Publications by authors named "Iga Wasilewska"

6 Publications

  • Page 1 of 1

-Deficient Zebrafish Reproduce Neurological and Inflammatory Symptoms of Niemann-Pick Type C Disease.

Front Cell Neurosci 2021 27;15:647860. Epub 2021 Apr 27.

Laboratory of Neurodegeneration, International Institute of Molecular and Cell Biology in Warsaw, Warsaw, Poland.

Niemann-Pick type C (NPC) disease is an autosomal recessive lysosomal storage disease that is caused by a mutation of the or gene, in which un-esterified cholesterol and sphingolipids accumulate mainly in the liver, spleen, and brain. Abnormal lysosomal storage leads to cell damage, neurological problems, and premature death. The time of onset and severity of symptoms of NPC disease are highly variable. The molecular mechanisms that are responsible for NPC disease pathology are far from being understood. The present study generated and characterized a zebrafish mutant that lacks Npc2 protein that may be useful for studies at the organismal, cellular, and molecular levels and both small-scale and high-throughput screens. Using CRISPR/Cas9 technology, we knocked out the zebrafish homolog of . Five-day-old mutants were morphologically indistinguishable from wildtype larvae. We found that live larvae exhibited stronger Nile blue staining. The larvae exhibited low mobility and a high anxiety-related response. These behavioral changes correlated with downregulation of the (mitochondrial calcium uniporter) gene, (calcineurin) gene, and genes that are involved in myelination ( and ). Histological analysis of adult zebrafish revealed that pathological changes in the nervous system, kidney, liver, and pancreas correlated with inflammatory responses (i.e., the upregulation of , κβ, and ; i.e., hallmarks of NPC disease). These findings suggest that the mutant zebrafish may be a model of NPC disease.
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http://dx.doi.org/10.3389/fncel.2021.647860DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111220PMC
April 2021

Knockout of Increases Calcium Oscillations in Neurons and Induces Hyperactive-Like Phenotype in Zebrafish Larvae.

Int J Mol Sci 2020 Aug 27;21(17). Epub 2020 Aug 27.

International Institute of Molecular and Cell Biology in Warsaw, Ks. Trojdena 4, 02-109 Warsaw, Poland.

Stromal interaction molecule (STIM) proteins play a crucial role in store-operated calcium entry (SOCE) as endoplasmic reticulum Ca sensors. In neurons, STIM2 was shown to have distinct functions from STIM1. However, its role in brain activity and behavior was not fully elucidated. The present study analyzed behavior in zebrafish () that lacked . The mutant animals had no morphological abnormalities and were fertile. RNA-sequencing revealed alterations of the expression of transcription factor genes and several members of the calcium toolkit. Neuronal Ca activity was measured in vivo in neurons that expressed the GCaMP5G sensor. Optic tectum neurons in fish had more frequent Ca signal oscillations compared with neurons in wildtype (WT) fish. We detected an increase in activity during the visual-motor response test, an increase in thigmotaxis in the open field test, and the disruption of phototaxis in the dark/light preference test in mutants compared with WT. Both groups of animals reacted to glutamate and pentylenetetrazol with an increase in activity during the visual-motor response test, with no major differences between groups. Altogether, our results suggest that the hyperactive-like phenotype of mutant zebrafish is caused by the dysregulation of Ca homeostasis and signaling.
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http://dx.doi.org/10.3390/ijms21176198DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503814PMC
August 2020

Knockout Induces Hyperactivity and Susceptibility to Seizures in Zebrafish Larvae.

Cells 2020 05 21;9(5). Epub 2020 May 21.

International Institute of Molecular and Cell Biology, 4 Ks. Trojdena Street, 02-109 Warsaw, Poland.

In neurons, stromal interaction molecule (STIM) proteins regulate store-operated Ca entry (SOCE) and are involved in calcium signaling pathways. However, STIM activity in neurological diseases is unclear and should be clarified by studies that are performed in vivo rather than in cultured cells in vitro. The present study investigated the role of neuronal Stim2b protein in zebrafish. We generated knockout zebrafish, which were fertile and had a regular lifespan. Using various behavioral tests, we found that zebrafish larvae were hyperactive compared with wild-type fish. The mutants exhibited increases in mobility and thigmotaxis and disruptions of phototaxis. They were also more sensitive to pentylenetetrazol and glutamate treatments. Using lightsheet microscopy, a higher average oscillation frequency and higher average amplitude of neuronal Ca oscillations were observed in larvae. RNA sequencing detected upregulation of the and genes and downregulation of the , , and genes. The latter gene encodes a protein that is involved in several processes that are involved in Ca homeostasis in neurons, including metabotropic glutamate receptors. We propose that Stim2b deficiency in neurons dysregulates SOCE and triggers changes in gene expression, thereby causing abnormal behavior, such as hyperactivity and susceptibility to seizures.
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http://dx.doi.org/10.3390/cells9051285DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7291033PMC
May 2020

Identification of Zebrafish Calcium Toolkit Genes and their Expression in the Brain.

Genes (Basel) 2019 03 18;10(3). Epub 2019 Mar 18.

International Institute of Molecular and Cell Biology in Warsaw, Trojdena 4, 02-109 Warsaw, Poland.

Zebrafish are well-suited for in vivo calcium imaging because of the transparency of their larvae and the ability to express calcium probes in various cell subtypes. This model organism has been used extensively to study brain development, neuronal function, and network activity. However, only a few studies have investigated calcium homeostasis and signaling in zebrafish neurons, and little is known about the proteins that are involved in these processes. Using bioinformatics analysis and available databases, the present study identified 491 genes of the zebrafish Calcium Toolkit (CaTK). Using RNA-sequencing, we then evaluated the expression of these genes in the adult zebrafish brain and found 380 hits that belonged to the CaTK. Based on quantitative real-time polymerase chain reaction arrays, we estimated the relative mRNA levels in the brain of CaTK genes at two developmental stages. In both 5 dpf larvae and adult zebrafish, the highest relative expression was observed for , which encodes a Golgi membrane protein. The present data on CaTK genes will contribute to future applications of zebrafish as a model for in vivo and in vitro studies of Ca signaling.
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http://dx.doi.org/10.3390/genes10030230DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6471419PMC
March 2019

Behavioral and electrophysiological changes in female mice overexpressing ORAI1 in neurons.

Biochim Biophys Acta Mol Cell Res 2019 07 16;1866(7):1137-1150. Epub 2019 Jan 16.

International Institute of Molecular and Cell Biology in Warsaw, 4 Ks. Trojdena Str., Warsaw 02-109, Poland.

Orai proteins form highly selective Ca release-activated channels (CRACs). They play a critical role in store-operated Ca entry (SOCE; i.e., the influx of external Ca that is induced by the depletion of endoplasmic reticulum Ca stores). Of the three Orai homologs that are present in mammals (Orai1-3), the physiological function of Orai1 is the best described. CRACs are formed by both homomeric assemblies and heteromultimers of Orais. Orai1 and Orai2 can form heteromeric channels that differ in conductivity during SOCE, depending on their Orai1-to-Orai2 ratio. The present study explored the potential consequences of ORAI1 overexpression in neurons where the dominant isoform is Orai2. We established the Tg(ORAI1)Ibd transgenic mouse line that overexpresses ORAI1 in brain neurons. We observed seizure-like symptoms in aged (≥15-month-old) female mice but not in males of the same age. The application of kainic acid and bicuculline to slices that were isolated from 8-month-old (±1 month) female Tg(ORAI1)Ibd mice revealed a significantly lower frequency of interictal bursts compared with samples that were isolated from wildtype mice. No differences were observed in male mice of a similar age. A battery of behavioral tests showed that context recognition decreased only in female transgenic mice. The phenotype that was observed in female mice suggests that ORAI1 overexpression may affect neuronal activity in a sex-dependent manner. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.
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http://dx.doi.org/10.1016/j.bbamcr.2019.01.007DOI Listing
July 2019

Overexpression of STIM1 in neurons in mouse brain improves contextual learning and impairs long-term depression.

Biochim Biophys Acta Mol Cell Res 2017 Jun 29;1864(6):1071-1087. Epub 2016 Nov 29.

International Institute of Molecular and Cell Biology in Warsaw, 4 Ks. Trojdena Str., 02-109 Warsaw, Poland. Electronic address:

STIM1 is an endoplasmic reticulum calcium sensor that is involved in several processes in neurons, including store-operated calcium entry. STIM1 also inhibits voltage-gated calcium channels, such as Ca1.2 and Ca3.1, and is thus considered a multifunctional protein. The aim of this work was to investigate the ways in which transgenic neuronal overexpression of STIM1 in FVB/NJ mice affects animal behavior and the electrophysiological properties of neurons in acute hippocampal slices. We overexpressed STIM1 from the Thy1.2 promoter and verified neuronal expression by quantitative reverse-transcription polymerase chain reaction, Western blot, and immunohistochemistry. Mature primary hippocampal cultures expressed STIM1 but exhibited no changes in calcium homeostasis. Basal synaptic transmission efficiency and short-term plasticity were comparable in slices that were isolated from transgenic mice, similarly as the magnitude of long-term potentiation. However, long-term depression that was induced by the glutamate receptor 1/5 agonist (S)-3,5-dihydroxyphenylglycine was impaired in STIM1 slices. Interestingly, transgenic mice exhibited a decrease in anxiety-like behavior and improvements in contextual learning. In summary, our data indicate that STIM1 overexpression in neurons in the brain perturbs metabotropic glutamate receptor signaling, leading to impairments in long-term depression and alterations in animal behavior. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.
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http://dx.doi.org/10.1016/j.bbamcr.2016.11.025DOI Listing
June 2017