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In natural systems, litter typically undergoes decomposition as species mixtures, with litter species composition and richness affecting decomposition processes and ecosystem functioning. In this study, we collected and measured leaf litter samples from 25 litter traps within a 1 ha permanent forest dynamics plot and 396 litter traps in three forest sites. Our objective was to investigate the temporal and spatial pattern of leaf litterfall in an old-growth evergreen broad-leaved forest. Mean annual leaf litterfall in the study area was 4.31 ± 0.46 Mg/ha. A total of 61 tree species were recorded in the leaf litter mixtures collected in the 1-ha forest plot, and 39, 48, and 37 tree species were found in the three forest sites. We noted that the abundance of the tree species in the leaf litter mixtures was best fitted as the preemption model. We observed a significant difference in litter species composition between the rain season and the dry season, as well as among the three forest sites, caused by the relative abundance of litterfall from evergreen and deciduous tree species in the two seasons, and the varying proportion of litterfall from dominant tree species in three forest sites. The species diversity of leaf litterfall also varied across different levels. Our study highlights the importance of the dominant tree species in producing leaf litters and determining the litter composition, and indicates that the studies on the leaf litter mixing effects in subtropical forests should be conducted at a high level of species richness, taking into account the relative abundance of litter species.

As an important hydrological ecosystem component, the glacier basin has great significance for climate and environment, and it is also linked to regional water sustainability. In this paper, the sampling and isotope analysis of glacial ice, ice-melt water, river water (river midstream and river downstream), groundwater (spring), and precipitation were carried out in a hydrological year of the Mingyong Glacier basin, which is located at the Meili Snow Mountains, Southeastern Tibetan Plateau. At the same time, the hydrograph separation of the recharge sources of the lower mountain pass is studied. The results show that the range of δD, δ18O, and d-excess (deuterium excess) in natural water bodies are significantly different, and the precipitation is the most obvious. The high values of δD and δ18O in the water samples all appeared in spring and summer, and the low values appeared in autumn and winter, while glacial ice showed opposite trends. Meanwhile, the local meteoric water line (LMWL) of the Mingyong Glacier basin is δD = 8.04δ18O + 13.06. The End-Member Mixing Analysis (EMMA) was adopted to determine the sources proportion of river water (river downstream) according to the δD, δ18O, and d-excess ratio relationships. The results showed that the proportion of ice-melt water, groundwater, and precipitation in the ablation period was 80.6%, 17.2%, and 2.2% as well as 19.2%, 73.1%, and 7.7% in the accumulation period, respectively. Ice-melt water has a higher conversion recharge rate to groundwater and indirectly recharges river water, especially in nonmonsoon seasons. In other words, the main recharge source of river water in the lower reaches of the Mingyong Glacier basin during the ablation period is ice-melt water. In the accumulation period, the main recharge source of river water in the lower reaches of the Mingyong Glacier basin is groundwater, while nearly half of the recharge of groundwater comes from ice-melt water. Therefore, regardless of the ablation period or the accumulation period, ice-melt water is sustainable and important to this region.

Toxic and major elements, such as As and Fe, in watersheds can significantly impact the surrounding water environment and ecosystem. Thus, in this study, we conducted an investigation into the origins and spatial distribution of typical toxic trace elements (As and Mn) and crustal major elements (Al, Fe, and Ti) in suspended particulate matter (SPM) across various glacial watersheds located at different elevations in the northeastern Tibetan Plateau (NETP) from June to July in 2017. The results revealed that the mean value of each element followed the order of abundance in the samples, with Al having the highest mean value at 21307 µg/L, followed by Fe at 13366 µg/L, Ti at 1520 µg/L, Mn at 245 µg/L, and As at 66.6 µg/L. Moreover, our study identified high content of these elements from the Dabanshan Snowpack, Laohugou Glacier No.12, and Yuzhufeng Glacier in the upper reaches of the basin, which were found to be 9.9, 10.2, and 19.4 times higher, respectively, than that of the upper reaches of the Heihe River. We found that As and Mn exhibited clear indications of anthropogenic influence on a local and regional scale. The calculated enrichment factor (EF) demonstrated a significant As enrichment (EF>100) in the Qiyi and Lenglongling Glaciers, possibly resulting in the release of upstream glacier melt and anthropogenic-derived As deposition. Our findings suggested that the upstream region was primarily linked to glacier meltwater discharge. In contrast, the middle and lower reaches of the basin exhibited a more pronounced influence from local human activities. Based on the findings, the water environment of the glacier watershed appears to be in good condition overall. However, the presence of elevated levels of As element in the water system can be traced back to both anthropogenic and natural factors. As a result, ensuring the safety of the water supply for nearby residents is a matter of utmost concern. This study provides a comprehensive examination of hydrochemical variations and the overall water environment of high-altitude glacier basins in the NETP, offering valuable insights into the topic.

In order to cope with environmental changes, plants constantly adjust their morphological characteristics in order to adapt to changing environment. In the present study, populations of Bombax ceiba from Mengla area and Yuanjiang area in Yunnan Province were selected as the research objects. Six tree structure factors, such as tree height and crown width, eight leaf trait factors, such as leaf area and leaf length, and several habitat factors, such as area topography, meteorology and soil nutrients, were measured. Structural equation model and variation decomposition method were applied to analyze the effects of various habitat factors on tree structure and leaf traits of B. ceiba, and to reveal its morphological responses to habitat heterogeneity. The results showed that there was a significant negative correlation between tree structure and leaf traits in the two study habitats (Mengla area and Yuanjiang area), and the correlation coefficient was −0.47 in Mengla area and −0.22 in Yuanjiang area. Both topographic and soil factors had positive effects on tree structure of the two habitats, and the topographic factors had a greater impact on tree structure than leaf traits. The main difference was that meteorological factors had a positive effect on tree structure of Mengla, but a negative effect on leaf traits, while Yuanjiang showed the opposite patterns. The variation analysis showed that the superposition of three environmental factors in Mengla area had a greater explanation power of tree structure and leaf traits than that in Yuanjiang area, and the topographic factors had the largest explanation power of tree structure in both areas, which reflected that fact that the characteristics of Mengla habitat imposed a greater influence on B. ceiba. The soil factors in Mengla area accounted for 20.1% of the leaf traits, while the meteorological factors in Yuanjiang area accounted for 11.6%. The results showed that leaf traits were sensitive to environmental differences. In general, the responses of B. ceiba to heterogeneous habitats is based on the specific performance of its resource utilization capacity. The research results can provide references for exploring the morphological responses of plants to heterogeneous habitats.

Abstract Alpine grasslands harbor diverse groups of flora and fauna, provide important ecosystem functions, and yield essential ecosystem goods and services, especially for the development of nature-based tourism. However, they are experiencing increasing anthropogenic perturbations such as tourist trampling. Although negative effects of tourist trampling on alpine vegetation have been frequently reported, previous studies have focused mainly on changes in taxonomic diversity after trampling, and rarely provide a mechanistic elucidation of trampling effects from a trait-based perspective. The present study evaluates the impacts of simulated trampling on taxonomic and functional diversity of a typical alpine grassland community in Shangri-La, China using a standardized protocol. The results showed that although taxonomic diversity was not statistically significantly affected by trampling, some functional attributes responded rapidly to trampling disturbance. Specifically, functional divergence decreased with an increase in trampling intensity, and characteristics of community-weighted mean trait values changed towards shorter species with reduced leaf area and lower leaf dry matter content. Such strong shifts in functional attributes may further affect ecosystem goods and services provided by alpine grasslands. Our inclusion of functional diversity in the analysis thus adds an important caution to previous studies predominantly focusing on taxonomic diversity, and it is urgent to keep alpine grasslands well managed and ecologically coherent so that their valuable functions and services can be safeguarded.

This study collected summer meltwater runoff samples from several glacier watersheds of the northeast Tibetan Plateau during June-July 2017, and measured the concentrations of 17 trace elements (Li, Be, Sc, V, Cr, Co, Ni, Cu, Zn, Ga, Rb, Mo, Cd, In, Sb, Cs, Ba) in meltwater suspended particulate matter (SPM), in order to reveal the elemental concentration, spatial distribution, and water quality in remote glacier watershed under regional anthropogenic activities. Results showed that, the concentration of heavy metal elements was relatively high in Yuzhufeng Glacier basin, ranging from 0.57 μg/L (In) to 1,551.6 μg/L (Ba), whereas in Qiyi Glacier basin it was the lowest, ranging from 0.02 to 85.05 μg/L; and relatively medium in other glacier watersheds, with total elemental concentration varying from 1,503.9 to 1726.2 μg/L. Moreover, enrichment factors (EFs) of SPM heavy metals showed significantly higher value in the downstream than that of upper glacier region of the watershed. Most heavy metals with low EFs mainly originated from crust dust, while others with higher EFs (e.g., Cd, Sb) probably originated from anthropogenic sources. Spatially, the EFs of heavy metals were higher in Yuzhufeng and Laohugou Glacier basins; while in other regions the EFs were relatively low, which may be caused by regional land-surface and atmospheric environmental differences surrounding the various glacier watersheds. Compared with other remote locations in global range, heavy metals level (e.g., Cu, Ni, and Zn) in this region is relatively higher. Meanwhile, we find that, though the water quality of the glacier basin in northeast Tibetan Plateau was relatively clean and pollution-free, it is still obviously affected by regional anthropogenic activities. Mining activities, transportation and natural weathering and erosion processes in the study areas have important effects on the content of heavy metal pollutants of river-water SPM in the glacier watershed. Moreover, backward air-mass trajectories demonstrated the potential atmospheric pollutants transport from the surrounding cities and suburbs, to deposit in the snowpack and glaciers, and then melted out and released into meltwater runoff. This study provides a new perspective on more complete view of heavy metals distribution in glacier watershed, and new understanding for the cryosphere water environment evaluation in the Tibetan Plateau region.

Summary: Nonalcoholic fatty liver disease (NAFLD) is a global health-care problem with limited therapeutic options. To obtain a cellular resolution of pathogenesis, 82,168 single-cell transcriptomes (scRNA-seq) across different NAFLD stages were profiled, identifying hepatocytes and 12 other non-parenchymal cell (NPC) types. scRNA-seq revealed insights into the cellular and molecular mechanisms of the disease. We discovered a dual role for hepatic stellate cells in gene expression regulation and in the potential to trans-differentiate into myofibroblasts. We uncovered distinct expression profiles of Kupffer cells versus monocyte-derived macrophages during NAFLD progression. Kupffer cells showed stronger immune responses, while monocyte-derived macrophages demonstrated a capability for differentiation. Three chimeric NPCs were identified including endothelial-chimeric stellate cells, hepatocyte-chimeric endothelial cells, and endothelial-chimeric Kupffer cells. Our work identified unanticipated aspects of mouse with NAFLD at the single-cell level and advanced the understanding of cellular heterogeneity in NAFLD livers.

Zoigê Plateau, China’s largest plateau marsh wetland, has experienced large-scale degradation of the marsh wetland and evolution of the wetland landscape pattern over the past 40 years due to climate warming and human activities. How exactly do the wetland landscape pattern characteristics change? How do climatic change and human activities affect the wetland evolution? These questions are yet to be systematically investigated. In order to investigate changes to the marsh wetland on the Zoigê Plateau, field investigations, spatial and statistical analysis were undertaken. Findings from our study indicate that from 1977–2016, the area of marsh wetland on the Plateau reduced by 56.54%, approximately 66,700 hm2 of marsh wetland has been lost. The centroids of both marsh and marshy meadow migrated and the landscape centroid migration behaviors were also correlated with the distribution and variation of the marsh wetland on different slopes. In addition, the number of marsh landscape patches initially increased before decreasing; the number of marshy meadow landscape patches also recorded an initial increase, followed by a decline before a final increase. As the effects of human activities weakened, the aggregation degrees of both marsh and marshy meadow increased. Overall, the fragmentation degree, diversity and fractal dimension of the marsh wetland all declined. An investigation into the driving factors affecting the Plateau area shows that the increase of annual average temperature was the natural factor while trenching and overgrazing were the main human factors resulting in wetland degradation. Results from this study provide basic data and theoretical foundation for the protection and restoration of marsh wetland in alpine regions.

Alpine grasslands provide essential ecosystem functions and services, yet they are increasingly subject to anthropogenic perturbations. Northwest Yunnan (NWY) is one of the most popular tourist destinations in China, and alpine grasslands of NWY are particularly susceptible to human recreational activities such as hiker trampling. However, studies that explicitly evaluate the effects of trampling disturbance on alpine communities of NWY are very limited, let alone research testing the responses of these communities to trampling from a functional trait perspective. The present research is performed in alpine grasslands within and outside of a core nature conservation area of Shangri-La, NWY of China, to study species- and community-level responses of alpine communities to simulated trampling through a functional trait-based approach. The results showed that although species-specific differences existed, alpine plants generally showed rapid morphological changes in response to trampling disturbance. At the community level, trampling disturbance caused strong shifts in taxonomy-based diversity metrics (e.g., species richness, Simpson’s index and Shannon’s index) and functional trait-based diversity metrics (e.g., functional richness, functional evenness, functional divergence, and community-weighted mean trait values). Specifically, increased trampling intensity led to a decrease in taxonomic diversity. Meanwhile, functional richness decreased, while functional evenness and functional divergence increased with an increase in trampling intensity. Also, characteristics of alpine community traits had changed towards those with shorter height, reduced leaf size and lower LDMC, and such strong shifts in taxonomic and functional diversity might further affect the functioning and value of alpine grasslands. Therefore, the protection of high-altitude natural grasslands with high sensitivity and vulnerability is urgent, especially as they are increasingly experiencing multiple environmental and climatic stressors nowadays.

Proteolytic bacterial communities, which mineralize organic nitrogen, play a key role in agricultural systems. In this study, alkaline metalloprotease (apr) gene fragments from proteolytic bacteria were investigated in bulk and rhizosphere paddy soil from four fields under organic management (for 2, 3, 5, and 9 years), and from one field under conventional management (for 2 years). We analyzed the abundance and structure of the proteolytic bacterial communities using real-time quantitative PCR and denaturing gradient gel electrophoresis. Our results showed that the abundance of proteolytic bacteria ranged from 1.57 × 108 to 8.02 × 108 copies/g of soil. In addition, the abundance of the proteolytic bacteria in the paddy soils under organic management was significantly higher than those in the paddy soil under conventional management. Moreover, the gene copy numbers in the rhizosphere soils were significantly higher than those in the bulk soils. The abundance of proteolytic bacteria tended to increase with the duration of organic management, with the highest abundance being found in the soil that had been under organic management for 5 years. However, the proteolytic bacteria communities in the paddy soils were not significantly affected by management practices. Phylogeny analysis showed that all gel bands obtained represented genes from Pseudomonas. Additionally, correlation analysis and canonical correspondence analysis showed that C/N, C, and N were important factors that influenced the abundance and community structure of the proteolytic bacteria. These results suggest that proteolytic bacteria are indicators in organic management systems, depolymerize organic N and hence maintain soil sustainability. Abbreviations: CM: conventional management; OM: organic management; DGGE: denaturing gradient gel electrophoresis; qPCR: real-time quantitative PCR detecting system; COFCC: China organic food certification center; CCA: canonical correspondence analysis

Reactive oxygen species (ROS) damage cell components, necessitating their clearance through autophagy. Here, the authors show that ROS can induce autophagy by triggering TRPML1 to release Ca2+from the lysosomal lumen, in turn activating the autophagy and lysosomal biogenesis regulator TFEB.

The Pettitt abrupt change test method based on ArcGIS was used to undertake change-point analysis on climatic (precipitation and potential evapotranspiration; 39 meteorological stations) and runoff data (27 hydrological stations) from 1954–2015 in the Nenjiang basin. The hydrological sensitivity analysis method was also used to calculate the influential component of climate change upstream, mid-stream, and downstream of the Nenjiang basin, as well as the effect of anthropogenic activities on runoff. Our results show that the upstream area has the highest contribution rate of climate change, followed by the mid-stream area; the downstream area has the lowest contribution rate. Studying climate change contribution rates in various sites in the Nenjiang basin, in addition to anthropogenic activities affecting runoff, can provide the foundation for the protection and utilization of basin water resources, as well as the conservation and restoration of wetlands.

Objectives: This study aims to investigate the expression patterns of mitochondrially encoded cytochrome c oxidase 1 (MT-CO1) in different organs and tissues of MRL/lpr mice aged six and 18 weeks. Materials and methods: Six-week-old female MRL/lpr mice (n=10) were considered young lupus model mice, and 18-week-old MRL/lpr mice (n=10) were considered old lupus model mice. Additionally, six-week-old (n=10) and 39-week-old (n=10) female Balb/c mice were used as the young and old controls, respectively. The messenger ribonucleic acid (mRNA) and protein expression levels of MT-CO1 in nine organs/tissues were detected via quantitative polymerase chain reaction (qPCR) and Western blot. Malondialdehyde (MDA) levels were determined with thiobarbituric acid colorimetry. The correlation coefficient of MT-CO1 mRNA levels and MDA levels in each organ/tissue at different ages was analyzed by Pearson correlation analysis. Results: The results showed that most non-immune organs/tissues (heart, lung, liver, kidneys, and intestines) showed increased MT-CO1 expression levels in younger MRL/lpr mice (p<0.05) and decreased MT-CO1 expression in older mice (p<0.05). Expression of MT-CO1 in the lymph nodes was low in younger mice but high in older mice. In other immune organs (spleen and thymus), MT-CO1 expression was low in older MRL/lpr mice. Lower mRNA expression and higher MDA levels were observed in the brains of MRL/lpr mice. However, all MRL/lpr mice showed higher MDA levels than Balb/c mice in every organ no matter younger or older MRL/lpr mice. Conclusion: Our study results suggest that lymphoid mitochondrial hyperfunction at organ level may be an important intrinsic pathogenesis in systemic lupus erythematosus activity, which may affect mitochondrial dysfunction in non-immune organs. [ABSTRACT FROM AUTHOR]

NASH is a chronic liver disease that affects 3%-6% of individuals and requires urgent therapeutic developments. Isolating the key cell types in the liver is a necessary step towards understanding their function and roles in disease pathogenesis. However, traditional isolation methods through gradient centrifugation can only collect one or a few cell types simultaneously and pose technical difficulties when applied to NASH livers. Taking advantage of identified cell surface markers from liver single-cell RNAseq, here we established the combination of gradient centrifugation and antibody-based cell sorting techniques to isolate five key liver cell types (hepatocytes, endothelial cells, stellate cells, macrophages and other immune cells) from a single mouse liver. This method yielded high purity of each cell type from healthy and NASH livers. Our five-in-one protocol simultaneously isolates key liver cell types with high purity under normal and NASH conditions, enabling for systematic and accurate exploratory experiments such as RNA sequencing. [ABSTRACT FROM AUTHOR]

Liver zonation characterizes the separation of metabolic pathways along the lobules and is required for optimal function. Wnt/β-catenin signaling controls metabolic zonation by activating genes in the perivenous hepatocytes, while suppressing genes in the periportal counterparts. We now demonstrate that glucagon opposes the actions of Wnt/β-catenin signaling on gene expression and metabolic zonation pattern. The effects were more pronounced in the periportal hepatocytes where 28% of all genes were activated by glucagon and inhibited by Wnt/β-catenin. The glucagon and Wnt/β-catenin receptors and their signaling pathways are uniformly distributed in periportal and perivenous hepatocytes and the expression is not regulated by the opposing signal. Collectively, our results show that glucagon controls gene expression and metabolic zonation in the liver through a counterplay with the Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Inactivating mutations in the insulin receptor results in extreme insulin resistance. The resulting hyperglycemia is very difficult to treat, and patients are at risk for early morbidity and mortality from complications of diabetes. We used the insulin receptor antagonist S961 to induce severe insulin resistance, hyperglycemia, and ketonemia in mice. Using this model, we show that glucagon receptor (GCGR) inhibition with a monoclonal antibody normalized blood glucose and β-hydroxybutyrate levels. Insulin receptor antagonism increased pancreatic β-cell mass threefold. Normalization of blood glucose levels with GCGR-blocking antibody unexpectedly doubled β-cell mass relative to that observed with S961 alone and 5.8-fold over control. GCGR antibody blockage expanded α-cell mass 5.7-fold, and S961 had no additional effects. Collectively, these data show that GCGR antibody inhibition represents a potential therapeutic option for treatment of patients with extreme insulin-resistance syndromes. [ABSTRACT FROM AUTHOR]

At the background of the experience economy becoming major economic form in place of the service economy, forest experience develops vigorously and becomes a new approach for people alleviating pressure, acquiring knowledge, and relaxation. On the basis of analyzing concept and characteristics of the forest experience, this paper analyzed current situation of forest experience by taking Kunming West Mountain Forest Park as an example. It found there are existing problems in subject image, development methods and degree, professional personnel and concept. In view of these problems, it came up with recommendations including determining subject image, designing special products, accelerating cultivation of professional personnel, and strengthening training of forest experience concept. It is expected to provide reference for multi-functional use theory of China and provide ideas for developing forest experience projects. [ABSTRACT FROM AUTHOR]

Neuronal polarization is pivotal for neural network formation during brain development. Axon differentiation is a hallmark of initial neuronal polarization. Here, we report that the leucine-rich repeat-containing protein netrin-G ligand-2 (NGL-2) as a polarity regulator that localizes asymmetrically in rat hippocampal neurons and is required for differentiation of the future axon. NGL-2 was associated with PAR complex, and this interaction resulted in local stabilization of axonal microtubules. Further study showed that the C terminal of NGL-2 binds to the PDZ domain of PAR6, and NGL-2 interacts with PAR3 and atypical PKCζ (aPKCζ), with PAR6 acting as a bridge or modifier. Then, NGL-2 regulates the local stabilization of microtubules and promotes axon differentiation by the aPKCζ/microtubule affinity-regulating kinase 2 pathway. These findings reveal the critical role of NGL-2 in regulating axon differentiation in rat hippocampal neurons and reveal a novel partner of the PAR complex. [ABSTRACT FROM AUTHOR]

Abstract: Many world natural heritages in China contain various protected areas, such as the nature reserve and the scenic park. In many heritage sites the heritage resource are not well protected and managed because of the existed problems in management, including lack of the chaotic of management aims, the complexity management structure, the nonconformity of management system. The effective management is the key to protect the world natural heritage. In order to seek the effective management model to manage the world natural heritage in China, it should learn from the international conversation system and protected manners of resources, the successful experiences of advanced national park management model in American and other countries. In this paper, taking the Jiajin Mountains Giant Panda Sanctuary as an example, the concept of national park management model was introduced to the world natural heritage in China. It examined the Jiajin Mountains Giant Panda Sanctuary national park management model by combining the Chinese national conditions. Firstly, the paper introduced the Jiajin Mountains giant panda sanctuary briefly. Secondly, it analyzed the conditions of protection and management in Jiajin Mountains Giant Panda Sanctuary and showed the existed problems of heritage management. Finally, it discussed the feasibility and necessity on national park managing the Jiajin Mountains Giant Panda Sanctuary. Therefore, it was carried on the useful exploration to constructer the national park management model of the world natural heritage with Chinese characteristic. [Copyright &y& Elsevier]

TRPML1 (mucolipin 1, also known as MCOLN1) is predicted to be an intracellular late endosomal and lysosomal ion channel protein that belongs to the mucolipin subfamily of transient receptor potential (TRP) proteins. Mutations in the human TRPML1 gene cause mucolipidosis type IV disease (ML4). ML4 patients have motor impairment, mental retardation, retinal degeneration and iron-deficiency anaemia. Because aberrant iron metabolism may cause neural and retinal degeneration, it may be a primary cause of ML4 phenotypes. In most mammalian cells, release of iron from endosomes and lysosomes after iron uptake by endocytosis of Fe3+-bound transferrin receptors, or after lysosomal degradation of ferritin–iron complexes and autophagic ingestion of iron-containing macromolecules, is the chief source of cellular iron. The divalent metal transporter protein DMT1 (also known as SLC11A2) is the only endosomal Fe2+ transporter known at present and it is highly expressed in erythroid precursors. Genetic studies, however, suggest the existence of a DMT1-independent endosomal and lysosomal Fe2+ transport protein. By measuring radiolabelled iron uptake, by monitoring the levels of cytosolic and intralysosomal iron and by directly patch-clamping the late endosomal and lysosomal membrane, here we show that TRPML1 functions as a Fe2+ permeable channel in late endosomes and lysosomes. ML4 mutations are shown to impair the ability of TRPML1 to permeate Fe2+ at varying degrees, which correlate well with the disease severity. A comparison of TRPML1-/- ML4 and control human skin fibroblasts showed a reduction in cytosolic Fe2+ levels, an increase in intralysosomal Fe2+ levels and an accumulation of lipofuscin-like molecules in TRPML1-/- cells. We propose that TRPML1 mediates a mechanism by which Fe2+ is released from late endosomes and lysosomes. Our results indicate that impaired iron transport may contribute to both haematological and degenerative symptoms of ML4 patients. [ABSTRACT FROM AUTHOR]

The resting membrane potential (Δψ) of the cell is negative on the cytosolic side and determined primarily by the plasma membrane's selective permeability to K+. We show that lysosomal Δψ is set by lysosomal membrane permeabilities to Na+ and H+, but not K+, and is positive on the cytosolic side. An increase in juxta-lysosomal Ca2+ rapidly reversed lysosomal Δψ by activating a large voltage-dependent and K+-selective conductance (LysoKVCa). LysoKVCa is encoded molecularly by SLO1 proteins known for forming plasma membrane BK channels. Opening of single LysoKVCa channels is sufficient to cause the rapid, striking changes in lysosomal Δψ. Lysosomal Ca2+ stores may be refilled from endoplasmic reticulum (ER) Ca2+ via ER-lysosome membrane contact sites. We propose that LysoKVCa serves as the perilysosomal Ca2+ effector to prime lysosomes for the refilling process. Consistently, genetic ablation or pharmacological inhibition of LysoKVCa, or abolition of its Ca2+ sensitivity, blocks refilling and maintenance of lysosomal Ca2+ stores, resulting in lysosomal cholesterol accumulation and a lysosome storage phenotype. [ABSTRACT FROM AUTHOR]

The mucolipin TRP (TRPML) proteins are a family of endolysosomal cation channels with genetically established importance in humans and rodent. Mutations of human TRPML1 cause type IV mucolipidosis, a devastating pediatric neurodegenerative disease. Our recent electrophysiological studies revealed that, although a TRPML 1-mediated current can only be recorded in late endosome and lysosome (LEL) using the lysosome patch clamp technique, a proline substitution in TRPML1 (TRPML1v432P) results in a large whole cell current. Thus, it remains unknown whether the large TRPML1v432P-mediated current results from an increased surface expression (trafficking), elevated channel activity (gating), or both. Here we performed systemic Pro substitutions in a region previously implicated in the gating of various 6 transmembrane cation channels. We found that several Pro substitutions displayed gain-of-function (GOF) constitutive activities at both the plasma membrane (PM) and endolysosomal membranes. Although wild-type TRPML1 and non-GOF Pro substitutions localized exclusively in LEL and were barely detectable in the PM, the GOF mutations with high constitutive activities were not restricted to LEL compartments, and most significantly, exhibited significant surface expression. Because lysosomal exocytosis is Ca2+dependent, constitutive Ca2+ permeability due to Pro substitutions may have resulted in stimulus-independent intralysosomal Ca2+ release, hence the surface expression and whole cell current of TRPML1. Indeed, surface staining of lysosome-associated membrane protein-1 (Lamp-1) was dramatically increased in cells expressing GOF TRPML1 channels. We conclude that TRPML1 is an inwardly rectifying, proton-impermeable, Ca2+ and Fe2+/Mn2+ duallypermeable cation channel that may be gated by unidentified cellular mechanisms through a conformational change in the cytoplasmic face of the transmembrane 5 (TM5). Furthermore, activation of TRPML1 in LEL may lead to the appearance of TRPML1 proteins at the PM. [ABSTRACT FROM AUTHOR]