Effects of Elevated Downstream Pressure and the Role of Smooth Muscle Cell Coupling through Connexin45 on Lymphatic Pacemaking
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Approval
2.2. Mice and Tamoxifen-Dependent Cre-Recombinase
2.3. Solutions and Chemicals
2.4. In Vivo Recording of Contractions of Popliteal Lymphatic Vessels
2.5. Vessel Isolation, Pressure Myography, and Data Acquisition
2.6. Assessment of Contractile Function of Isolated Lymphatic Vessels
2.7. Assessment of the Initiation and Propagation of Contraction Waves using Space Time Maps (STMs)
2.8. Calculation of Contractile and Pacemaking Parameters
2.9. Fluorescence Activated Cell Sorting (FACS) and End-point RT-PCR
2.10. Statistical Analysis
3. Results
3.1. In Vivo and Ex Vivo Assessement of the Initiation Sites of Spontaneous Contractions and Measurement of the Direction and Speed of the Associated Propagating Contraction Wave in Lymphatic Vessels
3.2. Lymphatic Contractions Display a Higher Probability of Initiating Downstream in the Lymphatic Network
3.3. Efficient Entrainment of Lymphatic Contractions Requires a Dominant Pacemaker and Strong Electrical Coupling between LMCs through Connexin 45-Containing Gap Junctions
3.4. Lymphatic Pacemaking can be Regulated by Localized Changes in Intraluminal Pressure
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Accession No. | Protein | Strand | Sequence | Amplicon (bp) |
---|---|---|---|---|
NM_080454 | Cx47 | s | AGC TCT GCC TTG TGC ATC TC | 216 |
as | CGT GTT GCA GGT GAA CTT GG | |||
NM_175452 | Cx47 | s | GAG AGG ATC AGC ATC CAG CC | 262 |
as | CGT GTT GCA GGT GAA CTT GG | |||
NM_008122 | Cx45 | s | GGT AAC AGG AGT TCT GGT GAA | 140 |
as | TCG AAA GAC AAT CAG CAC AGT | |||
NM_010288 | Cx43 | s | TGA GAG CCC GAA CTC TCC TT | 258 |
as | AGG CAG ACT GTT CAT CAC CC | |||
NM_008121 | Cx40 | s | CCA GAG CCT GAA GAA GCC AA | 143 |
NM_001271628 | as | CCG ATG ACT GTG GAG TGC TT | 200 | |
NM_008120 | Cx37 | s | GCT GCG CGC TAT TTA AGG C | 131 |
as | CAT GTT TCC AGG GCC TCT CT | |||
NM_001302497 | Cx32 | s | CAT GAG ACC ATA GGG GAG CTG | 291 |
as | ACG TGG GAG ATG GGG AAA AA | |||
NM_178596 | Cx30.2 | s | CGT CAT CTA CTC CAT GCA CCA | 316 |
as | GAC GGC GAA GTA GAA GAC CAC | |||
NM_001010937 | Cx30 | s | GAT CCC AAC GAG TGC CCT AAT | 430 |
as | CTG GAC ATC AGC AGC GGT AG | |||
NM_008125 | Cx26 | s | CAT TTC GGA CCA ACC CAG GA | 148 |
as | TGC CCC AAT CCA TCT TGT CC | |||
NM_008937 | Prox1 | s | GTA AGA CAT CAC CGC GTG C | 218 |
as | TCA TGG TCA GGC ATC ACT GG | |||
NM_009868 | VE-Cadherin | s | CTT CCT TAC TGC CCT CAT TGT | 313 |
as | CTG TTT CTC TCG GTC CAA GTT | |||
NM_008713 | e-NOS | s | CTG CCA CCT GAT CCT AAC TTG | 143 |
as | CAG CCA AAC ACC AAA GTC ATG | |||
NM_007392 | α-Actin | s | GAG CTA CGA ACT GCC TGA C | 129 |
as | CTG TTA TAG GTG GTT TCG TGG A | |||
NM_013607 | Myh11 | s | AAG CTG CGG CTA GAG GTC A | 238 |
as | CCC TCC CTT TGA TGG CTG AG |
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Castorena-Gonzalez, J.A.; Li, M.; Davis, M.J. Effects of Elevated Downstream Pressure and the Role of Smooth Muscle Cell Coupling through Connexin45 on Lymphatic Pacemaking. Biomolecules 2020, 10, 1424. https://doi.org/10.3390/biom10101424
Castorena-Gonzalez JA, Li M, Davis MJ. Effects of Elevated Downstream Pressure and the Role of Smooth Muscle Cell Coupling through Connexin45 on Lymphatic Pacemaking. Biomolecules. 2020; 10(10):1424. https://doi.org/10.3390/biom10101424
Chicago/Turabian StyleCastorena-Gonzalez, Jorge A., Min Li, and Michael J. Davis. 2020. "Effects of Elevated Downstream Pressure and the Role of Smooth Muscle Cell Coupling through Connexin45 on Lymphatic Pacemaking" Biomolecules 10, no. 10: 1424. https://doi.org/10.3390/biom10101424
APA StyleCastorena-Gonzalez, J. A., Li, M., & Davis, M. J. (2020). Effects of Elevated Downstream Pressure and the Role of Smooth Muscle Cell Coupling through Connexin45 on Lymphatic Pacemaking. Biomolecules, 10(10), 1424. https://doi.org/10.3390/biom10101424