Intravital Three-Dimensional Dynamic Pathology of Experimental Colitis in Living Mice Using Two-Photon Laser Scanning Microscopy

Morimoto, Yuhki; Toiyama, Yuji; Inoue, Yasuhiro; Araki, Toshimitsu; Uchida, Keiichi; Kimura, Kei; Mizoguchi, Akira; Kusunoki, Masato

doi:10.1007/s11605-011-1632-5

Cited by 12 publications

(11 citation statements)

References 14 publications

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“…As previously described [20]–[23], GFP mice were anesthetized with isofluorane 4 L/min (4%; Forane, Abbott, Japan). Body temperature was kept at 37°C using a heating pad.…”

Section: Methodsmentioning

confidence: 99%

“…The system allows us to capture high-magnification, high-resolution images of exteriorized living tissue, from the surface to several micrometers depth [18]–[23]. Previously, we have visualized in vivo real-time bacterial translocation in dextran sodium sulfate-induced colitis [18], thrombus formation in the laser-induced endothelium injury [19], three-dimensional steroid efficacy for DSS-induced colitis [20], colorectal liver metastatic formation [21], and chemotherapy response on the tumor microenvironment of colorectal liver metastases [22], [23].…”

Section: Introductionmentioning

confidence: 99%

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In Vivo Characterization of Neutrophil Extracellular Traps in Various Organs of a Murine Sepsis Model

et al. 2014

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Neutrophil extracellular traps (NETs) represent extracellular microbial trapping and killing. Recently, it has been implicated in thrombogenesis, autoimmune disease, and cancer progression. The aim of this study was to characterize NETs in various organs of a murine sepsis model in vivo and to investigate their associations with platelets, leukocytes, or vascular endothelium. NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils and anchored NETs that were anchored to neutrophils. Circulating cell-free NETs were characterized as fragmented or cotton-like structures, while anchored NETs were characterized as linear, reticular, membranous, or spot-like structures. In septic mice, both anchored and cell-free NETs were significantly increased in postcapillary venules of the cecum and hepatic sinusoids with increased leukocyte-endothelial interactions. NETs were also observed in both alveolar space and pulmonary capillaries of the lung. The interactions of NETs with platelet aggregates, leukocyte-platelet aggregates or vascular endothelium of arterioles and venules were observed in the microcirculation of septic mice. Microvessel occlusions which may be caused by platelet aggregates or leukocyte-platelet aggregates and heterogeneously decreased blood flow were also observed in septic mice. NETs appeared to be associated with the formation of platelet aggregates or leukocyte-platelet aggregates. These observational findings may suggest the adverse effect of intravascular NETs on the host during a sepsis.

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“…As previously described [20]–[23], GFP mice were anesthetized with isofluorane 4 L/min (4%; Forane, Abbott, Japan). Body temperature was kept at 37°C using a heating pad.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In Vivo Characterization of Neutrophil Extracellular Traps in Various Organs of a Murine Sepsis Model

et al. 2014

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“…However, such techniques rarely have been applied to the intestine, particularly in the context of infection (14,(24)(25)(26)(27)(28). Here we used a physiologically relevant oral infection model in conjunction with two-photon microscopy to reveal that neutrophils in the lumen of the small intestine are motile reservoirs of live T. gondii.…”

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confidence: 99%

Motile invaded neutrophils in the small intestine ofToxoplasma gondii-infected mice reveal a potential mechanism for parasite spread

Coombes

Charsar

Han

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

126

149

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Toxoplasma gondii infection occurs through the oral route, but we lack important information about how the parasite interacts with the host immune system in the intestine. We used two-photon laserscanning microscopy in conjunction with a mouse model of oral T. gondii infection to address this issue. T. gondii established discrete foci of infection in the small intestine, eliciting the recruitment and transepithelial migration of neutrophils and inflammatory monocytes. Neutrophils accounted for a high proportion of actively invaded cells, and we provide evidence for a role for transmigrating neutrophils and other immune cells in the spread of T. gondii infection through the lumen of the intestine. Our data identify neutrophils as motile reservoirs of T. gondii infection and suggest a surprising retrograde pathway for parasite spread in the intestine.neutrophil motility | dynamic imaging | gut | mucosal immunology T oxoplasma gondii infects around a third of humans worldwide and is widely dispersed in other warm-blooded hosts. Although clinical manifestations in the brain, eye, and developing fetus receive the most attention, T. gondii is an oral pathogen and first enters the body and establishes infection in the small intestine. Infection follows consumption of cyst-containing meat or oocyst-contaminated water and produce and is associated with the development of small intestinal pathology in a variety of nonhuman hosts (1). Most notably, experimental infection of C57BL/6 mice by the oral route results in an inflammation of the small intestine that shares immunological features with inflammatory bowel disease (2). This model is useful to further our understanding of host-pathogen interactions in the intestine and of common mechanisms underpinning the development of inflammatory bowel disease (3). Nevertheless, we have limited understanding of how and in which cells infection is established in the intestine, the extent to which the parasite replicates and spreads within the intestine, and how these factors contribute to the development of pathology (2, 4-9). The ability to label living parasites fluorescently and track them in the tissues of infected hosts provides an important tool for investigating these questions (10)(11)(12)(13)(14).Starting in the small intestine, T. gondii must travel long distances and surmount a variety of biological barriers to establish chronic infection in the brain. These barriers include the mucus, the intestinal epithelium, and the blood-brain barrier (7,15). Cells of the immune system are often highly motile and represent attractive transport vessels for pathogens seeking to reach and enter tissues while being protected from the external environment. Consequently, recent studies have focused on the role of immune cells in transporting parasites between tissues (4, 16-23). For example, cluster of differentiation 11b-positive (CD11b + ) cells have been implicated in the dissemination of T. gondii through the blood and across the blood-brain barrier (4, 19). Following oral infection, i...

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“…We also established a timeseries TPLSM technique consisting of multiple time points for intravital TPLSM observations (time-series TPLSM) in the same living mice over periods of days to months. The animals comprised a dextran sulfate sodium-induced colitis model [21] and a colorectal liver metastases model [22,23]. We evaluated in vivo and in real time the response to chemotherapy given for colorectal xenografts of liver metastatic tumor to study the dynamic interactions between metastatic tumor cells and host stromal cells in living mice [24].…”

Section: Introductionmentioning

confidence: 99%

In vivo optical pathology of paclitaxel efficacy on the peritoneal metastatic xenograft model of gastric cancer using two-photon laser scanning microscopy

et al. 2014

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Background We previously visualized in vivo responses to chemotherapy in a colorectal liver metastatic xenograft model using in vivo real-time and time-series intravital two-photon laser scanning microscopy (TPLSM). In this study, we established the method for evaluating the response of peritoneal xenografts to chemotherapy of metastatic gastric cancer using intravital TPLSM. Methods Red fluorescent protein-expressing gastric cancer cells (NUGC4) were inoculated into the peritoneal cavity of green fluorescent protein nude mice. Results Laparotomy revealed that 2 weeks after inoculation, macroscopic peritoneal metastatic nodules were formed. The first intravital TPLSM session revealed that they were composed of red tumor cell clusters and green surrounding stroma. Paclitaxel was administered intraperitoneally after the first TPLSM three times a week for 7 days in the treatment group. At the second laparotomy, there were significantly fewer and smaller nodules in the treated mice than in the controls. The second intravital TPLSM session showed tumor cell fragmentation, swelling, and nuclear condensation in the metastatic nodules-a response to chemotherapy. There were multinuclear tumor cells in the paclitaxel-treated living mice.Conclusions Our method may become a powerful tool for evaluating the efficacy of novel anti-gastric cancer drugs in a preclinical murine model with minimum interindividual variation.

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Intravital Three-Dimensional Dynamic Pathology of Experimental Colitis in Living Mice Using Two-Photon Laser Scanning Microscopy

Abstract: Intravital 3D tomographic visualization of experimental colitis using TPLSM in combination with the serosal-approaching method can provide dynamic pathology at a high magnification, which may be useful in evaluating treatment efficacy in the same living mice.

Cited by 12 publications

References 14 publications

In Vivo Characterization of Neutrophil Extracellular Traps in Various Organs of a Murine Sepsis Model

In Vivo Characterization of Neutrophil Extracellular Traps in Various Organs of a Murine Sepsis Model

Motile invaded neutrophils in the small intestine ofToxoplasma gondii-infected mice reveal a potential mechanism for parasite spread

In vivo optical pathology of paclitaxel efficacy on the peritoneal metastatic xenograft model of gastric cancer using two-photon laser scanning microscopy

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