Promoter-Specific Expression and Genomic Structure of IgLON Family Genes in Mouse

IgLON family is composed of five genes: Lsamp, Ntm, Opcml, Negr1, and Iglon5; encoding for five highly homologous neural adhesion proteins that regulate neurite outgrowth and synapse formation. In the current study we performed in silico analysis revealing that Ntm and Opcml display similar genomic structure as previously reported for Lsamp, characterized by two alternative promotors 1a and 1b. Negr1 and Iglon5 transcripts have uniform 5' region, suggesting single promoter. Iglon5, the recently characterized family member, shares high level of conservation and structural qualities characteristic to IgLON family such as N-terminal signal peptide, three Ig domains, and GPI anchor binding site. By using custom 5'-isoform-specific TaqMan gene-expression assay, we demonstrated heterogeneous expression of IgLON transcripts in different areas of mouse brain and several-fold lower expression in selected tissues outside central nervous system. As an example, the expression of IgLON transcripts in urogenital and reproductive system is in line with repeated reports of urogenital tumors accompanied by mutations in IgLON genes. Considering the high levels of intra-family homology shared by IgLONs, we investigated potential compensatory effects at the level of IgLON isoforms in the brains of mice deficient of one or two family members. We found that the lack of IgLONs is not compensated by a systematic quantitative increase of the other family members. On the contrary, the expression of Ntm 1a transcript and NEGR1 protein was significantly reduced in the frontal cortex of Lsamp-deficient mice suggesting that the expression patterns within IgLON family are balanced coherently. The actions of individual IgLONs, however, can be antagonistic as demonstrated by differential expression of Syp in deletion mutants of IgLONs. In conclusion, we show that the genomic twin-promoter structure has impact on both anatomical distribution and intra-family interactions of IgLON family members. Remarkable variety in the activity levels of 1a and 1b promoters both in the brain and in other tissues, suggests complex functional regulation of IgLONs by alternative signal peptides driven by 1a and 1b promoters.Peer reviewe

PGC-1 alpha Signaling Increases GABA(A) Receptor Subunit alpha 2 Expression, GABAergic Neurotransmission and Anxiety-Like Behavior in Mice

Peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1 alpha) is a master regulator of mitochondria biogenesis and cell stress playing a role in metabolic and degenerative diseases. In the brain PGC-1 alpha expression has been localized mainly to GABAergic interneurons but its overall role is not fully understood. We observed here that the protein levels of gamma-aminobutyric acid (GABA) type A receptor-alpha 2 subunit (GABAR alpha 2) were increased in hippocampus and brain cortex in transgenic (Tg) mice overexpressing PGC-1 alpha in neurons. Along with this, GABAR alpha 2 expression was enhanced in the hippocampus of the PGC-1 alpha Tg mice, as shown by quantitative PCR. Double immunostaining revealed that GABAR alpha 2 co-localized with the synaptic protein gephyrin in higher amounts in the striatum radiatum layer of the hippocampal CA1 region in the Tg compared with Wt mice. Electrophysiology revealed that the frequency of spontaneous and miniature inhibitory postsynaptic currents (mIPSCs) was increased in the CA1 region in the Tg mice, indicative of an augmented GABAergic transmission. Behavioral tests revealed an increase for anxiety-like behavior in the PGC-1 alpha Tg mice compared with controls. To study whether drugs acting on PPAR gamma can affect GABAR alpha 2, we employed pioglitazone that elevated GABAR alpha 2 expression in primary cultured neurons. Similar results were obtained using the specific PPAR gamma agonist, N-(2-benzoylphenyl)-O-[2-(methyl-2-pyridinylamino) ethyl]-L-tyrosine hydrate (GW1929). These results demonstrate that PGC-1 alpha regulates GABAR alpha 2 subunits and GABAergic neurotransmission in the hippocampus with behavioral consequences. This indicates further that drugs like pioglitazone, widely used in the treatment of type 2 diabetes, can influence GABAR alpha 2 expression via the PPAR gamma/PGC-1 alpha system.Peer reviewe

MEDAL (101079429)

The MEDAL consortium aims to reduce the recognised gap in access to expertise, training and research funding in Linguistics across Europe through a horizontal, network approach to engaging a new generation of researchers in empirically grounded, up-to-the-minute methodology. The consortium, comprising four highly acclaimed research organisations (Widening country member and coordinator, the University of Tartu, Estonia; Max Planck Institute for Psycholinguistics; Donders Institute at Radboud University, both in the Netherlands; University of Birmingham, UK) who will jointly lead a concerted move to increase the global competitiveness of Linguistics in Europe by bringing together world-class pioneers and early-career researchers, developing excellence in methodological training, and formulating a model of international collaboration on cross-linguistic, cross-modal, cross-disciplinary research. The project focuses on three clusters of empirically grounded methods which inform state-of-the-art language research: corpus studies, experimental methods, and computational modelling. The consortium will launch a multi-pronged effort to build better methodological expertise and awareness among the next generation of researchers, offer high-level training schools in the methods included in the project, and engage in the project’s collaborative Gold MEDAL Research project, in which early career researchers gain first-hand experience in conceptualising and designing a multi-method research programme, implementing it across a range of languages and modalities, and disseminating the results, leading to increased visibility for all partners. By 2025, the MEDAL consortium will have developed internal expertise, established a sustainable joint training and research programme and be poised for further collaboration, and the University of Tartu will be an internationally recognised Linguistics research excellence hub for Northern Europe. Project website https://medal.ut.ee Project Grant agreement ID: 101079429 https://cordis.europa.eu/project/id/101079429 The project is financed by the EU Horizon Europe Twinning project MEDAL (101079429) and the UK Research and Innovation organisation (101079429). Funded by the European Union. Views and opinions expressed are, however, those of the author(s) only and do not necessarily reflect those of the European Union or European Commission. Neither the European Union nor the granting authority can be held responsible for them

Neuropsühhiaatriliste häirete loommudelite kvantitatiivne uurimine: metaboolsed, käitumuslikud ja geneetilised profiilid

Väitekirja elektrooniline versioon ei sisalda publikatsiooneVaimuhaigused on oma olemuselt heterogeensed häired, mille esinemine sõltub geneetilisest ja keskkonnast tingitud soodumusest. Vaimuhaiguste järkjärgulist avaldumist varjutab tihti loomulik areng, kaasneb ennustamatu kulg, teadmata ravi kõrvaltoimed ning inimese olemust määratleva kõrgema ajutalitluse ja tundeelu märkimisväärne häirumine. Vaimuhaiguste polügeneetilise olemuse tõttu on neid keeruline omavahel eristada, diagnoosida ja õigeaegselt ravida. Vaimuhaiguse varajane avastamine ja asjakohane ravi võimaldab haiguse süvenemist ning sellega kaasuvaid kulusid ennetada. Selleks on vaja leida ja õppida ära kasutama haiguse kulgu kajastavaid bioloogilisi markereid. Neid markereid tuleb otsida nii molekulaarsel, sünapsite, raku, koe, närvivõrkude ja kõrgematel närvisüsteemi tasanditel. Selle hoomamatu ülesande teevad võimalikuks erinevad lihtsustatud mudelid, sealhulgas geneetiliselt muundatud või farmakoloogiliselt manipuleeritud hiiremudelid. Käesolev väitekiri uurib kolme erinevat tahku mudelit: PPARGC1α vahendatud mitokondriaalse energiametabolismi rolli inhibitoorses neurotransmissioonis, metaboolset eelsoodumust psühhoosilaadses seisundis ja neuraalse adhesioonimolekuliperekonna IgLON puudulikust. Nende eri tahku mudelite mõistmine viib meid lähemale vaimuhaiguste paremale mudeldamisele ja võimalike raviteede avastamisele. Töös uuritud kolme mudeli leiud saab kokku võtta järgnevalt. PPARGC1α (PGC1α) üleekspressioonimutant võimaldab meeleoluhäirete mudeldamist. 129Sv hiireliini geneetiline taust lihtsustab psühhoosi mudeldamisel eelsoodumusega populatsioonis. IgLONite avaldumise muster viitab nende potentsiaalile peegeldada ebakorrapärasusi aju arengus ja töös.Mental disorders are inherently heterogeneous conditions arising from various genetic and environmental insults. The gradual manifestation of mental disorder is often overshadowed by natural development; displays an unpredictable course, unknown side effects, and significant disturbances of cognition. The polygenetic nature of mental disorders makes them challenging to distinguish, diagnose and treat on time. Early detection and appropriate treatment of mental disorders are necessary to prevent the progression of symptoms and associated costs. Scientific research prioritises finding early biological markers that reflect the course of these disorders. These markers should be searched for on a molecular, synaptic, cellular, tissue, network and higher-order systems level. Different simplified models, including genetically modified or pharmacologically manipulated rodent models, make this task feasible. The following dissertation examines three simplified models: the role of mitochondrial energy metabolism in inhibitory neurotransmission, metabolic predisposition in psychosis, and the expression pattern of IglON neural cell adhesion molecules. Understanding these models takes us closer to understanding mental disorders and discovering therapeutic opportunities. The findings of these three models can be summarised as follows. PPARGC1α (PGC1α) overexpression mutant displays labile mood phenotype allowing modelling of mood-related disorders. The 129sv mouse strain reflects a predisposed population and simplifies the modelling of psychosis. The expression pattern of IgLONs refers to their potential to reflect irregularities in brain development and maintenance.https://www.ester.ee/record=b555542

Repeated Administration of D-Amphetamine Induces Distinct Alterations in Behavior and Metabolite Levels in 129Sv and Bl6 Mouse Strains

The main goal of the study was to characterize the behavioral and metabolomic profiles of repeated administration (for 11 days) of d-amphetamine (AMPH, 3 mg/kg i. p.), indirect agonist of dopamine (DA), in widely used 129S6/SvEvTac (129Sv) and C57BL/6NTac (Bl6) mouse strains. Acute administration of AMPH (acute AMPH) induced significantly stronger motor stimulation in Bl6. However, repeated administration of AMPH (repeated AMPH) caused stronger motor sensitization in 129Sv compared acute AMPH. Body weight of 129Sv was reduced after repeated saline and AMPH, whereas no change occurred in Bl6. In the metabolomic study, acute AMPH induced an elevation of isoleucine and leucine, branched chain amino acids (BCAA), whereas the level of hexoses was reduced in Bl6. Both BCAAs and hexoses remained on level of acute AMPH after repeated AMPH in Bl6. Three biogenic amines [asymmetric dimethylarginine (ADMA), alpha-aminoadipic acid (alpha-AAA), kynurenine] were significantly reduced after repeated AMPH. Acute AMPH caused in 129Sv a significant reduction of valine, lysophosphatidylcholines (lysoPC a C16:0, lysoPC a C18:2, lysoPC a C20:4), phosphatidylcholine (PC) diacyls (PC aa C34:2, PC aa C36:2, PC aa C36:3, PC aa C36:4) and alkyl-acyls (PC ae C38:4, PC ae C40:4). However, repeated AMPH increased the levels of valine and isoleucine, long-chain acylcarnitines (C14, C14:1-OH, C16, C18:1), PC diacyls (PC aa C38:4, PC aa C38:6, PC aa C42:6), PC acyl-alkyls (PC ae C38:4, PC ae C40:4, PC ae C40:5, PC ae C40:6, PC ae C42:1, PC ae C42:3) and sphingolipids [SM(OH)C22:1, SM C24:0] compared to acute AMPH in 129Sv. Hexoses and kynurenine were reduced after repeated AMPH compared to saline in 129Sv. The established changes probably reflect a shift in energy metabolism toward lipid molecules in 129Sv because of reduced level of hexoses. Pooled data from both strains showed that the elevation of isoleucine and leucine was a prominent biomarker of AMPH-induced behavioral sensitization. Simultaneously a significant decline of hexoses, citrulline, ADMA, and kynurenine occurred. The reduced levels of kynurenine, ADMA, and citrulline likely reflect altered function of N-methyl-D-aspartate (NMDA) and NO systems caused by repeated AMPH. Altogether, 129Sv strain displays stronger sensitization toward AMPH and larger variance in metabolite levels than Bl6

Data_Sheet_1_Repeated Administration of D-Amphetamine Induces Distinct Alterations in Behavior and Metabolite Levels in 129Sv and Bl6 Mouse Strains.docx

<p>The main goal of the study was to characterize the behavioral and metabolomic profiles of repeated administration (for 11 days) of d-amphetamine (AMPH, 3 mg/kg i. p.), indirect agonist of dopamine (DA), in widely used 129S6/SvEvTac (129Sv) and C57BL/6NTac (Bl6) mouse strains. Acute administration of AMPH (acute AMPH) induced significantly stronger motor stimulation in Bl6. However, repeated administration of AMPH (repeated AMPH) caused stronger motor sensitization in 129Sv compared acute AMPH. Body weight of 129Sv was reduced after repeated saline and AMPH, whereas no change occurred in Bl6. In the metabolomic study, acute AMPH induced an elevation of isoleucine and leucine, branched chain amino acids (BCAA), whereas the level of hexoses was reduced in Bl6. Both BCAAs and hexoses remained on level of acute AMPH after repeated AMPH in Bl6. Three biogenic amines [asymmetric dimethylarginine (ADMA), alpha-aminoadipic acid (alpha-AAA), kynurenine] were significantly reduced after repeated AMPH. Acute AMPH caused in 129Sv a significant reduction of valine, lysophosphatidylcholines (lysoPC a C16:0, lysoPC a C18:2, lysoPC a C20:4), phosphatidylcholine (PC) diacyls (PC aa C34:2, PC aa C36:2, PC aa C36:3, PC aa C36:4) and alkyl-acyls (PC ae C38:4, PC ae C40:4). However, repeated AMPH increased the levels of valine and isoleucine, long-chain acylcarnitines (C14, C14:1-OH, C16, C18:1), PC diacyls (PC aa C38:4, PC aa C38:6, PC aa C42:6), PC acyl-alkyls (PC ae C38:4, PC ae C40:4, PC ae C40:5, PC ae C40:6, PC ae C42:1, PC ae C42:3) and sphingolipids [SM(OH)C22:1, SM C24:0] compared to acute AMPH in 129Sv. Hexoses and kynurenine were reduced after repeated AMPH compared to saline in 129Sv. The established changes probably reflect a shift in energy metabolism toward lipid molecules in 129Sv because of reduced level of hexoses. Pooled data from both strains showed that the elevation of isoleucine and leucine was a prominent biomarker of AMPH-induced behavioral sensitization. Simultaneously a significant decline of hexoses, citrulline, ADMA, and kynurenine occurred. The reduced levels of kynurenine, ADMA, and citrulline likely reflect altered function of N-methyl-D-aspartate (NMDA) and NO systems caused by repeated AMPH. Altogether, 129Sv strain displays stronger sensitization toward AMPH and larger variance in metabolite levels than Bl6.</p