Increased store-operated Ca(2+) entry mediated by GNB5 and STIM1

Recent human genetic studies have shown that Gbeta5 is related to various clinical symptoms, such as sinus bradycardia, cognitive disability, and attention deficit hyperactivity disorder. Although the calcium signaling cascade is closely associated with a heterotrimeric G-protein, the function of Gbeta5 in calcium signaling and its relevance to clinical symptoms remain unknown. In this study, we investigated the in vitro changes of store-operated calcium entry (SOCE) with exogenous expression of Gbeta5. The cells expressing Gbeta5 had enhanced SOCE after depletion of calcium ion inside the endoplasmic reticulum. Gbeta5 also augmented Stim1- and Orai1-dependent SOCE. An ORAI1 loss-of-function mutant did not show inhibition of Gbeta5-induced SOCE, and a STIM1-ERM truncation mutant showed no enhancement of SOCE. These results suggested a novel role of GNB5 and Stim1, and provided insight into the regulatory mechanism of SOCE.ope

Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that Ca(2+) may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular Ca(2+) concentration ([Ca(2+)]i) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced [Ca(2+)]i increase with nonspecific Ca(2+) channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive [Ca(2+)]i elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a Ca(2+) chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that Ca(2+) may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream Ca(2+) regulation of the WNK-OSR1 pathway in intact cells.ope

DA-6034 Induces [Ca2+]i Increase in Epithelial Cells

DA-6034, a eupatilin derivative of flavonoid, has shown potent effects on the protection of gastric mucosa and induced the increases in fluid and glycoprotein secretion in human and rat corneal and conjunctival cells, suggesting that it might be considered as a drug for the treatment of dry eye. However, whether DA-6034 induces Ca2+ signaling and its underlying mechanism in epithelial cells are not known. In the present study, we investigated the mechanism for actions of DA-6034 in Ca2+ signaling pathways of the epithelial cells (conjunctival and corneal cells) from human donor eyes and mouse salivary gland epithelial cells. DA-6034 activated Ca2+-activated Cl- channels (CaCCs) and increased intracellular calcium concentrations ([Ca2+]i) in primary cultured human conjunctival cells. DA-6034 also increased [Ca2+]i in mouse salivary gland cells and human corneal epithelial cells. [Ca2+]i increase of DA-6034 was dependent on the Ca2+ entry from extracellular and Ca2+ release from internal Ca2+ stores. Interestingly, these effects of DA-6034 were related to ryanodine receptors (RyRs) but not phospholipase C/inositol 1,4,5-triphosphate (IP3) pathway and lysosomal Ca2+ stores. These results suggest that DA-6034 induces Ca2+ signaling via extracellular Ca2+ entry and RyRs-sensitive Ca2+ release from internal Ca2+ stores in epithelial cells.ope

Growth and Characterization of ZnO film by Metal Organic Chemical Vapor Deposition (MOCVD)

Maste

ZnO nanorod synthesis by a Metal Organic Chemical Vapor Deposition(MOCVD) and its electronic device applications

Docto

Visualization of pass-by noise by means of a line array of microphones affixed to the ground

This paper introduces the improved moving frame acoustic holography (MFAH) method and its application. MFAH allows us to visualize the noise generated by moving noise sources by employing a vertical line array of microphones affixed to the ground. The sound field generated by moving sources is different from that of stationary ones due to the movement of the sources. Therefore the measured sound pressure by the microphone on the ground has to be processed so that it cooperates the effect of the movement. This paper discusses the effect of moving noise sources on the obtained hologram by MFAH. This assures the applicability of MFAH to the visualization of moving sources. This paper also reviews the improved MFAH that can visualize a coherent narrow band noise and a pass-by noise. The practical applicability of the improved MFAH was demonstrated by visualizing tire noise during a pass-by test

Lysosomal Ca2+ Signaling is Essential for Osteoclastogenesis and Bone Remodeling

Lysosomal Ca2+ emerges as a critical component of receptor-evoked Ca2+ signaling and plays a crucial role in many lysosomal and physiological functions. Lysosomal Ca2+ release is mediated by the transient receptor potential (TRP) family member TRPML1, mutations that cause the lysosomal storage disease mucolipidosis type 4. Lysosomes play a key role in osteoclast function. However, nothing is known about the role of lysosomal Ca2+ signaling in osteoclastogenesis and bone metabolism. In this study, we addressed this knowledge gap by studying the role of lysosomal Ca2+ signaling in osteoclastogenesis, osteoclast and osteoblast functions, and bone homeostasis in vivo. We manipulated lysosomal Ca2+ signaling by acute knockdown of TRPML1, deletion of TRPML1 in mice, pharmacological inhibition of lysosomal Ca2+ influx, and depletion of lysosomal Ca2+ storage using the TRPML agonist ML-SA1. We found that knockdown and deletion of TRPML1, although it did not have an apparent effect on osteoblast differentiation and bone formation, markedly attenuated osteoclast function, RANKL-induced cytosolic Ca2+ oscillations, inhibited activation of NFATc1 and osteoclastogenesis-controlling genes, suppressed the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs), and markedly reduced the differentiation of bone marrow-derived macrophages into osteoclasts. Moreover, deletion of TRPML1 resulted in enlarged lysosomes, inhibition of lysosomal secretion, and attenuated the resorptive activity of mature osteoclasts. Notably, depletion of lysosomal Ca2+ with ML-SA1 similarly abrogated RANKL-induced Ca2+ oscillations and MNC formation. Deletion of TRPML1 in mice reduced the TRAP-positive bone surfaces and impaired bone remodeling, resulting in prominent osteopetrosis. These findings demonstrate the essential role of lysosomal Ca2+ signaling in osteoclast differentiation and mature osteoclast function, which play key roles in bone homeostasis. © 2016 American Society for Bone and Mineral Research.restrictio

Sound Visualization of Moving Noise Sources by using Moving Frame Acoustic Holography

This paper introduces moving frame acoustic holography (MFA H) method. The method was originally proposed to increase the aperture size and the spatial resolution of hologram by using a moving line array of microphones. The hologram of the scanned plane can be obtained by the relative coordinate transformation. The main advantage of this method is that it enables us to visualize the noise generated by moving noise sources by employing a vertical line array of microphones affixed to the ground. This paper demonstrates the practical applicability of MFAH for the visualization of moving noise sources and discusses the limitation of the method

TRPML1 as lysosomal fusion guard.

restrictio

Review of the Improved Moving Frame Acoustic Holography and Its Application to the Visualization of Moving Noise Sources

This paper reviews the improved moving frame acoustic holography (MFAH) method and its application. Moving frame acoustic holography was originally proposed to increase the aperture size and the spatial resolution of hologram by using a moving line array of microphones. The hologram of scanned plane can be obtained by assuming the sound field to be a product of spatial and temporal information. Although conventional MFAH was only applied to sinusoidal signals, it allows us to visualize the noise generated by moving noise sources by employing a vertical line array of microphones affixed to the ground. However, the sound field generated by moving sources becomes different from that of stationary ones due to the movement of the sources. Firstly, this paper introduces the effect of moving noise sources on the obtained hologram by MFAH and the applicability of MFAH to the visualization of moving sources. Secondly, this paper also reviews improved MFAH that can visualize a coherent narrow band noise and a pass-by noise. The practical applicability of the improved MFAH was demonstrated by visualizing tire noise during a pass-by test