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Global analysis of drug-regulated alternative expression of genes in mouse brain

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Korostynski M., Piechota M., Golda S., Przewlocki R.

Acta Neurobiologiae Experimentalis

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2013

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tom 73

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nr Suppl.1

Genomic response of neuronal cell to external stimuli includes expression of specific transcript isoforms, that use alternative transcription start sites and polyadenylation signals. We applied microarray profiling (Illumina Mouse WG-6) and next-generation sequencing (SOLID 5500xl) to screen for drug-induced activitydependent transcriptional events in the C57BL/6J mouse striatum. We compared effects on gene expression induced by psychoactive drugs with diverse neuropharmacological mechanisms of action (antidepressants, antipsychotics, anxiolytics, psychostimulants and opiods). Using whole-genome approach we identified a pool of transcripts that are regulated by the psychotropic drugs in mouse striatum (317 transcripts). We found that drug-responsive transcripts are organized into three main co-regulated gene expression networks. Furthermore, using Bowtie read aligner and Cufflinks algorithm we identified specific gene isoforms responsive to drug treatment. 58% among the drug-regulated transcripts were defined as alternative transcription events. To search for transcriptional factors that control alternative gene transcription in the brain we developed seqinterpreter web-based tool (http:// seqinterpreter.cremag.org). We found SRF, NPAS4 and GR as candidate regulatory factors. The complex program of regulation in gene transcription may further impact long-lasting alterations in brain function.

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New evidence for a role of mGluR7 in astrocyte survival: Possible implications for neuroprotection

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Jantas D., Lech T., Golda S., Pilc A., Lason W.

Acta Neurobiologiae Experimentalis

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2017

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tom 77

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nr Suppl.1

INTRODUCTION: A specific activation of metabotropic glutamatergic receptor subtype 7 (mGluR7) by its allosteric agonist AMN082 has been shown to protect neuronal cells against various detrimental factors. It is well established that some of subtypes of mGluRs (e.g., mGluR5 or mGluR3) engage glia cells to more efficiently protect neurons against various harmful stimuli. AIM(S): We aimed to study the role of mGluR7 in glia and neuronal cell survival. METHOD(S): We used primary cortical glia cell cultures and cerebellar granule neurons (CGNs)from mGluR7+/+ and mGluR7-/- C57Bl/6J mice which were exposed to various cell damaging factors (staurosporine (St), doxorubicin (Dox)and low potassium (LP)). MTT reduction, LDH release and caspase-3 activity biochemical assays were used for assessment of cell damage. The mRNA expression level of various subtypes of mGluRs was measured by qPCR. RESULTS: We showed the expression of mGluR7 in glia cell cultures and demonstrated the higher toxicity of St and Dox in mGluR7-/- glia cells when compared to wild type one. Moreover, we found a partial protection mediated by AMN082 against St and Dox in mGluR7+/+ glia cells. However, we did not find any differences in vulnerability of CGNs derived from mGluR7+/+ and mGluR7-/- animals to the cell damaging action of LP, St or Dox under standard treatment. Intriguingly, when we primed both types of CGNs by culturing them overnight in LP medium, we found significant higher toxic action of St and Dox in mGluR7‑/‑ CGNs. Finally, we confirmed neuroprotective properties of AMN082 in CGNs and showed that this effect is stimuli‑ and development-dependent. CONCLUSIONS: Our data obtained in isolated glia and neuronal cellular models showed a protective potential of mGluR7‑specific agonist AMN082 and pro‑survival role of mGluR7 in glia cells which together with its already known direct role on neuronal cells could suggest its higher efficacy under in vivo conditions. FINANCIAL SUPPORT: The study was supported by statutory funds for Institute of Pharmacology PAS and grant No NN405611638 from the Ministry of Science and Higher Education, Warsaw, Poland.

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Regulation of alternative gene expression in the mouse striatum in response to cocaine

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Zygmunt M., Rodriguez-Parkitna J., Golda S., Piechota M., Ficek J., Korostynski M.

Acta Neurobiologiae Experimentalis

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2015

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tom 75

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nr Supl.

BACKGROUND AND AIMS: Cocaine is a potent psychostimulant that increases levels of striatal dopamine and activates neuronal circuits controlling motivation and reward-based learning. Transcriptional response to cocaine includes expression of alternative gene isoforms and splicing variants. Unraveling the regulatory mechanisms that are involved in selection of active transcription start and termination sites provides novel insight into molecular basis of drug-induced brain plasticity. METHODS: We used next-generation sequencing (RNA-seq) to comprehensively map expression of genesin the mouse striatum. Total RNA and small RNA resequencing was performed in samples collected 1 h after acute treatment with 25 mg/kg cocaine. To identify transcripts responsive to drug treatment we used Tophat read-mapper and Cufflinks algorithm for FPKM quantification. The seqinterpreter online tool was used to search for key regulatory factors that control alternative gene transcription in the brain (http://seqinterpreter.cremag.org). RESULTS: In addition to increased expression of activity-regulated genes, different types of cocaine-inducible splicing variants and transcript biotypes were identified. Examples of different modalities of gene expression include alternative first exon (e.g. Stxbp1), alternative last exon (Hsph1), intron retention (Dnajb5), long noncoding RNA (Gm13889) and small RNA (Mir92b and Mir130a). In order to investigate neuron-type specificity of gene expression we have used fluorescence-activated cell sorting to isolate genetically labeled dopamine receptor 1 expressing neurons. CONCLUSIONS: Our results provide a comprehensive assessment of neuronal activity-induced gene expression at the level of individual transcriptional units rather than whole genes. Further experiments will explore differences in activity-regulated gene expression in discrete neuron types, i.e. the D1 or D2 expressing medium spiny neurons of the striatum. This work was supported by NCN grant SONATA 2011/03/D/ NZ3/01686.

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Molecular and behavioral patterns associated to stress responsivity and PTSD risk

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Szklarczyk K., Smutek M., Korostynski M., Solecki W., Golda S., Przewlocki R.

Acta Neurobiologiae Experimentalis

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2011

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tom 71

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nr S

Post-traumatic stress disorder (PTSD) is a chronic anxiety condition that develops as a result of a terrifying event. Clinical studies show that only about 10% of trauma-exposed people suffer from PTSD. Our research was focused on endophenotypes and molecular biomarkers of PTSD in an animal model. Differences in response to stress among inbred mouse strains (C57BL/6J, DBA/2J, SWR/J and 129P3/J) were compared: a single intense footshock was applied and ultrasonic vocalization during/after the stress was measured. Long-lasting effects were assessed 4-6 weeks after the traumatic event: conditioned and sensitized fear, social withdrawal, depressive-like behavior and susceptibility to drug addiction. SWR/J strain displayed the lowest conditioned fear, whereas sensitized fear was increased over time in C57BL/6J mice. Moreover, C57BL/6J strain exhibited increase in depressive-like behavior, while DBA/2J strain displayed increased social withdrawal. In addition, it was observed that exposition to traumatic stress increased sensitivity to rewarding properties of morphine in 129P3/J mice. Diverse long-lasting behavioral consequences of exposition to stress were associated with changes in basal and stress-induced profile of glucocorticoid receptor-dependent (GR) genes (e.g., Fkbp5 and Tsc22d3) in amygdala. Furthermore, our research showed that administration of GR antagonist disrupted reconsolidation of the traumatic event memory. Our research supports a model in which genetic factors are important for phenotypic variation in responsivity to stress. These genes may provide novel insight into mechanisms of stress-related disorders. This work was supported by Polish MSHE grants NN405 274137, N405 143238, IUVENTUS Plus and POIG De-Me-Ter 3.1

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Overexpression of biliverdin reductase enhances heme oxygenase expression, activates ERK1, JNK1 and p38 kinases and ensure partial protection against cellular stress

76%

Florczyk U., Golda S., Zieba A., Cisowski J., Jozkowicz A., Dulak J.

Acta Biochimica Polonica

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2007

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tom 54

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nr Supplement 4

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Optymalizacja otrzymywania wektorow AAV

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Jazwa A., Rutkowski A., Golda S., Rehhahn A., Jozkowicz A., Dulak J.

Biotechnologia

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2007

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nr 3

141-156

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Analysis of morphine-induced gene expression in the mouse hippocampus

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Barut J., Skupio U., Marut L., Tertil M., Golda S., Przewlocki R.

Acta Neurobiologiae Experimentalis

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2017

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tom 77

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nr Suppl.1

INTRODUCTION: Exposure to drugs of abuse initiates molecular alterations in the central nervous system that lead to an increased overall tenderness to addiction with subsequent drug exposures. These drug-induced alterations employ changes in gene expression, which may underlie the behavioral aberrancy that define a state of addiction. AIM(S): To identify the specific transcriptional alterations in different stages of morphine addiction in the hippocampus (Hip), brain region which play a role in the acquisition and extinction of memories associated with drug seeking behavior. METHOD(S): C57BL/6J male mice were injected twice daily for 3 weeks with morphine (increasing doses, 20-100 mg/kg i.p.). Animals were observed for spontaneous signs of withdrawal and behavior was measured in first and third week of abstinence. Morphine induced gene expression in the Hip was analyzed, using the qPCR technology. RESULTS: 24 h after chronic treatment we have observed spontaneous withdrawal syndrome and the peak of corticosterone levels in blood. Morphine-abstinent mice exhibited a variety of depression-like behaviors and cognitive deficits. Analyzes of Hip transcriptional responses to morphine indicated that most of genes regulated by morphine injection are GR-dependent, with a number of them being astrocyte‑specific (Gjb6, Plin4, Slc1a3, Gfap, Gja1). Analyzed genes clustered into few co-expressed groups, i.a. GR-dependent (Fkbp5, Tsc22d3, Zbtb16, Plin4) and activity-dependent (Fos, Fosl2) both upregulated in single and chronic exposure to morphine. Interestingly, 3-weeks abstinent mice didn’t exhibit any significant difference in transcription, but single dose of morphine (relapse) trigger sensitization of expression of some interesting genes (camk1g, Fosl2, Arc). CONCLUSIONS: Our results reveal that morphine induces drug‑specific transcriptional signatures in the Hip. Stress systems in Hip may modify the reward circuit through GR-dependent molecular pathways and this mechanism may be a fundamental for addiction therapy research. FINANCIAL SUPPORT: Polish National Science Centre Grant number 2013/08/A/NZ3/00848.

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HO-1 regulates expression of human MMP-1 and murine MMP-13 in vitro

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Golda S., Kabala P., Sokolowska M., Stopa M., Kotlinowski J., Sierpnowska A., Jozkowicz A., Boczkowski J., Dulak J.

Acta Biochimica Polonica

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2007

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tom 54

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nr Supplement 4

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Regulacja ekspresji genow w wektorach plazmidowych: system zalezny od doksycykliny i regulowany przez niedotlenienie

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Golda S., Kucharzewska P., Cisowski J., Florczyk U., Zagorska A., Jazwa A., Loboda A., Jozkowicz A., Dulak J.

Biotechnologia

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2007

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nr 3

82-97

Ograniczanie wyników

Global analysis of drug-regulated alternative expression of genes in mouse brain

New evidence for a role of mGluR7 in astrocyte survival: Possible implications for neuroprotection

Regulation of alternative gene expression in the mouse striatum in response to cocaine

Molecular and behavioral patterns associated to stress responsivity and PTSD risk

Overexpression of biliverdin reductase enhances heme oxygenase expression, activates ERK1, JNK1 and p38 kinases and ensure partial protection against cellular stress

Optymalizacja otrzymywania wektorow AAV

Analysis of morphine-induced gene expression in the mouse hippocampus

HO-1 regulates expression of human MMP-1 and murine MMP-13 in vitro

Regulacja ekspresji genow w wektorach plazmidowych: system zalezny od doksycykliny i regulowany przez niedotlenienie