Histological Study and LYVE-1 Immunolocalization of Mouse Mesenteric Lymph Nodes with "In Vivo Cryotechnique"

Bai, Yu; Wu, Bao; Terada, Nobuo; Ohno, Nobuhiko; Saitoh, Shu‐ichi; Saitoh, Yurika; Ohno, Shinichi

doi:10.1267/ahc.11002

Cited by 5 publications

(4 citation statements)

References 44 publications

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“…To further explore the contrast drainage, we dissected the smLN in live KX-anesthetized mice and performed ex vivo scanning. These scans confirmed Gd-DOTA uptake in small zones in the periphery of the smLN likely corresponding to the subcapsular areas in conjunction to afferent lymph vessels previously documented using other imaging and histological techniques 5,18,35,36 .…”

Section: Discussionsupporting

confidence: 83%

In vivo T1 mapping for quantifying glymphatic system transport and cervical lymph node drainage

Xue

Liu

Koundal

et al. 2020

Sci Rep

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Dynamic contrast-enhanced magnetic resonance imaging (MRI) for tracking glymphatic system transport with paramagnetic contrast such as gadoteric acid (Gd-DOTA) administration into cerebrospinal fluid (CSF) requires pre-contrast data for proper quantification. Here we introduce an alternative approach for glymphatic system quantification in the mouse brain via T1 mapping which also captures drainage of Gd-DOTA to the cervical lymph nodes. The Gd-DOTA injection into CSF was performed on the bench after which the mice underwent T1 mapping using a 3D spoiled gradient echo sequence on a 9.4 T MRI. In Ketamine/Xylazine (KX) anesthetized mice, glymphatic transport and drainage of Gd-DOTA to submandibular and deep cervical lymph nodes was demonstrated as 25-50% T1 reductions in comparison to control mice receiving CSF saline. To further validate the T1 mapping approach we also verified increased glymphatic transport of Gd-DOTA transport in mice anesthetized with KX in comparison with ISO. The novel T1 mapping method allows for quantification of glymphatic transport as well as drainage to the deep and superficial cervical lymph nodes. The ability to measure glymphatic transport and cervical lymph node drainage in the same animal longitudinally is advantageous and time efficient and the coupling between the two systems can be studied and translated to human studies. The glymphatic system (GS) in rodents was first described by Iliff and Nedergaard in 2012 as a brain-wide perivascular transit passageway for cerebrospinal fluid (CSF) facilitating waste clearance in an aquaporin 4 (AQP4) water channel dependent manner 1. Visualization and quantification of GS transport in whole mouse brain was performed ex vivo by first allowing tracers administered into CSF to circulate for a prefixed time interval (30 min-2 h) after which the rodents were euthanized and their brains examined for tracer uptake using fluorescence microscopy 1. In 2013, we introduced dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) as an in vivo method for tracking GS transport in the whole rodent brain 2 and later refined gadolinium based contrast administration into CSF and GS quantification 3,4. The DCE-MRI approach for brain-wide GS transport assessment has been widely used and adapted to different species including mice 5-7 , non-human primates 8 , and humans 9,10. Recently, the DCE-MRI approach for GS transport assessment was extended to studies in awake rats. The DCE-MRI GS method involves administering small molecular weight (MW) (e.g. gadopentetic acid MW 547 Da, gadoteric acid (Gd-DOTA), MW 559 Da) or large MW (e.g. GadoSpin P, MW 200 kDa) paramagnetic tracers into CSF via the cisterna magna (CM) 2,3 , cerebral lateral ventricles 11 or lumbar intrathecal route 9. Transport of the paramagnetic contrast molecule (a.k.a. 'solute') from CSF into brain parenchyma is tracked dynamically by a series of 3D T1-weighted MRI scans 1. The time series of 3D T1-weighted spoiled gradient echo (SPGR) brain images acquired before, during and after solute...

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Section: Discussionsupporting

confidence: 83%

In vivo T1 mapping for quantifying glymphatic system transport and cervical lymph node drainage

Xue

Liu

Koundal

et al. 2020

Sci Rep

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“…The sections from fishes and murine ovaries were incubated in 10% normal donkey serum, and then in anti-prox1 antibody (Abcam), or in a mixture of an antibody to Prox1 or LYVE-1 (Angiobio, Del Mar, CA, USA), another lymphatic endothelial molecule [3, 4, 16], together with an antibody to plakoglobin (PROGEN Biotechnik, Heidelberg, Germany) or CD31 (BD Biosciences Pharmingen, San Jose, CA, USA) at 4°C overnight. They were then treated with indocarbocyanine (Cy3)-conjugated donkey anti-rabbit IgG (Jackson ImmnoResearch, West Grove, PA, USA), or with a mixture of Cy3-conjugated donkey anti-rabbit IgG and fluorescein isothiocyanate (FITC)-conjugated donkey anti-mouse or rat IgG (Jackson ImmunoResearch) for 1 hr at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Immunohistochemical Demonstration of Lymphatic Vessels in Adult Zebrafish

Shimoda

Isogai

2012

Acta Histochem. Cytochem

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Morphological profiles of lymphatic vessels in adult zebrafish trunk and ovary were studied by immunohistochemistry and electron microscopy. The present immunohistochemistry for Prox1 was successful in demonstrating lymphatic vessels in zebrafish. The zebrafish trunk revealed two types of bilateral longitudinal lymphatic trunks draining lymph centripetally along dorsal aorta and posterior cardinal veins. Large honeycomb lymphatic sinus was further shown around common cardinal veins. In the zebrafish ovary, the lymphatic vessels, comprising endothelial cells only, encompassed arterioles in their lumen. This peculiar structure appeared to be conserved in vertebrates including mammals and might serve for control of blood temperature and tissue homeostasis. The present study is first to delineate lymphatic vessels in adult zebrafish by immunohistochemistry. Our immunohistochemical results showed usefulness of immunostaining for Prox1 not only for demonstration of lymphatic vessels in zebrafish, but also for examination of their function and dynamics in pathophysiological condition.

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“…Once mDCs enter the LN subcapsular sinus through the afferent lymphatics, the cells must overcome the barrier imposed by the subcapsular sinus floor that is lined by LYVE-1expressing lymphatic endothelial cells (83) and resident CD169 + macrophages (84). Published data indicate that mDCs overcome this barrier in a CCR7-dependent manner by responding to CCL21, particularly, the CCL21-Ser form, which is expressed in the subcapsular sinus and paracortex or T cell zone of the LN (54).…”

Section: Multiple Chemokine Receptors Mediate Entry Of Migratory Cdcs To Lnmentioning

confidence: 99%

Compartmentalization of dendritic cell and T‐cell interactions in the lymph node: Anatomy of T‐cell fate decisions

León

Lund

2019

Immunological Reviews

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Upon receiving cognate and co-stimulatory priming signals from antigen (Ag)-presenting dendritic cells (DCs) in secondary lymphoid tissues, naïve CD4 + T cells differentiate into distinct effector and memory populations. These alternate cell fate decisions, which ultimately control the T cell functional attributes, are dictated by programming signals provided by Ag-bearing DCs and by other cells that are present in the microenvironment in which T cell priming occurs. We know that DCs can be subdivided into multiple populations and that the various DC subsets exhibit differential capacities to initiate development of the different CD4 + T-helper populations. What is less well understood is why different sub-anatomic regions of secondary lymphoid tissues are colonized by distinct populations of Ag-presenting DCs and how the location of these DCs influences the type of T cell response that will be generated. Here we review how chemokine receptors and their ligands, which position allergen and nematode-activated DCs within different microdomains of secondary lymphoid tissues, contribute to the establishment of IL-4 committed follicular helper T (Tfh) and type 2 helper (Th2) cell responses.

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Histological Study and LYVE-1 Immunolocalization of Mouse Mesenteric Lymph Nodes with "In Vivo Cryotechnique"

Cited by 5 publications

References 44 publications

In vivo T1 mapping for quantifying glymphatic system transport and cervical lymph node drainage

In vivo T1 mapping for quantifying glymphatic system transport and cervical lymph node drainage

Immunohistochemical Demonstration of Lymphatic Vessels in Adult Zebrafish

Compartmentalization of dendritic cell and T‐cell interactions in the lymph node: Anatomy of T‐cell fate decisions

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