteins of which levels are increased in capacitated sperm represents that they could be involved in sperm structure. This study expands the molecular contribution to modulation of sperm functions and provides new insights into potential biological processes involving regulatory molecular machinery in capacitated sperm.

Breakthrough discoveries in RNA biology have led to a paradigm shift in our understanding of RNA-from passive intermediates to active regulators of gene expression. Technological innovations and deeper insights into RNA regulation have transformed the field, positioning RNA as a powerful tool for therapeutic development. Recent advances in RNA technologies have revolutionized medicine by enabling the precise targeting of specific mRNAs to modulate aberrant transcripts, correct genetic defects, and reprogram cellular behavior. This review provides an overview of the coordinated regulation of mRNA processing and its application to RNA-based therapeutics, including antisense oligonucleotides (ASOs), splice-switching oligonucleotides (SSOs), small interfering RNAs (siRNAs), and Aptamers. We focus on clinically approved RNA therapeutics, emphasizing their biological mechanisms such as RNA stability and splicing regulation. The expanding repertoire of RNA technologies underscores the translational potential of RNA biology and its growing clinical impact. Future developments are expected to yield highly specific, modular, and programmable RNA medicines capable of treating a wide range of previously intractable diseases.

Xanthoxyline, a plant-derived phytochemical, has anti-bacterial, anti-fungal, and anti-cancer activities. We intended to investigate the effect of xanthoxyline on the response to oxidative stress, aging, and Parkinson's disease. The effects of dietary supplementation with xanthoxyline on stress response and aging were examined in vivo using Caenorhabditis elegans as a model system. Genetic analysis using mutants, RNAi, and quantitative RT-PCR was performed to identify underlying mechanism involved in xanthoxyline-induced longevity. Animal disease models were employed to examine the effect of xanthoxyline on Parkinson's disease. Xanthoxyline increased resistance to the oxidative stress induced by H₂O₂. The mean lifespan of worms was significantly increased by supplementation with xanthoxyline. The lifespan-extending activity of xanthoxyline was not accompanied by reduced fertility. Xanthoxyline delayed the age-related decline in motility. Interestingly, the expression of two longevity-assuring genes, hsp-16.2, and sod-3, was increased by xanthoxyline supplementation. Genetic analysis suggested that lifespan extension by xanthoxyline was mediated by activation of autophagy and required DAF-16. In a model of Parkinson's disease, degeneration of dopaminergic neurons was prevented by supplementation with xanthoxyline, in a manner dependent on DAF-16. Taken together, we concluded that xanthoxyline exerts an anti-aging activity, possibly by activating the DAF-16-dependent stress response, and reduces the risk of Parkinson's disease, in a manner mediated by DAF-16. Xanthoxyline shows promise for the development of novel nutraceuticals against aging and Parkinson's disease.

RNA binding proteins (RBPs) play crucial roles in the post-transcriptional regulation of metabolic pathways. Although the RBP HuD has been extensively studied in pancreatic β-cells, its role in cellular metabolism remains poorly understood. In this study, we uncover a novel function of HuD in regulating fatty acid oxidation (FAO) in mouse insulinoma βTC6 cells. Through genetic knockdown and overexpression approaches, we demonstrate that HuD modulates the expression of long-chain acyl-CoA dehydrogenase (LCAD), a key enzyme in FAO, by binding to the 3'-untranslated region of its mRNA. Loss of HuD impaired FAO, leading to lipid droplet accumulation, elevated reactive oxygen species production, and increased lipotoxicity under lipid-stress conditions. These findings reveal a previously unrecognized role for HuD in maintaining fatty acid homeostasis and suggest that the HuD-LCAD regulatory axis may represent a promising therapeutic target for preserving β-cell integrity and function.

Mismatch repair (MMR) deficiency is a hallmark of microsatellite instability (MSI) in hereditary non-polyposis colorectal cancer, Lynch syndrome, contributing to resistance against conventional chemotherapy and posing a significant therapeutic challenge. In this study, we introduce UNI110, a novel small molecule derived from Baicalein, engineered for enhanced selectivity against MMR-deficient cancer cells. UNI110 exhibits a remarkable sevenfold increase in potency over Baicalein, demonstrating significantly lower IC50 values and heightened cytotoxic effects in MMR-deficient cell lines. Mechanistically, UNI110 selectively induces DNA damage in MMR-deficient cancer cells, ultimately resulting in cell death. Furthermore, UNI110 disrupts homologous recombination (HR) repair by inhibiting the MSH2-MSH3 complex, specifically blocking the interaction between MSH2 and EXO1, thereby impairing long-range end resection during double-strand break (DSB) repair. These findings establish UNI110 as a promising lead compound for the targeted treatment of MMR-deficient colorectal cancers, offering a potential breakthrough in overcoming chemotherapy resistance and improving patient outcomes.

The additional sex combs-like 1 (Asxl1) gene is a chromatin regulator involved in transcriptional activation and repression. While Asxl1 plays a crucial role in various organ development, its role in ocular development remains unclear. Here, we analyzed Asxl1 knockout (KO) mice and observed disrupted optic cup formation at embryonic day 10.5 (E10.5). RNA-seq of the E10.5 optic cup revealed dysregulation of Wnt signaling and early eye development genes. In further investigation using isolated cell from E10.5 retinal region, neuroepithelial stem cells from Asxl1 KO embryos exhibited impaired proliferation and spheroid formation. To elucidate the transcriptional mechanism by Asxl1 in optic cup formation, biochemical assays demonstrated that Asxl1 binds the LIM domain of Lhx2, facilitating repression of Wnt1, Wnt2, and Wnt8b. Following ChIP analysis showed that the gain of function of Asxl1 increased repressive histone marks (H3K27me3, H3K9me3) and reduced active marks (H3K4me3) at Lhx2-binding motifs within the cis-regulatory regions of canonical Wnt ligand genes. These findings establish Asxl1 as a key epigenetic regulator of optic cup development by modulating Lhx2-mediated Wnt signaling, providing insights into congenital eye disorders.