Investigating T Cell Receptor Signals In Situ by Two-Photon Microscopy of Thymocytes Expressing Genetic Reporters in Low-Density Chimeras

Fournier, Marilaine; Dong, Mengqi; Melichar, Heather J.

doi:10.1007/978-1-0716-0266-9_18

Cited by 2 publications

(1 citation statement)

References 48 publications

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“…Standard methods such as flow cytometry, immunohistochemistry, and imaging of excised tissue have been applied to study the thymus in preclinical mouse models but these are limited to ex vivo analysis. Ex vivo culture systems (Hong & Moore, 1996;Ramsdell et al, 2006) including thymic slices (Bhakta et al, 2005;Ehrlich et al, 2009;Fournier et al, 2020;Le Borgne et al, 2009;Markert et al, 1997;Zhou et al, 2020) and whole organ imaging (Campinoti et al, 2020;Dzhagalov et al, 2012) are very powerful systems for studying certain aspects of thymus biology such as thymocyte-stromal interactions (Ladi et al, 2008;Nakagawa et al, 2012) and T cell development (Le Borgne et al, 2009;Witt et al, 2005), but these techniques lack blood flow and may not fully recapitulate the in vivo situation. Due to a lack of viable intravital imaging methods for the native thymus, researchers have relied on thymus transplantation to optically accessible sites including the kidney capsule (Caetano et al, 2012;Gregorio et al, 2021;Iwai & Inaba, 2015;Morillon et al, 2015), anterior chamber of the eye (Oltra & Caicedo, 2018), and pinna of the mouse ear (Chen et al, 2013) for intravital microscopy, but transplantation may alter the native vascular connections and exposes the tissue to an aberrant environment (Caetano et al, 2012;Vollmann et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Intravital Two-Photon Microscopy of the Native Thymus

Seyedhassantehrani

Burns

Verrinder

et al. 2023

Preprint

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The thymus, a key organ involved in the adaptive immune system, is damaged by a variety of insults including cytotoxic preconditioning. This damage can lead to atrophy and potentially to changes in the hemodynamics of the thymic blood vascular system. Although the thymus has an innate ability to regenerate, the production of T cells relies on the trafficking of lymphoid progenitors from the bone marrow through the altered thymic blood vascular system. Our understanding of thymic blood vascular hemodynamics is limited due to technical challenges associated with accessing the native thymus in live mice. To overcome this challenge, we developed an intravital two-photon imaging method to visualize the native thymus in vivo and investigated functional changes to the vascular system following sublethal irradiation. We were able to quantify blood flow velocity and shear rate in cortical blood vessels and identified a subtle but significant increase in vessel diameter and barrier function ~24 hrs post-sublethal irradiation. Ex vivo whole organ imaging of optically cleared thymus lobes confirmed a disruption of the thymus vascular structure, resulting in an increase in blood vessel diameter and vessel area, and concurrent thymic shrinkage. This novel two-photon intravital imaging method enables a new paradigm for directly investigating the thymic microenvironment in vivo.

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Section: Introductionmentioning

confidence: 99%

Intravital Two-Photon Microscopy of the Native Thymus

Seyedhassantehrani

Burns

Verrinder

et al. 2023

Preprint

View full text Add to dashboard Cite

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Intravital two-photon microscopy of the native mouse thymus

Seyedhassantehrani,

Burns,

Verrinder

et al. 2024

PLoS ONE

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The thymus, a key organ in the adaptive immune system, is sensitive to a variety of insults including cytotoxic preconditioning, which leads to atrophy, compression of the blood vascular system, and alterations in hemodynamics. Although the thymus has innate regenerative capabilities, the production of T cells relies on the trafficking of lymphoid progenitors from the bone marrow through the altered thymic blood vascular system. Our understanding of thymic blood vascular hemodynamics is limited due to technical challenges associated with accessing the native thymus in live mice. To overcome this challenge, we developed an intravital two-photon imaging method to visualize the native thymus in vivo and investigated functional changes to the vascular system following sublethal irradiation. We quantified blood flow velocity and shear rate in cortical blood vessels and identified a subtle but significant increase in vessel leakage and diameter ~24 hrs post-sublethal irradiation. Ex vivo whole organ imaging of optically cleared thymus lobes confirmed a disruption of the thymus vascular structure, resulting in an increase in blood vessel diameter and vessel area, and concurrent thymic atrophy. This novel two-photon intravital imaging method enables a new paradigm for directly investigating the thymic microenvironment in vivo.

show abstract

Investigating T Cell Receptor Signals In Situ by Two-Photon Microscopy of Thymocytes Expressing Genetic Reporters in Low-Density Chimeras

Cited by 2 publications

References 48 publications

Intravital Two-Photon Microscopy of the Native Thymus

Intravital Two-Photon Microscopy of the Native Thymus

Intravital two-photon microscopy of the native mouse thymus

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