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Characterization of a novel transgenic rat model for imaging brain vascular dynamics in vivo using confocal endomicroscopy (686.27)
Author(s) -
Mayorov Dmitry,
Roloff Eva,
Alenitalia,
Bader Michael,
Teschemacher Anja,
Paton Julian,
Kasparov Sergey
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.686.27
Subject(s) - confocal microscopy , brainstem , confocal , pathology , endomicroscopy , anatomy , ex vivo , cortex (anatomy) , in vivo , medicine , biology , chemistry , neuroscience , microbiology and biotechnology , geometry , mathematics
We evaluated the feasibility of imaging brain vasculature in a transgenic rat model (L7543) which expressed a construct carrying the eGFP under the control of CAG promoter (Popova et al., Transgen. Res. 2008) using fiber endomicroscopy (CellVizio, Mauna Kea Technologies). eGFP expression were first evaluated in cortical and brainstem slices from L7543 rat using confocal microscopy (Leica SP5). Expression of eGFP in the cortex and brainstem was largely restricted to endothelial cells, although astrocytes in younger rats (up to P35) were also fluorescent. Next, the brainstem and cortex were imaged, without a contrasting agent, in anaesthetized P21‐P40 rats using a fiber probe‐based confocal laser endomicroscopy. Pial microvessels of the ventral or dorsal brainstem, or cortex were imaged using S‐1500 and Mini‐Z probes (1.5 and 0.94 mm tip diameter, respectively). The vascular contractility was visualized after application of a thromboxane agonist U46619 and sodium nitroprusside. Arterioles and veins were identified based on the eGFP expression profile. In addition, beveled probes S‐300/B and S‐650/B (0.3 and 0.65 mm tip diameter, respectively) were used to visualize vascular tree in Z‐dimension. Currently we are working to adapt this method for imaging in unanaesthetized rats. This approach should be useful to investigate changes in cerebral microcirculation in hypertension and other vascular pathologies. Grant Funding Source : Supported by British Heart Foundation