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Invariant asymmetry renews the lymphatic vasculature during homeostasis.

Connor AL, Kelley PM, Tempero RM - J Transl Med (2016)

Bottom Line: Interestingly, the morphology of tdT(+) lymphatic vasculature appeared relatively stable without significant remodeling during this time period.The results of these studies support a mechanism of invariant asymmetry to self renew the lymphatic vasculature during homeostasis.These original findings raise important questions related to the plasticity and self renewal properties that maintain the lymphatic vasculature during life.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosensory Genetics and Otolaryngology and Head and Neck Surgery, Boys Town National Research Hospital, 555 North 30th Street, Omaha, NE, 68131, USA.

ABSTRACT

Background: The lymphatic vasculature regulates tissue physiology and immunity throughout life. The self renewal mechanism that maintains the lymphatic vasculature during conditions of homeostasis is unknown. The purpose of this study was to investigate the cellular mechanism of lymphatic endothelial cell (LEC) self renewal and lymphatic vessel maintenance.

Methods: Inductive genetic techniques were used to label LECs with tandem dimer tomato (tdT) in adult mice. Two types of studies were performed, those with high dose inductive conditions to label nearly all the lymphatic vessels and studies with low dose inductive conditions to stochastically label individual clones or small populations of LECs. We coupled image guidance techniques and live fluorescence microscopy imaging with lineage tracing to track the fate of entire tdT(+) cutaneous lymphatic vessels or the behavior of individual or small populations of LECs over 11 months. We tracked the fate of 110 LEC clones and 80 small LEC populations (clusters of 2-7 cells) over 11 months and analyzed their behavior using quantitative techniques.

Results: The results of the high dose inductive studies showed that the lymphatic vessels remained tdT(+) over 11 months, suggesting passage and expression of the tdT transgene from LEC precursors to progenies, an intrinsic model of self- renewal. Interestingly, the morphology of tdT(+) lymphatic vasculature appeared relatively stable without significant remodeling during this time period. By following the behavior of labeled LEC clones or small populations of LECs individually over 11 months, we identified diverse LEC fates of proliferation, quiescence, and extinction. Quantitative analysis of this data revealed that the average lymphatic endothelial clone or small population remained stable in size despite diverse individual fates.

Conclusion: The results of these studies support a mechanism of invariant asymmetry to self renew the lymphatic vasculature during homeostasis. These original findings raise important questions related to the plasticity and self renewal properties that maintain the lymphatic vasculature during life.

No MeSH data available.


Related in: MedlinePlus

Visualizing cutaneous lymphatic vessels in Lyve1CreERT2tdT mice using live imaging. Low power live imaging epifluorescent microscopy of sedated Lyve1CreERT2tdT pinna reveals no detectable tdT labeling within the pinna (a). Using similar microscopy techniques, tdT+ lymphatic vessels were detected in the same animal 21 days following 4-OHT administration (b). The size standard is 0.5 mm
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Fig1: Visualizing cutaneous lymphatic vessels in Lyve1CreERT2tdT mice using live imaging. Low power live imaging epifluorescent microscopy of sedated Lyve1CreERT2tdT pinna reveals no detectable tdT labeling within the pinna (a). Using similar microscopy techniques, tdT+ lymphatic vessels were detected in the same animal 21 days following 4-OHT administration (b). The size standard is 0.5 mm

Mentions: We designed studies to investigate the remodeling of the cutaneous lymphatic vasculature longitudinally in Lyve1CreERT2tdT mice. We used bright field and fluorescence microscopy techniques to obtain images from the pinna of sedated Lyve1CreERT2tdT mice. Prior to 4-OHT administration, there was no detectable tdT within the Lyve1CreERT2tdT pinna. 3 weeks after induction, tdT fluorescence was readily detected using live imaging fluorescence microscopy (Fig. 1). The entire lymphatic vasculature within the animal was labeled with tdT; however, we elected to investigate the pinna as this tissue was accessible and provided the opportunity to obtain images using intravital microscopy with reasonable resolution when the pinna was gently positioned between two glass slides. Animal movement as a result of the work of breathing and cardiac contractility created continuous movement in sedated mice that made it difficult to obtain adequate images from other cutaneous regions such as the cervical or thoracic regions. The pinna contained two lymphatic plexuses, each deep to overlying epithelium. Thus, the image in Fig. 1b is a composite of two lymphatic plexuses.Fig. 1


Invariant asymmetry renews the lymphatic vasculature during homeostasis.

Connor AL, Kelley PM, Tempero RM - J Transl Med (2016)

Visualizing cutaneous lymphatic vessels in Lyve1CreERT2tdT mice using live imaging. Low power live imaging epifluorescent microscopy of sedated Lyve1CreERT2tdT pinna reveals no detectable tdT labeling within the pinna (a). Using similar microscopy techniques, tdT+ lymphatic vessels were detected in the same animal 21 days following 4-OHT administration (b). The size standard is 0.5 mm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4940917&req=5

Fig1: Visualizing cutaneous lymphatic vessels in Lyve1CreERT2tdT mice using live imaging. Low power live imaging epifluorescent microscopy of sedated Lyve1CreERT2tdT pinna reveals no detectable tdT labeling within the pinna (a). Using similar microscopy techniques, tdT+ lymphatic vessels were detected in the same animal 21 days following 4-OHT administration (b). The size standard is 0.5 mm
Mentions: We designed studies to investigate the remodeling of the cutaneous lymphatic vasculature longitudinally in Lyve1CreERT2tdT mice. We used bright field and fluorescence microscopy techniques to obtain images from the pinna of sedated Lyve1CreERT2tdT mice. Prior to 4-OHT administration, there was no detectable tdT within the Lyve1CreERT2tdT pinna. 3 weeks after induction, tdT fluorescence was readily detected using live imaging fluorescence microscopy (Fig. 1). The entire lymphatic vasculature within the animal was labeled with tdT; however, we elected to investigate the pinna as this tissue was accessible and provided the opportunity to obtain images using intravital microscopy with reasonable resolution when the pinna was gently positioned between two glass slides. Animal movement as a result of the work of breathing and cardiac contractility created continuous movement in sedated mice that made it difficult to obtain adequate images from other cutaneous regions such as the cervical or thoracic regions. The pinna contained two lymphatic plexuses, each deep to overlying epithelium. Thus, the image in Fig. 1b is a composite of two lymphatic plexuses.Fig. 1

Bottom Line: Interestingly, the morphology of tdT(+) lymphatic vasculature appeared relatively stable without significant remodeling during this time period.The results of these studies support a mechanism of invariant asymmetry to self renew the lymphatic vasculature during homeostasis.These original findings raise important questions related to the plasticity and self renewal properties that maintain the lymphatic vasculature during life.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosensory Genetics and Otolaryngology and Head and Neck Surgery, Boys Town National Research Hospital, 555 North 30th Street, Omaha, NE, 68131, USA.

ABSTRACT

Background: The lymphatic vasculature regulates tissue physiology and immunity throughout life. The self renewal mechanism that maintains the lymphatic vasculature during conditions of homeostasis is unknown. The purpose of this study was to investigate the cellular mechanism of lymphatic endothelial cell (LEC) self renewal and lymphatic vessel maintenance.

Methods: Inductive genetic techniques were used to label LECs with tandem dimer tomato (tdT) in adult mice. Two types of studies were performed, those with high dose inductive conditions to label nearly all the lymphatic vessels and studies with low dose inductive conditions to stochastically label individual clones or small populations of LECs. We coupled image guidance techniques and live fluorescence microscopy imaging with lineage tracing to track the fate of entire tdT(+) cutaneous lymphatic vessels or the behavior of individual or small populations of LECs over 11 months. We tracked the fate of 110 LEC clones and 80 small LEC populations (clusters of 2-7 cells) over 11 months and analyzed their behavior using quantitative techniques.

Results: The results of the high dose inductive studies showed that the lymphatic vessels remained tdT(+) over 11 months, suggesting passage and expression of the tdT transgene from LEC precursors to progenies, an intrinsic model of self- renewal. Interestingly, the morphology of tdT(+) lymphatic vasculature appeared relatively stable without significant remodeling during this time period. By following the behavior of labeled LEC clones or small populations of LECs individually over 11 months, we identified diverse LEC fates of proliferation, quiescence, and extinction. Quantitative analysis of this data revealed that the average lymphatic endothelial clone or small population remained stable in size despite diverse individual fates.

Conclusion: The results of these studies support a mechanism of invariant asymmetry to self renew the lymphatic vasculature during homeostasis. These original findings raise important questions related to the plasticity and self renewal properties that maintain the lymphatic vasculature during life.

No MeSH data available.


Related in: MedlinePlus