Mechanisms of Connexin-Related Lymphedema

Jorge A. Castorena-Gonzalez, Scott D. Zawieja, Min Li, R. Sathish Srinivasan, Alex Simon, Cor de Wit, Roger de la Torre, Luis A. Martinez-Lemus, Grant W. Hennig, Michael J. Davis

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

RATIONALE: Mutations in GJC2 and GJA1, encoding Cxs (connexins) 47 and 43, respectively, are linked to lymphedema, but the underlying mechanisms are unknown. Because efficient lymph transport relies on the coordinated contractions of lymphatic muscle cells (LMCs) and their electrical coupling through Cxs, Cx-related lymphedema is proposed to result from dyssynchronous contractions of lymphatic vessels.

OBJECTIVE: To determine which Cx isoforms in LMCs and lymphatic endothelial cells are required for the entrainment of lymphatic contraction waves and efficient lymph transport.

METHODS AND RESULTS: We developed novel methods to quantify the spatiotemporal entrainment of lymphatic contraction waves and used optogenetic techniques to analyze calcium signaling within and between the LMC and the lymphatic endothelial cell layers. Genetic deletion of the major lymphatic endothelial cell Cxs (Cx43, Cx47, or Cx37) revealed that none were necessary for the synchronization of the global calcium events that triggered propagating contraction waves. We identified Cx45 in human and mouse LMCs as the critical Cx mediating the conduction of pacemaking signals and entrained contractions. Smooth muscle-specific Cx45 deficiency resulted in 10- to 18-fold reduction in conduction speed, partial-to-severe loss of contractile coordination, and impaired lymph pump function ex vivo and in vivo. Cx45 deficiency resulted in profound inhibition of lymph transport in vivo, but only under an imposed gravitational load.

CONCLUSIONS: Our results (1) identify Cx45 as the Cx isoform mediating the entrainment of the contraction waves in LMCs; (2) show that major endothelial Cxs are dispensable for the entrainment of contractions; (3) reveal a lack of coupling between lymphatic endothelial cells and LMCs, in contrast to arterioles; (4) point to lymphatic valve defects, rather than contraction dyssynchrony, as the mechanism underlying GJC2- or GJA1-related lymphedema; and (5) show that a gravitational load exacerbates lymphatic contractile defects in the intact mouse hindlimb, which is likely critical for the development of lymphedema in the adult mouse.

Original languageEnglish (US)
Pages (from-to)964-985
Number of pages22
JournalCirculation Research
Volume123
Issue number8
DOIs
StatePublished - Sep 28 2018

Fingerprint

Connexins
Lymphedema
Muscle Cells
Lymph
Endothelial Cells
Connexin 43
Protein Isoforms
Optogenetics
Lymphatic Vessels
Calcium Signaling
Arterioles
Hindlimb
Smooth Muscle
Calcium
Mutation

Keywords

  • calcium signaling
  • connexins
  • lymph
  • lymphatic system
  • lymphedema
  • vascular disease

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Castorena-Gonzalez, J. A., Zawieja, S. D., Li, M., Srinivasan, R. S., Simon, A., de Wit, C., ... Davis, M. J. (2018). Mechanisms of Connexin-Related Lymphedema. Circulation Research, 123(8), 964-985. https://doi.org/10.1161/CIRCRESAHA.117.312576

Mechanisms of Connexin-Related Lymphedema. / Castorena-Gonzalez, Jorge A.; Zawieja, Scott D.; Li, Min; Srinivasan, R. Sathish; Simon, Alex; de Wit, Cor; de la Torre, Roger; Martinez-Lemus, Luis A.; Hennig, Grant W.; Davis, Michael J.

In: Circulation Research, Vol. 123, No. 8, 28.09.2018, p. 964-985.

Research output: Contribution to journalArticle

Castorena-Gonzalez, JA, Zawieja, SD, Li, M, Srinivasan, RS, Simon, A, de Wit, C, de la Torre, R, Martinez-Lemus, LA, Hennig, GW & Davis, MJ 2018, 'Mechanisms of Connexin-Related Lymphedema', Circulation Research, vol. 123, no. 8, pp. 964-985. https://doi.org/10.1161/CIRCRESAHA.117.312576
Castorena-Gonzalez JA, Zawieja SD, Li M, Srinivasan RS, Simon A, de Wit C et al. Mechanisms of Connexin-Related Lymphedema. Circulation Research. 2018 Sep 28;123(8):964-985. https://doi.org/10.1161/CIRCRESAHA.117.312576
Castorena-Gonzalez, Jorge A. ; Zawieja, Scott D. ; Li, Min ; Srinivasan, R. Sathish ; Simon, Alex ; de Wit, Cor ; de la Torre, Roger ; Martinez-Lemus, Luis A. ; Hennig, Grant W. ; Davis, Michael J. / Mechanisms of Connexin-Related Lymphedema. In: Circulation Research. 2018 ; Vol. 123, No. 8. pp. 964-985.
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AU - Zawieja, Scott D.

AU - Li, Min

AU - Srinivasan, R. Sathish

AU - Simon, Alex

AU - de Wit, Cor

AU - de la Torre, Roger

AU - Martinez-Lemus, Luis A.

AU - Hennig, Grant W.

AU - Davis, Michael J.

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N2 - RATIONALE: Mutations in GJC2 and GJA1, encoding Cxs (connexins) 47 and 43, respectively, are linked to lymphedema, but the underlying mechanisms are unknown. Because efficient lymph transport relies on the coordinated contractions of lymphatic muscle cells (LMCs) and their electrical coupling through Cxs, Cx-related lymphedema is proposed to result from dyssynchronous contractions of lymphatic vessels.OBJECTIVE: To determine which Cx isoforms in LMCs and lymphatic endothelial cells are required for the entrainment of lymphatic contraction waves and efficient lymph transport.METHODS AND RESULTS: We developed novel methods to quantify the spatiotemporal entrainment of lymphatic contraction waves and used optogenetic techniques to analyze calcium signaling within and between the LMC and the lymphatic endothelial cell layers. Genetic deletion of the major lymphatic endothelial cell Cxs (Cx43, Cx47, or Cx37) revealed that none were necessary for the synchronization of the global calcium events that triggered propagating contraction waves. We identified Cx45 in human and mouse LMCs as the critical Cx mediating the conduction of pacemaking signals and entrained contractions. Smooth muscle-specific Cx45 deficiency resulted in 10- to 18-fold reduction in conduction speed, partial-to-severe loss of contractile coordination, and impaired lymph pump function ex vivo and in vivo. Cx45 deficiency resulted in profound inhibition of lymph transport in vivo, but only under an imposed gravitational load.CONCLUSIONS: Our results (1) identify Cx45 as the Cx isoform mediating the entrainment of the contraction waves in LMCs; (2) show that major endothelial Cxs are dispensable for the entrainment of contractions; (3) reveal a lack of coupling between lymphatic endothelial cells and LMCs, in contrast to arterioles; (4) point to lymphatic valve defects, rather than contraction dyssynchrony, as the mechanism underlying GJC2- or GJA1-related lymphedema; and (5) show that a gravitational load exacerbates lymphatic contractile defects in the intact mouse hindlimb, which is likely critical for the development of lymphedema in the adult mouse.

AB - RATIONALE: Mutations in GJC2 and GJA1, encoding Cxs (connexins) 47 and 43, respectively, are linked to lymphedema, but the underlying mechanisms are unknown. Because efficient lymph transport relies on the coordinated contractions of lymphatic muscle cells (LMCs) and their electrical coupling through Cxs, Cx-related lymphedema is proposed to result from dyssynchronous contractions of lymphatic vessels.OBJECTIVE: To determine which Cx isoforms in LMCs and lymphatic endothelial cells are required for the entrainment of lymphatic contraction waves and efficient lymph transport.METHODS AND RESULTS: We developed novel methods to quantify the spatiotemporal entrainment of lymphatic contraction waves and used optogenetic techniques to analyze calcium signaling within and between the LMC and the lymphatic endothelial cell layers. Genetic deletion of the major lymphatic endothelial cell Cxs (Cx43, Cx47, or Cx37) revealed that none were necessary for the synchronization of the global calcium events that triggered propagating contraction waves. We identified Cx45 in human and mouse LMCs as the critical Cx mediating the conduction of pacemaking signals and entrained contractions. Smooth muscle-specific Cx45 deficiency resulted in 10- to 18-fold reduction in conduction speed, partial-to-severe loss of contractile coordination, and impaired lymph pump function ex vivo and in vivo. Cx45 deficiency resulted in profound inhibition of lymph transport in vivo, but only under an imposed gravitational load.CONCLUSIONS: Our results (1) identify Cx45 as the Cx isoform mediating the entrainment of the contraction waves in LMCs; (2) show that major endothelial Cxs are dispensable for the entrainment of contractions; (3) reveal a lack of coupling between lymphatic endothelial cells and LMCs, in contrast to arterioles; (4) point to lymphatic valve defects, rather than contraction dyssynchrony, as the mechanism underlying GJC2- or GJA1-related lymphedema; and (5) show that a gravitational load exacerbates lymphatic contractile defects in the intact mouse hindlimb, which is likely critical for the development of lymphedema in the adult mouse.

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