Ovariectomy and Subsequent Treatment with Estrogen Receptor Agonists Tune the Innate Immune System of the Hippocampus in Middle-Aged Female Rats

The innate immune system including microglia has a major contribution to maintenance of the physiological functions of the hippocampus by permanent monitoring of the neural milieu and elimination of tissue-damaging threats. The hippocampus is vulnerable to age-related changes ranging from gene expression to network connectivity. The risk of hippocampal deterioration increases with the decline of gonadal hormone supply. To explore the impact of hormone milieu on the function of the innate immune system in middle-aged female rats, we compared mRNA expression in the hippocampus after gonadal hormone withdrawal, with or without subsequent estrogen replacement using estradiol and isotype-selective estrogen receptor (ER) agonists. Targeted profiling assessed the status of the innate immune system (macrophage-associated receptors, complement, inhibitory neuronal ligands), local estradiol synthesis (P450 aromatase) and estrogen reception (ER). Results established upregulation of macrophage-associated (Cd45, Iba1, Cd68, Cd11b, Cd18, Fcgr1a, Fcgr2b) and complement (C3, factor B, properdin) genes in response to ovariectomy. Ovariectomy upregulated Cd22 and downregulated semaphorin3A (Sema3a) expression, indicating altered neuronal regulation of microglia. Ovariectomy also led to downregulation of aromatase and upregulation of ERa gene. Of note, analogous changes were observed in the hippocampus of postmenopausal women. In ovariectomized rats, estradiol replacement attenuated Iba1, Cd11b, Fcgr1a, C3, increased mannose receptor Mrc1, Cd163 and reversed Sema3a expression. In contrast, reduced expression of aromatase was not reversed by estradiol. While the effects of ERa agonist closely resembled those of estradiol, ERb agonist was also capable of attenuating the expression of several macrophage-associated and complement genes. These data together indicate that the innate immune system of the aging hippocampus is highly responsive to the gonadal hormone milieu. In ovariectomized female rats, estradiol replacement exerts potent immunomodulatory effects including attenuation of microglia sensitization, initiation of M2-like activation and modulation of complement expression by targeting hippocampal neurons and glial cells through ERa and ERb

Ovariectomy Alters Gene Expression of the Hippocampal Formation in Middle-Aged Rats

Ovarian hormones regulate the transcriptome of the hippocampus and modulate its functions. During 29 menopause, this complex signaling declines, leading to impaired learning and memory. This study was 30 undertaken to clarify the effects of long-term, surgical ovariectomy (OVX) on the rat hippocampal 31 transcriptome. At age of 13 months, intact control and ovariectomized groups were formed. All animals 32 were sacrificed 5 weeks after gonadectomy, hippocampal formations were dissected and processed for 33 transcriptome analysis. Microarray and PCR studies identified 252 and 61 genes, respectively, whose 34 expression was altered in the lack of ovarian hormones. Pathway analysis revealed impact on 35 neuroactive ligand-receptor interaction, endocannabinoid and estrogen signaling, among others. 36 Network and interaction analyses of proteins encoded by OVX-regulated genes revealed upregulation 37 of growth/ troph/transcription factor signaling assembly (Mdk, Fgf1, Igf2, Ngf, Ngfr, Ntf3, Ntrk1, Otx2, 38 Hif1a, Esr1, Nr4a3), peptides/peptide receptors (Cartp, Kl, Ttr, Gnrhr), neurotransmission (Grm1, 39 Gria4, Gls, Slc18a2, Kcj6) and genes serving immune functions (C3, Ccl2, Itgam, Il1b). Downregulated 40 clusters included neuropeptides and their receptors (Adcyap1, Cbln2, Cck, Cckbr, Crhr1 and 2, Oprd1, 41 Nts, Penk, Sstr1, Vip), neurotransmitter signaling (Htr2c, hrna3, Chrm4, Grm8, Hrh3, Slc17a6) and 42 potassium channels (Kcnk9, Kcnj9, Kcnma1, Kcnc2). Several transcription factors (Rxra, Thrb), solute 43 carriers and defense molecules (Apitd1, Bcl2, Clql3, Ilr3a, Sod1, Sncb) also underwent downregulation. 44 The findings indicate that surgical gonadectomy carried out at middle-age robustly changes the 45 hippocampal transcriptome that alters neurogenesis, synaptic plasticity, immune modulation causing 46 cognitive dysfunctions

Endokannabinoid szignalizáció szerepe a reprodukció hypothalamikus szintű szabályozásában = Endocannabinoid signaling in hypothalamic regulation of reproduction

A szaporodás idegrendszeri szabályozásában kulcs szerepet tölt be a gonadotropin-releasing hormone (GnRH) neuronrendszer. A rendszer működését perifériás hormonhatások és más agyi neuronhálózatok szabályozzák. Multidiszciplináris megközelítés alkalmazásával tanulmányoztuk a GnRH neuronrendszer kapcsolatait és szignál transzdukciós mechanizmusait, különös tekintettel a retrográd endokannabinoid szignalizáció szabályozó szerepére. Kísérleti eredményeinkről 24 tudományos közleményben adtunk számot, 96 összesített impakt értékkel. Feltártuk a hypothalamus kannabinoid receptor 1 (CB1) hírvivő RNS-t termelő idegsejtjeinek lokalizációját, valamint azok glutamáterg és GABA-erg fenotípusait. Igazoltuk, hogy a GnRH sejteken végződő GABA tartalmú idegvégződések CB1-t tartalmaznak, valamint bebizonyítottuk, hogy a GnRH idegsejtekből felszabaduló endokannabinoidok befolyásolják a GABA közvetítette információ átadást a GnRH neuronok felé. Feltártuk a ghrelin és endokannabinoid szignalizációs útvonalak kapcsolt jellegét a hypothalamusban. Igazoltuk a humán GnRH idegsejtek glutamát- és GABA-erg beidegzését. A GnRH neuronrendszer kisspeptinerg afferensei vonatkozásában új regulációs adatokat szolgáltattunk. Vizsgáltuk az ösztrogén szignalizáció szerepét a GnRH idegsejtek működésében, valamint az agykéregben. A GnRH idegsejtek működésének elmélyültebb tanulmányozására matematikai modellt alkottunk. Összegezve, eredményeink a reprodukció szabályozásának új mechanizmusait tárták fel. | Gonadotropin-releasing hormone (GnRH)-synthesizing neurons play a pivotal role in the central regulation of reproduction. Their operation depends on signaling by peripheral hormones and interactions with other neuronal circuits. By means of a multidisciplinary approach, the networking and signal transduction mechanisms of GnRH neurons were studied, with special reference to a putative retrograde endogenous cannabinoid signaling mechanism. The research results were published in 24 original papers representing a cumulative impact value of 96. Specifically, we mapped the hypothalamic distribution of cannabinoid receptor 1 (CB1) mRNA-expressing neurons and their GABA- and glutamatergic phenotypes, proved the presence of CB1 in neuronal afferents of GnRH neurons and characterized the impact of endocannabinoids liberated from GnRH neurons on the GABA-ergic signal transduction to GnRH cells. We provided evidence for the coupled nature of the ghrelin and the endocannabinoid signaling mechanisms. New GABA- and glutamatergic afferents of human GnRH neurons were also identified. In addition, novel regulatory mechanisms executed by kisspeptinergic circuits upon GnRH cells were revealed. We elucidated further characteristics of estradiol feedback effects to GnRH and cortical neurons. We established a mathematical model for the better understanding of GnRH cell performance. Collectively, our results shed light on novel mechanisms regulating reproduction at the hypothalamic level

Menopause leads to elevated expression of macrophage-associated genes in the aging frontal cortex: rat and human studies identify strikingly similar changes.

BACKGROUND The intricate interactions between the immune, endocrine and central nervous systems shape the innate immune response of the brain. We have previously shown that estradiol suppresses expression of immune genes in the frontal cortex of middle-aged ovariectomized rats, but not in young ones reflecting elevated expression of these genes in middle-aged, ovarian hormone deficient animals. Here, we explored the impact of menopause on the microglia phenotype capitalizing on the differential expression of macrophage-associated genes in quiescent and activated microglia. METHODS We selected twenty-three genes encoding phagocytic and recognition receptors expressed primarily in microglia, and eleven proinflammatory genes and followed their expression in the rat frontal cortex by real-time PCR. We used young, middle-aged and middle-aged ovariectomized rats to reveal age- and ovariectomy-related alterations. We analyzed the expression of the same set of genes in the postcentral and superior frontal gyrus of pre- and postmenopausal women using raw microarray data from our previous study. RESULTS Ovariectomy caused up-regulation of four classic microglia reactivity marker genes including Cd11b, Cd18, Cd45 and Cd86. The change was reversible since estradiol attenuated transcriptional activation of the four marker genes. Expression of genes encoding phagocytic and toll-like receptors such as Cd11b, Cd18, C3, Cd32, Msr2 and Tlr4 increased, whereas scavenger receptor Cd36 decreased following ovariectomy. Ovarian hormone deprivation altered the expression of major components of estrogen and neuronal inhibitory signaling which are involved in the control of microglia reactivity. Strikingly similar changes took place in the postcentral and superior frontal gyrus of postmenopausal women. CONCLUSIONS Based on the overlapping results of rat and human studies we propose that the microglia phenotype shifts from the resting toward the reactive state which can be characterized by up-regulation of CD11b, CD14, CD18, CD45, CD74, CD86, TLR4, down-regulation of CD36 and unchanged CD40 expression. As a result of this shift, microglial cells have lower threshold for subsequent activation in the forebrain of postmenopausal women

Evaluation of Digital Development Based on the International Digital Economy and Society Index 2020 Data

Digitalization and technological advances develop together with our society. The European Commission intends to monitor this development using the International Digital and Society Index (I-DESI) and provide an objective comparison of the participating countries. This comparison of the counties can be an important part of the roadmap of digital transformation for companies and other participants in the market. The aim of this study is to further analyze the 2020 data of I-DESI using multivariate statistical methods not included in the official report. Our objective is to show whether there are differences between the EU and non-EU countries (discriminant analysis, variance analysis), whether the dimensions of the I-DESI index are overlapping (correlation analysis, factor analysis), and whether different country groups can be formed (cluster analysis). Answering these questions, we can give a useful tool to companies for a more successful digital transformation

Distribution of type 1 cannabinoid receptor expressing neurons in the septal-hypothalamic region of the mouse. Colocalization with GABAergic and glutamatergic markers.

Type 1 cannabinoid receptor (CB1) is the principal mediator of retrograde endocannabinoid signaling in the brain. In this study, we addressed the topographic distribution and amino acid neurotransmitter phenotype of endocannabinoid-sensitive hypothalamic neurons in mice. The in situ hybridization detection of CB1 mRNA revealed high levels of expression in the medial septum (MS) and the diagonal band of Broca (DBB), moderate levels in the preoptic area and the hypothalamic lateroanterior (LA), paraventricular (Pa), ventromedial (VMH), lateral mammillary (LM) and ventral premammillary (PMV) nuclei, and low levels in many other hypothalamic regions including the suprachiasmatic (SCh) and arcuate (Arc) nuclei. This regional distribution pattern was compared to location of GABAergic and glutamatergic cell groups, as identified by the expression of glutamic acid decarboxylase 65 (GAD65) and type 2 vesicular glutamate transporter (VGLUT2) mRNAs, respectively. The MS, DBB and preoptic area showed overlaps between GABAergic and CB1-expressing neurons, whereas hypothalamic sites with moderate CB1 signals, including the LA, Pa, VMH, LM and PMV, were dominated by glutamatergic neurons. Low CB1 mRNA levels were also present in other glutamatergic and GABAergic regions. Dual-label in situ hybridization experiments confirmed the cellular co-expression of CB1 with both glutamatergic and GABAergic markers. In this report we provide a detailed anatomical map of hypothalamic glutamatergic and GABAergic systems whose neurotransmitter release is controlled by retrograde endocannabinoid signaling from hypothalamic and extrahypothalamic target neurons. This neuroanatomical information contributes to the understanding of the role that the endocannabinoid system plays in the regulation of endocrine and metabolic functions. J. Comp. Neurol., 2011. (c) 2011 Wiley-Liss, Inc

A nemzetközi digitális gazdaság és társadalom index 2020. évi adatainak statisztikai elemzése = Statistical analysis of 2020 data for the international digital economy and society index

A tanulmány célja az Európai Bizottság által készített nemzetközi digitális gazdaság és társadalom index (international digital economy and society index, I-DESI) dimenzióinak elemzése többváltozós statisztikai módszerekkel. A szerzők az EU- (Európai Unió) tagállamok és az EU-n kívüli országok adatainak szétválaszthatóságát diszkriminanciaanalízissel vizsgálják, a dimenziók értékeinek átlagát varianciaanalízissel hasonlítják össze. A dimenziók közötti korrelációt Pearsonféle és parciális korrelációs együtthatók alapján számítják, az összefüggéseket oksági láncon ábrázolják. Főkomponens-elemzés segítségével az öt dimenziót két főkomponensre vezetik vissza és az előző évi adatokhoz viszonyítva értelmezik. A kiugró adatokat a Mahalanobis-távolságokkal értékelik. Eredményeik alátámasztják az Európai Bizottság álláspontját a digitális gazdaság vizsgált dimenziónak szoros összefüggéséről, valamint azt, hogy nincs jelentős különbség az EU-tagállamok és az EU-n kívüli országok között. = The aim of this study is to analyse the dimensions of the international digital and society index (I-DESI) prepared by the European Commission using multivariate statistical methods. The distinction between EU (European Union) member states and non-EU countries states is examined through discriminant analysis and the means are compared by analysis of variance. Correlation between dimensions is studied by Pearson and partial correlation coefficients, and the relationships between dimensions are plotted on a causal chain. The five dimensions are traced back to two main components using factor analysis, and interpreted compared to the previous year’s data. Outliers are rated with Mahalanobis distances. Countries are then grouped using a hierarchical cluster analysis. The results support the European Commission’s view of the close link between the dimensions of the digital economy and no evidence of a significant difference is found between EU member states and non-EU actors

Glycinergic input to the mouse basal forebrain cholinergic neurons

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Hippocampal Gene Expression Is Highly Responsive to Estradiol Replacement in Middle-Aged Female Rats.

In the hippocampus, estrogens are powerful modulators of neurotransmission, synaptic plasticity and neurogenesis. In women, menopause is associated with increased risk of memory disturbances, which can be attenuated by timely estrogen therapy. In animal models of menopause, 17beta-estradiol (E2) replacement improves hippocampus-dependent spatial memory. Here, we explored the effect of E2 replacement on hippocampal gene expression in a rat menopause model. Middle-aged ovariectomized female rats were treated continuously for 29 days with E2 and then, the hippocampal transcriptome was investigated with Affymetrix expression arrays. Microarray data were analyzed by Bioconductor packages and web-based softwares, and verified with quantitative PCR. At standard fold change (FC) selection criterion, 156 genes responded to E2. All alterations but four were transcriptional activation. Robust activation (FC>10) occurred in the case of transthyretin, klotho, claudin 2, prolactin receptor, ectodin, coagulation factor V, insulin-like growth factor 2, Igfbp2 and sodium/sulfate symporter. Classification of the 156 genes revealed major groups including signaling (35 genes), metabolism (31 genes), extracellular matrix (17 genes) and transcription (16 genes). We selected 33 genes for further studies and all changes were confirmed by real-time PCR. The results suggest that E2 promotes retinoid, growth factor, homeoprotein, neurohormone and neurotransmitter signaling, changes metabolism, extracellular matrix composition, transcription, and induce protective mechanisms via genomic effects. We propose that these mechanisms contribute to effects of E2 on neurogenesis, neural plasticity and memory functions. Our findings provide further support for the rationale to develop safe estrogen receptor ligands for the maintenance of cognitive performance in postmenopausal women