Genetic code expansion, an emerging tool in the Ca2+ ion channel field.

Various methods for characterizing binding forces as well as for monitoring and remote control of ion channels are still emerging. A recent innovation is the direct incorporation of unnatural amino acids (UAAs) with corresponding biophysical or biochemical properties, which are integrated using genetic code expansion technology. Minimal changes to natural amino acids, which are achieved by chemical synthesis of corresponding UAAs, are valuable tools to provide insight into the contributions of physicochemical properties of side chains in binding events. To gain unique control over the conformational changes or function of ion channels, a series of light-sensitive, chemically reactive and posttranslationally modified UAAs have been developed and utilized. Here, we present the existing UAA tools, their mode of action, their potential and limitations as well as their previous applications to Ca2+-permeable ion channels.

A candidate gene of Alzheimer diseases was mutated in senescence-accelerated mouse prone (SAMP) 8 mice.

The senescence-accelerated mouse prone (SAMP) 8 strain exhibits age-related learning and memory deficits (LMD) at 2 months of age. We have found strong association of chromosome 12 locus with learning memory deficit (LMD) phenotype in SAMP8 strain. In the course of searching candidate gene, here we identified solute carrier family 24 sodium/potassium/calcium exchanger member 4 (Slc24a4) in SAMP8 chromosome 12 LMD possessing one single nucleotide polymorphism causing amino acid replacement of Threonine at 413 position with Methionine. Since SLC24A4 has been postulated as a candidate of late onset Alzheimer's diseases (LOAD), we further analyze the functional importance of this polymorphism. By expressing Slc24a4 protein in HEK293 cells, here we showed polymorphic SAMP8 type Slc24a4-T413 M causing significant loss of calcium ion (Ca2+) transporter activity in cells compared with that of wild type mouse (Slc24a4-WT). However, no study yet shows any functional association of human SLC24A4 polymorphism with the onset of LOAD pathogenesis. Thus, our present finding may further help to clarify the importance of this ion exchanger with age related cognitive dysfunction.

Fluorescence-based logic gate for sensing of Ca2+ and F- ions using PVP crowned chrysene nanoparticles in aqueous medium.

Polyvinyl pyrrolidone (PVP) crowned chrysene nanoparticles (CHYNPs) were prepared by using a reprecipitation method. Dynamic light scattering (DLS) and scanning electron microscope (SEM) studies indicate that the monodispersed spherical nanoparticles bear a negative charge on their surfaces. The bathochromic spectral shift in the UV-visible and fluorescence spectrum of CHYNPs from chrysene (CHY) in acetone solution supports the J- type aggregation of nanoparticles. The aggregation-induced enhanced emission of CHYNPs at 486 and 522 nm decreases by increasing the concentration of the Ca2+ ion solution. It can display an ON-OFF type fluorescence response with high selectivity towards Ca2+ ions aqueous medium. Furthermore, the in situ generated PVP-CHYNPs-Ca2+ ensemble could recover the quenched fluorescence upon the addition of fluoride anions resulting in an OFF-ON type sensor. The present method has a correlation coefficient R2 = 0.988 with a detection limit of 1.22 µg/mL for Ca2+ in the aqueous medium. The fluorescence changes of PVP crowned CHYNPs upon the addition of Ca2+ and F- can be utilized as an INHIBIT logic gate at the molecular level, using Ca2+ and F- chemical inputs and the fluorescence intensity signal as output.

Melatonin regulates splenocytes proliferation via IP3-dependent intracellular Ca2+ release in seasonally breeding bird, Perdicula asiatica.

Melatonin plays an important role in the immune regulation of birds. Both endogenous and exogenous melatonin modulates lymphocyte proliferation via its specific membrane receptors, Mel(1a), Mel(1b) and Mel(1c), though the mechanisms behind this process are poorly understood. We investigated the differences in melatonin membrane receptor Mel(1a), Mel(1b) and Mel(1c) expression by western blot and reverse transcription reaction and the in vitro effect of melatonin on the intracellular Ca(2+) concentration ([Ca2+]i) in splenocytes of the Indian Jungle Bush Quail, Perdicula asiatica. We used a non-selective melatonin receptor antagonist for Mel(1a) and Mel(1b), luzindole, and the selective Mel(1b) blocker, 4P-PDOT to check the specific role of melatonin receptor on ([Ca2+]i). The expression of Mel(1a), Mel(1b) and Mel(1c) receptors mRNA and protein was upregulated by melatonin (10(-7) M) with a significant high rise in ([Ca2+]i), which was differentially blocked by supplementation of antagonist, luzindole (10(-7) M) and 4P-PDOT (10(-7) M). Furthermore, we noted in vitro effect of melatonin and 2-aminoethoxydiphenyl borate (2-APB), a cell-permeable antagonist of inositol 1, 4, 5-trisphosphate (IP3) receptor to check the rise in ([Ca2+]i) through the IP3 pathway. Significantly low ([Ca2+]i) was noted in melatonin and 2-APB pretreated splenocytes when compared with splenocytes where 2-APB was absent. Thus, our data suggest that melatonin through its membrane receptor induced the elevation of ([Ca2+]i) via IP(3)-dependent pathway for splenocyte proliferation in P. asiatica.

Assessment of the biointeractivity of a novel vital pulp therapy agent derived from eggshell biowaste: An in vitro study.

This study aimed to assess the calcium (Ca2+) and hydroxyl (OH-) ion-releasing ability, namely the biointeractivity of eggshell-derived hydroxyapatite (ESDHA) in comparison with mineral trioxide aggregate (MTA) and calcium hydroxide (CH). ESDHA, MTA and CH samples (n = 10; 8 × 1.6 mm) were immersed in 10 mL of deionised water (37°C, pH 6.8). Ca2+ and OH- ion releases were detected in 1, 7 and 21 days. Scanning electron microscopy and Fourier transform infrared spectroscopy analyses were also conducted. IBM SPSS 20.0 was used for statistical analyses. The cumulative Ca2+ ions (56.22 ± 11.28 ppm) were detected as most significant in ESDHA (day 21; p < 0.05). The OH- ion values of the ESDHA group were statistically higher than MTA and CH (days 1 and 7; p < 0.05). ESDHA and CH showed a similar pattern with sharp peaks in Ca2+, oxygen and carbon elements. ESDHA being a sustainable material with a high ion-releasing ability may be a preferable alternative to the commercial vital pulp therapy agents.

Removal of Calcareous Concretions from Marine Archaeological Ceramics by Means of a Stimuli-Responsive Hydrogel.

The presence of calcareous concretions on the surface of marine archaeological ceramics is a frequently observed phenomenon. It is necessary to remove these materials when the deposits obscure the feature of ceramics. Unfortunately, calcareous concretions provide distinctive documentation of the burning history of ceramics. The interaction of acid solution or detachment of the deposit layers in physical ways leads to the loss of archeological information. To prevent the loss of archeological information and to achieve precise and gentle concretion removal, responsive hydrogel cleaning systems have been developed. The hydrogels synthesized are composed of networks of poly(vinyl acetate)/sodium alginate that exhibit desirable water retention properties, are responsive to Ca2+ ions, and do not leave any residues after undergoing cleaning treatment. Four distinct compositions were selected. The study of water retention properties involved quantifying the weight changes. The composition was obtained from Fourier transform infrared spectra. The microstructure was obtained from scanning electron microscopy. The mechanical properties were obtained from rheological measurements. To demonstrate both the efficiency and working mechanism of the selected hydrogels, a representative study of mocked samples is presented first. After selecting the most appropriate hydrogel composite, a cleaning process was implemented on the marine archaeological ceramics. This article demonstrates the advantages of stimuli-responsive hydrogels in controlling the release of acid solution release, thereby surpassing the limitations of traditional cleaning methods.

Bacterial production of recombinant contraceptive vaccine antigen from CatSper displayed on a human papilloma virus-like particle.

CatSper is a voltage dependent calcium ion channel present in the principal piece of sperm tail. It plays a crucial role in sperm hyperactivated motility and so in fertilization. Extracellular loops of mouse sperm CatSper were used to develop a vaccine to achieve protection from pregnancy. These loops were inserted at one of the three hypervariable regions of Human Papilloma Virus (HPV) capsid protein (L1). Recombinant vaccines were expressed in E.coli as inclusion body (IB), purified, refolded and assembled into virus-like particles (VLP) in vitro, and adsorbed on alum. Four vaccine candidates were tested in Balb/C mice. All the constructs proved immunogenic, one showed contraceptive efficacy. This recombinant contraceptive vaccine is a non-hormonal intervention and is expected to give long-acting protection from undesired pregnancies.

The Role of Calmodulin Binding Transcription Activator in Plants under Different Stressors: Physiological, Biochemical, Molecular Mechanisms of Camellia sinensis and Its Current Progress of CAMTAs.

Low temperatures have a negative effect on plant development. Plants that are exposed to cold temperatures undergo a cascade of physiological, biochemical, and molecular changes that activate several genes, transcription factors, and regulatory pathways. In this review, the physiological, biochemical, and molecular mechanisms of Camellia sinensis have been discussed. Calmodulin binding transcription activator (CAMTAs) by molecular means including transcription is one of the novel genes for plants' adaptation to different abiotic stresses, including low temperatures. Therefore, the role of CAMTAs in different plants has been discussed. The number of CAMTAs genes discussed here are playing a significant role in plants' adaptation to abiotic stress. The illustrated diagrams representing the mode of action of calcium (Ca2+) with CAMTAs have also been discussed. In short, Ca2+ channels or Ca2+ pumps trigger and induce the Ca2+ signatures in plant cells during abiotic stressors, including low temperatures. Ca2+ signatures act with CAMTAs in plant cells and are ultimately decoded by Ca2+sensors. To the best of our knowledge, this is the first review reporting CAMAT's current progress and potential role in C. sinensis, and this study opens a new road for researchers adapting tea plants to abiotic stress.

TRPP2 ion channels: The roles in various subcellular locations.

TRPP2 (PC2, PKD2 or Polycytin-2), encoded by PKD2 gene, belongs to the nonselective cation channel TRP family. Recently, the three-dimensional structure of TRPP2 was constructed. TRPP2 mainly functions in three subcellular compartments: endoplasmic reticulum, plasma membrane and primary cilia. TRPP2 can act as a calcium-activated intracellular calcium release channel on the endoplasmic reticulum. TRPP2 also interacts with other Ca2+ release channels to regulate calcium release, like IP3R and RyR2. TRPP2 acts as an ion channel regulated by epidermal growth factor through activation of downstream factors in the plasma membrane. TRPP2 binding to TRPC1 in the plasma membrane or endoplasmic reticulum is associated with mechanosensitivity. In cilium, TRPP2 was found to combine with PKD1 and TRPV4 to form a complex related to mechanosensitivity. Because TRPP2 is involved in regulating intracellular ion concentration, TRPP2 mutations often lead to autosomal dominant polycystic kidney disease, which may also be associated with cardiovascular disease. In this paper, we review the molecular structure of TRPP2, the subcellular localization of TRPP2, the related functions and mechanisms of TRPP2 at different sites, and the diseases related to TRPP2.

Shaped by leaky ER: Homeostatic Ca2+ fluxes.

At any moment in time, cells coordinate and balance their calcium ion (Ca2+) fluxes. The term 'Ca2+ homeostasis' suggests that balancing resting Ca2+ levels is a rather static process. However, direct ER Ca2+ imaging shows that resting Ca2+ levels are maintained by surprisingly dynamic Ca2+ fluxes between the ER Ca2+ store, the cytosol, and the extracellular space. The data show that the ER Ca2+ leak, continuously fed by the high-energy consuming SERCA, is a fundamental driver of resting Ca2+ dynamics. Based on simplistic Ca2+ toolkit models, we discuss how the ER Ca2+ leak could contribute to evolutionarily conserved Ca2+ phenomena such as Ca2+ entry, ER Ca2+ release, and Ca2+ oscillations.

Analysis of the mechanism of synaptic integration focusing on the charge held in the spine.

Successful synaptic integration is said to require that multiple excitatory postsynaptic potentials (EPSPs) occur almost simultaneously over a short period of time, so that they overlap and increase. However, if brain function is based on a chain of successful synaptic integrations, then constraints on the spacing of multiple EPSP generation must be released to allow for a higher probability of successful synaptic integration. This paper demonstrates that Ca2+ ions retained in spines after EPSP generation polarize spine neck fluid and dendritic fluid as a dielectric medium, that polarization is transmitted through dendrites to the cell body (soma), that polarization is enhanced by the addition of polarization from each spine, and that I propose that synaptic integration is successful when the membrane potential, as determined by the enhanced polarization and membrane capacitance, reaches the threshold of voltage-gated Na+ channels. Furthermore, the approach taken in this study suggests that a single neuron can integrate synapses for many combinations of synaptic inputs, that successful synaptic integration depends on spine neck capacitance and spine head size, and that spines farther from the soma are able to contribute to successful synaptic integration, and led to the elucidation of a number of important issues, including the fact that inhibitory post-synapses on dendrites suppress s effectively synaptic integration.

Investigation of the molecular mechanism underlying the inhibitory activities of ethanol extract of Bombyx mori pupa-incubated Cordyceps militaris fruiting bodies toward allergic rhinitis.

Cordyceps militaris has been widely studied for its various pharmacological activities such as antitumor, anti-inflammation, and immune regulation. The binding of an allergen to IgE-sensitized mast cells in nasal mucosa triggers allergic rhinitis. We found that oral administration of 300 mg/kg of the ethanol extract prepared from silkworm pupa-cultivated Cordyceps militaris fruiting bodies significantly alleviated the symptoms of ovalbumin-induced allergic rhinitis in mice, including sneeze/scratch, mast cell activation, eosinophil infiltration, and Syk activation. The treatment of ethanol extract significantly suppressed the release of ß-hexosaminidase (a degranulation marker) and mRNA expression levels of various cytokines, including IL-3, IL-10, and IL-13 in activated RBL2H3 cells. The ethanol extract and ß-sitostenone, which was purified from the extract, could respectively reduce the Ca2+ ion mobilization in activated RBL-2H3 cells. Furthermore, results collected from western immunoblotting demonstrated that ethanol extract significantly retarded Ca2+ ion mobilization-initiated signaling cascade, which provoked the expression of various allergic cytokines. Also, the extract incubation interfered with P38 as well as NF-kB activation and Nrf-2 translocation. Our study suggested that ethanol extract possessed some natural constituents which could inhibit immediate degranulation and de novo synthesis of allergic cytokines via inhibition of Ca2+ ion mobilization in mast cells in the nasal mucosa of allergic rhinitis mice.

'Targeting the feast of a sleeping beast': Nutrient and mineral dependencies of encysted Acanthamoeba castellanii.

Acanthamoeba spp. cause a corneal infection, Acanthamoeba keratitis (AK), and a cerebral infection, granulomatous amoebic encephalitis (GAE). Though aggressive chemotherapy has been able to kill the active trophozoite form of Acanthamoeba, the encysted form of this parasite has remained problematic to resist physiological concentrations of drugs. The emergence of encysted amoeba into active trophozoite form poses a challenge to eradicate this parasite. Acanthamoeba trophozoites have active metabolic machinery that furnishes energy in the form of ATPs by subjecting carbohydrates and lipids to undergo pathways including glycolysis and beta-oxidation of free fatty acids, respectively. However, very little is known about the metabolic preferences and dependencies of an encysted trophozoite on minerals or potential nutrients that it consumes to live in an encysted state. Here, we investigate the metabolic and nutrient preferences of the encysted trophozoite of Acanthamoeba castellanii and the possibility to target them by drugs that act on calcium ion dependencies of the encysted amoeba. The experimental assays, immunostaining coupled with bioinformatics tools show that the encysted Acanthamoeba uses diverse nutrient pathways to obtain energy in the quiescent encysted state. These findings highlight potential pathways that can be targeted in eradicating amoebae cysts successfully.

Lanthanum mediated rutin yellow-fluorescent carbon dots as multifaceted sensing probes for the detection of calcium ions in melanoma and plant cells.

Calcium (Ca2 +) as a signaling ion and intracellular second messenger plays a crucial role in living organisms for various cellular functions. In the present work, we have designed a novel yellow-fluorescent carbon dots (LERCDs) using lanthanum, ethylenediaminetetraacetic acid (EDTA), and rutin as precursors for the sensing of Ca2 + ions. In this sense, a combination of hydrothermal and reflux methods was adopted. The as-designed LERCDs display bright yellow color emission in the aqueous solutions with a high quantum yield (23.8%). The LERCDs showed excitation-independent emission property along with magnificent photostability and time stability. The LERCDs show potential fluorescence quenching response towards the Ca2+ ions in the concentration range of 0-25 µM with a detection limit of 2.19 µM. The LERCDs have studied for extracellular sensing of Ca2 + ions in both melanoma cell lines (A375) and onion epidermal cells by employing fluorescence microscopy. The LERCDs facilitate low cytotoxicity and superior biocompatibility features in A375 cells. The practicality of LERCDs was studied in biological samples like the human serum.

Calcium-Induced Activity and Folding of a Repeat in Toxin Lipase from Antarctic Pseudomonas fluorescens Strain AMS8.

It is hypothesized that the Ca2+ ions were involved in the activity, folding and stabilization of many protein structures. Many of these proteins contain repeat in toxin (RTX) motifs. AMS8 lipase from Antarctic Pseudomonas fluorescens strain AMS8 was found to have three RTX motifs. So, this research aimed to examine the influence of Ca2+ ion towards the activity and folding of AMS8 lipase through various biophysical characterizations. The results showed that CaCl2 increased lipase activity. The far-UV circular dichroism (CD) and Fourier-transform infrared (FTIR) analysis suggested that the secondary structure content was improved with the addition of CaCl2. Fluorescence spectroscopy analysis showed that the presence of CaCl2 increased protein folding and compactness. Dynamic light scattering (DLS) analysis suggested that AMS8 lipase became aggregated at a high concentration of CaCl2.The binding constant (Kd) value from the isothermal titration calorimetry (ITC) analysis proved that the Ca2+ ion was tightly bound to the AMS8 lipase. In conclusion, Ca2+ ions play crucial roles in the activity and folding of the AMS8 lipase. Calcium binding to RTX nonapeptide repeats sequences will induced the formation and folding of the RTX parallel ß-roll motif repeat structure.

Efficacy of pulsed electromagnetic fields and electromagnetic fields tuned to the ion cyclotron resonance frequency of Ca2+ on chondrogenic differentiation.

Previous studies provide strong evidence for the therapeutic effect of electromagnetic fields (EMFs) on different tissues including cartilage. Diverse exposure parameters applied in scientific reports and the unknown interacting mechanism of EMF with biological systems make EMF studies challenging. In 1985, Liboff proposed that when magnetic fields are tuned to the cyclotron resonance frequencies of critical ions, the motion of ions through cell membranes is enhanced, and thus biological effects appear. Such exposure system consists of a weak alternating magnetic field (B1 ) in the presence of a static magnetic field (B0 ) and depends on the relationship between the magnitudes of B0 and B1 and the angular frequency Ω. The purpose of the present study is to determine the chondrogenic potential of EMF with regards to pulsed EMF (PEMF) and the ion cyclotron resonance (ICR) theory. We used different stimulating systems to generate EMFs in which cells are either stimulated with ubiquitous PEMF parameters, frequently reported, or parameters tuned to satisfy the ICR for Ca2+ (including negative and positive control groups). Chondrogenesis was analysed after 3 weeks of treatment. Cell stimulation under the ICR condition showed positive results in the context of glycosaminoglycans and type II collagen synthesis. In contrast, the other electromagnetically stimulated groups showed no changes compared with the control groups. Furthermore, gene expression assays revealed an increase in the expression of chondrogenic markers (COL2A1, SOX9, and ACAN) in the ICR group. These results suggest that the Ca2+ ICR condition can be an effective factor in inducing chondrogenesis.

The Role of 17ß-Estradiol and Estrogen Receptors in Regulation of Ca2+ Channels and Mitochondrial Function in Cardiomyocytes.

Numerous epidemiological, clinical, and animal studies showed that cardiac function and manifestation of cardiovascular diseases (CVDs) are different between males and females. The underlying reasons for these sex differences are definitely multifactorial, but major evidence points to a causal role of the sex steroid hormone 17ß-estradiol (E2) and its receptors (ER) in the physiology and pathophysiology of the heart. Interestingly, it has been shown that cardiac calcium (Ca2+) ion channels and mitochondrial function are regulated in a sex-specific manner. Accurate mitochondrial function and Ca2+ signaling are of utmost importance for adequate heart function and crucial to maintaining the cardiovascular health. Due to the highly sensitive nature of these processes in the heart, this review article highlights the current knowledge regarding sex dimorphisms in the heart implicating the importance of E2 and ERs in the regulation of cardiac mitochondrial function and Ca2+ ion channels, thus the contractility. In particular, we provide an overview of in-vitro and in-vivo studies using either E2 deficiency; ER deficiency or selective ER activation, which suggest that E2 and ERs are strongly involved in these processes. In this context, this review also discusses the divergent E2-responses resulting from the activation of different ER subtypes in these processes. Detailed understanding of the E2 and ER-mediated molecular and cellular mechanisms in the heart under physiological and pathological conditions may help to design more specifically targeted drugs for the management of CVDs in men and women.

Quantitative Analysis of Tip Growth, Phototropic Responses, and Other Blue Light-Dependent Photoresponses of Vaucheria.

The coenocytic tip-growing alga Vaucheria exhibits positive and negative phototropism, apical expansion, polarotropism, and branch induction from the illuminated region of the cell, all of which are caused by blue light. The bending response of Vaucheria is a blue light-mediated growth response. Differently from diffuse-growing cells or organs, the apical hemispherical dome of the Vaucheria cell is the site of not only maximum growth activity but also the site of blue light perception. Thence the phototropic response is initiated by the bulging mechanism: that is, a quick shift of the growth center to the adjacent subapical flank region. Since tip growth is driven by localized exocytosis, both phototropic bending and branch induction are considered to be closely related blue light-responses. Here I describe first how to prepare a highly useful culture medium for most freshwater algae, to establish unialgal and axenic culture of Vaucheria, and then describe several simple illumination systems using ordinary and/or inverted microscopes for the measurements of tip growth and for analyses of phototropism, polarotropism, and blue light-induced branching. Brief information is also included concerning the nature and function of aureochrome, the newly discovered, ochrophyte-specific blue light receptor. Aureochrome mediates blue light-induced branching, but its role in the phototropic response is still not elucidated.

Screening of Novel Inhibitors Against Leishmania donovani Calcium ion Channel to Fight Leishmaniasis.

Leishmania is an intracellular protozoan parasite which causes Leishmaniasis, a global health problem affecting millions of people throughout 89 different countries in the world. The current treatment which includes use of amphotericin B, antimonials, and others has major drawbacks due to toxicity, resistance, and extraordinary high cost. So there is an urgent need of development of new drug targets to fight against leishmaniasis. In this regard we have selected Leishmania donovani Ca2+ ion channel (Ld-CC) as potential drug target. Ld-CC regulates concentration of Ca2+ ions which is involved in several functions like flagellar motion, mitochondrial oxidative metabolism and entry inside the macrophages. Since Ld-CC has not been characterised yet, we performed homology modelling of Leishmania donovani Ca2+ ion channel (Ld-CC) and docking studies of ligand library against this channel. 542 compound library of National Cancer Institute (NCI) diversity 3 dataset selected for screening studies. The ligands ZINC17287336 and ZINC29590262 were selected as best energy conformers because they show highest binding affinity towards its target (Ld-CC). They interact with the active site residues in the pocket of Ld-CC which suggests that the docked conformations are good and acceptable. Moreover, these two selected compounds also have relatively high binding affinity than nifedipine and verapamil, known human calcium channel blockers which had been reported to have mild anti-leishmanial activity. Among these two top screened inhibitors the ligand ZINC29590262 shows poor binding affinity towards the Human voltagedependent L-type calcium channel subunit alpha-1C in comparison to the Ld-CC. Therefore, we proposed this ligand as the best inhibitor which shows 40% more binding affinity with Ld-CC than the human-VDCC. These results suggest that our screened ligand ZINC29590262 could act as novel drug and may show much better antileishmanial activity.

The Potential Roles of Aquaporin 4 in Alzheimer's Disease.

Aquaporin 4 (AQP4) is the major water channel expressed in the central nervous system (CNS), and it is primarily expressed in astrocytes. It has been studied in various brain pathological conditions. However, the potential for AQP4 to influence Alzheimer's disease (AD) is still unclear. Research regarding AQP4 functions related to AD can be traced back several years and has gradually progressed toward a better understanding of the potential mechanisms. Currently, it has been suggested that AQP4 influences synaptic plasticity, and AQP4 deficiency may impair learning and memory, in part, through glutamate transporter-1 (GLT-1). AQP4 may mediate the clearance of amyloid beta peptides (Aß). In addition, AQP4 may influence potassium (K(+)) and calcium (Ca(2+)) ion transport, which could play decisive roles in the pathogenesis of AD. Furthermore, AQP4 knockout is involved in neuroinflammation and interferes with AD. To date, no specific therapeutic agents have been developed to inhibit or enhance AQP4. However, experimental results strongly emphasize the importance of this topic for future investigations.