Biology subjects

Zuberi, A. R.

Publications and source records attributed to Zuberi, A. R..

5 recordsLinked to original sources

bioRxiv2024

Association of cerebellar inflammation and neurodegeneration in a novel spinocerebellar ataxia type 13 mouse model

BackgroundNeuroinflammation is a recognized pathological characteristic of neurodegenerative diseases. Spinocerebellar ataxia 13 (SCA13) is a progressive neurodegenerative disease with no effective treatments. Our previous studies reported human mutations in KCNC3 gene are causative for SCA13. Human R423H allelic mutation induces early-onset neurodegeneration and aberrant intracellular retention of Epidermal Growth Factor Receptor (EGFR) in drosophila. However, the neurodegeneration and inflammatory response induced by the R424H allele are unknown in a mammalian model of disease. MethodIn this study, a single Kcnc3 R424H mutation (Analogous to the human SCA13 R423H isoform) transgenic mice were created using CRISPR/Cas 9 technique. Motor function (gait, tremor, coordination and balance) and cerebellar volume (scanned and imaged with 7T MRI) of the R424H transgenic mice were evaluated at multiple timepoints. Neurodegeneration (Purkinje cells loss) as well as cerebellar (astroglia, microglia and macrophage activation) and peripheral (plasma cytokines levels) inflammatory responses were examined and analyzed. ResultThe R424H transgenic mice showed marked neurological motor dysfunction with high-frequency tremor, aberrant gait, and short latency to fall in Rotarod testing at 3 and 6 months of age. Abnormal spontaneous firing was recorded in electrophysiology of Purkinje cells. Pathological changes in our R424H transgenic mice included progressive Purkinje cell degeneration and cerebellar atrophy. Over-active microglia, astrocytes, and macrophages were observed in the cerebella of transgenic mice. Pearson correlation analyses indicated that the number of Calbindin positive cells, a Purkinje cell marker, showed a strong inverse correlation with the positive cell number of EGFR, phosphorylated EGFR (pEGFR), and CD68. The expression of EGFR/pEGFR was positively correlated with CD68 and Glial Fibrillary Acidic Protein. ConclusionTransgenic R424H mice provide a novel SCA13 model showing significant motor deficits, Purkinje cells loss, cerebellar inflammation, and atrophy. Our study suggests that the activation of inflammatory immune cells (astroglia, microglia and macrophages) and strong expression of EGFR/ pEGFR signal in these immune cells are associated with Purkinje cell loss in the cerebellum. This abnormal neuroinflammation may play a significant role in the aggressive procession of neurodegeneration.

neuroscience↗

bioRxiv2024

In-utero rescue of neurological dysfunction in a mouse model of Wiedemann-Steiner syndrome

Wiedemann-Steiner syndrome (WDSTS) is a rare genetic cause of intellectual disability primarily caused by heterozygous loss of function variants in the gene encoding the histone methyltransferase KMT2A. Prior studies have shown successful postnatal amelioration of disease phenotypes for Rett, Rubinstein-Taybi and Kabuki syndromes, related Mendelian disorders of the epigenetic machinery. To explore whether the neurological phenotype in WDSTS is treatable in-utero, we created a novel mouse model carrying a loss of function variant in between two loxP sites. Kmt2a+/LSL mice demonstrate core features of WDSTS including growth retardation, craniofacial abnormalities, and hypertrichosis as well as hippocampal memory defects. The neurological phenotypes show rescue upon restoration of KMT2A in-utero following breeding to a nestin-Cre. Together, our data provide a novel mouse model to explore the therapeutic window in WDSTS. Our work suggests that WDSTS has a window of opportunity extending at least until the mid-point of in-utero development, making WDSTS an ideal candidate for future therapeutic strategies. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/604339v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1b913caorg.highwire.dtl.DTLVardef@1c23ce3org.highwire.dtl.DTLVardef@12b8a25org.highwire.dtl.DTLVardef@17a9a4e_HPS_FORMAT_FIGEXP M_FIG Graphical abstract C_FIG

genetics↗

bioRxiv2024

Valine and Inflammation Drive Epilepsy in a Mouse Model of ECHS1 Deficiency

ECHS1 Deficiency (ECHS1D) is a rare and devastating pediatric disease that currently has no defined treatments. This disorder results from missense loss-of-function mutations in the ECHS1 gene that result in severe developmental delays, encephalopathy, hypotonia, and early death. ECHS1 enzymatic activity is necessary for the beta-oxidation of fatty acids and the oxidation of branched-chain amino acids within the inner mitochondrial matrix. The pathogenesis of disease remains unknown, however it is hypothesized that disease is driven by an accumulation of toxic metabolites from impaired valine oxidation. To expand our knowledge on disease mechanisms, a novel mouse model of ECHS1D was generated that possesses a disease-associated knock-in (KI) allele and a knock-out (KO) allele. To investigate the behavioral phenotype, a battery of testing was performed at multiple time points, which included assessments of learning, motor function, endurance, sensory responses, and anxiety. Neurological abnormalities were assessed using wireless telemetry EEG recordings, pentylenetetrazol (PTZ) seizure induction, and immunohistochemistry. Metabolic perturbations were measured within the liver, serum, and brain using mass spectrometry and magnetic resonance spectroscopy. To test disease mechanisms, mice were subjected to disease pathway stressors and then survival, body weight gain, and epilepsy were assessed. Mice containing KI/KI or KI/KO alleles were viable with normal development and survival, and the presence of KI and KO alleles resulted in a significant reduction in ECHS1 protein. ECHS1D mice displayed reduced exercise capacity and pain sensation. EEG analysis revealed increased slow wave power that was associated with perturbations in sleep. ECHS1D mice had significantly increased epileptiform EEG discharges, and were sensitive to seizure induction, which resulted in death of 60% of ECHS1D mice. Under basal conditions, brain structure was grossly normal, although histological analysis revealed increased microglial activation in aged ECHS1D mice. Increased dietary valine only affected ECHS1D mice, which significantly exacerbated seizure susceptibility and resulted in death. Lastly, acute inflammatory challenge drove regression and early lethality in ECHS1D mice. In conclusion, we developed a novel model of ECHS1D that may be used to further knowledge on disease mechanisms and to develop therapeutics. Our data suggests altered metabolic signaling and inflammation may contribute to epilepsy in ECHS1D, and these alterations may be attributed to impaired valine metabolism.

neuroscience↗

bioRxiv2024

Plural molecular and cellular mechanisms of pore domain KCNQ2 encephalopathy

KCNQ2 variants in children with neurodevelopmental impairment are difficult to assess due to their heterogeneity and unclear pathogenic mechanisms. We describe a child with neonatal-onset epilepsy, developmental impairment of intermediate severity, and KCNQ2 G256W heterozygosity. Analyzing prior KCNQ2 channel cryoelectron microscopy models revealed G256 as a node of an arch-shaped non-covalent bond network linking S5, the pore turret, and the ion path. Co-expression with G256W dominantly suppressed conduction by wild-type subunits in heterologous cells. Ezogabine partly reversed this suppression. G256W/+ mice have epilepsy leading to premature deaths. Hippocampal CA1 pyramidal cells from G256W/+ brain slices showed hyperexcitability. G256W/+ pyramidal cell KCNQ2 and KCNQ3 immunolabeling was significantly shifted from axon initial segments to neuronal somata. Despite normal mRNA levels, G256W/+ mouse KCNQ2 protein levels were reduced by about 50%. Our findings indicate that G256W pathogenicity results from multiplicative effects, including reductions in intrinsic conduction, subcellular targeting, and protein stability. These studies provide evidence for an unexpected and novel role for the KCNQ2 pore turret and introduce a valid animal model of KCNQ2 encephalopathy. Our results, spanning structure to behavior, may be broadly applicable because the majority of KCNQ2 encephalopathy patients share variants near the selectivity filter.

neuroscience↗

bioRxiv2020

A Drosophila natural variation screen identifies NKCC1 as a substrate of NGLY1 deglycosylation and a modifier of NGLY1 deficiency

N-Glycanase 1 (NGLY1) is a cytoplasmic deglycosylating enzyme. Loss-of-function mutations in the NGLY1 gene cause NGLY1 deficiency, which is characterized by developmental delay, seizures, and a lack of sweat and tears. To model the phenotypic variability observed among patients, we crossed a Drosophila model of NGLY1 deficiency onto a panel of genetically diverse strains. The resulting progeny showed a phenotypic spectrum from 0-100% lethality. Association analysis on the lethality phenotype as well as an evolutionary rate covariation analysis generated lists of modifying genes, providing insight into NGLY1 function and disease. The top association hit was Ncc69 (human NKCC1/2), a conserved ion transporter. Analyses in NGLY1 -/- mouse cells demonstrated that NKCC1 is misglycosylated and has reduced function, making it only the second confirmed NGLY1 enzymatic substrate. The misregulation of this ion transporter may explain the observed defects in secretory epithelium function in NGLY1 deficiency patients.

genomics↗