Biomedical subjects

Dominik Oliver

Publications and source records attributed to Dominik Oliver.

7 recordsLinked to original sources

PubMed2005

Ca2+-independent activation of BKCa channels at negative potentials in mammalian inner hair cells.

The defining characteristic of large-conductance Ca(2)(+)- and voltage-activated K(+) channels (BK(Ca)) is their allosteric activation by two distinct stimuli, membrane depolarization and cytosolic Ca(2)(+) ions. In this allosteric gating, increasing cytosolic Ca(2)(+) concentration ([Ca(2)(+)](i)) shifts the depolarization required for channel opening into the physiological voltage range. In fact, according to present knowledge, elevation of [Ca(2)(+)](i) to micromolar levels is the only means to activate BK(Ca) at membrane potentials below 0 mV. We recorded BK(Ca)-mediated currents from auditory inner hair cells (IHCs) in acutely isolated organs of Corti using the patch-clamp technique in whole-cell and excised patch configuration. In inside-out and outside-out patches, activation of BK(Ca) channels from IHCs showed the prototypic sensitivity to increased [Ca(2)(+)](i). However, channel activation at 0 [Ca(2)(+)](i) occurred at unusually negative potentials (half-maximal activation (V(h)) around 0 mV), indicating that a large fraction of the channels can be activated at physiological voltages without elevated [Ca(2)(+)](i). In intact IHCs, the activation curve of BK(Ca) currents recorded in whole-cell configuration exhibited a V(h) of -42 mV together with a high voltage dependence (slope factor of 10 mV) and submillisecond onset of current. Surprisingly, this activation was independent of changes in local [Ca(2)(+)](i) as shown by experiments that interfered with Ca(2)(+) influx through voltage-gated Ca(2)(+) (Cav) channels, release of Ca(2)(+) from internal stores, or intracellular buffer capacity. This behaviour is not due to beta-subunits of BK(Ca) (BKbeta), as genetic inactivation of the beta-subunit expressed in IHCs, KCNMB1, did not affect BK(Ca) gating. We conclude that the BK(Ca) channel protein in IHCs may be modified in order to rapidly activate and deactivate at resting [Ca(2)(+)](i). Our results suggest that BK(Ca) may function as a purely voltage-gated K(+) channel with exceptionally rapid activation kinetics, challenging the view that both increased cytosolic Ca(2)(+) and depolarization are generally required for activation of BK(Ca).

Animals↗

PubMed2005

Cod106, a novel synaptic protein expressed in sensory hair cells of the inner ear and in CNS neurons.

The exquisite performance of the highly specialized mammalian inner ear requires a multitude of specific proteins. Yet, only a subset of these proteins has been identified and studied in detail. Here, we describe a novel gene expressed in the organ of Corti that encodes a membrane-associated protein of 106 kDa. The new protein, termed Cod106, lacks sequence homology to characterized gene products. As shown by in situ hybridization, it is expressed in auditory and vestibular hair cells, as well as in distinct sets of CNS neurons with particularly high abundance in hippocampus and cerebellum. Immunohistochemistry detected Cod106 at the basal, synaptic pole of cochlear outer hair cells and vestibular hair cells. In cultured hippocampal neurons, Cod106 immunofluorescence co-localized with the postsynaptic density protein 95 (PSD95), indicating a postsynaptic localization. Cell-type specificity and subcellular localization may be consistent with an involvement of Cod106 in synaptic processes.

Amino Acid Sequence↗

PubMed2004

Protein kinase CK2 is coassembled with small conductance Ca(2+)-activated K+ channels and regulates channel gating.

Small conductance Ca(2+)-activated K+ channels (SK channels) couple the membrane potential to fluctuations in intracellular Ca2+ concentration in many types of cells. SK channels are gated by Ca2+ ions via calmodulin that is constitutively bound to the intracellular C terminus of the channels and serves as the Ca2+ sensor. Here we show that, in addition, the cytoplasmic N and C termini of the channel protein form a polyprotein complex with the catalytic and regulatory subunits of protein kinase CK2 and protein phosphatase 2A. Within this complex, CK2 phosphorylates calmodulin at threonine 80, reducing by 5-fold the apparent Ca2+ sensitivity and accelerating channel deactivation. The results show that native SK channels are polyprotein complexes and demonstrate that the balance between kinase and phosphatase activities within the protein complex shapes the hyperpolarizing response mediated by SK channels.

Adenosine Triphosphate↗

PubMed2004

Functional conversion between A-type and delayed rectifier K+ channels by membrane lipids.

Voltage-gated potassium (Kv) channels control action potential repolarization, interspike membrane potential, and action potential frequency in excitable cells. It is thought that the combinatorial association between distinct alpha and beta subunits determines whether Kv channels function as non-inactivating delayed rectifiers or as rapidly inactivating A-type channels. We show that membrane lipids can convert A-type channels into delayed rectifiers and vice versa. Phosphoinositides remove N-type inactivation from A-type channels by immobilizing the inactivation domains. Conversely, arachidonic acid and its amide anandamide endow delayed rectifiers with rapid voltage-dependent inactivation. The bidirectional control of Kv channel gating by lipids may provide a mechanism for the dynamic regulation of electrical signaling in the nervous system.

Animals↗

PubMed2003

Resting potential and submembrane calcium concentration of inner hair cells in the isolated mouse cochlea are set by KCNQ-type potassium channels.

Cochlear inner hair cells (IHCs) transduce sound-induced vibrations into a receptor potential (RP) that controls afferent synaptic activity and, consequently, frequency and timing of action potentials in the postsynaptic auditory neurons. The RP is thought to be shaped by the two voltage-dependent K+ conductances, I(K,f) and I(K,s), that are carried by large-conductance Ca2+- and voltage-dependent K+ (BK)- and K(V)-type K+ channels. Using whole-cell voltage-clamp recordings in the acutely isolated mouse cochlea, we show that IHCs display an additional K+ current that is active at the resting membrane potential (-72 mV) and deactivates on hyperpolarization. It is potently blocked by the KCNQ-channel blockers linopirdine and XE991 but is insensitive to tetraethylammonium and 4-aminopyridine, which inhibit I(K,f) and I(K,s), respectively. Single-cell PCR and immunocytochemistry showed expression of the KCNQ4 subunit in IHCs. In current-clamp experiments, block of the KCNQ current shifted the resting membrane potential by approximately 7 to -65 mV and led to a significant activation of BK channels. Using BK channels as an indicator for submembrane intracellular Ca2+ concentration ([Ca2+]i), it is shown that the shift in IHC resting potential observed after block of the KCNQ channels leads to an increase in [Ca2+]i to values > or =1 microm. In conclusion, KCNQ channels set the resting membrane potential of IHCs in the isolated organ of Corti and thus maintain [Ca2+]i at low levels. Destabilization of the resting potential and increase in [Ca2+]i, as may result from impaired KCNQ4 function in IHCs, provide a novel explanation for the progressive hearing loss (DFNA2) observed in patients with defective KCNQ4 genes.

Animals↗

PubMed2003

Solution structure and function of the "tandem inactivation domain" of the neuronal A-type potassium channel Kv1.4.

Cumulative inactivation of voltage-gated (Kv) K(+) channels shapes the presynaptic action potential and determines timing and strength of synaptic transmission. Kv1.4 channels exhibit rapid "ball-and-chain"-type inactivation gating. Different from all other Kvalpha subunits, Kv1.4 harbors two inactivation domains at its N terminus. Here we report the solution structure and function of this "tandem inactivation domain" using NMR spectroscopy and patch clamp recordings. Inactivation domain 1 (ID1, residues 1-38) consists of a flexible N terminus anchored at a 5-turn helix, whereas ID2 (residues 40-50) is a 2.5-turn helix made up of small hydrophobic amino acids. Functional analysis suggests that only ID1 may work as a pore-occluding ball domain, whereas ID2 most likely acts as a "docking domain" that attaches ID1 to the cytoplasmic face of the channel. Deletion of ID2 slows inactivation considerably and largely impairs cumulative inactivation. Together, the concerted action of ID1 and ID2 may promote rapid inactivation of Kv1.4 that is crucial for the channel function in short term plasticity.

Amino Acid Sequence↗

PubMedDate not supplied

Prestin, the motor protein of outer hair cells.

Prestin is a gene recently cloned from mammalian cochlear outer hair cells (OHC) using a single cell type, outer minus inner hair cell, specific suppressive subtractive hybridization procedure. The localization and gene expression profile of the prestin protein fits the pattern of OHC's development of electromotility. When prestin is abundantly expressed in normally nonmotile kidney cells, nonlinear capacitance and motility that are normally only seen in OHCs can be recorded. Furthermore, both nonlinear capacitance and motility can be reduced by salicylate, a well-known inhibitor of electromotility. These data suggest that prestin is the motor protein of OHCs. Amino acid sequence and gene structure analysis indicate that prestin is the fifth member of a newly discovered anion transport family (SLC26) that includes PDS, DRA and DTDST, which are chloride-iodide transporters, Cl-/HCO3- exchangers or sulfate transporters. Prestin shares overall structure similarity with this anion transporter family. Recently, intracellular anions (chloride or bicarbonate) were found to be essential for OHC electromotility and prestin's function.

Animals↗