Zebrafish Carrying pycr1 Gene Deficiency Display Aging and Multiple Behavioral Abnormalities

Aging is a natural process that internal gene control and external stimuli mediate. Clinical data pointed out that homozygotic or heterozygotic mutation in the pyrroline-5-carboxylate reductase 1 (PYCR1) gene in humans caused cutis laxa (ARCL) disease, with progeroid appearance, lax and wrinkled skin, joint laxity, osteopenia, and mental retardation phenotypes. In this study, we aimed to generate pycr1 knockout (KO) zebrafish and carried out biochemical characterizations and behavior analyses. Marked apoptosis and senescence were detected in pycr1 KO zebrafish, which started from embryos/larvae stage. Biochemical assays showed that adult pycr1 KO fish have significantly reduced proline and extracellular matrix contents, lowered energy, and diminished superoxide dismutase (SOD) and telomerase activity when compared to the wild type fish, which suggested the pycr1 KO fish may have dysfunction in mitochondria. The pycr1 KO fish were viable; however, displayed progeria-like phenotype from the 4 months old and reach 50% mortality around six months old. In adult stage, we found that pycr1 KO fish showed reduced locomotion activity, aggression, predator avoidance, social interaction interest, as well as dysregulated color preference and circadian rhythm. In summary, we have identified multiple behavioral alterations in a novel fish model for aging with pycr1 gene loss-of-function by behavioral tests. This animal model may not only provide a unique vertebrate model to screen potential anti-aging drugs in the future, but also be an excellent in vivo model towards a better understanding of the corresponding behavioral alterations that accompany aging.

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Age-dependent emergence of neurophysiological and behavioral abnormalities in progranulin-deficient mice

Alzheimer s Research & Therapy ◽

10.1186/s13195-019-0540-x ◽

2019 ◽

Vol 11 (1) ◽

Author(s):

Dávid Nagy ◽

Lauren Herl Martens ◽

Liza Leventhal ◽

Angela Chen ◽

Craig Kelley ◽

...

Keyword(s):

Frontotemporal Dementia ◽

Neurodegenerative Disorder ◽

Neuronal Loss ◽

Loss Of Function ◽

Behavioral Alterations ◽

Response Latencies ◽

Reward Seeking ◽

Behavioral Abnormalities ◽

Age Dependent

Abstract Background Loss-of-function mutations in the progranulin gene cause frontotemporal dementia, a genetic, heterogeneous neurodegenerative disorder. Progranulin deficiency leads to extensive neuronal loss in the frontal and temporal lobes, altered synaptic connectivity, and behavioral alterations. Methods The chronological emergence of neurophysiological and behavioral phenotypes of Grn heterozygous and homozygous mice in the dorsomedial thalamic—medial prefrontal cortical pathway were evaluated by in vivo electrophysiology and reward-seeking/processing behavior, tested between ages 3 and 12.5 months. Results Electrophysiological recordings identified a clear age-dependent deficit in the thalamocortical circuit. Both heterozygous and homozygous mice exhibited impaired input-output relationships and paired-pulse depression, but evoked response latencies were only prolonged in heterozygotes. Furthermore, we demonstrate firstly an abnormal reward-seeking/processing behavior in the homozygous mice which correlates with previously reported neuroinflammation. Conclusion Our findings indicate that murine progranulin deficiency causes age-dependent neurophysiological and behavioral abnormalities thereby indicating their validity in modeling aspects of human frontotemporal dementia.

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Cytoplasmic accumulation of FUS triggers early behavioral alterations linked to cortical neuronal hyperactivity and defects in inhibitory synapses

10.1101/2020.06.09.141556 ◽

2020 ◽

Author(s):

Jelena Scekic-Zahirovic ◽

Inmaculada Sanjuan-Ruiz ◽

Vanessa Kan ◽

Salim Megat ◽

Pierre De Rossi ◽

...

Keyword(s):

Neuronal Activity ◽

Rna Binding ◽

Gene Mutations ◽

Neuronal Loss ◽

Inhibitory Neurons ◽

Inhibitory Synapses ◽

Behavioral Alterations ◽

Behavioral Abnormalities ◽

Cytoplasmic Accumulation

AbstractGene mutations causing cytoplasmic mislocalization of the RNA-binding protein FUS, lead to severe forms of amyotrophic lateral sclerosis (ALS). Cytoplasmic accumulation of FUS is also observed in other diseases, with unknown consequences. Here, we show that cytoplasmic mislocalization of FUS drives behavioral abnormalities in knock-in mice, including locomotor hyperactivity and alterations in social interactions, in the absence of widespread neuronal loss. Mechanistically, we identified a profound increase in neuronal activity in the frontal cortex of Fus knock-in mice in vivo. Importantly, RNAseq analysis suggested involvement of defects in inhibitory neurons, that was confirmed by ultrastructural and morphological defects of inhibitory synapses and increased synaptosomal levels of mRNAs involved in inhibitory neurotransmission. Thus, cytoplasmic FUS triggers inhibitory synaptic deficits, leading to increased neuronal activity and behavioral phenotypes. FUS mislocalization may trigger deleterious phenotypes beyond motor neuron impairment in ALS, but also in other neurodegenerative diseases with FUS mislocalization.

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SIRT1 contributes to telomere maintenance and augments global homologous recombination

The Journal of Cell Biology ◽

10.1083/jcb.201005160 ◽

2010 ◽

Vol 191 (7) ◽

pp. 1299-1313 ◽

Cited By ~ 156

Author(s):

Jose A. Palacios ◽

Daniel Herranz ◽

Maria Luigia De Bonis ◽

Susana Velasco ◽

Manuel Serrano ◽

...

Keyword(s):

Homologous Recombination ◽

Telomerase Activity ◽

Histone Deacetylation ◽

Telomere Maintenance ◽

Loss Of Function ◽

Entire Genome ◽

Telomere Shortening ◽

Telomere Biology ◽

Sir Complex

Yeast Sir2 deacetylase is a component of the silent information regulator (SIR) complex encompassing Sir2/Sir3/Sir4. Sir2 is recruited to telomeres through Rap1, and this complex spreads into subtelomeric DNA via histone deacetylation. However, potential functions at telomeres for SIRT1, the mammalian orthologue of yeast Sir2, are less clear. We studied both loss of function (SIRT1 deficient) and gain of function (SIRT1super) mouse models. Our results indicate that SIRT1 is a positive regulator of telomere length in vivo and attenuates telomere shortening associated with aging, an effect dependent on telomerase activity. Using chromatin immunoprecipitation assays, we find that SIRT1 interacts with telomeric repeats in vivo. In addition, SIRT1 overexpression increases homologous recombination throughout the entire genome, including telomeres, centromeres, and chromosome arms. These findings link SIRT1 to telomere biology and global DNA repair and provide new mechanistic explanations for the known functions of SIRT1 in protection from DNA damage and some age-associated pathologies.

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Moxifloxacin-Induced Visual Hallucinations, Alterations in Mood and Behavior, and Hyperglycemia

Journal of Pharmacy Practice ◽

10.1177/0897190019830567 ◽

2019 ◽

Vol 33 (3) ◽

pp. 368-371 ◽

Cited By ~ 1

Author(s):

Burak Uz

Keyword(s):

Blood Glucose ◽

Illicit Drug ◽

Sudden Increase ◽

Visual Hallucinations ◽

Drug Reactions ◽

Behavioral Alterations ◽

Illicit Drug Abuse ◽

And Behavior ◽

Neuropsychiatric Adverse Effects ◽

External Stimuli

Visual hallucinations are sensory perceptions that occur without external stimuli. Moxifloxacin-induced visual hallucinations are very rarely reported (<0.1%). We describe a 66-year-old woman, without any known neuropsychiatric disorder or illicit drug abuse, who experienced complex visual hallucinations, mood, and behavioral alterations secondary to peroral 1 dose of moxifloxacin treatment that persisted for approximately 36 hours. In addition, a sudden increase in her blood glucose level was noted which also improved a few days after discontinuation of moxifloxacin treatment. Although very rare, it should be kept in mind that moxifloxacin may coincidentally induce neuropsychiatric adverse effects and hyperglycemia. Clinicians should be alert to recognize such uncommon offending adverse drug reactions.

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A combination of cyclophosphamide, interleukin-2 allow CD4+ T cells converted to Tregs to control scurfy syndrome

Blood ◽

10.1182/blood.2020009187 ◽

2021 ◽

Author(s):

Marianne Delville ◽

Florence Bellier ◽

Juliette Leon ◽

Roman Klifa ◽

Sabrina Lizot ◽

...

Keyword(s):

T Cells ◽

Autoimmune Disease ◽

Adoptive Transfer ◽

Cd4 T Cells ◽

Lentiviral Vector ◽

Interleukin 2 ◽

Foxp3 Expression ◽

In Vivo Model ◽

Loss Of Function

Immunodysregulation polyendocrinopathy enteropathy X-linked (IPEX) syndrome is caused by mutations in FOXP3, which lead to the loss of function of regulatory T cells (Treg) and the development of autoimmune manifestations early in life. The selective induction of a Treg program in autologous CD4+ T cells by FOXP3 gene transfer is a promising approach for curing IPEX. We have established a novel in vivo assay of Treg functionality, based on adoptive transfer of these cells into scurfy mice (an animal model of IPEX) and a combination of cyclophosphamide conditioning and interleukin-2 treatment. This model highlighted the possibility of rescuing scurfy disease after the latter's onset. By using this in vivo model and an optimized lentiviral vector expressing human Foxp3 and as a reporter a truncated form of the 5 low-affinity nerve growth factor receptor (DLNGFR), we demonstrated that the adoptive transfer of FOXP3-transduced scurfy CD4+ T cells enabled the long-term rescue of scurfy autoimmune disease. The efficiency was similar to that seen with wild-type Treg. After in vivo expansion, the converted CD4FOXP3 cells recapitulated the transcriptomic core signature for Treg. These findings demonstrate that FOXP3 expression converts CD4+ T cells into functional Treg capable of controlling severe autoimmune disease.

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MicroRNA regulation of the MRN complex impacts DNA damage, cellular senescence and angiogenic signaling

10.1101/132258 ◽

2017 ◽

Author(s):

Cristina Espinosa-Diez ◽

RaeAnna Wilson ◽

Namita Chatterjee ◽

Clayton Hudson ◽

Rebecca Ruhl ◽

...

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Targeted Delivery ◽

Genotoxic Stress ◽

Telomerase Activity ◽

Loss Of Function ◽

Mrn Complex ◽

Vegf Signaling

AbstractMicroRNAs contribute to biological robustness by buffering cellular processes from external perturbations. Here we report an unexpected link between DNA damage response and angiogenic signaling that is buffered by two distinct microRNAs. We demonstrate that genotoxic stress-induced miR-494 and miR-99b inhibit the DNA repair machinery by targeting the MRE11a-RAD50-NBN (MRN) complex. Functionally, gain and loss of function experiments show that miR-494 and miR-99b affect telomerase activity, activate p21 and Rb pathways and diminish angiogenic sproutingin vitroandin vivo. Genetic and pharmacological disruption of VEGFR-2 signaling and the MRN complex reveal a surprising co-dependency of these pathways in regulating endothelial senescence and proliferation. Vascular-targeted delivery of miR-494 decreases both growth factor-induced and tumor angiogenesis in mouse models. Mechanistically, disruption of the MRN complex induced CD44, a known driver of senescence and regulator of VEGF signaling in addition to suppressing IL-13 a stimulator of VEGF signaling. Our work identifies a putative miR-facilitated mechanism by which endothelial cells can be insulated against VEGF signaling to facilitate the onset of senescence and highlight the potential of targeting DNA repair to disrupt pathological angiogenesis.

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Protrudin-deficient mice manifest depression-like behavior with abnormalities in activity, attention, and cued fear-conditioning

Molecular Brain ◽

10.1186/s13041-020-00693-3 ◽

2020 ◽

Vol 13 (1) ◽

Cited By ~ 1

Author(s):

Michiko Shirane ◽

Hirotaka Shoji ◽

Yutaka Hashimoto ◽

Hiroyuki Katagiri ◽

Shizuka Kobayashi ◽

...

Keyword(s):

Fear Conditioning ◽

Neuronal Development ◽

Physiological Role ◽

Loss Of Function ◽

Reduced Body ◽

Intermediate Phenotypes ◽

Behavioral Abnormalities ◽

And Behavior ◽

Deficient Mice

Abstract Protrudin is a protein that resides in the membrane of the endoplasmic reticulum and is highly expressed in the nervous system. Although mutations in the human protrudin gene (ZFYVE27, also known as SPG33) give rise to hereditary spastic paraplegia (HSP), the physiological role of the encoded protein has been largely unclear. We therefore generated mice deficient in protrudin and subjected them to a battery of behavioral tests designed to examine their intermediate phenotypes. The protrudin-deficient mice were found to have a reduced body size and to manifest pleiotropic behavioral abnormalities, including hyperactivity, depression-like behavior, and deficits in attention and fear-conditioning memory. They exhibited no signs of HSP, however, consistent with the notion that HSP-associated mutations of protrudin may elicit neural degeneration, not as a result of a loss of function, but rather as a result of a gain of toxic function. Overall, our results suggest that protrudin might play an indispensable role in normal neuronal development and behavior.

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Cytoplasmic FUS triggers early behavioral alterations linked to cortical neuronal hyperactivity and inhibitory synaptic defects

Nature Communications ◽

10.1038/s41467-021-23187-9 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Jelena Scekic-Zahirovic ◽

Inmaculada Sanjuan-Ruiz ◽

Vanessa Kan ◽

Salim Megat ◽

Pierre De Rossi ◽

...

Keyword(s):

Frontal Cortex ◽

Neuronal Activity ◽

Rna Binding ◽

Gene Mutations ◽

Neuronal Loss ◽

Progressive Increase ◽

Behavioral Alterations ◽

Behavioral Abnormalities ◽

Morphological Defects

AbstractGene mutations causing cytoplasmic mislocalization of the RNA-binding protein FUS lead to severe forms of amyotrophic lateral sclerosis (ALS). Cytoplasmic accumulation of FUS is also observed in other diseases, with unknown consequences. Here, we show that cytoplasmic mislocalization of FUS drives behavioral abnormalities in knock-in mice, including locomotor hyperactivity and alterations in social interactions, in the absence of widespread neuronal loss. Mechanistically, we identified a progressive increase in neuronal activity in the frontal cortex of Fus knock-in mice in vivo, associated with altered synaptic gene expression. Synaptic ultrastructural and morphological defects were more pronounced in inhibitory than excitatory synapses and associated with increased synaptosomal levels of FUS and its RNA targets. Thus, cytoplasmic FUS triggers synaptic deficits, which is leading to increased neuronal activity in frontal cortex and causing related behavioral phenotypes. These results indicate that FUS mislocalization may trigger deleterious phenotypes beyond motor neuron impairment in ALS, likely relevant also for other neurodegenerative diseases characterized by FUS mislocalization.

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The METTL3/MALAT1/PTBP1/USP8/TAK1 axis promotes pyroptosis and M1 polarization of macrophages and contributes to liver fibrosis

Cell Death Discovery ◽

10.1038/s41420-021-00756-x ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Bo Shu ◽

Ying-Xia Zhou ◽

Hao Li ◽

Rui-Zhi Zhang ◽

Chao He ◽

...

Keyword(s):

Liver Fibrosis ◽

Western Blot ◽

Rna Stability ◽

Interferon Γ ◽

In Vivo Model ◽

Loss Of Function ◽

M1 Polarization ◽

The Impact

AbstractPro-inflammatory M1 macrophages, via activating hepatic stellate cells, contribute to liver fibrosis. In this study, we examined the mechanism and the significance of a signaling axis, METTL3/MALAT1/PTBP1/USP8/TAK1, in regulating pyroptosis and M1 polarization of hepatic macrophages. Liver fibrosis model was established in vivo by CCl4 treatment; M1 polarization was induced in vitro by treating macrophages with lipopolysaccharide or interferon γ. Expressions of METTL3, MALAT1, PTBP1, USP8, and TAK1 were measured by RT-PCR and/or Western blot in Kupffer cells (KCs) isolated from in vivo model or in vitro activated macrophages. Macrophage phenotypes including inflammation (RT-qPCR analysis of a panel of proinflammatory cytokines and ELISA on productions of interleukin (IL)−1β and IL-18) and pyroptosis (Western blot of NLRP3, Caspase-1, and GSDMD) were investigated. The impact of METTL3 on m6A methylation of MALAT1 was examined by methylated RNA immunoprecipitation (RIP), the interaction between PTBP1 and MALAT1 or USP8 mRNA by combining RNA pull-down, RIP, and RNA stability assays, and the crosstalk between USP8 and TAK1 by co-immunoprecipitation and protein degradation assays. Functional significance of individual component of METTL3/MALAT1/PTBP1/USP8/TAK1 axis was assessed by combining gain-of-function and loss-of-function approaches. In KCs isolated from in vivo liver fibrosis model or in vitro M1-polarized macrophages, METTL3 was up-regulated, and sequentially, it increased MALAT1 level via m6A methylation, which promoted USP8 mRNA degradation through the interaction with PTBP1. Reduced USP8 expression regulated the ubiquitination and protein stability of TAK1, which promoted pyroptosis and inflammation of macrophages. The signaling cascade METTL3/MALAT1/PTBP1/USP8/TAK1, by essentially stimulating pyroptosis and inflammation of macrophages, aggravates liver fibrosis. Therefore, targeting individual components of this axis may benefit the treatment of liver fibrosis.

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