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1

Addicted brain mapping and imaging using light-sheet fluorescence microscopy

100%
Stefaniuk M.
Acta Neurobiologiae Experimentalis
|
2019
|
tom 79
|
nr Suppl.1
Addiction is a disease that affects circuits and brain areas involved in reward, stress, and self‑control. Continued substance abuse can lead to dependence that is associated with withdrawal symptoms when drug availability is ceased, and increased intake upon relapse. With many brain structures involved in the disease, the whole-brain analysis is a promising strategy for gaining a deeper insight into events leading to addiction development and its particular aspects, such as relapse. We used a mouse model of alcohol addiction to perform whole-brain analysis of the activity during addiction‑like behaviors. Mice were trained to drink ethanol in “drinking in the dark” paradigm adapted to the IntelliCage system. Afterwards, the animals were deprived of alcohol and tested for relapse. Whole hemispheres of each mouse were subjected to optical clearing, c-Fos staining, light-sheet imaging, and analyzed for c-Fos expression to identify brain areas involved in relapse.
2

RNA sequencing as a powerful tool in searching for genes influencing health and performance traits of horses

63%
Stefaniuk M., Ropka-Molik K.
Journal of Applied Genetics
|
2016
|
tom 57
|
nr 2
3

Manipulation of Ttyh1 gene expression influences cell morphology in neuroblastoma and hippocampal neurons in vitro

63%
Stefaniuk M., Lukasiuk K.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 3
Ttyh1 gene is a member of tweety family of putative large conductance maxi-Cl- channels. According to our previous data, expression of Ttyh1 mRNA in the brain is localized in neurons and is increased following status epilepticus in the animal model of epileptogenesis. The function of Ttyh1 has not been elucidated. Here we aimed at characterization of Ttyh1 by overexpressing or silencing it in neuroblastoma cell line or in hippocampal neurons in vitro. In hippocampal neurons transfected with plasmid coding Ttyh1-EGFP fused protein, Ttyh1-EGFP was present in dots along the neurites and at the ends of new formed projections. Similar Ttyh1-EGFP localization was observed in neuroblastoma cells. Overexpression of Ttyh1 in hippocampal neurons in vitro induced formation of new, often branched projections as soon as after 24 h. Even more intense branching was observed when cells were transfected with Ttyh1 7 or 14DIV. Ttyh1 silencing in hippocampal neurons in vitro using siRNA increased the number and length of primary dendrites. In addition, it affected cell morphology causing abnormal pattern of MAP2 distribution. Since manipulation of Ttyh1 expression infl uences cell morphology and distribution of cytoskeleton elements, we propose that Ttyh1 is involved in cytoskeleton functions. It is tempting to suggest, that by infl uencing the neurite growth and ramifi cation, Ttyh1 participates in aberrant network formation and by this – the development of epilepsy. Supported by MNiSW N N301 162135
4

Wyścigi a gen szybkości

51%
Bielewicz M., Sosinska A., Wozowicz W., Stefaniuk M.
Konie i Rumaki
|
2014
|
nr 06
5

The impact of matrix metalloproteinase 9 on alcohol-addiction related mouse behaviour

51%
Sakharchuk O., Stefaniuk M., Radwanska K., Kaczmarek L.
Acta Neurobiologiae Experimentalis
|
2013
|
tom 73
|
nr Suppl.1
Alcohol addiction is a chronic, psychiatric disease defined by compulsive alcohol drinking and seeking. Long-term alcohol intake induces aberrant synaptic plasticity in the amygdala and striatum as well as enhanced c-Fos expression in the central nucleus of amygdala (CeAmy). Interestingly, human studies have implicated matrix metalloproteinase 9 (MMP-9), whose gene is regulated by c-Fos, in alcohol addiction. Notably, recently critical role of MMP-9 in reward-driven learning as well as synaptic plasticity has been revealed. In the present study we aimed at elucidating a role of MMP-9 in alcohol addiction in mice. First, we analyzed the effects of MMP-9 levels on the dendritic spine morphology in the CeAmy of C57BL/6 wild type (WT) and mice lacking MMP-9 (MMP-9 KO). Next, to verify the role of MMP-9 in alcohol addiction we subjected WT, MMP-9 KO and heterozygous mice to behavioral tests in the Intellicage system, previously shown to be suitable to investigated addictive behaviors. Dendritic spine analysis of the CeAmy revealed that MMP-9 KO mice have longer and thinner dendritic spines than WT mice. Preliminary data analysis from the Intellicages showed there were no differences between MMP-9-KO and WT mice during first hour and first day activity, as well as in neophobia. Currently we are investigating the pattern of development of the alcohol addiction, to evaluate the role of MMP-9 in aversive aspects of this behavior.
6

MMP-9 contributes to alcohol-induced synaptic plasticity in central amygdala

51%
Beroun A., Stefaniuk M., Kaczmarek L.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: Matrix metalloprotease 9 (MMP-9) is an extracellularly operating protease shown to play the key role in the morphological reorganization of dendritic spines as well as specific forms of learning and memory. Here we focus on a pathological form of plasticity - synaptic plasticity of alcohol addiction. AIM(S): The aim of this study was to investigate the role of MMP-9 in functional and structural synaptic plasticity during development of alcohol addiction. METHOD(S): Mice, housed in the IntelliCage system, were constantly monitored for alcohol consumption and motivation to reward. Spine morphology was evaluated by confocal imaging. We then performed whole-cell patch clamp recordings to test the strength of synapses as well as formation of silent synapses in the central amygdala. RESULTS: MMP-9 KO mice display lower motivation towards ethanol compared to wild type mice (WT). Moreover, in central amygdala, chronic alcohol drinking produced alterations in dendritic spine shape of both WT and KO but interestingly, more pronounced changes were observed in WT high alcohol consumers. To test how altered structural plasticity affects functionality of synaptic connections, we performed electrophysiological analysis of the strength of glutamatergic synapses in central amygdala. We discovered that alcohol consumption elevates the number of silent synapses (neonatal-like, immature synapses considered as substrates for increased plasticity). MMP-9 KO mice, however, showed no such synaptic adaptations neither after alcohol drinking nor subsequent withdrawal. CONCLUSIONS: These data suggest that MMP-9 is involved in synaptic plasticity associated with alcohol addiction. The change of spine morphology together with elevated silent synapse number might represent ongoing circuitry reorganization that primes neurons for enhanced learning, which might lead to compulsive ethanol use. FINANCIAL SUPPORT: This study is supported by the National Science Centre grant Sonata (2015/19/D/ NZ4/03701).
7

MMP-9 activity in animal model of traumatic brain injury

51%
Pijet B., Stefaniuk M., Rejmak-Kozicka E., Kaczmarek L.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: Epilepsy in 20% of cases, develops as an effect of traumatic brain injury (TBI). Recent evidences indicate important role of extracellular matrix metalloproteinase-9 (MMP-9) in neuronal circuitry remodeling and synaptic plasticity. AIM(S): The aim of the present study was to evaluate the MMP-9 activity changes, dendritic spines density after brain injury and the influence of MMP‑9 expression level on spontaneous seizures appearance after TBI. METHOD(S): We used Controlled Cortical Impact (CCI) as a model of TBI in animals with altered MMP-9 levels (lacking: MMP-9 KO; overexpressing: MMP-9 OE) and their WT siblings. 12 weeks after CCI animals were subjected to continuous video-EEG monitoring. To verify MMP-9 changes after TBI: gel zymography and in situ hybridization were used. For dendritic spine alterations staining using liophilic dye DiI were performed. RESULTS: TBI resulted in progressive cortex (Cx) degeneration during 30 days after TBI. This effect was MMP-9 dependent. In MMP-9 KO animals degeneration volume degree was significantly smaller compared to wildtype siblings and MMP-9 OE mice. Gel zymography analysis showed time-associated elevation of MMP-9 activity in ipsilateral Cx and Hp, also in the thalamus samples during 3 days after CCI. Moreover, in situ hybridization showed increase of MMP-9 mRNA expression which reached the peak 6 hours post-CCI. Density of the dendritic spines measured 7 and 14 d after CCI was significantly decreased in ipsilateral Cx and CA1. EEG recordings with threshold test showed decreased seizure latency in MMP-9 OE mice while increased in MMP-9 KO. Interestingly total seizure number was the highest in MMP-9 OE animals. CONCLUSIONS: We described the correlation between TBI and MMP-9 activity and action post trauma. We indicated that MMP-9 might be an important factor for major dendritic spine reshaping, observed after brain injury, which in consequence may lead to altered sensibility of neuronal circuits to trigger seizures. FINANCIAL SUPPORT: This work was supported by PBS Program founded by The National Centre of Research and Development.
8

Szybka metoda oznaczania substancji alkoholowo nierozpuszczalnych [AIS] w mrozonkach i konserwach zielonego groszku

51%
Stefaniuk M., Scigalski A., Swiderski A.
Przemysł Fermentacyjny i Owocowo-Warzywny
|
1995
|
tom 39
|
nr 02
22-24
Standardowa metoda amerykańska (metoda A) oznaczania substancji alkoholowo nierozpuszczalnych, które decydują o przydatności zielonego grochu na mrożonki i konserwy ( AO AC 1984, s. 608), oparta jest na ekstrakcji pulpy wodnej grochu 80-procentowym alkoholem oraz wielokrotnym przemywaniu alkoholem próbek na sączku i bezpośrednim wyliczaniu procentowej zawartości substancji nierozpuszczalnych w alkoholu. Metoda ta zostala porównana z metodą wodno-etanolową (metoda B) zmodyfikowaną w następujący sposób: sączenie zastąpiono wirowaniem próbek, uzyskując supernatant części rozpuszczalnych w etanolu. Procent wagowy składników rozpuszczalnych w etanolu (a) i nierozpuszczalnych w etanolu (y) obliczono ze wzoru: y = [100(s — a)]:[100 — a], gdzie s = procent wagowy suchej masy. W drugiej modyfikacji nazwanej metodą etanolową (metoda C) pulpę wodną zastąpiono pulpą etanolową. Procent wagowy suchej masy (s) oraz procent wagowy części nierozpuszczalnych w etanolu (y) obliczono jak w metodzie B. Z otrzymanych danych wynika, że pomiędzy tymi metodami (Ai В oraz A i C) brak jest istotnych różnic, co potwierdza przydatność obu modyfikacji jako szybszych, tańszych i nadających się do badań seryjnych.
9

Hyperphagic obesity in adult dicer CKO mice: the role of hypothalamic AgRP/NPY neurons

51%
Hajdukiewicz K., Stefaniuk M., Grinievich V., Konopka W.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: Obesity is a worldwide disease of complex etiology. The main appetite regulatory center is located within the brain, in the hypothalamus. A putative mechanism responsible for the obesity phenotype involves microRNA interplay between feeding regulatory elements of the hypothalamic AgRP/NPY-expressing, POMC-producing and probably other arcuate nucleus neurons. Dicer is a key enzyme in microRNA processing. In Dicer’s absence, there is a pronounced lack of mature microRNAs and a disturbed regulation of translation. AIM(S): We want to observe how massive, spatially and temporally defined, loss of microRNAs impacts metabolism and obesity outcome. METHOD(S): We injected rAAV-coding Cre recombinase under the AgRP specific promoter into the arcuate nucleus of mice with a Cre- dependent Dicer sequence. As NPY is a known appetite stimulator, to determine whether NPY is the key player in this phenotype, we induced Dicer loss in NPY knockout (KO) mice via Tamoxifen IP injections. RESULTS: Our preliminary data show that administration of AAV-AgRP- Cre construct leads to visible weight gain, correlated with increased food intake. This phenotypic effect is AAV‑dose‑dependent and is likely accompanied by an imbalance between anorexygenic and orexigenic neuropeptide levels. However, NPY KO mice with massive microRNA loss gradually put on weight, though with different kinetics as compared to Dicer CKO mice. CONCLUSIONS: Our approach demonstrates that microRNA loss in a subpopulation of arcuate nucleus neurons has a very pronounced effect on the central regulation of metabolism, expressed by weight gain as well as hyperphagy. Moreover, it is likely the mechanism involves an extensive system of complex relationships because loss of single-gene coding of the main orexigenic neuropeptide (NPY) does not inhibit weight gain. Nevertheless, the exact mechanism underlying this phenomenon has not yet been elucidated.
10

Choroby genetycze. JEB

51%
Stefaniuk M., Michalska A., Kmiecik B.
Konie i Rumaki
|
2016
|
nr 01
11

Analiza sezonów rozrodowych w Stadninie Koni Iwno za lata 1992-2012

51%
Podstawski Z., Stefaniuk M., Sadlek J., Kulisa M.
Wiadomości Zootechniczne
|
2014
|
tom 52
|
nr 4
12

The methodology of magnetotelluric investigation in heterogeneous media

51%
Miecznik J., Stefaniuk M., Klitynski W.
Bulletin of the Polish Academy of Sciences. Earth Sciences
|
1995
|
tom 43
|
nr 2
13

Rapid estimation of major sugars in honey. Analytical considerations

45%
Samotus B., Scigalski A., Stefaniuk M., Swiderski A., Dorre E.
Zeszyty Naukowe Akademii Rolniczej w Krakowie. Ogrodnictwo
|
1995
|
tom 22
97-106
14

Applicability of Folin reagent for the ascorbic acid determination in vitamin preparation, fruits and vegetables

45%
Samotus B., Dorre E., Swiderski A., Stefaniuk M., Scigalski A.
Acta Agraria et Silvestria. Series Agraria
|
1994
|
tom 32
85-93
The applicability of Folin reagent developed by Tsap [12] for the determination of pure ascorbic acid solutions was checked for more complex solutions as vitamin preparations, fruits and vegetables. The AA can be determined in vitamin preparations without any modification. However it must be modified in the case of plant extracts (fruits and vegetables) as follows: one sample is determined after strong alkalization of the sample, another one without alkalization. The result of AA concentration can be achieved by subtraction of these two determinations. Good results were obtained for lettuce, parsley (tops), Brussels sprouts, lemon, orange, grapefruit, chives, kiwifruit and spinach.
15

Stadniny i Stada Ogierów Gniezno

45%
Podstawki Z., Stefaniuk M., Kaczmarczyk J.
Konie i Rumaki
|
2015
|
nr 12
16
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Co jest zapisane w chromosomie Y o udomowieniu koni?

44%
Stefaniuk M.
Wszechświat
|
2013
|
tom 114
|
nr 10-12
17

Historia udomowienia - czyli kronika pisana kopytem

39%
Pietrzyk K., Ryszka M., Musial J., Niezgodzka M., Solawa J., Stefaniuk M.
Konie i Rumaki
|
2014
|
nr 07
18

Evaluation of minimally invasive muscle biopsy method for genetic analysis in horse

39%
Stefaniuk M., Ropka-Molik K., Piorkowska K., Bereta A., Szpar P., Czerwonka Z., Podstawski Z.
Annals of Animal Science
|
2015
|
tom 15
|
nr 3
19

Identification of a new haplotype within the promoter region of the MSTN gene in horses from five of the most common breeds in Poland

39%
Stefaniuk M., Kaczor U., Augustyn R., Gurgul A., Kulisa M., Podstawski Z.
Folia Biologica
|
2014
|
tom 62
|
nr 3
20

Whole-brain mapping of neuroplasticity in different experimental paradigms in mice - a computational perspective

39%
Bednarek S., Jermakow N., Stefaniuk M., Pawlowska M., Wojcik D. K., Majka P.
Acta Neurobiologiae Experimentalis
|
2019
|
tom 79
|
nr Suppl.1
INTRODUCTION: Imaging of entire brains at cellular resolution, enabled by light-sheet fluorescence microscopy (LSFM) and optical tissue clearing, offers insights into neural activity at a high magnification while preserving the brain‑wide context. AIM(S): We propose a set of open-source computational tools that address three fundamental challenges associated with the analysis of LSFM images of entire rodent brains, namely: management of voluminous imaging data, alignment to a reference atlas, and object detection and localization. METHOD(S): The data for each brain, such as multichannel acquisitions and spatial information, are compressed and stored in an HDF5-based container as a pyramid of resolutions to facilitate and standardize data access and manipulation. Unlike most other alignment approaches, our pipeline is not only guided by standard similarity metrics such as mutual information, but also utilizes Deep Convolutional Neural Networks to generate label maps corresponding to specific brain structures such as main white matter tracts or dentate gyrus. This step significantly increases the accuracy and robustness of the registration procedure. The c‑Fos‑positive nuclei are identified and quantified with the help of another neural network trained on synthetic data, generated to simulate the original nuclei which eliminated the laborious process of manual image annotation. The software was applied to investigate c-Fos-mediated neuroplasticity in iDISCO-cleared brains in experimental paradigms of appetitive and aversive learning and alcohol addiction. RESULTS: Voxel-wise statistical analysis revealed brain areas involved in the neuroplasticity of alcohol addiction and appetitive or aversive learning in mice. CONCLUSIONS: We demonstrate the ability of our software to combine efficient data management, accurate atlas alignment, and object detection to facilitate LSFM analyses. FINANCIAL SUPPORT: ERA‑NET NEURON/17/2017 grant from NCBR, G2631 grant from NCN.
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