Abstract
The permeability of the lymphatic vasculature is tightly regulated to prevent the excessive leakage of lymph into the tissues, which has profound consequences for edema, immune responses, and lipid absorption. Dysregulated lymphatic permeability is associated with several diseases, including life-threatening chylothorax and pleural effusion that occur in patients with congenital lymphedema and lymphatic malformations. Due to a growing interest in uncovering new mechanisms regulating lymphatic vascular permeability, we recently pioneered methods to quantify this aspect of lymphatic function. Here, we detail our ex vivo method to determine the permeability of mouse collecting lymphatic vessels from direct measurements of solute flux. This method is modified from a similar ex vivo assay that we described for studying the contractile function of murine collecting lymphatic vessels. Since this method also uses the mouse as a model, it enables powerful genetic tools to be combined with this physiological assay to investigate signaling pathways regulating lymphatic vascular permeability.
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References
Landis EM (1926) The capillary pressure in frog mesentery as determined by micro-injection methods. Am J Phys 75:548–570
Landis EM (1927) Micro-injection studies of capillary permeability. II. The relation between capillary pressure and the rate at which fluid passes through the walls of single capillaries. Am J Phys 82:217–238
Michel CC, Mason JC, Curry FE, Tooke JE, Hunter PJ (1974) A development of the Landis technique for measuring the filtration coefficient of individual capillaries in the frog mesentery. Q J Exp Physiol Cogn Med Sci 59(4):283–309. https://doi.org/10.1113/expphysiol.1974.sp002275
Huxley VH, Curry FE, Adamson RH (1987) Quantitative fluorescence microscopy on single capillaries: alpha-lactalbumin transport. Am J Phys 252(1 Pt 2):H188–H197. https://doi.org/10.1152/ajpheart.1987.252.1.H188
Rumbaut RE, Huxley VH (2002) Similar permeability responses to nitric oxide synthase inhibitors of venules from three animal species. Microvasc Res 64(1):21–31. https://doi.org/10.1006/mvre.2002.2394
Yuan Y, Chilian WM, Granger HJ, Zawieja DC (1993) Permeability to albumin in isolated coronary venules. Am J Phys 265(2 Pt 2):H543–H552. https://doi.org/10.1152/ajpheart.1993.265.2.H543
Castorena-Gonzalez JA, Scallan JP, Davis MJ (2018) Methods for assessing the contractile function of mouse lymphatic vessels ex vivo. Methods Mol Biol 1846:229–248. https://doi.org/10.1007/978-1-4939-8712-2_15
Jannaway M, Scallan JP (2021) VE-cadherin and vesicles differentially regulate lymphatic vascular permeability to solutes of various sizes. Front Physiol 12:687563
Zawieja SD, Castorena-Gonzalez JA, Scallan JP, Davis MJ (2018) Differences in L-type Ca(2+) channel activity partially underlie the regional dichotomy in pum** behavior by murine peripheral and visceral lymphatic vessels. Am J Physiol Heart Circ Physiol 314(5):H991–H1010. https://doi.org/10.1152/ajpheart.00499.2017
To KHT, Gui P, Li M, Zawieja SD, Castorena-Gonzalez JA, Davis MJ (2020) T-type, but not L-type, voltage-gated calcium channels are dispensable for lymphatic pacemaking and spontaneous contractions. Sci Rep 10(1):70. https://doi.org/10.1038/s41598-019-56953-3
Scallan JP, Hill MA, Davis MJ (2015) Lymphatic vascular integrity is disrupted in type 2 diabetes due to impaired nitric oxide signalling. Cardiovasc Res 107(1):89–97. https://doi.org/10.1093/cvr/cvv117
Scallan JP, Huxley VH (2010) In vivo determination of collecting lymphatic vessel permeability to albumin: a role for lymphatics in exchange. J Physiol 588(Pt 1):243–254. https://doi.org/10.1113/jphysiol.2009.179622
Bingaman S, Huxley VH, Rumbaut RE (2003) Fluorescent dyes modify properties of proteins used in microvascular research. Microcirculation 10(2):221–231. https://doi.org/10.1038/sj.mn.7800186
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Jannaway, M., Scallan, J.P. (2024). Lymphatic Vascular Permeability Determined from Direct Measurements of Solute Flux. In: Tharakan, B. (eds) Vascular Hyperpermeability. Methods in Molecular Biology, vol 2711. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3429-5_3
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DOI: https://doi.org/10.1007/978-1-0716-3429-5_3
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