Stretch-activated cation channel from larval bullfrog skin

Stanley D Hillyard; Niels J Willumsen; Mario B Marrero

doi:10.1242/jeb.040568

Stretch-activated cation channel from larval bullfrog skin

Stanley D Hillyard, Niels J Willumsen, Mario B Marrero

Molecular Integrative Physiology

7 Citations (Scopus)

Abstract

Cell-attached patches from isolated epithelial cells from larval bullfrog skin revealed a cation channel that was activated by applying suction (-1 kPa to -4.5 kPa) to the pipette. Activation was characterized by an initial large current spike that rapidly attenuated to a stable value and showed a variable pattern of opening and closing with continuing suction. Current-voltage plots demonstrated linear or inward rectification and single channel conductances of 44-56 pS with NaCI or KCl Ringer's solution as the pipette solution, and a reversal potential (-V_p) of 20-4OmV. The conductance was markedly reduced with N-methyl-D-glucamide (NMDG)-CI Ringer's solution in the pipette. Neither amiloride nor ATP, which are known to stimulate an apical cation channel in Ussing chamber preparations of larval frog skin, produced channel activation nor did these compounds affect the response to suction. Stretch activation was not affected by varying the pipette concentrations of Ca²⁺ between 0 mmoll^-1 and 4 mmoll^-1 or by varying pH between 6.8 and 8.0. However, conductance was reduced with 4 mmoll^-1 Ca²⁺. Western blot analysis of membrane homogenates from larval bullfrog and larval toad skin identified proteins that were immunoreactive with mammalian TRPC1 and TRPC5 (TRPC, canonical transient receptor potential channel) antibodies while homogenates of skin from newly metamorphosed bullfrogs were positive for TRPC1 and TRPC3/6/7 antibodies. The electrophysiological response of larval bullfrog skin resembles that of a stretch-activated cation channel characterized in Xenopus oocytes and proposed to be TRPC1. These results indicate this channel persists in all life stages of anurans and that TRP isoforms may be important for sensory functions of their skin.

Original language	English
Journal	Journal of Experimental Biology
Volume	213
Issue number	Pt 10
Pages (from-to)	1782-7
Number of pages	6
ISSN	0022-0949
DOIs	https://doi.org/10.1242/jeb.040568
Publication status	Published - 15 May 2010

Keywords

Animals
Blotting, Western
Calcium
Epithelial Cells
Hydrogen-Ion Concentration
Ion Channel Gating
Larva
Potassium Chloride
Rana catesbeiana
Skin
Sodium Chloride
Stress, Mechanical
TRPC Cation Channels

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10.1242/jeb.040568

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title = "Stretch-activated cation channel from larval bullfrog skin",

abstract = "Cell-attached patches from isolated epithelial cells from larval bullfrog skin revealed a cation channel that was activated by applying suction (-1 kPa to -4.5 kPa) to the pipette. Activation was characterized by an initial large current spike that rapidly attenuated to a stable value and showed a variable pattern of opening and closing with continuing suction. Current-voltage plots demonstrated linear or inward rectification and single channel conductances of 44-56 pS with NaCI or KCl Ringer's solution as the pipette solution, and a reversal potential (-Vp) of 20-4OmV. The conductance was markedly reduced with N-methyl-D-glucamide (NMDG)-CI Ringer's solution in the pipette. Neither amiloride nor ATP, which are known to stimulate an apical cation channel in Ussing chamber preparations of larval frog skin, produced channel activation nor did these compounds affect the response to suction. Stretch activation was not affected by varying the pipette concentrations of Ca2+ between 0 mmoll-1 and 4 mmoll-1 or by varying pH between 6.8 and 8.0. However, conductance was reduced with 4 mmoll-1 Ca2+. Western blot analysis of membrane homogenates from larval bullfrog and larval toad skin identified proteins that were immunoreactive with mammalian TRPC1 and TRPC5 (TRPC, canonical transient receptor potential channel) antibodies while homogenates of skin from newly metamorphosed bullfrogs were positive for TRPC1 and TRPC3/6/7 antibodies. The electrophysiological response of larval bullfrog skin resembles that of a stretch-activated cation channel characterized in Xenopus oocytes and proposed to be TRPC1. These results indicate this channel persists in all life stages of anurans and that TRP isoforms may be important for sensory functions of their skin.",

keywords = "Animals, Blotting, Western, Calcium, Epithelial Cells, Hydrogen-Ion Concentration, Ion Channel Gating, Larva, Potassium Chloride, Rana catesbeiana, Skin, Sodium Chloride, Stress, Mechanical, TRPC Cation Channels",

author = "Hillyard, {Stanley D} and Willumsen, {Niels J} and Marrero, {Mario B}",

year = "2010",

month = may,

day = "15",

doi = "10.1242/jeb.040568",

language = "English",

volume = "213",

pages = "1782--7",

journal = "Journal of Experimental Biology",

issn = "0022-0949",

publisher = "The/Company of Biologists Ltd.",

number = "Pt 10",

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TY - JOUR

T1 - Stretch-activated cation channel from larval bullfrog skin

AU - Hillyard, Stanley D

AU - Willumsen, Niels J

AU - Marrero, Mario B

PY - 2010/5/15

Y1 - 2010/5/15

N2 - Cell-attached patches from isolated epithelial cells from larval bullfrog skin revealed a cation channel that was activated by applying suction (-1 kPa to -4.5 kPa) to the pipette. Activation was characterized by an initial large current spike that rapidly attenuated to a stable value and showed a variable pattern of opening and closing with continuing suction. Current-voltage plots demonstrated linear or inward rectification and single channel conductances of 44-56 pS with NaCI or KCl Ringer's solution as the pipette solution, and a reversal potential (-Vp) of 20-4OmV. The conductance was markedly reduced with N-methyl-D-glucamide (NMDG)-CI Ringer's solution in the pipette. Neither amiloride nor ATP, which are known to stimulate an apical cation channel in Ussing chamber preparations of larval frog skin, produced channel activation nor did these compounds affect the response to suction. Stretch activation was not affected by varying the pipette concentrations of Ca2+ between 0 mmoll-1 and 4 mmoll-1 or by varying pH between 6.8 and 8.0. However, conductance was reduced with 4 mmoll-1 Ca2+. Western blot analysis of membrane homogenates from larval bullfrog and larval toad skin identified proteins that were immunoreactive with mammalian TRPC1 and TRPC5 (TRPC, canonical transient receptor potential channel) antibodies while homogenates of skin from newly metamorphosed bullfrogs were positive for TRPC1 and TRPC3/6/7 antibodies. The electrophysiological response of larval bullfrog skin resembles that of a stretch-activated cation channel characterized in Xenopus oocytes and proposed to be TRPC1. These results indicate this channel persists in all life stages of anurans and that TRP isoforms may be important for sensory functions of their skin.

AB - Cell-attached patches from isolated epithelial cells from larval bullfrog skin revealed a cation channel that was activated by applying suction (-1 kPa to -4.5 kPa) to the pipette. Activation was characterized by an initial large current spike that rapidly attenuated to a stable value and showed a variable pattern of opening and closing with continuing suction. Current-voltage plots demonstrated linear or inward rectification and single channel conductances of 44-56 pS with NaCI or KCl Ringer's solution as the pipette solution, and a reversal potential (-Vp) of 20-4OmV. The conductance was markedly reduced with N-methyl-D-glucamide (NMDG)-CI Ringer's solution in the pipette. Neither amiloride nor ATP, which are known to stimulate an apical cation channel in Ussing chamber preparations of larval frog skin, produced channel activation nor did these compounds affect the response to suction. Stretch activation was not affected by varying the pipette concentrations of Ca2+ between 0 mmoll-1 and 4 mmoll-1 or by varying pH between 6.8 and 8.0. However, conductance was reduced with 4 mmoll-1 Ca2+. Western blot analysis of membrane homogenates from larval bullfrog and larval toad skin identified proteins that were immunoreactive with mammalian TRPC1 and TRPC5 (TRPC, canonical transient receptor potential channel) antibodies while homogenates of skin from newly metamorphosed bullfrogs were positive for TRPC1 and TRPC3/6/7 antibodies. The electrophysiological response of larval bullfrog skin resembles that of a stretch-activated cation channel characterized in Xenopus oocytes and proposed to be TRPC1. These results indicate this channel persists in all life stages of anurans and that TRP isoforms may be important for sensory functions of their skin.

KW - Animals

KW - Blotting, Western

KW - Calcium

KW - Epithelial Cells

KW - Hydrogen-Ion Concentration

KW - Ion Channel Gating

KW - Larva

KW - Potassium Chloride

KW - Rana catesbeiana

KW - Skin

KW - Sodium Chloride

KW - Stress, Mechanical

KW - TRPC Cation Channels

U2 - 10.1242/jeb.040568

DO - 10.1242/jeb.040568

M3 - Journal article

C2 - 20435829

SN - 0022-0949

VL - 213

SP - 1782

EP - 1787

JO - Journal of Experimental Biology

JF - Journal of Experimental Biology

IS - Pt 10

ER -

Stretch-activated cation channel from larval bullfrog skin

Abstract

Keywords

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