Põletiku ja dieedi mõju Bl6 and 129Sv hiireliinide metaboolsele profiilile ja valitud geneetilistele parameetritele

Väitekirja elektrooniline versioon ei sisalda publikatsiooneHiirte geneetiline ülesehitus ühtib suuresti inimesega ning seetõttu on nad asendamatud mudelorganismid nii ravimitööstuses kui ka meditsiinilistes alusteadustes. Hiiri kasutatakse laialdaselt inimeste haiguste modelleerimiseks, et mõista haiguste algpõhjusi ja olemust. Kuid ka ravimite väljatöötamisel nende efektiivsuse ja ohutuse hindamisel. Teaduslaborites kasutatavates hiireliinides on alleelid reeglina homosügootses olekus. Selline geneetiline homogeensus vähendab tulemuste varieeruvust ja hõlbustab uuringute läbiviimist. Samas põhjustab see ka igale hiireliinile unikaalse fenotüübi tekke, mis mõjutab katsetulemusi. Eksperimentide planeerimisel on oluline valida katse eesmärkide jaoks sobilik hiireliin ning sellest lähtuvalt on oluline põhjalikult kaardistada hiireliinide funktsionaalseid erinevusi. Käesoleva doktoritöö eesmärk oli võrrelda kahte hiireliini, Bl6 ja 129Sv, pöörates erilist tähelepanu põletikule ja metaboolse sündroomi tekkele. Meie tulemused näitavad, et Bl6 hiired tulevad paremini toime nii põletiku kui kõrge rasvasisaldusega dieedi põhjustatud negatiivsete mõjudega. Seevastu 129Sv hiired on tundlikumad immuunaktivatsioonile ning suurema vastuvõtlikkusega metaboolse sündroomi tekkele, mis on mõlemad psühhiaatriliste häirete olulised tunnused. Kokkuvõttes võib väita, et Bl6 hiired sobivad paremini agressiivsuse ja sõltuvuskäitumise uurimiseks, seevastu 129Sv hiired sobivad paremini raskemate psühhiaatriliste häiretega seotud endofenotüüpide modelleerimiseks.The genetic makeup of mice largely corresponds to that of humans, which is why they are irreplaceable model organisms in both the pharmaceutical industry and biomedical research. Mice are often used as model organisms for human diseases to understand the causes and nature of disease. But they are also used in drug development for efficacy and safety testing. Mouse lines used in research laboratories are genetically homogeneous. Such genetic homogeneity reduces variability of results and facilitates experimentation. However, different inbred lines carry different fixed alleles and therefore exhibit different phenotypic characteristics, which affects experimental results. Choosing the right inbred strain for a particular experiment is an important consideration when working with laboratory mice and thus it is important to thoroughly map the functional differences between mouse lines. In this dissertation, two mouse lines, Bl6 and 129Sv, were compared with particular emphasis on inflammation and metabolic syndrome. Our results show that Bl6 mice are better able to cope with the negative effects of inflammation and a high-fat diet. In contrast, 129Sv mice are more sensitive to immune activation and more susceptible to metabolic syndrome, both important features of psychiatric disorders. In summary, Bl6 mice are better suited for studying aggressiveness and addictive behaviors, whereas 129Sv mice are better suited for modeling endophenotypes associated with more severe psychiatric disorders.https://www.ester.ee/record=b554956

Lsamp geeni/valgu roll täiskasvanud hiire hipokampaalses neurogeneesis

Limbilise süsteemiga seotud membraanvalku kodeeriva geeni (Lsamp) polümorfisme seostatakse ärevus- ning psühhiaatriliste häiretega. Varasemad katselooma ja in vitro uuringud viitavad seostele LSAMP valgu ja närvisüsteemi plastilisuse vahel. Plastilisusele viitab kõrgenenud Lsamp ekspressiooni tase rikastatud keskkonnas elavate hiirte hipokampuses. Lisaks sellele on Lsamp osutunud paljude vähitüüpide puhul tuumorsupressorgeeniks. Käesolevas magistritöös uuriti LSAMP valgu võimalikku osalust hipokampaalses neurogeneesis. Katse tulemused näitavad, et rikastatud keskkonnas elavatel Lsamppuudulikkusega hiirtel on hammaskäärus rohkem uusi neuroneid kui nende metsiktüüpi pesakonnakaaslastel

Microglia contribute to social behavioral adaptation to chronic stress

Microglial activation has been regarded mainly as an exacerbator of stress response, a common symptom in psychiatric disorders. This study aimed to determine whether microglia contribute to adaptive response of the brain and behavior toward stress using a mild and adaptive stress model - chronic restraint stress (CRS) - with wild type (WT) and CX3CR1-GFP (CX3CR1[G]) mice and human schizophrenia patients' data. Our results revealed that CRS did not exacerbate anxiety and depressive-like behaviors, but instead strengthened social dominance and short-term spatial learning in WT mice. Compared to WT and CX3CR1(+/G) heterozygous mice, CX3CR1(G/G) homozygotes were subordinate in social interaction before and after CRS. Microglia in WT mice underwent a series of region-specific changes involving their phagocytosis of presynaptic vesicular glutamate transporter 2 protein, contacts with synaptic elements, CD206(+)microglial proportion, and gene expressions such as Cx3cr1. By contrast, CX3CR1-deficient microglia showed decreased CD206(+) while increased MHCII+ subpopulations and hypo-ramification in the hippocampus, as well as sensitized polarization and morphological change in response to CRS. Furthermore, CD206(+) microglial abundancy was positively correlated with social dominancy and microglial ramification in CX3CR1-GFP mice. Moreover, CX3CR1 mRNA level was reduced in CRS-treated mouse brains and showed a smaller interactome with other brain genes in the dorsal-lateral prefrontal cortices of patients with schizophrenia. Our findings overall highlight microglia and its receptor CX3CR1 as key contributors in regulation of social behavioral adaptation to chronic stress.Peer reviewe

Lipopolysaccharide-Induced Strain-Specific Differences in Neuroinflammation and MHC-I Pathway Regulation in the Brains of Bl6 and 129Sv Mice

Many studies have demonstrated significant mouse-strain-specific differences in behavior and response to pathogenic and pharmacological agents. This study seeks to characterize possible differences in microglia activation and overall severity of neuroinflammation in two widely used mouse strains, C57BL/6NTac (Bl6) and 129S6/SvEvTac (129Sv), in response to acute lipopolysaccharide (LPS) administration. Locomotor activity within the open field arena revealed similar 24 h motor activity decline in both strains. Both strains also exhibited significant bodyweight loss due to LPS treatment, although it was more severe in the Bl6 strain. Furthermore, LPS induced a hypothermic response in Bl6 mice, which was not seen in 129Sv. We found that 24 h LPS challenge significantly increased the inflammatory status of microglia in 129Sv mice. On the other hand, we observed that, under physiological conditions, microglia of Bl6 seemed to be in a higher immune-alert state. Gene and protein expression analysis revealed that LPS induces a significantly stronger upregulation of MHC-I-pathway-related components in the brain of Bl6 compared to 129Sv mice. The most striking difference was detected in the olfactory bulb, where we observed significant LPS-induced upregulation of MHC-I pathway components in Bl6 mice, whereas no alterations were observed in 129Sv. We observed significant positive correlations between bodyweight decline and expressions of MHC-I components in the olfactory bulbs of Bl6 mice and the frontal cortex of 129Sv, highlighting different brain regions most affected by LPS in these strains. Our findings suggest that the brains of Bl6 mice exist in a more immunocompetent state compared to 129Sv mice

Impact of a High-Fat Diet on the Metabolomics Profile of 129S6 and C57BL6 Mouse Strains

Different inbred mouse strains vary substantially in their behavior and metabolic phenotype under physiological and pathological conditions. The purpose of this study was to extend the knowledge of distinct coping strategies under challenging events in two differently adapting mouse strains: C57BL/6NTac (Bl6) and 129S6/SvEvTac (129Sv). Thus, we aimed to investigate possible similarities and differences in the body weight change, behavior, and several metabolic variables in Bl6 and 129Sv strains in response to high-fat diet (HFD) using the AbsoluteIDQ p180 kit. We found that 9 weeks of HFD induced a significant body weight gain in 129Sv, but not in Bl6 mice. Besides that, 129Sv mice displayed anxiety-like behavior in the open-field test. Metabolite profiling revealed that 129Sv mice had higher levels of circulating branched-chain amino acids, which were even more amplified by HFD. HFD also induced a decrease in glycine, spermidine, and t4-OH-proline levels in 129Sv mice. Although acylcarnitines (ACs) dominated in baseline conditions in 129Sv strain, this strain had a significantly stronger AC-reducing effect of HFD. Moreover, 129Sv mice had higher levels of lipids in baseline conditions, but HFD caused more pronounced alterations in lipid profile in Bl6 mice. Taken together, our results show that the Bl6 line is better adapted to abundant fat intake

Lipopolysaccharide-Induced Strain-Specific Differences in Neuroinflammation and MHC-I Pathway Regulation in the Brains of Bl6 and 129Sv Mice

Many studies have demonstrated significant mouse-strain-specific differences in behavior and response to pathogenic and pharmacological agents. This study seeks to characterize possible differences in microglia activation and overall severity of neuroinflammation in two widely used mouse strains, C57BL/6NTac (Bl6) and 129S6/SvEvTac (129Sv), in response to acute lipopolysaccharide (LPS) administration. Locomotor activity within the open field arena revealed similar 24 h motor activity decline in both strains. Both strains also exhibited significant bodyweight loss due to LPS treatment, although it was more severe in the Bl6 strain. Furthermore, LPS induced a hypothermic response in Bl6 mice, which was not seen in 129Sv. We found that 24 h LPS challenge significantly increased the inflammatory status of microglia in 129Sv mice. On the other hand, we observed that, under physiological conditions, microglia of Bl6 seemed to be in a higher immune-alert state. Gene and protein expression analysis revealed that LPS induces a significantly stronger upregulation of MHC-I-pathway-related components in the brain of Bl6 compared to 129Sv mice. The most striking difference was detected in the olfactory bulb, where we observed significant LPS-induced upregulation of MHC-I pathway components in Bl6 mice, whereas no alterations were observed in 129Sv. We observed significant positive correlations between bodyweight decline and expressions of MHC-I components in the olfactory bulbs of Bl6 mice and the frontal cortex of 129Sv, highlighting different brain regions most affected by LPS in these strains. Our findings suggest that the brains of Bl6 mice exist in a more immunocompetent state compared to 129Sv mice

Dopamine System, NMDA Receptor and EGF Family Expressions in Brain Structures of Bl6 and 129Sv Strains Displaying Different Behavioral Adaptation

C57BL/6NTac (Bl6) and 129S6/SvEvTac (129Sv) mice display different coping strategies in stressful conditions. Our aim was to evaluate biomarkers related to different adaptation strategies in the brain of male 129Sv and Bl6 mice. We focused on signaling pathways related to the dopamine (DA) system, N-methyl-D-aspartate (NMDA) receptor and epidermal growth factor (EGF) family, shown as the key players in behavioral adaptation. Mice from Bl6 and 129Sv lines were divided into either home cage controls (HCC group) or exposed to repeated motility testing and treated with saline for 11 days (RMT group). Distinct stress responses were reflected in severe body weight loss in 129Sv and the increased exploratory behavior in Bl6 mice. Besides that, amphetamine caused significantly stronger motor stimulation in Bl6. Together with the results from gene expression (particularly Maob), this study supports higher baseline activity of DA system in Bl6. Interestingly, the adaptation is reflected with opposite changes of DA markers in dorsal and ventral striatum. In forebrain, stress increased the gene expressions of Egf-Erbb1 and Nrg1/Nrg2-Erbb4 pathways more clearly in 129Sv, whereas the corresponding proteins were significantly elevated in Bl6. We suggest that not only inhibited activity of the DA system, but also reduced activity of EGF family and NMDA receptor signaling underlies higher susceptibility to stress in 129Sv. Altogether, this study underlines the better suitability of 129Sv for modelling neuropsychiatric disorders than Bl6

High-Fat Diet Induces Pre-Diabetes and Distinct Sex-Specific Metabolic Alterations in Negr1-Deficient Mice

In the large GWAS studies, NEGR1 gene has been one of the most significant gene loci for body mass phenotype. The purpose of the current study was to clarify the role of NEGR1 in the maintenance of systemic metabolism, including glucose homeostasis, by using both male and female Negr1−/− mice receiving a standard or high fat diet (HFD). We found that 6 weeks of HFD leads to higher levels of blood glucose in Negr1−/− mice. In the glucose tolerance test, HFD induced phenotype difference only in male mice; Negr1−/− male mice displayed altered glucose tolerance, accompanied with upregulation of circulatory branched-chain amino acids (BCAA). The general metabolomic profile indicates that Negr1−/− mice are biased towards glyconeogenesis, fatty acid synthesis, and higher protein catabolism, all of which are amplified by HFD. Negr1 deficiency appears to induce alterations in the efficiency of energy storage; reduced food intake could be an attempt to compensate for the metabolic challenge present in the Negr1−/− males, particularly during the HFD exposure. Our results suggest that the presence of functional Negr1 allows male mice to consume more HFD and prevents the development of glucose intolerance, liver steatosis, and excessive weight gain