Alternative splice isoforms of small conductance calcium-activated SK2 channels differ in molecular interactions and surface levels

Small conductance Ca²⁺-sensitive potassium (SK2) channels are voltage-independent, Ca²⁺-activated ion channels that conduct potassium cations and thereby modulate the intrinsic excitability and synaptic transmission of neurons and sensory hair cells. In the cochlea, SK2 channels are functionally coupled to the highly Ca²⁺ permeant α9/10-nicotinic acetylcholine receptors (nAChRs) at olivocochlear postsynaptic sites. SK2 activation leads to outer hair cell hyperpolarization and frequency-selective suppression of afferent sound transmission. These inhibitory responses are essential for normal regulation of sound sensitivity, frequency selectivity, and suppression of background noise. However, little is known about the molecular interactions of these key functional channels. Here we show that SK2 channels co-precipitate with α9/10-nAChRs and with the actin-binding protein α-actinin-1. SK2 alternative splicing, resulting in a 3 amino acid insertion in the intracellular 3′ terminus, modulates these interactions. Further, relative abundance of the SK2 splice variants changes during developmental stages of synapse maturation in both the avian cochlea and the mammalian forebrain. Using heterologous cell expression to separately study the 2 distinct isoforms, we show that the variants differ in protein interactions and surface expression levels, and that Ca²⁺ and Ca²⁺-bound calmodulin differentially regulate their protein interactions. Our findings suggest that the SK2 isoforms may be distinctly modulated by activity-induced Ca²⁺ influx. Alternative splicing of SK2 may serve as a novel mechanism to differentially regulate the maturation and function of olivocochlear and neuronal synapses.     

Is there modal opponency in the thermal senses?

1. 1.The sensations evoked by pairs of distinct thermal stimuli applied to the back of the hand were studied in 17 volunteer subjects. Four stimulus combinations were used; neutral-cold (NC), neutral-neutral (NN), neutral-warm (NW), and cold-warm (CW).

2. 2.The subjects were first asked to estimate the magnitude of the thermal sensations evoked by the thermal stimuli. On average, the four pairs were reported as increasing magnitude in the following order: NC, CW, NN, and NW, seeming to suggest that the subjects experienced the cold-warm combination as a composite sensation of cold and warmth intermediate between pure cold and pure warmth.

3. 3.When asked only to detect the presence of a cold stimulus, the subjects performed as well for the CW combination as for the CN combination. This second result indicates that the reported composite magnitude of CW does not result from a true opponency of cold and warmth but from a cognitive combination of distinct sensations of cold and warmth.

The introduction ofMimorista pulchellalis [Lepidoptera: Pyraustidae] into South Africa for the biological control of jointed cactus,Opuntia aurantiaca. 2. Field evaluation

The mothMimorista pulchellalis was monitored over 2 years after liberation in a jointed cactus (Opuntia aurantiaca) infestation in South Africa. Moth and cactus densities were estimated using a system of randomly-assigned quadrats and the impact of the moth on the cactus population quantified. Moths appeared adapted to survive on the etiolated form of jointed cactus plants, killing approximately 1% of the increment in small plants annually. Large plants were also attacked but damage was negligible. The moths occurred in low numbers throughout the study period and generally went through 3 generations in a year.

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Résumé La pyraleMimorista pulchellalis a été surveillée durant les deux années qui suivirent sa libération dans une infestation de figuier de Barbarie en Afrique du Sud. Les densités de pyrale et de cactus ont été estimées en utilisant un système de quadrats distribués au hasard et l'impact de la pyrale sur la population de cactus a été quantifiée. Les pyrales semblaient adaptées à survivre sur la forme étiolée des plants de cactus tuant approximativement 1% de l'accroissement annuel des petits plants. Les gros plants étaient aussi attaqués, mais les dégats étaient négligeables. La pyrale existait en faibles nombres durant toute la période d'étude et elle présentait généralement 3 générations par an.

     

IND综合评价的逻辑结构及成药性评价

基于各学科信息作出安全性、有效性和质量可控性方面的综合评价是新药研究评价的灵魂，是一个多学科、多组织参与的综合决策过程。根据新药评价研究的实际工作经验，介绍一个新药临床试验综合评价的逻辑性、结构性的工作思考模式，有助于将不同专业的技术信息和技术结论有机整合，作出科学决策。     

Ciliary receptors associated with the mouth and pharynx of Acoela (Acoelomorpha): a comparative ultrastructural study

The ultrastructure and distribution of receptor cells near the mouth and (where present) the pharynx of Hofstenia miamia, Proporus bermudensis, Conaperta thela, and Convoluta convoluta (Acoela) were investigated by transmission electron microscopy and confocal laser scanning microscopy of specimens stained with a fluorescence marker for actin. Five types of monociliary receptors were identified: (1) non‐collared receptors with a single long and narrow ciliary rootlet; (2) non‐collared receptors with a wide main ciliary rootlet and a smaller posterior rootlet; (3) non‐collared receptors with a single wide and hollow ciliary rootlet with a granulated core; (4) Collar (?) receptors with obliquely radial filament bundles in the cell apex and with a single hollow ciliary rootlet composed of numerous strand‐like elements; and (5) Collar receptors lacking a striated rootlet but with a granular body (swallow's nest rootlet). While H. miamia bears the first two receptor types, P. bermudensis has receptors of type 1, 3 and 5, and Cona. thela and Conv. convoluta have receptors of type 3, 4 and 5. The density of receptors is generally highest at the anterior body tip, regardless of where the mouth is located. Most receptor types occur scattered over the whole body but type 2 receptors of H. miamia are restricted to the pharynx and mouth region. The lack of a common receptor type specific for the mouth and pharynx of the investigated species points to an independent origin of the pharynges in Hofsteniidae and in Proporidae and of the mouth tube in Convolutidae. Moreover, the homology of the so‐called collar receptors in Acoela with typical collar receptors in other invertebrates is questioned.     

The ultrastructure of the so-called olfactory setae on the antennula ofDaphnia magna Straus (Crustacea,Cladocera)

A group of nine sensory setae is found on the tip of the antennula ofDaphnia magna in both sexes. Inside a seta four dendrites are situated, each with one receptor cilium. The receptor cilia extend through a liquor space into the exterior part of the seta. The exterior part of the liquor space is divided from the interior part by a knob-like thickening of the innermost layer of the epicuticle, the basal bead. The basal bead narrows the liquor space and the receptor cilia. The interior part of the liquor space is surrounded by five sheath cells, the exterior part by a thin cuticle. In the exterior part the receptor cilia branch partly and reach a terminal pellet on the tip of the seta. The terminal pellet is a thickened part of the epicuticle. It is permeable to several dissolved substances. It is the exterior part of the receptor that projects over the tip of the antennula and seems to be the entire seta. During the premoult the fifth sheath cell builds up the articulation of the seta, the fourth the basal bead, and the third the shaft of the seta. The first sheath cell forms the cuticular sheath. The organ seems to be a chemoreceptor, but the adequate stimulus is as yet unknown.     

Patch-clamp study of isolated taste receptor cells of the frog

Summary Taste discs were dissected from the tongue ofR. ridibunda and their cells dissociated by a collagenase/low Ca/mechanical agitation protocol. The resulting cell suspension contained globular epithelial cells and, in smaller number, taste receptor cells. These were identified by staining properties and by their preserved apical process, the tip of which often remained attached to an epithelial (associated) cell. When the patch pipette contained 110mm KCl and the cells were superfused with NaCl Ringer's during whole-cell recording, the mean zero-current potential of 22 taste receptor cells was –65.2 mV and the slope resistance 150 to 750 M. Pulse-depolarization from a holding voltage of –80 mV activated a transient TTX-blockable inward Na current. Activation became noticeable at –25 mV and was half-maximal at –8 mV. Steady-state inactivation was half-maximal at –67 mV and complete at –50 mV. Peak Na current averaged –0.5 nA/cell. The Ca-ionophore A23187 shifted the activation and inactivation curve to more negative voltages. Similar shifts occurred when the pipette Ca was raised. External Ni (5mm) shifted the activation curve towards positive voltages by 10 mV. Pulse depolarization also activated outward K currents. Activation was slower than that of Na current and inactivation slower still. External TEA (7.5mm) and 4-aminopyridine (1mm) did not block, but 5mm Ba blocked the K currents. K-tail currents were seen on termination of depolarizing voltage pulses. A23187 shifted theI _K(V)-curve to more negative voltages. Action potentials were recorded when passing pulses of depolarizing outward current. Of the frog gustatory stimulants, 10mm Ca caused a reversible 5-to 10-mV depolarization in the current-clamp mode. Quinine (0.1mm, bitter) produced a reversible depolarization accompanied by a full block of Na current and, with slower time-course, a partial block of K currents. Cyclic AMP (5mm in the external solution or 0.5 m in the pipette) caused reversible depolarization (to –40 to –20 mV) due to partial blockage of K currents, but only if ATP was added to the pipette solution. Similar responses were elicited by stimulating the adenylate cyclase with forskolin. Blockage of cAMP-phosphodiesterase enhanced the response to cAMP. These results suggest that cAMP may be one of the cytosolic messengers in taste receptor cells. Replacement of ATP by AMP-PNP in the pipette abolished the depolarizing response to cAMP. Inclusion of ATP--S in the pipette caused slow depolarization to –40 to –20 mV, due to partial blockage of K currents. Subsequently, cAMP was without effect. The remaining K currents were blockable by Ba. These results suggest that cAMP initiates phosphorylation of one set of K channels to a nonconducting conformation.     

Fast axon activity and the motor pattern in cockroach legs during swimming

Abstract Electromyographic recordings were made from muscles that extend the trochanter/femur of each of the six legs of American cockroaches, Periplaneta americana (L.), while the insects swam in water. The recordings showed two novel features. (1) During swimming, muscle activity in different legs was coordinated in the alternating tripod pattern commonly seen during free walking on land, not in the pattern of synchronous leg pairs common to other large terrestrial insects in water. (2) Fast axons were usually recruited along with slow axons, even when the insect swam at a moderate pace. Fast axon activity always started after the middle of the slow axon burst in intact insects, but vanished from most bursts in the stump of the leg after amputation of the femur. The alternating tripod pattern was maintained even after amputation. Possible causes of fast axon recruitment are discussed.     

A practical test for in vitro evaluation of photosensitization: Assessment with hematoporphyrin derivative (HPD)

A simple procedure is described for the determination of the photosensitizing potency of drugs, using three leukemic cell lines, two of lymphocytic origin, L1210 and P388 and one of erythroid type, Friend-745. The procedure allows one to investigate several aspects of the photosensitization properties of tested compounds such as cellular localization and direct (trypan blue exclusion) or delayed (clonogenicity) photomediated toxicities.The method was assessed using crude hematoporphyrin derivative (HPD) as well as dihematoporphyrin ether (DHE) or commercially available Photofrin II. Results were compared to those obtained with normal cells, e.g spleen lymphocytes and erythropoietic stem cells (CFU-e), and discussed in the light of the relative response of normal versus transformed cells.Abbreviations DHE Dihematoporphyrin Ether - FCS Fetal Calf Serum - HPD Hematoporphyrin Derivative - PDT Photodynamic Therapy