Voditelj:
Prof.dr.sc. Mario Vukšić
Izvanredni profesor neuroznanosti & anatomije
Hrvatski institut za istraživanje mozga
Medicinski fakultet Sveučilišta u Zagrebu
Šalata 12, 10000 Zagreb
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Laboratorij za konfokalnu mikroskopiju osnovan je 2008. godine a koristite ga svi znanstvenici zaposleni na Hrvatskom institutu za istraživanje mozga. Laboratorij je opremljen laserskim konfokalnim mikroskopom tvrtke Zeiss tipa LSM 510-META. Uređaj koristi lasere: Argon 458/477/488/514nm, HeNe 543 nm, HeNe laser 633 nm te posjeduje i META detektor koji omogućava razlikovanje različitih preklapajućih emisijskih signala. Sistem je nadograđen na invertni mikroskop Axiovert 200M tvrtke Zeiss koji se također može koristiti za standardnu fluorescenciju te posjeduje slijedeće objektive: 10x i 20x zračne; te 40x i 63x uljne.
Trenutno se uređaj koristi za: snimanje pojedinačnih konfokalnih slika, trodimenzionalnu rekonstrukciju pojedinačnih živčanih stanica koristeći konfokalni mikroskop i Neurolucida sustav, dokazivanje prisutnosti i raspodjele različitih fluorescentnom bojom označenih struktura.
Istraživački interesi:
Procesi oporavka nakon moždane ozljede i prekidanja veza, plastičnost dendrita i dendritičkih trnova nakon lezije, pojava izrastanja aksonskih mladica.
Primjeri aplikacije:
Konfokalna slagalina prikazuje GFP-pozitivne zrnate stanice u girus dentatusu transgeničnog Thy-1 miša I trodimenzionalna rekonstrukcija iste stanice koristeći Neurolucida sustav.
(Vuksic i sur., Hippocampus 2008)
Entorinalna denervacija u transgeničnog Thy-1 miša uzrokuje brze i prolazne promjene gustoće dendritičkih trnova na zrnatim stanicama girus dentatusa.
(Vuksic i sur., Experimental Neurology 2011)
Head:
Mario Vukšić, MD, PhD
Associate Professor of Neuroscience & Anatomy
Croatian Institute for Brain Research
School of Medicine University of Zagreb
Šalata 12, 10000 Zagreb, Croatia
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The Laboratory of Confocal Microscopy was established in 2008. This is an open laboratory available to all scientists from Croatian Institute for Brain Research. The Lab basic equipment is a laser scanning microscope Zeiss LSM 510-META. The appliance uses Argon laser 458/477/488/514nm, HeNe laser 543 nm, HeNe laser 633 nm and the META detector which can spectrally separate overlapping emission signals. The system was coupled to inverted microscope Zeiss Axiovert 200M which is also equipped for standard epi-fluorescence imaging. The following objectives are available with the system: 10x air, 20x air, 40x oil and 63x oil.
Current applications include: single confocal images, 3D-reconstruction of single neurons using confocal microscopy and Neurolucida system, colocalization and distribution of various fluorescently-tagged proteins.
Research interests:
Reorganizational processes following brain trauma and deafferentation, lesion-induced plasticity of dendrites and dendritic spines; axonal sprouting.
Example application:
Confocal image stack showing GFP-expressing granule cell in the fascia dentata of the Thy-1 GFP transgenic mouse and 3D reconstruction of the same cell using Neurolucida system
(Vuksic et al., Hippocampus 2008)
Entorhinal denervation of the Thy-1 GFP transgenic mouse induces fast and transient changes in spine densities of granule cells in the fascia dentata.
(Vuksic et al., Experimental Neurology 2011)
- Slade N, Zorić A, Horvat B, Vukšić M, Kostović I, Poljak L (2015) Suppression of Smad-1 mRNA expression level by Smad-2 likely control dichotomy of NF-κB and Smads mediated activation. Immunobiology. 220(1):48-53
- Kostović I, Sedmak G, Vukšić M, Judaš M (2015) The relevance of human fetal subplate zone for developmental neuropathology of neuronal migration disorders and cortical dysplasia. CNS Neuroscience & Therapeutics 21(2):74-82.
- Kosi N, Alić I, Kolačević M, Vrsaljko N, Jovanov Milošević N, Sobol M, Philimonenko A, Hozak P, Gajović S, Pochet R, Mitrečić D. (2015): Nop2 is expressed during proliferation of neural stem cells and in adult mouse and human brain. Brain Research 1597:65-76.
- Jovanov-Milošević N, Petrović D, Sedmak G, Vukšić M, Hof PR, Šimić G (2012) Human fetal tau protein isoform: Focus on possibilities for Alzheimer’s disease treatment. The International Journal of Biochemistry & Cell Biology 44:1290-1294.
- Kapuralin K, Van Ginneken C, Curlin M, Timmermans JP, Gajovic S. (2012): Neurons and a subset of interstitial cells of Cajal in the enteric nervous system highly express Stam2 gene. Anatomical Record-Advances in integrative anatomy and evolutionary biology 295:113-20.
- Curlin M, Kapuralin K, Muro AF, Baralle FE, Chowdhury K, Gajović S. (2012): Stam2 expression pattern during embryo development. Gene Expression Patterns 12: 68-76.
- Vukšić M, Del Turco D, Vlachos A, Schuldt G, Müller CM, Schneider G, Deller T (2011) Unilateral entorhinal denervation leads to long-lasting dendritic alterations of mouse hippocampal granule cells. Experimental Neurology 230(2):176–185.
- Vukšić M, Petanjek Z, Kostović I (2011) Development of prefrontal layer III pyramids in infants with Down syndrome. Translational Neuroscience 2(3):225-232.
- Ghebremedhin E, Rosenberger A, Rüb U, Vukšić M, Berhe T, Bickeböller H, de Vos RAI, Thal DR, Deller T (2010) Inverse relationship between cerebrovascular lesions and severity of Lewy body pathology in patients with Lewy body disease. Journal of Neuropathology & Experimental Neurology 29(5):442-448.
- Banfić H, Višnjić D, Mise N, Balakrishnan S, Deplano S, Korchev YE, Domin J (2009) Epidermal growth factor stimulates translocation of the class II phosphoinositide 3-kinase PI3K-C2beta to the nucleus. Biochemical Journal 422(1):53-60.
- Vukšić M, Del Turco D, Bas-Orth C, Burbach GJ, Feng G, Müller CM, Schwarzacher SW, Deller T (2008). 3D-Reconstruction and functional properties of GFP-positive and GFP-negative granule cells in the fascia dentata of the Thy1-GFP mouse. Hippocampus 18(4):364-75.
- Schwarzacher SW*, Vukšić M*, Haas CA, Burbach GJ, Sloviter RS and Deller T (2006) Neuronal hyperactivity induces astrocytic expression of neurocan in the adult rat hippocampus. Glia 53:704-714. *equally contribution
- Ghebremedhin E, Del Tredici K, Vukšić M, Rüb U, Thal DR, Burbach GJ, Rosenberger A, Bickeböller H, Deller T, Rob VAI, Jansen Steur ENH, Braak H (2006) Relationship of ApoE and age at onset to Parkinson’s disease neuropathology. Journal of Neuropathology & Experimental Neurology 65:116-123.
- Kienzler F, Jedlicka P, Vukšić M, Deller T, Schwarzacher SW (2006) Excitotoxic hippocampal neuron loss following sustained electrical stimulation of the perforant pathway in the mouse. Brain Research 1085:195-198.
- Gierga K, Bürk K, Bauer M, Diaz Orozco G, Auburger G, Schultz C, Vukšić M, Schöls L, de Los RAI, Braak H, Deller T, Rüb U (2005) Involvement of the cranial nerves and their nuclei in spinocerebellar ataxia type 2 (SCA2). Acta Neuropathologica 109:617-631.
- Crljen V, Visnjić D, Banfić H (2004) Presence of different phospholipase C isoforms in the nucleus and their activation during compensatory liver growth. FEBS Lett 571(1-3):35-42.
