title: A Novel Model of Intravital Platelet Imaging Using CD41-ZsGreen1 Transgenic Rats.
abstract: Platelets play pivotal roles in both hemostasis and thrombosis. Although models of intravital platelet imaging are available for thrombosis studies in mice, few are available for rat studies. The present effort aimed to generate fluorescent platelets in rats and assess their dynamics in a rat model of arterial injury. We generated CD41-ZsGreen1 transgenic rats, in which green fluorescence protein ZsGreen1 was expressed specifically in megakaryocytes and thus platelets. The transgenic rats exhibited normal hematological and biochemical values with the exception of body weight and erythroid parameters, which were slightly lower than those of wild-type rats. Platelet aggregation, induced by 20 μM ADP and 10 μg/ml collagen, and blood clotting times were not significantly different between transgenic and wild-type rats. Saphenous arteries of transgenic rats were injured with 10% FeCl3, and the formation of fluorescent thrombi was evaluated using confocal microscopy. FeCl3 caused time-dependent increases in the mean fluorescence intensity of injured arteries of vehicle-treated rats. Prasugrel (3 mg/kg, p.o.), administered 2 h before FeCl3, significantly inhibited fluorescence compared with vehicle-treated rats (4.5 ± 0.4 vs. 14.9 ± 2.4 arbitrary fluorescence units at 30 min, respectively, n = 8, P = 0.0037). These data indicate that CD41-ZsGreen1 transgenic rats represent a useful model for intravital imaging of platelet-mediated thrombus formation and the evaluation of antithrombotic agents.
authors.affiliation: Biological Research Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Mizuno
authors.affiliation: Biological Research Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan.
authors.forename: Atsuyuki
authors.initials: A
authors.lastname: Tomizawa
authors.affiliation: Biological Research Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan.
authors.forename: Kousaku
authors.initials: K
authors.lastname: Ohno
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, United States of America.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Jakubowski
authors.affiliation: Biological Research Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan.
authors.forename: Atsuhiro
authors.initials: A
authors.lastname: Sugidachi
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000502:physiology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000097:blood* / Q000235:genetics*; D000069416:Intravital Microscopy / Q000379:methods*; D008297:Male; D008533:Megakaryocytes / Q000502:physiology; D023421:Models, Animal; D010974:Platelet Aggregation; D040201:Platelet Membrane Glycoprotein IIb / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D055647:Rats, Transgenic / Q000097:blood* / Q000235:genetics*; D011994:Recombinant Proteins / Q000097:blood / Q000235:genetics
chemical_list: D040201:Platelet Membrane Glycoprotein IIb; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins