Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Furosemide (Furo) effect on urine pH. Urine pH of wild-type (WT; A) and β4-knockout (β4-KO; B) on day −1, 1, 4, 7, and 11 of control (Ctrl) and Furo treatments or on day −1, 1, 4, and 7 of Furo + acetazolamide (Actz) treatment. *P < 0.05 vs. HK + Ctrl water, #P < 0.05 vs. Day –1, analyzed with two-way repeated-measures ANOVA with a post hoc Holm-Sidak test (P < 0.001 for both treatment and days with interaction P < 0.01). n = 12–14 for WT Ctrl and Furo; N = 7 – 8 for β4-KO Ctrl and Furo; N = 4 for WT Furo + Actz; n = 3 for β4-KO Furo + Actz.

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques: Knock-Out

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Blood measurements after 11 days (or 7 days for HK + Furo Actz) of treatment

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques:

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Furosemide (Furo) effect on plasma [K+] and renal K+ clearance in mice on regular diet (RD) and high-K+ diet (HK). A and B: plasma [K+] and renal K+ clearance of WT on RD on day 11 of control (Ctrl) or Furo treatments. *P < 0.05 vs. RD + Ctrl analyzed with Student’s t-test; n = 4 for Ctrl, n = 6 for Furo. C and D: plasma [K+] and renal K+ clearance of WT and β4-knockout (β4-KO) on day 11 of Ctrl and Furo treatment or day 7 of Furo + acetazolamide (Actz) treatment. *P < 0.05 vs. HK + Ctrl; #P < 0.05 vs. HK + Furo, analyzed with two-way ANOVA with a post hoc Tukey test (P < 0.001 for treatment in plasma [K+]; P < 0.01 for treatment in renal K+ clearance); n = 12 – 14 for WT HK + Ctrl and HK + Furo; n = 7 – 8 for β4-KO HK + Ctrl and HK + Furo; n = 4 for WT HK + Furo Actz; n = 3 for β4-KO HK + Furo Actz.

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques: Knock-Out

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Furosemide (Furo) effect on urinary K+ excretion (UKV̇) normalized to kidney weight in mice on high-K diet (HK). UKV̇ of wild-type (WT; A) and β4-knockout (β4-KO; B) on day −1, 1, 4, 7, and 11 of control (Ctrl) and Furo treatment or on day −1, 1, 4, and 7 of Furo + acetazolamide (Actz) treatment. *P < 0.05 vs. HK + Ctrl; #P < 0.05 vs. day −1, analyzed with two-way repeated measures ANOVA (P < 0.05 for drug treatment in WT) with a post hoc Holm-Sidak test; n = 12–14 for WT HK + Ctrl and HK + Furo; n = 7–8 for β4-KO HK + Ctrl and HK + Furo; n = 4 for WT HK + Furo Actz; n = 3 for β4-KO HK + Furo Actz.

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques: Knock-Out

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Metabolic cage measurements after 11 days (or 7 days for HK + Furo Actz) of treatment

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques:

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Correlation between renal K+ clearance and urine pH in wild-type (WT; A) and β4-knockout (β4-KO; B) on high-K+ diet (HK). ●, HK + control (Ctrl) group; ○, HK + furosemide (Furo) group; ▲, HK + Furo acetazolamide (Actz) group; △, HKCl group.

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques: Knock-Out

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Furosemide (Furo) effect on large conductance Ca2+-activated K+ (BK)-β4 expression in the kidney. A: BK-β4 expression in the kidney medulla of wild-type (WT) on high-K diet (HK) treated with control (Ctrl) or Furo water; n = 4/group. B: BK-β4 expression in the kidney cortex of WT on HK treated with Ctrl vs. Furo water. *P < 0.05 vs. HK + Ctrl analyzed with Student’s t-test; n = 7/group. C: BK-β4 expression in the kidney cortex of WT on HK treated with Ctrl vs. Furo acetazolamide (Actz) water; n = 4/group.

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques: Expressing

Journal: American Journal of Physiology - Renal Physiology

Article Title: Furosemide reduces BK-αβ4-mediated K + secretion in mice on an alkaline high-K + diet

doi: 10.1152/ajprenal.00223.2018

Figure Lengend Snippet: Furosemide concentrations in urine and plasma of WT and BK-β4-KO

Article Snippet: Primary antibodies included anti-BK-β4 (rabbit polyclonal, diluted 1:500; Alomone Laboratories, Jerusalem, Israel), anti-NHE3 (mouse monoclonal, diluted 1:200; Invitrogen, Carlsbad, CA), and anti-V-ATPase B1 (rabbit polyclonal, diluted 1:200; GeneTex, Irvine, CA).

Techniques:

Journal: Nature neuroscience

Article Title: BK channels modulate pre- and postsynaptic signaling at reciprocal synapses in retina

doi: 10.1038/nn.2302

Figure Lengend Snippet: A17s varicosities express rapidly inactivating BK channels that are functionally coupled to L-type VGCCs. ( a ) Depolarizing voltage steps (from −90 to −30mV; 100ms) in an A17 elicited a rapidly inactivating outward current (with potassium-based internal solution) that was blocked by iberiotoxin (100 nM) ( b ) Paired depolarizing steps delivered at varying intervals revealed the time course of recovery from inactivation. ( c ) The transient outward current also was blocked by the L-type VGCC blocker, isradipine (10 µM). ( d ) Immunohistochemical techniques reveal BK (red) channel expression throughout the inner plexiform layer of a slice. Anti-PKCα was used to label RBCs (blue) and the dendrites of a single A17 were filled with neurobiotin (green) through the patch pipette. Scale bar = 10 µm. ( e,f ) A higher-magnification view of sublamina 5 in the IPL indicates that BK ( e ) and β2 subunit ( f ) puncta are localized to A17 varicosities that are adjacent to RBC terminals. Scale bars = 5 µm. ( g,h ) Antibody staining for PKC (blue) and BK (red, g ) or β2 (red, h ) in whole-mount retina illustrates the clustering of channels antibody around RBC terminals. Scale bars = 2 µm.

Article Snippet: Slices were then incubated for two hours in the standard solution containing an antibody to PKC-α (mouse, Santa Cruz Biotechnology, Santa Cruz, CA; 1:100) and an antibody to BK or BK β2 subunit (rabbit, Alomone, Jerusalem, Israel; 1:100).

Techniques: Immunohistochemical staining, Expressing, Transferring, Staining