Dynamic pathology for leukocyte-platelet formation in sepsis model.
BACKGROUND
This study aimed to evaluate the dynamic pathology for in vivo real-time leukocyte-endothelium-platelet aggregation in a mouse model of sepsis.


METHODS
A lipopolysaccharide-induced model of sepsis was analyzed in green fluorescent protein transgenic mice using two-photon laser-scanning microscopy (TPLSM). The real-time process of leukocyte-endothelium-platelet complex (LEPC) formation was assessed in vivo using blood flow dynamics.


RESULTS
TPLSM allowed direct visualization of LEPC formation at the single-platelet level. Leukocytes rolling number and speed, blood flow velocity, and shear rate gradually decreased with time during the acute phase of sepsis compared with those in the control groups. The number of adherent leukocytes and platelet counts gradually increased over time in the septic group. In the septic group, microcirculatory dysfunction was seen in the postcapillary venules before the capillaries.


CONCLUSIONS
In vivo real-time imaging and analysis of LEPC formation can be achieved with little inter-experimental variation using TPLSM. In the acute phase of sepsis, new treatment strategies should target the postcapillary venules because their LEPC formation and blood flow dynamics start to change before those in the capillaries.
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan. Electronic address: koikyon@clin.medic.mie-u.ac.jp.
Yuhki
Y
Koike
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Koji
K
Tanaka
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Minako
M
Kobayashi
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Yuji
Y
Toiyama
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Yasuhiro
Y
Inoue
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Yasuhiko
Y
Mohri
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Keiichi
K
Uchida
Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Akira
A
Mizoguchi
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Masato
M
Kusunoki
D000818:Animals; D001792:Blood Platelets / Q000473:pathology* / Q000502:physiology; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000473:pathology* / Q000503:physiopathology; D007962:Leukocytes / Q000473:pathology* / Q000502:physiology; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008833:Microcirculation; D018613:Microscopy, Confocal; D018805:Sepsis / Q000473:pathology* / Q000503:physiopathology
Green fluorescent protein; Leukocyte–endothelium–platelet interaction; Two-photon laser-scanning microscopy