(A) RT-PCR showing the absence of PKD2 transcript in isolated myocytes from tamoxifen-treated Pkd2fl/fl:myh11-cre/ERT2 mice. (B) Western blots illustrating the effect of tamoxifen-treatment in Pkd2fl…
Ethidium bromide gel illustrating PCR products in vasculature of C57BL/6J (WT) mice and tamoxifen-treated Pkd2fl/fl and Pkd2fl/fl:myh11cre/ERT2 mice.
(A) Western blots illustrating PKD2 protein was lower in mesenteric arteries of tamoxifen-treated Pkd2fl/fl:myh11-cre/ERT2 mice, whereas other proteins were similar. (B) Mean data for proteins in …
(A) Western blots illustrating Angiotensin II type one receptor (AT1R), Piezo1, α1-adrenergic receptor A (α1A), α1-adrenergic receptor B (α1B), α1-adrenergic receptor D (α1D) and G protein-coupled …
(A) Original telemetric blood pressure recordings from Pkd2 smKO and Pkd2fl/fl mice. (B) Mean systolic and diastolic blood pressures in Pkd2fl/fl (n = 11) and Pkd2 smKO (n = 12) mice. * indicates p<0…
(A) Mean arterial blood pressure (MAP) in Pkd2 smKO and Pkd2fl/fl mice (n = 6 per group). * indicates p<0.05 versus Pkd2fl/fl. (B) Mean data of locomotor activity, n = 6 per group.
(A) Representative traces illustrating diameter responses to intravascular pressure in gastrocnemius arteries of Pkd2fl/fl and Pkd2 smKO mice. (B) Mean data for myogenic tone in gastrocnemius …
(A) Mean passive diameter at 80 mmHg of first-order gastrocnemius arteries (G) and third-, fourth-and fifth-order mesenteric arteries (M) (Pkd2fl/fl: G, n = 5; M3rdn = 4; M4thn = 5; M5thn = 5 and Pkd…
(A) Representative traces of microelectrode impalements under indicated conditions illustrating that pressure-induced depolarization is attenuated in gastrocnemius arteries of Pkd2 smKO mice. …
(A) Representative I-V relationships recorded between −100 and +100 mV in the same hindlimb artery myocytes of Pkd2fl/fl or Pkd2 smKO mice in control and PE (10 µM). (B) Mean data for current …
(A) Mean vasoconstriction over a range of pressures in resistance-size mesenteric arteries from Pkd2fl/fl (n = 7) and Pkd2 smKO (n = 9) mice. (B) Original recordings of concentration-dependent, …
(A) Mean myogenic tone at 80 mmHg illustrating that myogenic tone is similar in third-, fourth-and fifth-order mesenteric arteries and unaltered by PKD2 knockout (Pkd2fl/fl: 3rd n = 4; 4th n = 5; 5th…
(A) Representative traces of microelectrode impalements illustrating that phenylephrine (PE, 1 µM)-induced depolarization is attenuated in mesenteric arteries of Pkd2 smKO mice. Scale bars: Y = 10 …
(A) Representative I-V relationships from the same isolated mesenteric artery myocytes of Pkd2fl/fl or Pkd2 smKO mice in isosmosmotic (300 mOsm), hyposmotic (250 mOsm) and hyposmotic (250 mOsm) + Gd3…
(A) Telemetric blood pressure time course showing the development of angiotensin II-induced hypertension in Pkd2fl/fl (n = 6) and Pkd2 smKO mice (n = 9). Osmotic minipumps containing either saline …
(A) Mean phenylephrine-induced vasoconstriction in pressurized (80 mmHg) mesenteric arteries from angiotensin II-treated mice (Pkd2fl/fl, n = 7–8; Pkd2 smKO, n = 8).
Pkd2fl/fl | Pkd2 smKO | |
---|---|---|
Plasma hormones (pg/ml) | ||
Angiotensin II | 202.5 ± 17.2 (n = 16) | 204.7 ± 12.1 (n = 15) |
Aldosterone | 341.0 ± 18.2 (n = 16) | 365.6 ± 14.0 (n = 10) |
ANP | 107.2 ± 9.4 (n = 18) | 118.3 ± 12.3 (n = 18) |
Plasma electrolytes (mM) | ||
Na+ | 142.3 ± 1.0 (n = 7) | 152.0 ± 5.5 (n = 6) |
K+ | 6.4 ± 0.4 (n = 7) | 6.8 ± 0.3 (n = 6) |
Cl- | 78.8 ± 0.7 (n = 7) | 84.1 ± 4.3 (n = 6) |
Urine electrolytes (mM) | ||
Na+ | 136.9 ± 9.3 (n = 15) | 147.0 ± 16.2 (n = 15) |
K+ | 563.2 ± 30.2 (n = 15) | 548.6 ± 37.6 (n = 15) |
Cl- | 455.4 ± 24.7 (n = 15) | 479.1 ± 42.0 (n = 15) |
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus) | Pkd2fl/fl | John Hopkins PKD Core | PMID 20862291 | Mice with Pkd2 gene flanked by loxP regions |
Strain, strain background (M. musculus) | Myh11-cre/ERT2 | Jackson Laboratories. PMID 18084302 | Stock # 019079 RRID:IMSR_JAX:019079 | Mice with tamoxifen-inducible Cre recombinase that is expressed under the smooth muscle myosin heavy polipeptide 11 (Myh11) promoter. |
Strain, strain background (M. musculus) | Pkd2fl/fl:Myh11-cre/ERT2 | This paper | Mouse line created in-house by mating Pkd2fl/fl with Myh11-cre/ERT2. Mice with inducible smooth muscle-specific deletion of PKD2. | |
Antibody | anti PKD2 (rabbit polyclonal) | Baltimore PKD Core | Rabbit mAB 3374 CT-14/4, | IF 1:200 dilution |
Antibody | anti PKD2 (mouse monoclonal) | Santa Cruz | Cat. # sc-100415 RRID:AB_1127284 | WB 1:100 dilution, IF 1:100 dilution |
Antibody | anti PKD1 (mouse monoclonal) | Santa Cruz | Cat. # sc-130554 RRID:AB_2163355 | WB 1:100 dilution |
Antibody | anti CaV1.2 (mouse monoclonal) | UC Davis/NIH NeuroMab | Cat# 73–053. RRID:AB_10672290 | WB 1:100 dilution |
Antibody | anti ANO1 (rabbit monoclonal) | Abcam | Cat. # ab64085 RRID:AB_1143505 | WB 1:100 dilution |
Antibody | anti TRPC6 (rabbit polyclonal) | Abcam | Cat. # ab62461 RRID:AB_956401 | WB 1:1000 dilution |
Antibody | anti TRPM4 (rabbit polyclonal) | Abcam | Cat. # ab106200 RRID:AB_10865760 | WB 1:500 dilution |
Antibody | anti Actin (mouse monoclonal) | Millipore Sigma | Cat. # MAB1501 RRID:AB_2223041 | WB 1:5000 dilution |
Antibody | Alexa 555 secondary antibodies (anti rabbit and anti mouse) | Thermofisher | Cat. # A-21429 (RRID:AB_141761) and # A-31570 (RRID:AB_2536180) | IF 1:400 dilution |
Other | Nuclear staining (DAPI) | Thermofisher | Cat. # 3571 RRID:AB_2307445 | IF 1:1000 dilution |
Commercial assay or kit | EZ-Link Sulfo-NHS -LC-LC-Biotin | Thermofisher | Cat. # 21338 | |
Commercial assay or kit | EZ-Link Maleimide-PEG2-Biotin | Thermofisher | Cat. # 21901BID | |
Commercial assay or kit | Mouse Angiotensin II ELISA kit | Elabscience | Cat. # E-EL-M2612 | |
Commercial assay or kit | Mouse Atrial Natriuretic Peptide ELISA kit | Elabscience | Cat. # E-EL-M0166 | |
Commercial assay or kit | Mouse Aldosterone ELISA kit | Mybiosource | Cat. # MBS775626 | |
Chemical compound, drug | Angiotensin II | Sigma-Aldrich | Cat. # A9525 |
Primers used for RT PCR.
The PKD2 forward primer recognizes nucleotides in exon 9 and 10 and the reverse primer was aligned with a sequence in exon 13.