Normal Mice Research Articles

Aberrant outputs of glutamatergic neurons in deep cerebellar nuclei mediate dystonic movements.

Dystonia, characterized by repetitive twisting movements or abnormal postures, has been linked to the deep cerebellar nuclei (DCN). However, the specific roles of distinct neuronal populations within the DCN in driving dystonic behaviors remain unclear. This study explores the contributions of three distinct groups of DCN neurons in an animal model of paroxysmal dystonia harboring a mutation in the proline-rich transmembrane protein 2 (Prrt2) gene. We observed sustained calcium activity elevation across glutamatergic, glycinergic, and GABAergic inferior olive (IO)-projecting neurons within the DCN during episodes of dystonia in Prrt2-mutant mice. However, only the optogenetic activation of DCN glutamatergic neurons, but not glycinergic or GABAergic IO-projecting neurons, elicited dystonia-like behaviors in normal mice. Selective ablation of DCN glutamatergic neurons effectively eliminated aberrant cerebellar DCN outputs and alleviated dystonia attacks in both Prrt2-associated and kainic acid-induced dystonia mouse models. Collectively, our findings highlight the pivotal role of aberrant activation of DCN glutamatergic neurons in the neuropathological mechanisms underlying cerebellar-originated dystonia.

Mechanisms of LncRNA FTX in Regulating Islet Function of Pregnant Mice Born With Low-Protein Diet-Induced Intrauterine Growth Retardation.

Glucose metabolism during pregnancy in adult females born with intrauterine growth restriction (IUGR) remains inadequately understood. This study aims to investigate how LncRNA FTX regulates islet function during pregnancy in F1 female mice born with IUGR (F1 IUGR pregnant mice). A pregnant mouse model was established using F1 female mice born with IUGR (F1 IUGR pregnant mouse model). Intraperitoneal glucose tolerance test (IPGTT), immunohistochemistry (IHC) staining, quantitative real-time PCR (qPCR) were performed in both F1 IUGR and normal mice during pregnancy and non-pregnancy periods. RNA-sequencing was conducted on islets from F1 IUGR and normal pregnant mice. Insulin-related gene expression analysis, cell proliferation, and apoptosis assessment were performed in TC6 cells following FTX knockdown or overexpression. A luciferase reporter assay was conducted to validate the molecular interactions. F1 IUGR pregnant mice exhibited a smaller increase in insulin-staining area and lower upregulation of insulin-related gene expression levels compared to normal pregnant mice. There were 1,007 differentially expressed lncRNAs between F1 IUGR and normal pregnant islets; among these, FTX was down-regulated during pregnancy, although its downregulation in F1 IUGR pregnant mice was less pronounced than in normal pregnant mice. FTX was closely related to cell proliferation activity, apoptosis, insulin-related transcription factor expression. The pten/PI3K/AKT pathway was also regulated by FTX. Luciferase reporter assay confirmed FTX acted as a competing endogenous RNA (CeRNA) to target pten by sponging miR-22-3p. LncRNA FTX regulates islet function during pregnancy in F1 mice born with IUGR via the miR-22-3p/pten axis.

Docosahexaenoic Acid and Nervonic Acid Synergically Enhance Cognitive Memory in Normal Mice via Brain Fatty Acids Remodeling

ABSTRACTDocosahexaenoic acid (DHA) and nervonic acid (NA) are the main components of brain gray and white matter, respectively. Preclinical and clinical studies suggest the effects of DHA or NA supplementation on memory improvement in aging and Alzheimer's disease models. However, it is not yet clear whether dietary supplementation with DHA and NA can enhance memory by increasing their levels in gray and white matter. The present study aimed to examine the spatial memory and brain white and gray matter lipidomic profiles in adolescent mice with or without a DHA and NA‐enriched diet. Spatial memory and conditional fear memory were evaluated in adolescent mice using the Morris Water Maze and passive avoidance test beginning at 7‐week of age and mice were killed at 13‐week of age to allow for the dissection of brain tissues. Combined low dose treatment of DHA (600 mg/kg) and NA (94 mg/kg) synergically enhanced spatial memory and fear memory function, and the effects were better than that of DHA intervention. While, high dose of DHA and NA treatment was detrimental to long‐term spatial memory and fear memory, corresponding to the decreased levels of serotonin (5‐HT) in the cortex and hippocampus, and imbalanced fatty acids in the white and gray matter. In addition, c‐Fos positive neurons were increased in low dose DHA treated mice comparing to that of control mice. Lipidomic analyses suggest levels of NA, dihomo‐γ‐linolenic acid, EPA, DHA, and linoleic acid were increased in the white matter post low dose of DHA + NA treatment, as well as increased levels of dihomo‐γ‐linolenic acid, EPA, and DHA in the gray matter compared with control mice. Understanding how DHA and NA supplementation during the adolescent periods affects cognitive function in a dose‐specific manner has important implications for determining the dietary requirements of DHA and NA. The present study reported for the first time that DHA and NA combination regulate spatial memory in a dose‐specific manner and provides evidence that further research needs to consider how different ratios of DHA and NA regulate the fatty acid composition in white and gray matter, thereby influencing memory function.

Rap2a promotes cardiac fibrosis and exacerbates myocardial infarction through the TNIK/Merlin/YAP axis

Myocardial fibrosis constitutes the primary pathological characteristic of myocardial infarction (MI). The activation and proliferation of myocardial fibroblasts serve as crucial factors in the process of the development of fibrosis in the myocardium. Our research delved into the role that Rap2a plays in cardiac function as well as myocardial fibrosis, while its effects on cardial fibroblasts (CFs) proliferation, migration, and phenotypic transformation were also explored. Examination of the GEO database showed a notable increase in the expression of Rap2a within myocardial tissue from mice with MI compared to normal mice. Rap2a deficiency relieves MI in mice and restrains the phenotypic transition, proliferation, and migration of CFs. The absence of Rap2a mitigates MI in mice. Besides, it curbs the growth of CFs, restricts their movement, and prevents them from undergoing phenotypic conversion. Rap2a can bind to TNIK in myocardial fibroblasts and enhance TNIK expression; Merlin/YAP signaling pathway was assessed as a downstream target of TNIK to further elucidate the regulatory mechanism through which Rap2a influences cardiomyocytes. In conclusion, this study provides evidence that Rap2a promotes myocardial fibrosis through mediating the myofibroblast transformation, proliferation, and migration of CFs via the TNIK/Merlin/YAP pathway, thereby exacerbating symptoms of myocardial infarction.

Design and synthesis of azole derivatives of echinocystic acid as α-glucosidase inhibitors with hypoglycemic activity.

Design and synthesis of azole derivatives of echinocystic acid as α-glucosidase inhibitors with hypoglycemic activity.

Gut flora-derived succinate exacerbates Allergic Airway Inflammation by promoting protein succinylation.

Gut flora-derived succinate exacerbates Allergic Airway Inflammation by promoting protein succinylation.

CXCL13-neutralizing Antibody Alleviate Chronic Skeletal Muscle Degeneration in a Mouse Model.

Skeletal muscle degeneration is a common effect of chronic muscle injuries, including fibrosis and fatty infiltration, which is the replacement of preexisting parenchymal tissue by extracellular matrix proteins and abnormal invasive growth of fibroblasts and adipocytes. This remodeling limits muscle function and strength, eventually leading to reduced quality of life for those affected. Chemokines play a major role in the regulation of immunocyte migration, inflammation, and tissue remodeling and are implicated in various fibrotic and degenerative diseases. In this study, we aimed to investigate the role of the B-cell chemokine CXCL13 in the gastrocnemius muscle of the Achilles tendon rupture model mouse. We hypothesize that CXCL13 may promote fibrosis and aggravate skeletal muscle degeneration. We performed RNA sequencing and bioinformatics analysis of gastrocnemius muscle from normal and model mice to identify differentially expressed genes and signal pathways related to skeletal muscle degeneration and fibrosis. Our results show that CXCL13 is highly expressed in chronically degenerating skeletal muscle. Furthermore, CXCL13-neutralising antibodies with therapeutic potential were observed to inhibit fibrosis and adipogenesis in vivo and in vitro. Our study reveals the underlying therapeutic implications of CXCL13 inhibition for clinical intervention in skeletal muscle degeneration, thereby improving patient prognosis.

PIM kinase inhibition counters resistance to radiotherapy and chemotherapy in human prostate cancer.

PIM kinase inhibition counters resistance to radiotherapy and chemotherapy in human prostate cancer.

TWEAK regulates the functions of hair follicle stem cells via the Fn14-Wnt/β-catenin-CXCR4 signalling axis.

Hair follicle stem cells (HFSCs) are crucial for maintaining cutaneous functions under various pathological conditions, including wounds. Tumour necrosis factor-like weak inducer of apoptosis (TWEAK) interacts with its receptor, fibroblast growth factor-inducible 14 (Fn14), and plays a role in the development and tissue repair of skin diseases. This study aims to elucidate the effects of TWEAK/Fn14 signalling on HFSCs and the associated mechanisms. The expressions of HFSC markers, including K19, integrin β1 and K15, were analysed via immunohistochemistry in normal and Fn14-deficient mouse skin. Primary HFSCs were cultured in vitro and then treated with TWEAK or a chemokine (CXC motif) (CXCR) 4 inhibitor. The phenotype markers and secreted cytokines of HFSCs were assessed via immunofluorescence analysis, Western blotting and real-time polymerase chain reaction. Our results showed that both Fn14 and CXCR4 were highly expressed in hair follicles. Fn14 deficiency led to a decrease in the expression levels of K19 and CD34. Exogenous TWEAK enhanced the expression of K15, K19, integrin β1, tumour necrosis factor receptor type 2 and CXCR4 in cultured HFSCs. Additionally, TWEAK induced the proliferation, migration and cytokine production in HFSCs. Furthermore, the Wnt/β-catenin signalling pathway was upregulated in HFSCs upon TWEAK stimulation, and inhibitors of β-catenin or CXCR4 suppressed the effects of TWEAK on the differentiation and secretory functions of HFSCs. In conclusion, TWEAK/Fn14 interaction regulates the expression of differentiation markers and secretory functions of HFSCs in vitro. Wnt/β-catenin signalling or CXCR4 activation mediates the effects of TWEAK on HFSCs. Targeting the Fn14-Wnt/β-catenin-CXCR4 signalling axis may offer a potential approach for managing HFSC-related skin diseases, such as wounds.

Adrenocorticotropic hormone and its receptor as a novel testicular system involves in the development of spermatogenesis.

Adrenocorticotropic hormone and its receptor as a novel testicular system involves in the development of spermatogenesis.

Antimicrobial and anticancer activity of Streptomyces ambofaciens (Myt 8) and S. globisporus ONU 1019 (Myt 11) secondary metabolites isolated from the Odesa Bay, the Black Sea: An in vitro study.

Antimicrobial and anticancer activity of Streptomyces ambofaciens (Myt 8) and S. globisporus ONU 1019 (Myt 11) secondary metabolites isolated from the Odesa Bay, the Black Sea: An in vitro study.

GSK621 ameliorates lipid accumulation via AMPK pathways and reduces oxidative stress in hepatocytes in vitro and in obese mice in vivo.

GSK621 ameliorates lipid accumulation via AMPK pathways and reduces oxidative stress in hepatocytes in vitro and in obese mice in vivo.

The emerging role of IL-22 as a potential radiosensitivity biomarker for radiation-induced intestinal injury.

The emerging role of IL-22 as a potential radiosensitivity biomarker for radiation-induced intestinal injury.

3D printing of GelMA/nanohydroxyapatite/melanin nanoparticles composite hydrogel scaffolds for bone regeneration through immunomodulation.

3D printing of GelMA/nanohydroxyapatite/melanin nanoparticles composite hydrogel scaffolds for bone regeneration through immunomodulation.

Goat Milk-Derived Extracellular Vesicles Alleviate Colitis Potentially Through Improved Gut Microbiota in Mice

Ulcerative colitis (UC) is characterized clinically by intestinal inflammation and gut microbiota dysbiosis. The consumption of biologics, although effective in inflammation control, may lead to adverse effects and is inconvenient for at-home administration. Goat milk-derived extracellular vesicles (GMEVs) have been proposed as a supplement to prevent intestinal inflammation. However, their therapeutic potential for colitis remains elusive. This study aimed to explore the preventive effect of GMEVs on colitis and its underlying mechanisms through the microbiota-immune axis using a dextran sodium sulfate (DSS)-induced colitis mouse model. We found that a pre-treatment of 20 mg/kg/d GMEVs effectively prevented body weight loss, colon shortening, the depletion of colonic goblet cells, and the disappearance of crypts, while enhancing the intestinal mucosal barrier. Consistent with these phenotypes, GMEV pre-treatment increased levels of IL-22 and IL-10 and decreased levels of IL-1β, TNF-α, IL-6, and iNOS. However, GMEVs themselves had no effect on normal mice. Paralleling the alleviation of intestinal inflammation, GMEV pre-treatment also restored the reduction in unclassified Muribaculaceae, Dubosiella, and Lactobacillus and suppressed the expansion of Alistipes and Proteobacteria following DSS treatment. Additionally, GMEV intake significantly downregulated the expression of proteins in the NF-κB signaling pathway induced by DSS. In summary, GMEVs could prevent colitis by regulating intestinal inflammation, the intestinal mucosal barrier, gut microbiota, organ damage, and the immune microenvironment. This study demonstrated that GMEVs have potential application prospects for UC prevention.

Simply crushed zizyphi spinosi semen prevents neurodegenerative diseases and reverses age-related cognitive decline in mice.

Neurodegenerative diseases are age-related disorders characterized by the cerebral accumulation of amyloidogenic proteins, and cellular senescence underlies their pathogenesis. Thus, it is necessary for preventing these diseases to remove toxic proteins, repair damaged neurons, and suppress cellular senescence. As a source for such prophylactic agents, we selected zizyphi spinosi semen (ZSS), a medicinal herb used in traditional Chinese medicine. Oral administration of ZSS hot water extract ameliorated Aβ and tau pathology and cognitive impairment in mouse models of Alzheimer's disease and frontotemporal dementia. Non-extracted ZSS simple crush powder showed stronger effects than the extract and improved α-synuclein pathology and cognitive/motor function in Parkinson's disease model mice. Furthermore, when administered to normal aged mice, the ZSS powder suppressed cellular senescence, reduced DNA oxidation, promoted brain-derived neurotrophic factor expression and neurogenesis, and enhanced cognition to levels similar to those in young mice. The quantity of known active ingredients of ZSS, jujuboside A, jujuboside B, and spinosin was not proportional to the nootropic activity of ZSS. These results suggest that ZSS simple crush powder is a promising dietary material for the prevention of neurodegenerative diseases and brain aging.

Caffeine-Boosted Silver Nanoparticles Target Breast Cancer Cells by Triggering Oxidative Stress, Inflammation, and Apoptotic Pathways.

Caffeine-Boosted Silver Nanoparticles Target Breast Cancer Cells by Triggering Oxidative Stress, Inflammation, and Apoptotic Pathways.

Anti-ferroptosis effects of undescribed abietane diterpenoids from Prunella vulgaris subsp. hispida (Benth.) Hultén

A phytochemical investigation of Prunella vulgaris subsp. Hispida (Benth.) Hultén. led to eight diterpenoids including two previously undescribed compounds. Prunelida A (1) is a 20-nor-abietane diterpenoid, and prunelida B (2) is a rearranged C-17(15→14)-abeo-abietane diterpenoid. Their structural characterisation were performed based on comprehensive spectroscopic analyses including HRESIMS, NMR and calculated ECD methods. The potential of all compounds to inhibit ferroptosis were evaluated in normal mouse hippocampal neuronal (HT22) cells. Compound 4 showed significant inhibitory effects with an EC50 value of 0.77 ± 0.08 μM. Moreover, HT22 cells intracellular reactive oxygen species (ROS) accumulation levels treat with 4 has been assayed.

Expression Distribution of Keratins in Normal and Pathological Corneas and the Regulatory Role of Krt17 on Limbal Stem Cells.

This study aimed to compare variations in keratins (KRTs), particularly stress KRTs, under normal and pathological conditions, with a specific focus on investigating the role of KRT17. This research examined changes in KRT and limbal stem cell (LSC) markers in normal and various pathological corneas using mRNA and proteomic sequencing data alongside single-cell sequencing data from normal mouse corneas. The effects of the Krt17 recombinant protein and specific small interfering RNA on the clonal formation and proliferation of human limbal epithelial cells (HLECs) were investigated. mRNA sequencing was conducted on Krt17 knockdown HLECs, and the findings were validated using qPCR, immunofluorescence staining, neutrophil chemotaxis, and herpes simplex virus 1 infection assay. The STRING database was used to predict Krt17's interacting proteins. Various KRTs in the corneal epithelia exhibited differences in expression levels and patterns. Under pathological conditions, stress KRTs Krt17 and Krt16 were upregulated, while differentiation-related Krt12 was downregulated, and the expression of the LSC markers Krt17, Krt14, and IFITM3 were commonly increased. Supplementation and intervention experiments confirmed that Krt17 promotes clonal formation and proliferation in HLECs. Krt17 knockdown resulted in the upregulation of genes related to inflammation and defense responses, while downregulating molecules associated with differentiation pathways. Krt17 knockdown promoted neutrophil chemotaxis and alleviated herpes simplex virus 1 infection in HLECs. KRTs play a crucial role in the homeostasis and pathological regulation of the corneal epithelium. The stress Krt17 located in LSCs is involved in regulating the stemness, proliferation, and differentiation of LSCs, as well as immune and defense responses.

Abstract 4312: CTR9 controls the mammary luminal lineage and susceptibility to carcinogen/hormone-induced breast tumors

CTR9 encodes a key component of the human RNA polymerase II (RNAPII)-associated factor complex (hPAFc). CTR9 loss-of-function variants are predisposed to rare myeloid malignancies and Wilms tumors. Although CTR9 promotes the growth of estrogen receptor (ER)-positive breast cancer cells and controls the expression of 90% of ER-target genes, the roles of CTR9 in mammary gland development and breast cancer etiology in vivo remain unknown. About 0.1 % of breast cancer patients were found to harbor loss of function CTR9 mutations based on TCGA data. Analyzing normal mouse mammary gland scRNA-seq data reveals that CTR9 is highly expressed in luminal cells, which is confirmed by immunofluorescence staining in human breast tumors. To study the effects of the loss-of-function of CTR9 on mammary gland development, we generated a mammary gland-specific CTR9 knockout (KO) mouse using MMTV-Cre and CTR9 loxP system. CTR9 depletion from mammary epithelial cells was validated using qPCR, Western blot, and IHC. Loss of CTR9 in C57B/6 female mice led to increased duct branching and elongation during puberty and increased alveologenesis during pregnancy. This observation coincides with the detection of expanded mammary stem cells by flow cytometry in the 6-week-old mouse mammary glands. When treated with DMBA and progestin (MPA) to induce mammary tumors, both wild-type (WT) and CTR9 null mice began to develop tumors 6 weeks after the last DMBA treatment, but CTR9 null mice exhibited a markedly enhanced susceptibility to carcinogen/hormone-induced mammary tumors as compared with the WT counterparts. To detect changes during the early stage of tumorigenesis, we profiled the stem and luminal progenitor cell populations in the mammary gland 4-week after the last DMBA treatment. Remarkably, CTR9 depletion enhanced the stem cell and luminal progenitor populations while decreasing the mature luminal cells. CTR9 KO mammary gland had elevated Ki67 levels, decreased ER and PR, decreased mature luminal cells, and increased expression of KRT15, a luminal progenitor marker. The CTR9 null mammary gland expressed lower levels of KDM5A and higher levels of H3K4me3 as compared to the wild-type mice. Taken together, our findings establish that CTR9 depletion blocks the terminal differentiation of luminal cell lineage possibly through epigenetic regulation of histone methylation, and renders the mammary glands more susceptible to carcinogen/hormone-induced tumors. Citation Format: Gui Ma, Ang Gao, Mingshan Hu, Ngai Ting Chan, Yidan Wang, Wei Xu. CTR9 controls the mammary luminal lineage and susceptibility to carcinogen/hormone-induced breast tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4312.