Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BK Ca channel regulates calcium oscillations induced by alpha-2-macroglobulin in human myometrial smooth muscle cells

doi: 10.1073/pnas.1516863113

Figure Lengend Snippet: Summary of Ca 2+ imaging experiments

Article Snippet: Some of the Ca 2+ recordings were performed in the presence of the BK Ca selective blocker paxilline (500 nM; Tocris Biosciences) or opener NS11021 (500 nM; Glixx Laboratories), which were added to the cells after the oscillations were observed.

Techniques: Imaging

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BK Ca channel regulates calcium oscillations induced by alpha-2-macroglobulin in human myometrial smooth muscle cells

doi: 10.1073/pnas.1516863113

Figure Lengend Snippet: Effect of BK Ca modulators on α 2 M*-induced Ca 2+ oscillations

Article Snippet: Some of the Ca 2+ recordings were performed in the presence of the BK Ca selective blocker paxilline (500 nM; Tocris Biosciences) or opener NS11021 (500 nM; Glixx Laboratories), which were added to the cells after the oscillations were observed.

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BK Ca channel regulates calcium oscillations induced by alpha-2-macroglobulin in human myometrial smooth muscle cells

doi: 10.1073/pnas.1516863113

Figure Lengend Snippet: BKCa channel modulates α2M*-induced Ca2+ oscillations. Representative live-cell Ca2+ imaging recordings of Fura-2-AM–loaded hMSMCs are shown. The specific BKCa blocker paxilline (Pax) (A and C, □) or opener NS11021 (B and D, ♢) was added after OXT (▲) and α2M* (●). (C and D) No α2M* was added.

Article Snippet: Some of the Ca 2+ recordings were performed in the presence of the BK Ca selective blocker paxilline (500 nM; Tocris Biosciences) or opener NS11021 (500 nM; Glixx Laboratories), which were added to the cells after the oscillations were observed.

Techniques: Imaging

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BK Ca channel regulates calcium oscillations induced by alpha-2-macroglobulin in human myometrial smooth muscle cells

doi: 10.1073/pnas.1516863113

Figure Lengend Snippet: Neither a BKCa channel blocker nor an opener changes Ca2+ levels in the presence of inactive α2M and OXT. Representative live-cell Ca2+ imaging recordings of Fura-2-AM–loaded hMSMCs. The specific BKCa blocker paxilline (A, Pax, □) or opener NS11021 (B, ♢) was added after OXT (▲) and inactive α2M (○). a.u., arbitrary units.

Article Snippet: Some of the Ca 2+ recordings were performed in the presence of the BK Ca selective blocker paxilline (500 nM; Tocris Biosciences) or opener NS11021 (500 nM; Glixx Laboratories), which were added to the cells after the oscillations were observed.

Techniques: Imaging

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BK Ca channel regulates calcium oscillations induced by alpha-2-macroglobulin in human myometrial smooth muscle cells

doi: 10.1073/pnas.1516863113

Figure Lengend Snippet: Working model of Ca2+ dynamics regulation by the functional interaction between BKCa and α2M*-LRP1 in hMSMCs. (A) OXT bound to its receptor induces production of IP3, which binds its receptor and leads to release of Ca2+ from the sarcoplasmic reticulum. (Insets) Release is observed as a peak in [Ca2+]i. (B) Release of Ca2+ from the sarcoplasmic reticulum activates SOCs. In the presence of α2M* bound to LRP1, SOC activation induces a moderate transient Ca2+ influx and activation of BKCa channels. BKCa channel activation leads to K+ efflux, which facilitates further influx of Ca2+ through SOCs. This hyperpolarizes the membrane, increasing the driving force for Ca2+ entry and inducing the oscillatory rise in [Ca2+]i, as shown in the Inset. (C) Blocking BKCa with paxilline prevents K+ efflux, so [Ca2+]i is maintained at low levels. (D) In the presence of the BKCa opener NS11021, K+ efflux is increased, thereby increasing the Ca2+ influx rate to maintain the electrochemical gradient. DAG, diacylglycerol; Gαq/11, α subunit of protein Gq/11; Gβγ, β and γ subunits of protein G; IP3R, IP3 receptor; OTR, OXT receptor; PIP2, phosphatidylinositol biphosphate; PLC, phospholipase C.

Article Snippet: Some of the Ca 2+ recordings were performed in the presence of the BK Ca selective blocker paxilline (500 nM; Tocris Biosciences) or opener NS11021 (500 nM; Glixx Laboratories), which were added to the cells after the oscillations were observed.

Techniques: Functional Assay, Activation Assay, Membrane, Blocking Assay

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Patch clamp studies on smooth muscle cells from rat corpus cavernosum. Increasing concentrations of NS11021 (10−8, 10−7, 10−6 M) stimulate membrane conductances which were then significantly inhibited by IbTX. (A) Typical experimental recordings from smooth muscle cell (membrane capacitance 17 pF) consequently stimulated with increasing concentrations of NS11021 and, then IbTX (0.1 µM). Lower insert in (A) shows voltage ramp protocol used in these experiments. (B) shows average (n= 5, Cm = 19 ± 1 pF) current–voltage recordings of IbTX-sensitive currents under control conditions and after cumulative increase of NS11021. The current–voltage curves achieved by subtraction of resting IbTX-insensitive current obtained at the end of each experiment. *, ***P < 0.05, 0.001 versus control IbTX-sensitive current by two-way anova.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques: Patch Clamp

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Patch clamp studies on smooth muscle cells from rat corpus cavernosum. Low concentrations (10−8 and 10−7 M) of NS11021 stimulate only IbTX-sensitive potassium current (see also Figure 1). Pretreatment with 0.1 µM IbTX inhibits initial of membrane conductances with low concentrations (10−8 and 10−7 M) of NS11021. A high concentration NS11021 (10−6 M) stimulates, however, another outward rectifying conductance which is sensitive to tetraethylammonium (TEA, 1 mM). (A) Representative experimental recordings (membrane capacitance 25 pF) and (B) averaged results (n= 5, Cm = 16 ± 3 pF) shown as current–voltage curves. ###P < 0.001 versus control current by two-way anova. ***P < 0.001 versus current in the presence of 10−6 M NS11021 by two-way anova.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques: Patch Clamp, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Average effect of NS11021 (10−6 M) on the current–voltage relationships of voltage-clamped human umbilical vein endothelial cells (HUVECs, n= 5).

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Effects of NS11021 on [Ca2+]i and tension in rat intracavernous artery. Effect of NS11021 in (A) phenylephrine (PE, 1 µM)- and (B) 125 mM K+-contracted arteries. (C) Average data showing the decrease in [Ca2+]i and tension in PE-and K-PSS-contracted preparations in response to cumulative administration of NS11021 (n= 4). *P < 0.05 by two-way anova.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Average NS11021 (A, B) and sildenafil (C, D) induced relaxation in phenylephrine (PE)- contracted and high potassium (125 mM K+)-contracted intracavernous artery (A, C, n= 4–6) and corpus cavernosum strips (B, D, n= 5–6) isolated from rat penis. *P < 0.05 by two-way anova.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques: Isolation

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Average effect of (A) NS11021, (B) sildenafil, and vehicle on electrical field stimulation-evoked neurogenic contractions in rat corpus cavernosum strips (n= 3–4).

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Average NS11021 (A, n= 4) and sildenafil (B, n= 3) induced relaxations in phenylephrine (PE, 0.5 µM)-contracted human corpus cavernosum strips in the presence and absence of iberiotoxin (IbTX, 0.1 µM). *P < 0.05 by two-way anova.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: (A) Original trace of mean arterial pressure and intra-cavernous pressure recordings obtained in male Wistar rat anaesthetized with pentobarbital. NS11021 (0.1 and 1 mg·kg−1) was administered (↓) after establishing a maximal (10 Hz, 1 ms, 6 V) and submaximal response (10 Hz, 1 ms, 0.6 to 1.55 V) to cavernous nerve stimulation for 30 seconds. Submaximal stimulation was repeated 3, 13 and 23 min after drug administration. Upper trace shows the mean arterial pressure (MAP), lower trace represents the intra-cavernous pressure (ICP). (B) Represent a magnification of an erectile response induced by electrical field stimulation (EFS) of the cavernous nerve. The plateau response considered for calculation of the peak intracavernous pressure (PICP) during the erectile response. Erectile response was calculated by PICP(mm Hg)/MAP(mm Hg) × 100.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: NS11021, a novel opener of large-conductance Ca 2+ -activated K + channels, enhances erectile responses in rats

doi: 10.1111/j.1476-5381.2009.00404.x

Figure Lengend Snippet: Average effect of vehicle (A), sildenafil (B) and NS11021 (C) on mean arterial pressure and erectile response obtained in male Wistar rats anaesthetized with pentobarbital. Drugs or vehicle were administered after establishing the maximal (10 Hz, 1 ms, 6 V) and submaximal responses (10 Hz, 1 ms, 0.6 to 1.55 V) to cavernous nerve stimulation for 30 s. Submaximal stimulation was repeated 3, 13 and 23 min after drug or vehicle administration. (D) Effect of intravenous tetraethylammonium (TEA, 6 mg·kg−1) pre-treatment on mean arterial pressure, erectile response and on effect of 1 mg·kg−1 NS11021. Upper traces show the mean arterial pressure (MAP), bars represent the erectile response ((PICP/MAP)*100), where PICP is the peak intracavernous pressure. *P < 0.05 compared with submaximal control response using one-way anova followed by Dunnett's test.

Article Snippet: The following drugs were used: NS11021 (NeuroSearch A/S, Ballerup, Denmark), sildenafil (gift from Pfizer Ltd.), ADMA, IbTX, phenylephrine hydrochloride, propranolol hydrochloride and tetraethylammonium chloride (Sigma, St. Louis, MO, USA).

Techniques: