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Cooperability
| Project leader |
Gajovic Srecko |
| Project co-leader: |
Prof. dr. Jasna Križ |
| Administering organization: |
School of Medicine University of Zagreb
Šalata 3, HR-10000 Zagreb, Croatia
www.mef.hr
Prof. dr. Nada Čikeš, Dean
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| Partner Institution/Company: |
Laval University, Faculty of Medicine |
| Grant type: |
1B |
| Project title: |
Regeneration and plasticity after ischemic brain damage studied on innovative transgenic mouse models |
| Project summary: |
Brain diseases highly contribute to the total burden of diseases in the society. Among them stroke represents the major health problem, as it causes long term consequences on brain and impairs the abilities of the patient. Moreover no efficient treatment is available.
In mouse the ischemic injury corresponding to the stroke in humans can be induced by middle cerebral artery occlusion. In order to get insight in the molecular events accompanying the ischemic injury, the innovative transgenic mouse models were created. The monitoring of gene activity was achieved by introduction of bioluminescent and/or fluorescent reporters under the influence of the promoter of the investigated genes. Unlike current animal studies that are based on single end point data, these mice provide new powerful analytical tools for real time monitoring of gene activity in living animals.The axonal regeneration and sprouting after ischemia would be analyzed using mice carrying dual reporter system including luciferase and green fluorescent protein under the transcriptional control of GAP-43 promoter. Immune/inflammatory response of microglia will be assessed with the transgenic mice with the same dual reporter system under the control of TLR-2 promoter.
The novel mouse models generated by Canadian partner and their facility for in vivo imaging of the introduced reporters would be combined with the already existing laboratory platform of the Croatian partner which enables morphological and behavioral analysis of brain phenotype changes in transgenic mice. The Croatian facility would provide assessment of the restoration of behavioral deficits, synaptogenesis after ischemia would be revealed by electron microscopy, and gene expression/induction studies using confocal microscopy of fluorescent reporters.
The complementary activities of both partners would enable to study inflammation, repair and regeneration after ischemia in time dependent manner. These would be used to clarify the influence of the postischemic inflammation on the regeneration and plasticity. The common experimental platform would be used for screening purposes of the neuroprotective and/or regenerative treatments of ischemia, and its usage would outlast the proposed project. In addition, the correlative in vivo imaging with light and electron microscopy would indicate the path for further development of novel transgenic mouse models.
The benefits for Croatian science and society include the transfer of knowledge and application of innovative genetic technologies, which would enable insight in regeneration and plasticity after ischemia and resulted with the ready-to-use platform for treatment screening in order to attack a major health problem of the society – stroke.
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| Hrvatski sažetak: |
Bolesti mozga znatno doprinose ukupnom teretu bolesti u društvu. Među njima moždani udar predstavlja glavni zdravstveni problem, jer uzrokuje dugotrajne posljedice na mozak i smanjuje sposobnosti oboljeloga. Štoviše, ne postoji djelotvorna terapija moždanog udara.
U miša se ozljeda ishemijom koja odgovara moždanom udaru postiže zatvaranjem središnje arterije mozga (a. cerebri media). Kako bi se dobio uvid u molekularna zbivanja koja prate ishemijsku ozljedu, dobiveni su inovativni transgenični mišji modeli. Praćenje djelovanja gena postiže se uvođenjem bioluminiscentnih i/ili fluorescentnih biljega pod utjecajem promotora proučavanih gena. Za razliku od istraživanjima na životinjama u kojima se podaci prikupljaju na kraju pokusa, ovi miševiomogućuju novi snažni analitički alat za praćenje u stvarnom vremenu genske djelatnosti u živim životinjama. Obnavljanje aksona i izrastanje novih nakon ishemije pratit će se na miševima koji posjeduju dvostruki reporterski sustav, luciferazu i zelenu fluorescentnu bjelančevinu pod transkripcijskom kontrolo GAP-43 promotora. Imunosni/upalni odgovor mikroglije bit će ustanovljen pomoću transgeničnog miša koji posjeduje isti dvostruki reporterski sustav pod kontrolom TLR-2 promotora.
Novi mišji modeli koji su dobiveni kod kanadskog partnera i njihova oprema za in vivo praćenje ubačenih reportera kombinirat će se s većpostojećom laboratorijskom platformom hrvatskog partnera koja omogućuje morfološkom i analizom ponašanja proučavanje fenotipa mozga transgeničnih miševa. Hrvatska strana doprinijet će ustanovljavanjem oporavka deficita ponašanja, sinaptogeneza nakon ishemije pokazat će se elektronskom mikroskopijom, a genska ekspresija/indukcija će se proučavati konfokalnom mikroskopijom fluorescentnih reportera.
Komplementarne aktivnosti oba partnera omogućit će proučavanje upale, popravka i obnove nakon ishemije ovisno o proteklom vremenu. To ćeomogućiti razjašnjavanje utjecaja postishemijske upale na obnovu i plastičnost mozga. Zajednička eksperimentalna platforma upotrijebit će se za ispitivanje neuroprotektivne i/ili regnerativne terapije ishemije, a njena upotreba nastavit će se i poslije predloženog projekta. Korelacija između in vivo oslikavanja, svjetlosne i elektronske mikroskopije ukazat će na pravac razvoja novih transgeničnih mišjih modela.
Dobrobit za hrvatsku znanost i društvo uključuje prijenos znanja i primjenu inovativne genetske tehnologije, što će omogućiti uvid u obnovu i plastičnost nakon ishemije i rezultirati spremnom za upotrebu platformom za procjenu terapija kako bi se razriješio glavni zdravstveni problem društva – moždani udar.
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| Amount requested from UKF: |
1.460.000 HRK |
| Amount of matching funding: |
511.000 HRK |
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