Pre-collecting lymphatic vessels form detours following obstruction of lymphatic flow and function as collecting lymphatic vessels

Autoři: Kimi Asano aff001;  Yukari Nakajima aff003;  Kanae Mukai aff003;  Tamae Urai aff004;  Mayumi Okuwa aff003;  Junko Sugama aff005;  Chizuko Konya aff006;  Toshio Nakatani aff003
Působiště autorů: Department of Clinical Nursing, Graduate Course of Nursing Science, Division of Health Sciences, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan aff001;  School of Nursing, Kanazawa Medical University, Uchinada, Japan aff002;  Faculty of Health Sciences, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, Japan aff003;  Faculty of Nursing, Toyama Prefectural University, Toyama, Japan aff004;  Advanced Health Care Science Research Unit, Innovative Integrated Bio-Research Core, Institute for Frontier Science Initiative, Kanazawa University, Kanazawa, Japan aff005;  Faculty of Nursing, Ishikawa Prefectural Nursing University, Kahoku, Japan aff006
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article



Previously, we showed that lymphatic vessels (LVs) formed detours after lymphatic obstruction, contributing to preventing lymphedema. In this study, we developed detours using lymphatic ligation in mice and we identified the detours histologically.

Methods and results

Under anesthesia, both hindlimbs in mice were subcutaneously injected with Evans blue dye to detect LVs. We tied the right collecting LV on the abdomen that passes through the inguinal lymph node (LN) at two points. The right and left sides comprised the operation and sham operation sides, respectively. Lymphography was performed to investigate the lymph flow after lymphatic ligation until day 30, using a near-infrared fluorescence imaging system. Anti-podoplanin antibody and 5-ethynyl-2’-deoxyuridine (EdU) were used to detect LVs and lymphangiogenesis. Within 30 days, detours had developed in 62.5% of the mice. Detours observed between two ligation sites were enlarged and irregular in shape. Podoplanin+ LVs, which were located in the subcutaneous tissue of the upper panniculus carnosus muscle, connected to collecting LVs at the upper portion from the cranial ligation site and at the lower portion from the caudal ligation site. EdU+ cells were not observed in these detours. The sham operation side showed normal lymph flow and did not show enlarged pre-collecting LVs until day 30.


Detours after lymphatic ligation were formed not by lymphangiogenesis but through an enlargement of pre-collecting LVs that functioned as collecting LVs after lymphatic ligation. Further studies are required to explore the developmental mechanism of the lymphatic detour for treatment and effective care of lymphedema in humans.

Klíčová slova:

Abdomen – Histology – Immunostaining – Lymph – Lymphedema – Muscle tissue – Surgical and invasive medical procedures – Subcutaneous tissue


1. Executive Committee. The diagnosis and treatment of peripheral lymphedema: 2016 consensus document of the International Society of Lymphology. Lymphology. 2016;49: 170–184. 29908550

2. Taghian NR, Miller CL, Jammallo LS, O’Toole J, Skolny MN. Lymphedema following breast cancer treatment and impact on quality of life: a review. Crit Rev Oncol Hematol. 2014;92: 227–234. doi: 10.1016/j.critrevonc.2014.06.004 Epub 2014 Jul 2. 25085806

3. Moshiri M, Katz DS, Boris M, Yung E. Using lymphoscintigraphy to evaluate suspected lymphedema of the extremities. Am J Roentgenol. 2002;178: 405–412. doi: 10.2214/ajr.178.2.1780405 11804905

4. Lohrmann C, Foeldi E, Speck O, Langer M. High-resolution MR lymphangiography in patients with primary and secondary lymphedema. AJR Am J Roentgenol. 2006;187: 556–561. doi: 10.2214/AJR.05.1750 16861563

5. Mikami T, Hosono M, Yabuki Y, Yamamoto Y, Yasumura K, Sawada H, et al. Classification of lymphoscintigraphy and relevance to surgical indication for lymphaticovenous anastomosis in upper limb lymphedema. Lymphology. 2011;44: 155–167. 22458117

6. Lu Q, Delproposto Z, Hu A, Tran C, Liu N, Li Y, et al. MR lymphography of lymphatic vessels in lower extremity with gynecologic oncology-related lymphedema. PLoS One. 2012;7: e50319. doi: 10.1371/journal.pone.0050319 Epub 2012 Nov 28. 23209708

7. Szuba A, Chacaj A, Koba-Wszedybyl M, Hawro R, Jasinski R, Tarkowski R, et al. Upper extremity lymphedema after axillary lymph node dissection: prospective lymphoscintigraphic evaluation. Lymphology. 2016;49: 44–56. 29906360

8. Neligan PC, Kung TA, Maki JH. MR lymphangiography in the treatment of lymphedema. J Surg Oncol. 2017;115: 18–22. doi: 10.1002/jso.24337 Epub 2016 Jul 4. Review. 27377990

9. Sarri AJ, Tinois da Silva E, Vieira RADC, Koga KH, Cação PHM, Sarri VC, et al. Lymphoscintigraphy detecting alterations of upper limb lymphatic flow following early sentinel lymph node biopsy in breast cancer. Breast Cancer (Dove Med Press). 2017;9: 279–285. doi: 10.2147/BCTT.S131407 eCollection 2017. 28458580

10. Abe R. A study on the pathogenesis of postmastectomy lymphedema. Tohoku J Exp Med. 1976;118: 163–171. doi: 10.1620/tjem.118.163 982430

11. Maegawa J, Mikami T, Yamamoto Y, Satake T, Kobayashi S. Types of lymphoscintigraphy and indications for lymphaticovenous anastomosis. Microsurgery. 2010;30: 437–442. doi: 10.1002/micr.20772 20878726

12. Pecking AP, Albérini JL, Wartski M, Edeline V, Cluzan RV. Relationship between lymphoscintigraphy and clinical findings in lower limb lymphedema (LO): toward a comprehensive staging. Lymphology. 2008;41: 1–10. 18581953

13. Ikomi F, Yokoyama Y, Ogiwara N, Sasaki K, Mizuno R, Ohhashi T. Recanalization of the collecting lymphatics in rabbit hind leg. Microcirculation. 2006;13: 365–376. doi: 10.1080/10739680600745810 16815822

14. Blum KS, Proulx ST, Luciani P, Leroux JC, Detmar M. Dynamics of lymphatic regeneration and flow patterns after lymph node dissection. Breast Cancer Res Treat. 2013;139: 81–86. doi: 10.1007/s10549-013-2537-7 Epub 2013Apr 24. 23613202

15. Takeno Y, Fujimoto E. Alterations of lymph flow after lymphadenectomy in rats revealed by real time fluorescence imaging system. Lymphology. 2013;46: 12–19. 23930437

16. Takeno Y, Arita H, Oshima C, Mawaki A, Nakanishi K, Kurono F, et al. Long-term follow-up of collateral pathways established after lymphadenectomy in rats. Lymphology. 2017;50: 131–135. 30234249

17. Suami H, Yamashita S, Soto-Miranda MA, Chang DW. Lymphatic territories (lymphosomes) in a canine: an animal model for investigation of postoperative lymphatic alterations. PLoS One. 2013;8: e69222. doi: 10.1371/journal.pone.0069222 Print 2013. 23894435

18. Suami H, Schaverien MV. Swine hind limb model for supermicrosurgical lymphaticovenular anastomosis training. J Plast Reconstr Aesthet Surg. 2016;69: 723–725. doi: 10.1016/j.bjps.2016.01.002 Epub 2016 Jan 18. 26826844

19. Kwon S, Price RE. Characterization of internodal collecting lymphatic vessel function after surgical removal of an axillary lymph node in mice. Biomed Opt Express. 2016;7: 1100–1115. doi: 10.1364/BOE.7.001100 eCollection 2016 Apr 1. 27446639

20. Komatsu E, Nakajima Y, Mukai K, Urai T, Asano K, Okuwa M, et al. Lymph drainage during wound healing in a hindlimb lymphedema mouse model. Lymphat Res Biol. 2017;15: 32–38. doi: 10.1089/lrb.2016.0026 Epub 2017 Feb 2. 28151088

21. Nakajima Y, Asano K, Mukai K, Urai T, Okuwa M, Sugama J, et al. Near-infrared fluorescence imaging directly visualizes lymphatic drainage pathways and connections between superficial and deep lymphatic systems in the mouse hindlimb. Sci Rep. 2018;8: 7078. doi: 10.1038/s41598-018-25383-y 29728629

22. Yamaji Y, Akita S, Akita H, Miura N, Gomi M, Manabe I, et al. Development of a mouse model for the visual and quantitative assessment of lymphatic trafficking and function by in vivo imaging. Sci Rep. 2018;8: 5921. doi: 10.1038/s41598-018-23693-9 29651026

23. Alitalo K, Tammela T, Petrova TV. Lymphangiogenesis in development and human disease. Nature. 2005;438: 946–953. doi: 10.1038/nature04480 16355212

24. Norrmén C, Tammela T, Petrova TV, Alitalo K. Biological basis of therapeutic lymphangiogenesis. Circulation. 2011;123: 1335–1351. doi: 10.1161/CIRCULATIONAHA.107.704098 21444892

25. Suami H, Pan WR, Taylor GI. Changes in the lymph structure of the upper limb after axillary dissection: radiographic and anatomical study in a human cadaver. Plast Reconstr Surg. 2007;120: 982–991. doi: 10.1097/01.prs.0000277995.25009.3e 17805128

26. Shimamura K, Nakatani T, Ueda A, Sugama J, Okuwa M. Relationship between lymphangiogenesis and exudates during the wound-healing process of mouse skin full-thickness wound. Wound Repair Regen. 2009;17: 598–605. doi: 10.1111/j.1524-475X.2009.00512.x 19614925

27. Tammela T, Saaristo A, Holopainen T, Lyytikkä J, Kotronen A, Pitkonen M, et al. Therapeutic differentiation and maturation of lymphatic vessels after lymph node dissection and transplantation. Nat Med. 2007;13: 1458–1466. Epub 2007 Dec 2. doi: 10.1038/nm1689 18059280

28. Limsirichaikul S, Niimi A, Fawcett H, Lehmann A, Yamashita S, Ogi T. A rapid non-radioactive technique for measurement of repair synthesis in primary human fibroblasts by incorporation of ethynyl deoxyuridine (EdU). Nucleic Acids Res. 2009;37: e31. 19179371

29. Breiteneder-Geleff S, Soleiman A, Kowalski H, Horvat R, Amann G, Kriehuber E, et al. Angiosarcomas express mixed endothelial phenotypes of blood and lymphatic capillaries: podoplanin as a specific marker for lymphatic endothelium. Am J Pathol. 1999;154: 385–394. doi: 10.1016/S0002-9440(10)65285-6 10027397

30. Norrmén C, Ivanov K, Cheng J, Zangger N, Delorenzi M, Jaquet M, et al. FOXC2 controls formation and maturation of lymphatic collecting vessels through cooperation with NFATc1. J Cell Biol. 2009;185: 439–457. doi: 10.1083/jcb.200901104 19398761

31. Bouta EM, Wood RW, Brown EB, Rahimi H, Ritchlin CT, Schwarz EM. In vivo quantification of lymph viscosity and pressure in lymphatic vessels and draining lymph nodes of arthritic joints in mice. J Physiol. 2014;592: 1213–1223. doi: 10.1113/jphysiol.2013.266700 Epub 2014 Jan 13. 24421350

32. Kretz O, Kubik S. Anatomy of the lymphatics system and the terminal vascular bed. In: Földi M, Földi M, editors. Földi’s textbook of lymphology for physicians and lymphedema therapists. New York: Urban & Fischer; 2012. pp. 120–121.

33. Sacchi G, Weber E, Aglianò M, Raffaelli N, Comparini L. The structure of superficial lymphatics in the human thigh: precollectors. Anat Rec. 1997;247: 53–62. doi: 10.1002/(SICI)1097-0185(199701)247:1<53::AID-AR8>3.0.CO;2-G 8986303

34. Lonza Walkersville Inc. Certificate of analysis. Walkersville: Lonza Walkersville Inc.; 2018:1. Accessed September 5, 2018.

Článek vyšel v časopise


2020 Číslo 1
Nejčtenější tento týden