Dataset Information

Cloning and distribution of Ca2+-activated K+ channels in lobster Panulirus interruptus.

ABSTRACT: Large conductance Ca(2+)-activated potassium (BK) channels play important roles in controlling neuronal excitability. We cloned the PISlo gene encoding BK channels from the spiny lobster, Panulirus interruptus. This gene shows 81-98% sequence identity to Slo genes previously found in other organisms. We isolated a number of splice variants of the PISlo cDNA within Panulirus interruptus nervous tissue. Sequence analysis indicated that there are at least seven alternative splice sites in PISlo, each with multiple alternative segments. Using immunohistochemistry, we found that the PISlo proteins are distributed in the synaptic neuropil, axon and soma of stomatogastric ganglion (STG) neurons.

SUBMITTER: Ouyang Q

PROVIDER: S-EPMC2940416 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature

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Cloning and distribution of Ca2+-activated K+ channels in lobster Panulirus interruptus.

Ouyang Q Q Patel V V Vanderburgh J J Harris-Warrick R M RM

Neuroscience 20100802 3

Large conductance Ca(2+)-activated potassium (BK) channels play important roles in controlling neuronal excitability. We cloned the PISlo gene encoding BK channels from the spiny lobster, Panulirus interruptus. This gene shows 81-98% sequence identity to Slo genes previously found in other organisms. We isolated a number of splice variants of the PISlo cDNA within Panulirus interruptus nervous tissue. Sequence analysis indicated that there are at least seven alternative splice sites in PISlo, ea ...[more]

PMID: 20682332

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Project description:Merkel cell-neurite complexes are highly sensitive touch receptors comprising epidermal Merkel cells and sensory afferents. Based on morphological and molecular studies, Merkel cells are proposed to be mechanosensory cells that signal afferents via neurotransmission; however, functional studies testing this hypothesis in intact skin have produced conflicting results. To test this model in a simplified system, we asked whether purified Merkel cells are directly activated by mechanical stimulation. Cell shape was manipulated with anisotonic solution changes and responses were monitored by Ca2+ imaging with fura-2. We found that hypotonic-induced cell swelling, but not hypertonic solutions, triggered cytoplasmic Ca2+ transients. Several lines of evidence indicate that these signals arise from swelling-activated Ca2+-permeable ion channels. First, transients were reversibly abolished by chelating extracellular Ca2+, demonstrating a requirement for Ca2+ influx across the plasma membrane. Second, Ca2+ transients were initially observed near the plasma membrane in cytoplasmic processes. Third, voltage-activated Ca2+ channel (VACC) antagonists reduced transients by half, suggesting that swelling-activated channels depolarize plasma membranes to activate VACCs. Finally, emptying internal Ca2+ stores attenuated transients by 80%, suggesting Ca2+ release from stores augments swelling-activated Ca2+ signals. To identify candidate mechanotransduction channels, we used RT-PCR to amplify ion-channel transcripts whose pharmacological profiles matched those of hypotonic-evoked Ca2+ signals in Merkel cells. We found 11 amplicons, including PKD1, PKD2, and TRPC1, channels previously implicated in mechanotransduction in other cells. Collectively, these results directly demonstrate that Merkel cells are activated by hypotonic-evoked swelling, identify cellular signaling mechanisms that mediate these responses, and support the hypothesis that Merkel cells contribute to touch reception in the Merkel cell-neurite complex.

Dataset Information

Cloning and distribution of Ca2+-activated K+ channels in lobster Panulirus interruptus.

Publications

Cloning and distribution of Ca2+-activated K+ channels in lobster Panulirus interruptus.

Similar Datasets

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