Structured Review

Biochrom mmol ∙ l 1 glutamine
Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .
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Images

1) Product Images from "The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+"

Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

Journal: PLoS ONE

doi: 10.1371/journal.pone.0193519

Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .
Figure Legend Snippet: Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

Techniques Used: Expressing, Blocking Assay, Activation Assay, Permeability

Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na +  and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1  CaCl 2  bath solution. Ca 2+  had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+  d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na +  of 132 ± 4 pS, for Ca 2+  of 21 ± 3 pS, and for NMDG +  of 36 ± 3 pS. The conductance for NH 4 +  was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+  in the bath.
Figure Legend Snippet: Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

Techniques Used: Transferring, Blocking Assay

Effects of menthol and Mg 2+  on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i  in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i  (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i  in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i  in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i  /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p
Figure Legend Snippet: Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

Techniques Used:

Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l  −1  MgCl 2 , c) Na-gluconate solution without MgCl 2  and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).
Figure Legend Snippet: Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

Techniques Used: Expressing, Transferring

2) Product Images from "The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+"

Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

Journal: PLoS ONE

doi: 10.1371/journal.pone.0193519

Effects of menthol and Mg 2+  on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i  in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i  (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i  in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i  in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i  /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p
Figure Legend Snippet: Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

Techniques Used:

Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na +  and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1  CaCl 2  bath solution. Ca 2+  had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+  d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na +  of 132 ± 4 pS, for Ca 2+  of 21 ± 3 pS, and for NMDG +  of 36 ± 3 pS. The conductance for NH 4 +  was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+  in the bath.
Figure Legend Snippet: Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

Techniques Used: Transferring, Blocking Assay

Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .
Figure Legend Snippet: Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

Techniques Used: Expressing, Blocking Assay, Activation Assay, Permeability

Response to various agonists of bTRPV3. Cells were filled with a CsCl pipette solution and stimulated with the continuous pulse protocol I. a) Original recording of a cell expressing bTRPV3, showing the response to application of menthol, thymol (both 1 mmol∙l −1 ), and 2-APB (0.3 mmol∙l −1 ) in NaCl bath solution. In response to 2-APB, the current rose dramatically, requiring an interruption of the measurement to readjust the gain (arrow). b) Original recording of a control cell transfected with the empty vector.
Figure Legend Snippet: Response to various agonists of bTRPV3. Cells were filled with a CsCl pipette solution and stimulated with the continuous pulse protocol I. a) Original recording of a cell expressing bTRPV3, showing the response to application of menthol, thymol (both 1 mmol∙l −1 ), and 2-APB (0.3 mmol∙l −1 ) in NaCl bath solution. In response to 2-APB, the current rose dramatically, requiring an interruption of the measurement to readjust the gain (arrow). b) Original recording of a control cell transfected with the empty vector.

Techniques Used: Transferring, Expressing, Transfection, Plasmid Preparation

Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l  −1  MgCl 2 , c) Na-gluconate solution without MgCl 2  and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).
Figure Legend Snippet: Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

Techniques Used: Expressing, Transferring

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Cell Culture:

Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+
Article Snippet: The HA-Strep-bTRPV3 construct was then subcloned into pIRES2-AcGFP1 (Clontech Laboratories, Mountain View, CA, USA) via NheI and EcoRI restriction sites and into pcDNA5/TO (Life Technologies, Darmstadt, Germany) via the restriction sites HindIII and KpnI. .. Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany). .. Expression in both vector systems (pIRES2-AcGFP1 and pcDNA5/TO) was confirmed by immunoblotting ( ).

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Transfection:

Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+
Article Snippet: The HA-Strep-bTRPV3 construct was then subcloned into pIRES2-AcGFP1 (Clontech Laboratories, Mountain View, CA, USA) via NheI and EcoRI restriction sites and into pcDNA5/TO (Life Technologies, Darmstadt, Germany) via the restriction sites HindIII and KpnI. .. Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany). .. Expression in both vector systems (pIRES2-AcGFP1 and pcDNA5/TO) was confirmed by immunoblotting ( ).

Modification:

Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+
Article Snippet: The HA-Strep-bTRPV3 construct was then subcloned into pIRES2-AcGFP1 (Clontech Laboratories, Mountain View, CA, USA) via NheI and EcoRI restriction sites and into pcDNA5/TO (Life Technologies, Darmstadt, Germany) via the restriction sites HindIII and KpnI. .. Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany). .. Expression in both vector systems (pIRES2-AcGFP1 and pcDNA5/TO) was confirmed by immunoblotting ( ).

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Article Snippet: To evaluate the cytotoxicity of bio-nanocomposite scaffolds, the 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Aldrich) assay based on extraction method was used . .. Initially, Human fibro-blast cells (HuGu) cells which were taken from gums of a 45-year old female in the Iranian National Center for Biological Resources were cultured in a fresh growth medium containing 89% high-glucose Dulbecco’s Modified Eagle’s Medium (DMEM), 2 mmol.L −1 glutamine (Biochrom, UK), 15% FBS and 1% penicillin (Merck, Germany). ..

Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+
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    Biochrom mmol ∙ l 1 glutamine
    Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .
    Mmol ∙ L 1 Glutamine, supplied by Biochrom, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmol ∙ l 1 glutamine/product/Biochrom
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    86
    Biochrom glutamine
    Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .
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    Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Blocking Assay, Activation Assay, Permeability

    Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na +  and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1  CaCl 2  bath solution. Ca 2+  had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+  d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na +  of 132 ± 4 pS, for Ca 2+  of 21 ± 3 pS, and for NMDG +  of 36 ± 3 pS. The conductance for NH 4 +  was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+  in the bath.

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Transferring, Blocking Assay

    Effects of menthol and Mg 2+  on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i  in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i  (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i  in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i  in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i  /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques:

    Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l  −1  MgCl 2 , c) Na-gluconate solution without MgCl 2  and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Article Snippet: HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Transferring

    Effects of menthol and Mg 2+  on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i  in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i  (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i  in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i  in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i  /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Effects of menthol and Mg 2+ on intracellular calcium [Ca 2+ ] i . a) Average of ten original recordings of control HEK-293 cells, showing a significant increase of Δ [Ca 2+ ] i in the 200 s interval after application of menthol (1 mmol∙l −1 ) (95% confidence interval in grey), with the timeline identical to that in b), showing the average [Ca 2+ ] i (± 95%) of eleven recordings of HEK-293 cells overexpressing bTRPV3. While resting levels of [Ca 2+ ] i in bTRPV3 cells were not significantly different from those of control cells, menthol led to a significantly higher Δ [Ca 2+ ] i in these cells (p = 0.003). c) Boxplots comparing the slopes (Δ [Ca 2+ ] i /Δ t) of the graphs in (a and b) in a 50 s interval at the beginning of the measurement and in the 50 s interval after application of menthol (differing letters above the boxes indicate p

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques:

    Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na +  and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1  CaCl 2  bath solution. Ca 2+  had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+  d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na +  of 132 ± 4 pS, for Ca 2+  of 21 ± 3 pS, and for NMDG +  of 36 ± 3 pS. The conductance for NH 4 +  was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+  in the bath.

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of one patch from a bTRPV3 cell with NH 4 Cl in the pipette. a) Channel events are clearly visible in NaCl bath solution in response to pulse protocol III both at negative potentials and positive potentials, reflecting influx of Na + and efflux of NH 4 + . b) same patch in 72.5 mmol∙l −1 CaCl 2 bath solution. Ca 2+ had a clear blocking effect on the current level at positive potentials (efflux of NH 4 + ). c) Detail from (b), showing single-channel events at negative potentials, reflecting influx of Ca 2+ d) same patch in NMDGCl bath solution. e) Insert showing single-channel events at negative potential level, suggesting influx of NMDG + . f) Current-voltage plot of unitary current amplitudes from this individual patch, fitted with the current formulation of the Goldman-Hodgkin-Katz equation. The fit yields a conductance for Na + of 132 ± 4 pS, for Ca 2+ of 21 ± 3 pS, and for NMDG + of 36 ± 3 pS. The conductance for NH 4 + was similar in NaCl and NMDGCl solution (250 ± 3 pS or 264 ± 3 pS, respectively) but dropped to 88 ± 4 pS with high Ca 2+ in the bath.

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Transferring, Blocking Assay

    Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+  (10 mmol∙l −1 ) and replacement by Ca-gluconate 2  (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+  is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1  Ca 2+  to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1  Ca-gluconate 2  solution was 7.1 mV, revealing permeability to Ca 2+ .

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Original recordings of two bTRPV3 cells filled with NMDG-gluconate solution. a) As before, replacement of chloride by gluconate resulted in a decrease in outward current at positive potentials, suggesting expression of chloride channels. Both addition of Ca 2+ (10 mmol∙l −1 ) and replacement by Ca-gluconate 2 (60 mmol∙l −1 ) had a strong blocking effect, with washout to levels higher than before. The current could be partially blocked by Mg 2+ , arguing against a rupture of the seal. Note the current kinetics with amplitude decreasing after a hyperpolarising pulse suggesting that Mg 2+ is being drawn into the channel pore by the negative membrane voltage. Conversely, a depolarising voltage pulse leads to a relief of the block. In both cases, the currents appear curved in contrast to the linear kinetics of currents in pure NMDG-gluconate solution. b) Second cell, showing a spontaneous activation of inward and outward current amplitude by a factor of about four between the first set of pulses in NMDG-gluconate EGTA solution (1) and a second set (2) that was run 100 seconds later. Current amplitude continued to increase both after addition of 10 mmol∙l −1 Ca 2+ to the NMDG-gluconate solution, and after replacement of NMDG-gluconate by Ca-gluconate 2 . Both the current kinetics and the tail currents indicate a partial voltage-dependent block. The arrows above the first set of pulses indicate the interval used for the determination of mean current amplitude for the current-voltage plot, which is depicted in c), showing rectification as a further sign of a voltage-dependent block. The reversal potential in 60 mmol∙l −1 Ca-gluconate 2 solution was 7.1 mV, revealing permeability to Ca 2+ .

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Blocking Assay, Activation Assay, Permeability

    Response to various agonists of bTRPV3. Cells were filled with a CsCl pipette solution and stimulated with the continuous pulse protocol I. a) Original recording of a cell expressing bTRPV3, showing the response to application of menthol, thymol (both 1 mmol∙l −1 ), and 2-APB (0.3 mmol∙l −1 ) in NaCl bath solution. In response to 2-APB, the current rose dramatically, requiring an interruption of the measurement to readjust the gain (arrow). b) Original recording of a control cell transfected with the empty vector.

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Response to various agonists of bTRPV3. Cells were filled with a CsCl pipette solution and stimulated with the continuous pulse protocol I. a) Original recording of a cell expressing bTRPV3, showing the response to application of menthol, thymol (both 1 mmol∙l −1 ), and 2-APB (0.3 mmol∙l −1 ) in NaCl bath solution. In response to 2-APB, the current rose dramatically, requiring an interruption of the measurement to readjust the gain (arrow). b) Original recording of a control cell transfected with the empty vector.

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Transferring, Expressing, Transfection, Plasmid Preparation

    Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l  −1  MgCl 2 , c) Na-gluconate solution without MgCl 2  and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Journal: PLoS ONE

    Article Title: The bovine TRPV3 as a pathway for the uptake of Na+, Ca2+, and NH4+

    doi: 10.1371/journal.pone.0193519

    Figure Lengend Snippet: Inside-out measurements from cells expressing bTRPV3 in symmetrical solutions. Panels (a to d) show recordings from the same patch and identical scaling in a) symmetrical NaCl solution, b) NaCl solution with 1.5 mmol∙l −1 MgCl 2 , c) Na-gluconate solution without MgCl 2 and d) return to symmetrical NaCl solution. The shift in the baseline at 0 mV pipette potential in (c) reflects the impact of the liquid junction potential. e) Patch from a second cell in symmetrical NH 4 Cl solution. For clarity, this graph only shows currents at every other voltage step of pulse protocol III. f) Current-voltage plot of the unitary current amplitudes of the data from (a to e) (linear fit).

    Article Snippet: Cell culture and transient transfection of bTRPV3 HEK-293 cells were cultivated in Dulbecco’s modified Eagle’s medium supplemented with 10% (vol/vol) fetal bovine serum, 4 mmol∙l-1 glutamine and 100 units∙ml-1 of both penicillin and streptomycin (Biochrom, Berlin, Germany).

    Techniques: Expressing, Transferring