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Isolation of Human Lymphatic Endothelial Cells by Multi-parameter Fluorescence-activated Cell Sorting

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Published on: May 1, 2015

NFATc1 regulates lymphatic endothelial development.

Rishikesh M Kulkarni¹, James M Greenberg, Ann L Akeson

¹Division of Pulmonary Biology, MLC7009, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH 45229, USA.

Mechanisms of Development

|February 24, 2009

Summary

This summary is machine-generated.

The calcineurin-NFAT pathway, particularly NFATc1, is crucial for lymphatic development and patterning. This signaling pathway regulates lymphatic endothelial cell specification and lymphangiogenesis, impacting lymphatic vessel formation.

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Murine Dermal Lymphatic Endothelial Cell Isolation

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Published on: July 21, 2023

Area of Science:

Molecular Biology
Developmental Biology
Immunology

Background:

NFATc1 is a key transcription factor involved in T-cell differentiation, cardiac development, and bone remodeling.
Lymphatic development is a complex process involving lymphatic endothelial cell (LEC) specification, sprouting, and patterning.
Calcineurin-NFAT signaling is a critical pathway in cellular development and immune responses.

Purpose of the Study:

To investigate the role of calcineurin-NFAT signaling in lymphatic development and patterning.
To determine the specific NFAT family member involved in lymphangiogenesis.
To elucidate the molecular mechanisms by which NFATc1 regulates lymphatic endothelial gene expression.

Main Methods:

Analysis of NFATc1 null mice and embryos treated with calcineurin inhibitors (cyclosporine-A).
In vivo studies using a murine model of injury-induced lymphangiogenesis.
In vitro experiments with human microvascular endothelial cells using siRNA knockdown and reporter assays.
Immunohistochemistry to assess the localization of NFATc1 and lymphatic markers (Prox-1, VEGFR-3, podoplanin).

Main Results:

NFATc1 is expressed in LECs during lymphatic development and is colocalized with lymphatic markers.
NFATc1 null mice exhibit defects in lymph sac formation and lymphatic patterning.
Inhibition of calcineurin or NFATc1 reduces lymphatic marker expression and impairs lymphangiogenesis.
NFATc1 directly activates lymphatic-specific gene promoters, indicating its role as a transcriptional regulator.

Conclusions:

The calcineurin-NFAT pathway, with NFATc1 as the primary effector, plays a critical role in lymphatic development, patterning, and lymphangiogenesis.
NFATc1 regulates the expression of key lymphatic genes, including Prox-1, VEGFR-3, and podoplanin.
Targeting the calcineurin-NFAT pathway may offer therapeutic potential for modulating lymphangiogenesis.