Joanna Goscik

The Effect of Acute and Chronic Social Stress on the Hippocampal Transcriptome in Mice

Psychogenic stress contributes to the formation of brain pathology. Using gene expression microarrays, we analyzed the hippocampal transcriptome of mice subjected to acute and chronic social stress of different duration. The longest period of social stress altered the expression of the highest number of genes and most of the stress-induced changes in transcription were reversible after 5 days of rest. Chronic stress affected genes involved in the functioning of the vascular system (Alas2, Hbb-b1, Hba-a2, Hba-a1), injury response (Vwf, Mgp, Cfh, Fbln5, Col3a1, Ctgf) and inflammation (S100a8, S100a9, Ctla2a, Ctla2b, Lcn2, Lrg1, Rsad2, Isg20). The results suggest that stress may affect brain functions through the stress-induced dysfunction of the vascular system. An important issue raised in our work is also the risk of the contamination of brain tissue samples with choroid plexus. Such contamination would result in a consistent up- or down-regulation of genes, such as Ttr, Igf2, Igfbp2, Prlr, Enpp2, Sostdc1, 1500015O10RIK (Ecrg4), Kl, Clic6, Kcne2, F5, Slc4a5, and Aqp1. Our study suggests that some of the previously reported, supposedly specific changes in hippocampal gene expression, may be a result of the inclusion of choroid plexus in the hippocampal samples.

Novel candidate genes for alcoholism — transcriptomic analysis of prefrontal medial cortex, hippocampus and nucleus accumbens of Warsaw alcohol-preferring and non-preferring rats

OBJECTIVE Animal models provide opportunity to study neurobiological aspects of human alcoholism. Changes in gene expression have been implicated in mediating brain functions, including reward system and addiction. The current study aimed to identify genes that may underlie differential ethanol preference in Warsaw High Preferring (WHP) and Warsaw Low Preferring (WLP) rats. METHODS Microarray analysis comparing gene expression in nucleus accumbens (NAc), hippocampus (HP) and medial prefrontal cortex (mPFC) was performed in male WHP and WLP rats bred for differences in ethanol preference. RESULTS Differential and stable between biological repeats expression of 345, 254 and 129 transcripts in NAc, HP and mPFC was detected. Identified genes and processes included known mediators of ethanol response (Mx2, Fam111a, Itpr1, Gabra4, Agtr1a, LTP/LTD, renin-angiotensin signaling pathway), toxicity (Sult1c2a, Ces1, inflammatory response), as well as genes involved in regulation of important addiction-related brain systems such as dopamine, tachykinin or acetylcholine (Gng7, Tac4, Slc5a7). CONCLUSIONS The identified candidate genes may underlie differential ethanol preference in an animal model of alcoholism. COMMENT Names of genes are written in italics, while names of proteins are written in standard font. Names of human genes/proteins are written in all capital letters. Names of rodent genes/proteins are written in capital letter followed by small letters.

Brief Communication: Maternal Plasma Autoantibodies Screening in Fetal Down Syndrome

Imbalance in the metabolites levels which can potentially be related to certain fetal chromosomal abnormalities can stimulate mothers immune response to produce autoantibodies directed against proteins. The aim of the study was to determine the concentration of 9000 autoantibodies in maternal plasma to detect fetal Down syndrome. Method. We performed 190 amniocenteses and found 10 patients with confirmed fetal Down syndrome (15th–18th weeks of gestation). For the purpose of our control we chose 11 women without confirmed chromosomal aberration. To assess the expression of autoantibodies in the blood plasma, we used a protein microarray, which allows for simultaneous determination of 9000 proteins per sample. Results. We revealed 213 statistically significant autoantibodies, whose expression decreased or increased in the study group with fetal Down syndrome. The second step was to create a classifier of Down syndrome pregnancy, which includes 14 antibodies. The predictive value of the classifier (specificity and sensitivity) is 100%, classification errors, 0%, cross-validation errors, 0%. Conclusion. Our findings suggest that the autoantibodies may play a role in the pathophysiology of Down syndrome pregnancy. Defining their potential as biochemical markers of Down syndrome pregnancy requires further investigation on larger group of patients.

Sphingolipids as a new factor in the pathomechanism of preeclampsia – Mass spectrometry analysis

Objective(s) and design The aim of the study was to analyse a panel of 11 sphingolipids in plasma and three blood fractions (platelet-poor plasma, platelets and red blood cells) of women with mild preeclampsia. Materials and methods We recruited 21 women between 25–40 weeks gestation with diagnosed mild preeclampsia to the study group and 36 healthy women with uncomplicated pregnancies, who corresponded with the study group according to gestational age, to the control group. To assess the concentration of 11 sphingolipids in the blood plasma and blood fractions, we used ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC/MS/MS). Results We showed a significant increase in the concentration of eight sphingolipids in the plasma of women with preeclampsia in comparison to the control group: Sph (p = 0.0032), S1P (p = 0.0289), C20-Cer (p < 0.0001), C18-Cer (p < 0.0001), C16-Cer (p = 0.012), C18:1-Cer (p = 0.003), C22-Cer (p = 0.0071), and C24:1-Cer (p = 0.0085). Conclusion We showed that selected sphingolipids, especially C20-Cer and C18-Cer, are totally new factors in the pathomechanism of PE and that these bioactive lipids may play an important role in apoptosis and autophagy.

Angiogenic factor screening in women with mild preeclampsia – New and significant proteins in plasma

HighlightsIncrease in the concentration of 8 proteins in the plasma of women with preeclampsia.IFN‐&ggr;, IL‐6, LIF, Hb‐EGF, HGF, IP‐10, leptin, PDGF‐BB.Significant decrease in the concentration of 3 proteins in the plasma of women with preeclampsia.VEGF, PlGF and follistatin. Introduction The aim of this study was to analyse a panel of 60 angiogenic factors (pro‐angiogenic and antiangiogenic) in the plasma of women with mild preeclampsia. Materials and Methods We recruited 21 women between 25 and 40 weeks gestation with diagnosed mild preeclampsia into the study group and 27 healthy women with uncomplicated pregnancies of corresponding gestational age to that of the study to the control group. We used a quantitative protein macroarray method that allowed for analysis of 60 angiogenic proteins per sample simultaneously. Results We showed a statistically significant increase in the concentration of 8 proteins, interferon gamma (IFN‐&ggr;), interleukin 6 (IL‐6), leukaemia inhibitory factor (LIF), heparin‐binding EGF‐like growth factor (HB‐EGF), hepatocyte growth factor (HGF), C‐X‐C motif chemokine 10 (IP‐10), leptin and platelet‐derived growth factor BB (PDGF‐BB), as well as a significant decrease in the concentration of 3 proteins, vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and follistatin, in the plasma of women with preeclampsia. Conclusion Based on our findings, it seems that protein factors may play an important role in the pathogenesis of preeclampsia, and there are many proteins that have not been studied in PE to date. There are no previous studies assessing the LIF, follistatin, HGF, HB‐EGF and PDGF‐BB concentrations in the plasma of women with PE; therefore, our obtained results indicate that these proteins are new factors that can play an important role in the pathomechanisms of PE.

Pregnancy at Advanced Maternal Age Affects Behavior and Hippocampal Gene Expression in Mouse Offspring

Abstract There is growing evidence that advanced maternal age is a risk factor for neurological and neuropsychiatric disorders in offspring. However, it remains unclear whether the altered brain programming induced by advanced maternal age is mediated by pre- or postnatal factors. Here, a mouse model was used to investigate whether pregnancy at advanced age may provoke behavioral and brain gene expression changes in offspring. Swiss Albino mice conceived by 3-month-old males and either 15–18-month-old (n = 11) or 3-month-old control females (n = 5), were delivered by cesarean section, fostered after birth by 3-month-old dams and subjected to a battery of behavioral tests. Furthermore, genome-wide mRNA expression was analyzed in the hippocampi of 4-month-old males offspring using microarrays. Offspring conceived by old mothers exhibited increased ultrasound vocalization activity during separation from the foster mother, increased anxiety-like behaviors in adult life, and altered patterns of hippocampal gene expression, compared to controls. These effects were not reversed by the postnatal maternal care provided by the young foster mothers, suggesting that the altered brain programming is already established at birth, consistent with prenatal effects related to maternal aging.

Maternal Plasma Metabolomic Profiles in Spontaneous Preterm Birth: Preliminary Results

Objective To profile maternal plasma metabolome in spontaneous preterm birth. Method In this retrospective case-control study, we have examined plasma of patient with preterm birth (between 22 and 36 weeks of pregnancy (n = 57)), with threatened preterm labor (between 23 and 36 weeks of pregnancy (n = 49)), and with term delivery (n = 25). Plasma samples were analysed using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) in positive and negative polarity modes. Results We found 168 differentially expressed metabolites that were significantly distinct between study groups. We determined 51 metabolites using publicly available databases that could be subdivided into one of the five groups: amino acids, fatty acids, lipids, hormones, and bile acids. PLS-DA models, verified by SVM classification accuracy, differentiated preterm birth and term delivery groups. Conclusions Maternal plasma metabolites are different between term and preterm parturitions. Part of them may be related with preterm labor, while others may be affected by gestational age or the beginning of labor. Metabolite profile can classify preterm or term delivery groups raising the potential of metabolome as a biomarker to identify high-risk pregnancies. Metabolomic studies are also a tool to detect individual compounds that may be further tested in targeted researches.

Maternal plasma angiogenic and inflammatory factor profiling in foetal Down syndrome

Objective and design Angiogenic factors are proteins that are related to certain foetal chromosomal abnormalities. The aim of this study was to determine the concentration of 60 angiogenic factors in the plasma of women with offspring possessing trisomy 21/Down syndrome (DS). Method After analysing karyotyping results, we selected 20 patients with foetuses possessing DS, and for the control group, we selected 28 healthy patients with uncomplicated pregnancies who delivered healthy newborns at term (i.e., 15–18 weeks of gestation). To assess the concentration of proteins in the blood plasma, we used a protein macroarray which enabled simultaneous determination of 60 angiogenic factors per sample. Results We observed a statistically significant increase in the concentration of these five angiogenic and inflammatory factors: TGFb1 (p = 0.039), angiostatin (p = 0.0142), I-309 (p = 0.0476), TGFb3 (p = 0.0395), and VEGF-D (p = 0.0173)—compared to concentrations in patients with healthy foetuses. Conclusion Our findings suggest that angiogenic factors may play role in DS pathogenesis.

Developmental sequelae of advanced paternal age on fetal brain and placenta in mice

Stem cells of the intestinal epithelium are located in a distinct compartment known as the crypt of Liberkühn. While matrigel or synthetic hydrogel-based system support growth of isolated crypts as 3D organoids, the morphology and stability of the organoids does not resemble normal intestinal epithelium. Moreover, lack of native ECM components and ECM positional context render both of the systems unfit for investigating the impact of age of the underlying ECM in stem cell maintenance. Here, we demonstrate the reconstruction of the proper intestinal stem cell niche by seeding FACS sorted single Lgr5+ intestinal stem cells or cultured organoids on decellularized mouse intestinal extracellular matrix scaffold ex vivo. The stem cells form distinct and stable crypt and villous units according to the former crypt and villous ‘ECM mould’. Using the model, we have discovered that regenerative capacity of young stem cell is declined when they are cultured on ‘old ECM’ suggesting that age of ECM transcriptomically rewires the functions of young stem cell. In fact, our RNAseq data shows that stem cell signature genes are specifically suppressed in both young stem cells that are cultured on old ECM and freshly isolated old crypts. Moreover, TGFβ pathway is specifically enriched both in the young cells that are cultured in old ECM and old isolated crypts suggesting that reduced regeneration of old mouse intestine may be due to alterations in the TGFβ signaling. In fact, the gradient of TGFβ signaling, being lower in the crypt, is lost in the old small intestine. These findings suggest that chronic TGFβ might underlie reduced regenerative capacity of old small intestine. In summary, we have developed a novel organotypic culture system for small intestinal stem cells based on tissue’s native ECM and discovered a de-regulated TGF β signaling underlies reduced regeneration in the old mouse small intestine.