Control of P2X2 Channel Permeability by the Cytosolic Domain

ATP-gated P2X channels are the simplest of the three families of transmitter-gated ion channels. Some P2X channels display a time- and activation-dependent change in permeability as they undergo the transition from the relatively Na+-selective I1 state to the I2 state, which is also permeable to organic cations. We report that the previously reported permeability change of rat P2X2 (rP2X2) channels does not occur at mouse P2X2 (mP2X2) channels expressed in oocytes. Domain swaps, species chimeras, and point mutations were employed to determine that two specific amino acid residues in the cytosolic tail domain govern this difference in behavior between the two orthologous channels. The change in pore diameter was characterized using reversal potential measurements and excluded field theory for several organic ions; both rP2X2 and mP2X2 channels have a pore diameter of ~11 Å in the I1 state, but the transition to the I2 state increases the rP2X2 diameter by at least 3 Å. The I1 to I2 transition occurs with a rate constant of ~0.5 s^-1. The data focus attention on specific residues of P2X2 channel cytoplasmic domains as determinants of permeation in a state-specific manner

Psychophysiology

Abstract Joint performance can lead to the synchronization of physiological processes among group members during a shared task. Recently, it has been shown that synchronization is indicative of subjective ratings of group processes and task performance. However, different methods have been used to quantify synchronization, and little is known about the effects of the choice of method and level of analysis (individuals, dyads, or triads) on the results. In this study, participants performed a decision-making task in groups of three while physiological signals (heart rate and electrodermal activity), positive affective behavior, and personality traits were measured. First, we investigated the effects of different levels of analysis of physiological synchrony on affective behavior. We computed synchrony measures as (a) individual contributions to group synchrony, (b) the average dyadic synchrony within a group, and (c) group-level synchrony. Second, we assessed the association between physiological synchrony and positive affective behavior. Third, we investigated the moderating effects of trait anxiety and social phobia on behavior. We discovered that the effects of physiological synchrony on positive affective behavior were particularly strong at the group level but nonsignificant at the individual and dyadic levels. Moreover, we found that heart rate and electrodermal synchronization showed opposite effects on group members' display of affective behavior. Finally, trait anxiety moderated the relationship between physiological synchrony and affective behavior, perhaps due to social uncertainty, while social phobia did not have a moderating effect. We discuss these results regarding the role of different physiological signals and task demands during joint action

Using Touch Technology to Foster Storytelling in the Preschool Classroom

This practitioner research explores ways children engage in literacy learning through storytelling with the use of touch technology in a VPK (Voluntary Pre-Kindergarten) classroom. With access to diverse touch technology devices but no experience using these technologies, a VPK teacher explored strategies to use the resources to enhance literacy learning in the classroom with the support of a professional learning community (PLC). The PLC consisted of a master’s student, university faculty, school director, and a technology liaison. The implementation of this study took place over three weeks, and every week children created a different story. Collected data include photographs, student voice recordings, anecdotal notes, and a reflective journal. The three weeks of implementation data showed how touch technology provided a new modality of learning representation for young children in my classroom. The findings suggest that multiliteracies complemented traditional literacy, storytelling enhanced children’s communication, and touch technology functionality went beyond literacy skills

Functional Amyloid Formation within Mammalian Tissue

Amyloid is a generally insoluble, fibrous cross-β sheet protein aggregate. The process of amyloidogenesis is associated with a variety of neurodegenerative diseases including Alzheimer, Parkinson, and Huntington disease. We report the discovery of an unprecedented functional mammalian amyloid structure generated by the protein Pmel17. This discovery demonstrates that amyloid is a fundamental nonpathological protein fold utilized by organisms from bacteria to humans. We have found that Pmel17 amyloid templates and accelerates the covalent polymerization of reactive small molecules into melanin—a critically important biopolymer that protects against a broad range of cytotoxic insults including UV and oxidative damage. Pmel17 amyloid also appears to play a role in mitigating the toxicity associated with melanin formation by sequestering and minimizing diffusion of highly reactive, toxic melanin precursors out of the melanosome. Intracellular Pmel17 amyloidogenesis is carefully orchestrated by the secretory pathway, utilizing membrane sequestration and proteolytic steps to protect the cell from amyloid and amyloidogenic intermediates that can be toxic. While functional and pathological amyloid share similar structural features, critical differences in packaging and kinetics of assembly enable the usage of Pmel17 amyloid for normal function. The discovery of native Pmel17 amyloid in mammals provides key insight into the molecular basis of both melanin formation and amyloid pathology, and demonstrates that native amyloid (amyloidin) may be an ancient, evolutionarily conserved protein quaternary structure underpinning diverse pathways contributing to normal cell and tissue physiology

Proprotein convertase cleavage liberates a fibrillogenic fragment of a resident glycoprotein to initiate melanosome biogenesis

Lysosome-related organelles are cell type–specific intracellular compartments with distinct morphologies and functions. The molecular mechanisms governing the formation of their unique structural features are not known. Melanosomes and their precursors are lysosome-related organelles that are characterized morphologically by intralumenal fibrous striations upon which melanins are polymerized. The integral membrane protein Pmel17 is a component of the fibrils and can nucleate their formation in the absence of other pigment cell–specific proteins. Here, we show that formation of intralumenal fibrils requires cleavage of Pmel17 by a furin-like proprotein convertase (PC). As in the generation of amyloid, proper cleavage of Pmel17 liberates a lumenal domain fragment that becomes incorporated into the fibrils; longer Pmel17 fragments generated in the absence of PC activity are unable to form organized fibrils. Our results demonstrate that PC-dependent cleavage regulates melanosome biogenesis by controlling the fibrillogenic activity of a resident protein. Like the pathologic process of amyloidogenesis, the formation of other tissue-specific organelle structures may be similarly dependent on proteolytic activation of physiological fibrillogenic substrates

Collapse-and-revival dynamics of strongly laser-driven electrons

The relativistic quantum dynamics of an electron in an intense single-mode quantized electromagnetic field is investigated with special emphasis on the spin degree of freedom. In addition to fast spin oscillations at the laser frequency, a second time scale is identified due to the intensity dependent emissions and absorptions of field quanta. In analogy to the well-known phenomenon in atoms at moderate laser intensity, we put forward the conditions of collapses and revivals for the spin evolution in laser-driven electrons starting at feasible $10^{18}$ W/cm$^2$.Comment: 18 pages, 4 figure

Two specific mutations are prevalent causes of recessive retinitis pigmentosa in North American patients of Jewish ancestry.

PURPOSE: Retinitis pigmentosa is a Mendelian disease with a very elevated genetic heterogeneity. Most mutations are responsible for less than 1% of cases, making molecular diagnosis a multigene screening procedure. In this study, we assessed whether direct testing of specific alleles could be a valuable screening approach in cases characterized by prevalent founder mutations. METHODS: We screened 275 North American patients with recessive/isolate retinitis pigmentosa for two mutations: an Alu insertion in the MAK gene and the p.Lys42Glu missense in the DHDDS gene. All patients were unrelated; 35 reported Jewish ancestry and the remainder reported mixed ethnicity. RESULTS: We identified the MAK and DHDDS mutations homozygously in only 2.1% and 0.8%, respectively, of patients of mixed ethnicity, but in 25.7% and 8.6%, respectively, of cases reporting Jewish ancestry. Haplotype analyses revealed that inheritance of the MAK mutation was attributable to a founder effect. CONCLUSION: In contrast to most mutations associated with retinitis pigmentosa-which are, in general, extremely rare-the two alleles investigated here cause disease in approximately one-third of North American patients reporting Jewish ancestry. Therefore, their screening constitutes an alternative procedure to large-scale tests for patients belonging to this ethnic group, especially in time-sensitive situations.Genet Med 17 4, 285-290

Changes in readthrough acetylcholinesterase expression modulate amyloid-beta pathology

Alzheimer's disease has long been known to involve cholinergic deficits, but the linkage between cholinergic gene expression and the Alzheimer's disease amyloid pathology has remained incompletely understood. One known link involves synaptic acetylcholinesterase (AChE-S), shown to accelerate amyloid fibrils formation. Here, we report that the ‘Readthrough' AChE-R splice variant, which differs from AChE-S in its 26 C-terminal residues, inversely exerts neuroprotective effects from amyloid β (Aβ) induced toxicity. In vitro, highly purified AChE-R dose-dependently suppressed the formation of insoluble Aβ oligomers and fibrils and abolished Aβ toxicity to cultured cells, competing with the prevalent AChE-S protein which facilitates these processes. In vivo, double transgenic APPsw/AChE-R mice showed lower plaque burden, fewer reactive astrocytes and less dendritic damage than single APPsw mice, inverse to reported acceleration of these features in double APPsw/AChE-S mice. In hippocampi from Alzheimer's disease patients (n = 10), dentate gyrus neurons showed significantly elevated AChE-R mRNA and reduced AChE-S mRNA. However, immunoblot analyses revealed drastic reductions in the levels of intact AChE-R protein, suggesting that its selective loss in the Alzheimer's disease brain exacerbates the Aβ-induced damages and revealing a previously unforeseen linkage between cholinergic and amyloidogenic event

Multiphoton Ionization as Time-Dependent Tunneling

A new semiclassical approach to ionization by an oscillating field is presented. For a delta-function atom, an asymptotic analysis is performed with respect to a quantity h, defined as the ratio of photon energy to ponderomotive energy. This h appears formally equivalent to Planck's constant in a suitably transformed Schroedinger equation and allows semiclassical methods to be applicable. Systematically, a picture of tunneling wave packets in complex time is developped, which by interference account for the typical ponderomotive features of ionization curves. These analytical results are then compared to numerical simulations and are shown to be in good agreement.Comment: 36 pages (also printable half size), uuencoded compressed tarred Latex file with 9 Postscript figures included automaticall