Intravital imaging of the cellular dynamics of LysM-positive cells in a murine corneal suture model

Ueta, Mayumi; Koga, Ayaka; Kikuta, Junichi; Yamada, Katsushi; Kojima, Sachi; Shinomiya, Katsuhiko; Ishii, Masaru; Kinoshita, Shigeru

doi:10.1136/bjophthalmol-2015-307024

Cited by 7 publications

(10 citation statements)

References 9 publications

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“…The setup of Steven et al (54), investigating leukocyte transmigration into lymphatic vessels in the peripheral cornea and limbus, was not temperature controlled, which as we demonstrate, is necessary to visualize central CL kinetics in a noninvasive manner, and furthermore, did not use transgenic mice. More recently, 2 studies have used MPM to investigate endogenously, fluorescently labeled neutrophil kinetics on the ocular surface of mice (69,70). Whereas the setup used by Ueta et al (69) for analyzing the cornea likely maintains the correct ocular surface temperature required for analysis of CLs, it is highly invasive, as the entire eye is dislocated to immobilize the cornea and compensate/ correct for the various image stability issues (e.g., drift and ocular pulsations) and positions the mouse on its back for the entire length of imaging inside a dark, 37°C box.…”

Section: Discussionmentioning

confidence: 99%

“…More recently, 2 studies have used MPM to investigate endogenously, fluorescently labeled neutrophil kinetics on the ocular surface of mice (69,70). Whereas the setup used by Ueta et al (69) for analyzing the cornea likely maintains the correct ocular surface temperature required for analysis of CLs, it is highly invasive, as the entire eye is dislocated to immobilize the cornea and compensate/ correct for the various image stability issues (e.g., drift and ocular pulsations) and positions the mouse on its back for the entire length of imaging inside a dark, 37°C box. Furthermore, the nature of vessels and corneal nerves in this preparation is unknown.…”

Section: Discussionmentioning

confidence: 99%

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Kinetics of corneal leukocytes by intravital multiphoton microscopy

et al. 2018

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Corneal immune privilege is integral in maintaining the clear avascular window to the foreign world. The presence of distinct populations of corneal leukocytes (CLs) in the normal cornea has been firmly established. However, their precise function and kinetics remain, as of yet, unclear. Through intravital multiphoton microscopy (IV-MPM), allowing the means to accumulate critical spatial and temporal cellular information, we provide details for long-term investigation of CL morphology and kinetics under steady state and following inflammation. Significant alterations in size and morphology of corneal CD11c dendritic cells (DCs) were noted following acute sterile inflammation, including cell volume (4364.4 ± 489.6 vs. 1787.6 ± 111.0 μm, P < 0.001) and sphericity (0.82 ± 0.01 vs. 0.42 ± 0.02, P < 0.001) compared with steady state. Furthermore, IV-MPM analyses revealed alterations in both the CD11c DC and major histocompatibility complex class II (MHC)-II mature antigen-presenting cell population kinetics during inflammation, including track displacement length (CD11c: 16.57 ± 1.41 vs. 4.64 ± 0.56 μm, P < 0.001; MHC-II: 9.03 ± 0.37 vs. 4.09 ± 0.39, P < 0.001) and velocity (CD11c: 1.91 ± 0.07 μm/min vs. 1.73 ± 0.1302 μm/min; MHC-II: 2.97 ± 0.07 vs. 1.62 ± 0.08, P < 0.001) compared with steady state. Our results reveal in vivo evidence of sessile CL populations exhibiting dendritic morphology under steady state and increased velocity of spherical leukocytes following inflammation. IV-MPM represents a powerful tool to study leukocytes in corneal diseases in context.-Seyed-Razavi, Y., Lopez, M. J., Mantopoulos, D., Zheng, L., Massberg, S., Sendra, V. G., Harris, D. L., Hamrah, P. Kinetics of corneal leukocytes by intravital multiphoton microscopy.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Kinetics of corneal leukocytes by intravital multiphoton microscopy

et al. 2018

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“…Around these non‐motile‐infected dendritic cells, neutrophils converge by directed migration and aggregate at the Listeria foci to wall off any potential spread of bacteria . More recent intravital imaging studies have extended the list of organs where interstitial neutrophil swarming behavior has been observed, including brain tissue after traumatic injury, brains with amyloid Aβ plaque depositions, sutured corneas, and intestinal tissue infected by parasites . To date, extravascular neutrophil swarms have been reported for various conditions of sterile inflammation and infection with bacteria, fungi, and parasites .…”

Section: Swarming Of Mouse Neutrophilsmentioning

confidence: 99%

“…More recent intravital imaging studies have extended the list of organs where interstitial neutrophil swarming behavior has been observed, including brain tissue after traumatic injury, 36 brains with amyloid Aβ plaque depositions, 37 sutured corneas, 38 and intestinal tissue infected by parasites. 39 To date, extravascular neutrophil swarms have been reported for various conditions of sterile inflammation 28,29,34,[36][37][38][40][41][42][43] and infection with bacteria, 28,34,35,41,42,[44][45][46] fungi, 27 and parasites. 29,33,39 In accordance with the biological sentinel function of neutrophils in monitoring our body for tissue injury and microbial invasion, neutrophil swarms robustly form in diverse tissues and inflammatory settings.…”

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confidence: 99%

Neutrophil swarming: an essential process of the neutrophil tissue response

Kienle

Lämmermann

2016

Immunological Reviews

178

186

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Neutrophil infiltration into inflamed and infected tissues is a fundamental process of the innate immune response. While neutrophil interactions with the blood vessel wall have been intensely studied over the last decades, neutrophil dynamics beyond the vasculature have for a long time remained poorly investigated. Recent intravital microscopy studies of neutrophil populations directly at the site of tissue damage or microbial invasion have changed our perspective on neutrophil responses within tissues. Swarm-like migration patterns of neutrophils, referred to as 'neutrophil swarming', have been detected in diverse tissues under conditions of sterile inflammation and infection with various pathogens, including bacteria, fungi, and parasites. Current work has begun to unravel the molecular pathways choreographing the sequential phases of highly coordinated chemotaxis followed by neutrophil accumulation and the formation of substantial neutrophil clusters. It is now clear that intercellular communication among neutrophils amplifies their recruitment in a feed-forward manner, which provides them with a level of self-organization during neutrophil swarming. This review will summarize recent developments and current concepts on neutrophil swarming, an important process of the neutrophil tissue response with a critical role in maintaining the balance between host protection and inflammation-driven tissue destruction.

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“…LysM-positive neutrophils are located around the limbal vessels in the periphery, but are not found in the central cornea [60]. Similarly, mast cells are found in the corneal limbus and conjunctival parenchyma, but not in the central cornea of adult mice.…”

Section: Immune Cells In the Resting Corneamentioning

confidence: 99%

Wound-Healing Studies in Cornea and Skin: Parallels, Differences and Opportunities

Bukowiecki

Hos

Cursiefen

et al. 2017

IJMS

136

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The cornea and the skin are both organs that provide the outer barrier of the body. Both tissues have developed intrinsic mechanisms that protect the organism from a wide range of external threats, but at the same time also enable rapid restoration of tissue integrity and organ-specific function. The easy accessibility makes the skin an attractive model system to study tissue damage and repair. Findings from skin research have contributed to unravelling novel fundamental principles in regenerative biology and the repair of other epithelial-mesenchymal tissues, such as the cornea. Following barrier disruption, the influx of inflammatory cells, myofibroblast differentiation, extracellular matrix synthesis and scar formation present parallel repair mechanisms in cornea and skin wound healing. Yet, capillary sprouting, while pivotal in proper skin wound healing, is a process that is rather associated with pathological repair of the cornea. Understanding the parallels and differences of the cellular and molecular networks that coordinate the wound healing response in skin and cornea are likely of mutual importance for both organs with regard to the development of regenerative therapies and understanding of the disease pathologies that affect epithelial-mesenchymal interactions. Here, we review the principal events in corneal wound healing and the mechanisms to restore corneal transparency and barrier function. We also refer to skin repair mechanisms and their potential implications for regenerative processes in the cornea.

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Intravital imaging of the cellular dynamics of LysM-positive cells in a murine corneal suture model

Abstract: Our results showed that corneal sutures may elicit the continuous infiltration of neutrophils.

Cited by 7 publications

References 9 publications

Kinetics of corneal leukocytes by intravital multiphoton microscopy

Kinetics of corneal leukocytes by intravital multiphoton microscopy

Neutrophil swarming: an essential process of the neutrophil tissue response

Wound-Healing Studies in Cornea and Skin: Parallels, Differences and Opportunities

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