Enhanced activation of an amino-terminally truncated isoform of the voltage-gated proton channel HVCN1 enriched in malignant B cells

The final published version can be found here: http://dx.doi.org/10.1073/pnas.1411390111M.C. is the recipient of a Bennett Fellowship from Leukaemia and Lymphoma Research (ref. 12002). M.A.B. is supported by a GlaxoSmithKline Oncology–Biotechnology and Biological Sciences Research Council Collaborative Awards in Science and Engineering PhD studentship. This work was supported by National Institutes of Health Grants GM087507 and GM102336 (to T.E.D.)

A Voltage-Gated H+ Channel Underlying pH Homeostasis in Calcifying Coccolithophores

Marine coccolithophorid phytoplankton are major producers of biogenic calcite, playing a significant role in the global carbon cycle. Predicting the impacts of ocean acidification on coccolithophore calcification has received much recent attention and requires improved knowledge of cellular calcification mechanisms. Uniquely amongst calcifying organisms, coccolithophores produce calcified scales (coccoliths) in an intracellular compartment and secrete them to the cell surface, requiring large transcellular ionic fluxes to support calcification. In particular, intracellular calcite precipitation using HCO3− as the substrate generates equimolar quantities of H+ that must be rapidly removed to prevent cytoplasmic acidification. We have used electrophysiological approaches to identify a plasma membrane voltage-gated H+ conductance in Coccolithus pelagicus ssp braarudii with remarkably similar biophysical and functional properties to those found in metazoans. We show that both C. pelagicus and Emiliania huxleyi possess homologues of metazoan Hv1 H+ channels, which function as voltage-gated H+ channels when expressed in heterologous systems. Homologues of the coccolithophore H+ channels were also identified in a diversity of eukaryotes, suggesting a wide range of cellular roles for the Hv1 class of proteins. Using single cell imaging, we demonstrate that the coccolithophore H+ conductance mediates rapid H+ efflux and plays an important role in pH homeostasis in calcifying cells. The results demonstrate a novel cellular role for voltage gated H+ channels and provide mechanistic insight into biomineralisation by establishing a direct link between pH homeostasis and calcification. As the coccolithophore H+ conductance is dependent on the trans-membrane H+ electrochemical gradient, this mechanism will be directly impacted by, and may underlie adaptation to, ocean acidification. The presence of this H+ efflux pathway suggests that there is no obligate use of H+ derived from calcification for intracellular CO2 generation. Furthermore, the presence of Hv1 class ion channels in a wide range of extant eukaryote groups indicates they evolved in an early common ancestor

Comparison of proton channel, phagocyte oxidase, and respiratory burst levels between human eosinophil and neutrophil granulocytes.

Robust production of reactive oxygen species (ROS) by phagocyte NADPH oxidase (phox) during the respiratory burst (RB) is a characteristic feature of eosinophil and neutrophil granulocytes. In these cells the voltage-gated proton channel (Hv1) is now considered as an ancillary subunit of the phox needed for intense ROS production. Multiple sources reported that the expression of phox subunits and RB is more intensive in eosinophils than in neutrophils. In most of these studies the eosinophils were not isolated from healthy individuals, and a comparative analysis of Hv1 expression had never been carried out. We performed a systematic comparison of the levels of essential phox subunits, Hv1 expression and ROS producing capacity between eosinophils and neutrophils of healthy individuals. The expression of phox components was similar, whereas the amount of Hv1 was approximately 10-fold greater in eosinophils. Furthermore, Hv1 expression correlated with Nox2 expression only in eosinophils. Additionally, in confocal microscopy experiments co-accumulation of Hv1 and Nox2 at the cell periphery was observed in resting eosinophils but not in neutrophils. While phorbol-12-myristate-13-acetate-induced peak extracellular ROS release was approximately 1.7-fold greater in eosinophils, oxygen consumption studies indicated that the maximal intensity of the RB is only approximately 1.4-fold greater in eosinophils. Our data reinforce that eosinophils, unlike neutrophils, generate ROS predominantly extracellularly. In contrast to previous works we have found that the two granulocyte types display very similar phox subunit expression and RB capacity. The large difference in Hv1 expression suggests that its support to intense ROS production is more important at the cell surface

Torpor, arousal and activity of hibernating greater horseshoe bats (Rhinolophus ferrumequinum)

1. Patterns of torpor, arousal, and activity in free-living greater horseshoe bats, Rhinolophus ferrumequinum, were investigated during the hibernation period by using temperature-sensitive radio-transmitters. 2. Torpor bouts varied between 0.1 - 11.8 days, with individual means ranging from 1.3 - 7.4 days. Torpor bout duration decreased with increasing ambient temperature. 3. Activity duration varied from 37 minutes – 54 hours 24 minutes, with individual means ranging from 2:29 to 8:58 hours. Activity duration increased with ambient temperatures above approximately 10oC. 4. Ten of 11 bats synchronised their arousals with dusk. The circadian rhythm of one bat showed a free-running pattern over a period of about five weeks. Arousals were more highly synchronised, and closer to dusk, in individuals with lower body condition. 5. That bats forage in mild weather is supported by the strong synchronisation of arousals with dusk, especially in bats with low body condition. 6. Patterns of torpor and subsequent activity are consistent with predictions that torpor lasts until a critical metabolic or water imbalance is achieved. Because metabolism and water loss are temperature-dependent, torpor bout duration decreases with increasing temperature. The imbalance is corrected during subsequent activity, which is relatively constant in duration until a temperature threshold of 10oC, above which increasing levels of foraging lead to longer activity bouts

The galaxies missed by Hubble and ALMA: the contribution of extremely red galaxies to the cosmic census at 3<z<8

Using deep JWST imaging from JADES, JEMS and SMILES, we characterize optically-faint and extremely red galaxies at $z>3$ that were previously missing from galaxy census estimates. The data indicate the existence of abundant, dusty and post-starburst-like galaxies down to $10^8$M$_\odot$, below the sensitivity limit of Spitzer and ALMA. Modeling the NIRCam and HST photometry of these red sources can result in extreme, high values for both stellar mass and star formation rate (SFR); however, including 7 MIRI filters out to 21$\mu$m results in decreased mass (median 0.6 dex for log$_{10}$M$^*$/M$_{\odot}>$10), and SFR (median 10$\times$ for SFR$>$100 M$_{\odot}$/yr). At $z>6$, our sample includes a high fraction of little red dots (LRDs; NIRCam-selected dust-reddened AGN candidates). We significantly measure older stellar populations in the LRDs out to rest-frame 3$\mu$m (the stellar bump) and rule out a dominant contribution from hot dust emission, a signature of AGN contamination to stellar population measurements. This allows us to measure their contribution to the cosmic census at $z>3$, below the typical detection limits of ALMA ($L_{\rm IR}<10^{12}L_\odot$). We find that these sources, which are overwhelmingly missed by HST and ALMA, could effectively double the obscured fraction of the star formation rate density at $4<z<6$ compared to some estimates, showing that prior to JWST, the obscured contribution from fainter sources could be underestimated. Finally, we identify five sources with evidence for Balmer breaks and high stellar masses at $5.5<z<7.7$. While spectroscopy is required to determine their nature, we discuss possible measurement systematics to explore with future data.Comment: submitted to AAS Journals, comments welcome

Field and Laboratory Studies Provide Insights into the Meaning of Day-Time Activity in a Subterranean Rodent (Ctenomys aff. knighti), the Tuco-Tuco

South American subterranean rodents (Ctenomys aff. knighti), commonly known as tuco-tucos, display nocturnal, wheel-running behavior under light-dark (LD) conditions, and free-running periods >24 h in constant darkness (DD). However, several reports in the field suggested that a substantial amount of activity occurs during daylight hours, leading us to question whether circadian entrainment in the laboratory accurately reflects behavior in natural conditions. We compared circadian patterns of locomotor activity in DD of animals previously entrained to full laboratory LD cycles (LD12∶12) with those of animals that were trapped directly from the field. In both cases, activity onsets in DD immediately reflected the previous dark onset or sundown. Furthermore, freerunning periods upon release into DD were close to 24 h indicating aftereffects of prior entrainment, similarly in both conditions. No difference was detected in the phase of activity measured with and without access to a running wheel. However, when individuals were observed continuously during daylight hours in a semi-natural enclosure, they emerged above-ground on a daily basis. These day-time activities consisted of foraging and burrow maintenance, suggesting that the designation of this species as nocturnal might be inaccurate in the field. Our study of a solitary subterranean species suggests that the circadian clock is entrained similarly under field and laboratory conditions and that day-time activity expressed only in the field is required for foraging and may not be time-dictated by the circadian pacemaker

Inhibition of HERG1 K+ channel protein expression decreases cell proliferation of human small cell lung cancer cells

HERG (human ether-à-go-go-related gene) K+ currents fulfill important ionic functions in cardiac and other excitable cells. In addition, HERG channels influence cell growth and migration in various types of tumor cells. The mechanisms underlying these functions are still not resolved. Here, we investigated the role of HERG channels for cell growth in a cell line (SW2) derived from small cell lung cancer (SCLC), a malignant variant of lung cancer. The two HERG1 isoforms (HERG1a, HERG1b) as well as HERG2 and HERG3 are expressed in SW2 cells. Inhibition of HERG currents by acute or sustained application of E-4031, a specific ERG channel blocker, depolarized SW2 cells by 10–15 mV. This result indicated that HERG K+ conductance contributes considerably to the maintenance of the resting potential of about −45 mV. Blockage of HERG channels by E-4031 for up to 72 h did not affect cell proliferation. In contrast, siRNA-induced inhibition of HERG1 protein expression decreased cell proliferation by about 50%. Reduction of HERG1 protein expression was confirmed by Western blots. HERG current was almost absent in SW2 cells transfected with siRNA against HERG1. Qualitatively similar results were obtained in three other SCLC cell lines (OH1, OH3, H82), suggesting that the HERG1 channel protein is involved in SCLC cell growth, whereas the ion-conducting function of HERG1 seems not to be important for cell growth