KCNK5 channels mostly expressed in cochlear outer sulcus cells are indispensable for hearing

International audienceIn the cochlea, K þ is essential for mechano-electrical transduction. Here, we explore cochlear structure and function in mice lacking K þ channels of the two-pore domain family. A profound deafness associated with a decrease in endocochlear potential is found in adult Kcnk5 À / À mice. Hearing occurs around postnatal day 19 (P19), and completely disappears 2 days later. At P19, Kcnk5 À / À mice have a normal endolymphatic [K þ ] but a partly lowered endocochlear potential. Using Lac-Z as a gene reporter, KCNK5 is mainly found in outer sulcus Claudius', Boettcher's and root cells. Low levels of expression are also seen in the spiral ganglion, Reissner's membrane and stria vascularis. Essential channels (KCNJ10 and KCNQ1) contributing to K þ secretion in stria vascularis have normal expression in Kcnk5 À / À mice. Thus, KCNK5 channels are indispensable for the maintenance of hearing. Among several plausible mechanisms, we emphasize their role in K þ recycling along the outer sulcus lateral route

Role of TASK2 Potassium Channels Regarding Volume Regulation in Primary Cultures of Mouse Proximal Tubules

Several papers reported the role of TASK2 channels in cell volume regulation and regulatory volume decrease (RVD). To check the possibility that the TASK2 channel modulates the RVD process in kidney, we performed primary cultures of proximal convoluted tubules (PCT) and distal convoluted tubules (DCT) from wild-type and TASK2 knockout (KO) mice. In KO mice, the TASK2 coding sequence was in part replaced by the lac-Z gene. This allows for the precise localization of TASK2 in kidney sections using β-galactosidase staining. TASK2 was only localized in PCT cells. K+ currents were analyzed by the whole-cell clamp technique with 125 mM K-gluconate in the pipette and 140 mM Na-gluconate in the bath. In PCT cells from wild-type mice, hypotonicity induced swelling-activated K+ currents insensitive to 1 mM tetraethylammonium, 10 nM charybdotoxin, and 10 μM 293B, but blocked by 500 μM quinidine and 10 μM clofilium. These currents were increased in alkaline pH and decreased in acidic pH. In PCT cells from TASK2 KO, swelling-activated K+ currents were completely impaired. In conclusion, the TASK2 channel is expressed in kidney proximal cells and could be the swelling-activated K+ channel responsible for the cell volume regulation process during osmolyte absorptions in the proximal tubules

Étude fonctionnelle des souris dont les gènes KCNE1 et TWIK1 ont été invalidés

De tous les canaux ioniques, les canaux K+ forment une classe où la diversité structurale et fonctionnelle est certainement la plus importante. Ils interviennent dans un nombre considérable de processus physiologiques. Les canaux K+ sont répartis en trois familles selon leurs propriétés structurales et fonctionnelles, et 67 gènes sont connus à ce jour. Les caractéristiques de la plupart de ces canaux sont bien établies in vitro, mais leur rôle physiologique n'est pas toujours connu. L'invalidation de gènes chez la souris est un outil qui peut permettre de déterminer le rôle d'un gène donné in vitro. Nous avons utlisé cette technique pour étudier la fonction des sous-unités KCNE1 et TWIK1. Les souris kcne1 -/- ont dèjà permis de comprendre la fonction des courants Iks dans l'oreille interne. Ces souris présentent en effet une surdité identique à celle des patients atteints du syndrome de Jervell et Lange-Nielsen. Des mutations de gènes humains kcne1 et kcnq1 ont été associées à ce syndrome cardio-auditif. L'étude de la fonction cardiaque des souris kcne1 -/- a révélé que l'intervalle QT de l'électrocardiogramme n'est pas très différent de celui des souris sauvages. Par contre, la sous-unité KCNE1 permet l'adaptation de l'intervalle QT en fonction des battements du coeur. D'autre part, les souris kcne1 -/- présentent une légère hypokaliémie associée à un hyperaldostéronisme (...). La seconde partie de ce travail concerne l'étude de la sous-unité TWIK1. Ce canal est fortement exprimé dans le système nerveux central et dans de nombreux tissus épithéliaux. (...) L'analyse des électrorétinogrammes montre que les propriétés fonctionnelles de la rétine sont atténuées chez les souris kcne1 -/-.NICE-BU Sciences (060882101) / SudocSudocFranceF

The multifaceted phenotype of the knockout mouse for the KCNE1 potassium channel gene

Mutations of the KCNE1 gene (IsK, minK) are related to hereditary forms of cardiac arrhythmias, so-called long QT syndromes (LQT). Here we review the phenotype of a mouse model for the recessive form of LQT known as Jervell and Lange-Nielsen syndrome. KCNE1 knockout mice exhibit an enhanced QT-RR adaptability, which is probably part of the pathophysiological mechanism leading to life-threatening tachyarrhythmia in patients. Like patients, knockout mice are deaf and show vestibular symptoms due to an impaired endolymph production. Knockout mice show urinary and fecal salt wasting and volume depletion. The renal phenotype is due to diminished reabsorption of Na+and glucose. The mice are hypokalemic and have increased aldosterone levels. Besides volume depletion, aldosterone is elevated via a set-point shift for sensing of extracellular K+in aldosterone-secreting glomerulosa cells, which physiologically express KCNE1. In conclusion, KCNE1 knockout leads to a complex phenotype resulting from direct loss of KCNE1 and compensatory mechanisms. Murine KCNE1 physiology could be helpful for the pathophysiological understanding and perhaps gene-specific treatment of long QT patients.</jats:p

Identification of different receptor types for toxic phospholipases A2 in rabbit skeletal muscle

AbstractOxyuranus scutellatus scutellatus toxins I (OS1) and 2 (OS2) are two phospholipase A2s (PLA2) isolated from the venom of the Australian Taipan snake. Their iodinated derivatives have been used to characterize PLA2 binding sites on rabbit skeletal muscle. Competition and cross-linking experiments indicate that 125I-labelled OS2 binding sites in rabbit skeletal muscle in vivo are distributed into two classes of receptors. One class binds OS2 and OS1 and is insensitive to the bee venom PLA2. It is composed of a 180 kDa binding protein. This class of PLA2 receptor is expressed at a high level in rabbit myotube membranes. The other class of PLA2 receptor identified with 125I-OS2 also binds with high affinity the bee venom PLA2 but not OS1 and is composed of major polypeptides of 34.48 and 82 kDa. This second class of receptor is similar to the one found in brain membranes. The density of the two classes of receptors varies during muscle development