AQP Genes Research Articles

Effect of heat stress on the hypothalamic expression profile of water homeostasis-associated genes in low- and high-water efficient chicken lines.

With climate change, selection for water efficiency and heat resilience are vitally important. We undertook this study to determine the effect of chronic cyclic heat stress (HS) on the hypothalamic expression profile of water homeostasis-associated markers in high (HWE)- and low (LWE)-water efficient chicken lines. HS significantly elevated core body temperatures of both lines. However, the amplitude was higher by 0.5-1°C in HWE compared to their LWE counterparts. HWE line drank significantly less water than LWE during both thermoneutral (TN) and HS conditions, and HS increased water intake in both lines with pronounced magnitude in LWE birds. HWE had better feed conversion ratio (FCR), water conversion ratio (WCR), and water to feed intake ratio. At the molecular level, the overall hypothalamic expression of aquaporins (AQP8 and AQP12), arginine vasopressin (AVP) and its related receptor AVP2R, angiotensinogen (AGT), angiotensin II receptor type 1 (AT1), and calbindin 2 (CALB2) were significantly lower; however, CALB1 mRNA and AQP2 protein levels were higher in HWE compared to LWE line. Compared to TN conditions, HS exposure significantly increased mRNA abundances of AQPs (8,12), AVPR1a, natriuretic peptide A (NPPA), angiotensin I-converting enzyme (ACE), CALB1 and 2, and transient receptor potential cation channel subfamily V member 1 and 4 (TRPV1 and TRPV4) as well as the protein levels of AQP2, however it decreased that of AQP4 gene expression. A significant line by environment interaction was observed in several hypothalamic genes. Heat stress significantly upregulated AQP2 and SCT at mRNA levels and AQP1 and AQP3 at both mRNA and protein levels, but it downregulated that of AQP4 protein only in LWE birds. In HWE broilers, however, HS upregulated the hypothalamic expression of renin (REN) and AVPR1b genes and AQP5 proteins, but it downregulated that of AQP3 protein. The hypothalamic expression of AQP (5, 7, 10, and 11) genes was increased by HS in both chicken lines. In summary, this is the first report showing improvement of growth performances in HWE birds. The hypothalamic expression of several genes was affected in a line- and/or environment-dependent manner, revealing potential molecular signatures for water efficiency and/or heat tolerance in chickens.

Exogenous Easily Extractable Glomalin-Related Soil Protein Induces Differential Response in Plant Growth of Tea Plants via Regulating Water Channel Protein Expression

Glomalin, a glycoprotein secreted by arbuscular mycorrhizal fungi (AMFs), exhibits multiple beneficial functions in regard to plant growth. However, the roles and regulatory mechanisms of exogenous easily extractable glomalin-related soil protein (EE-GRSP) in water and their effects on the quality of tea plants (Camellia sinensis (L.) O. Ktze.) remain unclear. The present study aimed to investigate the effects of a quarter-strength exogenous EE-GRSP solution (1/4 EE-GRSP), half-strength exogenous EE-GRSP solution (1/2 EE-GRSP), three-quarter-strength exogenous EE-GRSP solution (3/4 EE-GRSP), and full-strength exogenous EE-GRSP solution (full EE-GRSP) on plant growth, the root system architecture, leaf water status, and the tea quality of tea seedlings, along with examining the changes in the relative expression of water channel proteins (AQPs) in tea plants. The results indicated that exogenous EE-GRSP of different strengths had different effects on both the growth performance (height, leaf numbers, and biomass) and root architecture parameters of tea seedlings, and the best positive effects on plant growth and the root architecture appeared under the three-quarter-strength exogenous EE-GRSP treatment. Similarly, the exogenous EE-GRSP application also differently affected tea quality indicators, in which only the quarter- and half-strength exogenous EE-GRSP solutions significantly increased most of the indicators, including carbohydrates, tea polyphenols, total amino acids, catechins, and flavonoids. Moreover, the half- and three-quarter-strength exogenous EE-GRSP treatments significantly increased the leaf relative water content (LRWC), but all of the exogenous EE-GRSP treatments significantly decreased the leaf water potential (LWP). Furthermore, the expression of AQP genes in the root system of tea plants was related to the strength of the exogenous EE-GRSP treatments, and different genes were significantly up-regulated or down-regulated under the treatment of exogenous EE-GRSP at different strengths. Moreover, the correlation analysis showed that most of the relative expression of AQPs was significantly and positively correlated with tea plant growth, the root architecture, and the leaf relative water content, but negatively correlated with tea quality indicators; however, the expression of CsNIPs and CsSIPs was markedly and negatively correlated with plant growth performance. Therefore, we speculated that the application of exogenous EE-GRSP could facilitate plant growth and improve the quality indirectly by regulating the expression of root AQPs, thus ameliorating the water uptake and nutrient accumulation in tea plants.

LbKAT3 may assist in mycorrhizal potassium uptake, and overexpression of LbKAT3 may promote potassium, phosphorus, and water transport from arbuscular mycorrhizal fungi to the host plant.

Potassium plays important roles in most plant physiological processes. Arbuscular mycorrhizal (AM) fungi promote plant water and mineral nutrient acquisition to promote plant growth. However, few studies have focused on the effect of AM colonization on potassium uptake by the host plant. In this study, the effects of an AM fungus (Rhizophagus irregularis) and potassium concentration (0, 3, or 10 mM K+) on Lycium barbarum were evaluated. A split-root test with L. barbarum seedlings was conducted, and the potassium uptake capacity of LbKAT3 was verified in yeast. A tobacco line overexpressing LbKAT3 was generated and mycorrhizal functions under two potassium concentrations (0.2 and 2 mM K+) were studied. Inoculation of R. irregularis and application of potassium increased the dry weight, and potassium and phosphorus contents of L. barbarum, and increased the colonization rate and arbuscule abundance of R. irregularis. In addition, the expression of LbKAT3 and AQP genes in L. barbarum was upregulated. Inoculation of R. irregularis induced LbPT4, Rir-AQP1, and Rir-AQP2 expression, and application of potassium upregulated the expression of these genes. Inoculation with the AM fungus locally regulated the expression of LbKAT3. Inoculation of R. irregularis improved the growth, and potassium and phosphorus contents, and induced NtPT4, Rir-AQP1, and Rir-AQP2 expression in tobacco overexpressing LbKAT3 under both potassium concentrations. Overexpression of LbKAT3 in tobacco improved the growth, potassium accumulation, and AM colonization, and upregulated the expression of NtPT4 and Rir-AQP1 in mycorrhizal tobacco. The results suggest that LbKAT3 may assist in mycorrhizal potassium uptake, and overexpression of LbKAT3 may promote potassium, phosphorus, and water transport from the AM fungus to tobacco.

Genome-Wide Identification, Phylogenetic Analysis and Expression Pattern Profiling of the Aquaporin Family Genes in Leuciscus waleckii

Aquaporin (Aqp) is a transmembrane-specific channel for small molecules that help in regulating homeostasis in fishes when adapting to changing environments, but its role in Amur ide’s response to alkaline stress is yet to be revealed. Therefore, the purpose of this study is to investigate the response of the Aqp gene exposed to alkaline water in Amur ide (Leuciscus waleckii) using a genome-transcriptional assay. Based on the results, we classified the Aqps of the L. waleckii (LwAqps) genome and analyzed its transcriptional expression profile and genetic evolution under carbonate alkalinity stress. A total of 18 Aqp genes were identified in four grades in L. waleckii. The highest Aqp gene expression was found in the gill and kidney of L. waleckii from the Wusuli River (WSL) in comparison to those in the Dali Lake (DL), whereas aqp3a, -3ap1, -7, and -9a expressions were found at intensively higher levels in the gill rather than in the kidneys and livers. The experiment of L. waleckii under alkalinity stress (carbonate alkalinity 50 mmol·L−1) and its recovery showed that the expressions of aqp0a, -3a, -3ap1, -7, -8aa, and -9a were upregulated in alkaline water and downregulated in freshwater. We identified 1460 single nucleotide polymorphism (SNP) markers in the Aqp genes. The average value of Fst of SNP markers in the CDS region was 0.177 ± 0.256, and the first 5% SNPs were identified at aqp3a and -11b. Residue Ser66 does not bring about an overall change in the three-dimensional structure of Aqp3a, but may change the penetration of solutes across the membrane. This indicates that Aqp genes are involved in the response of L. waleckii to alkaline stress, and aqp3a is one of the key genes involved in regulating L. waleckii’s adaptation to alkaline environments.

Association of AQP4 single nucleotide polymorphisms (rs335929 and rs2075575) with Parkinson’s disease: A case-control study

ObjectiveThe glymphatic system plays an important role in brain waste removal and is functionally and structurally dependent on astrocyte aquaporin-4 (AQP4). Genetic variation in the AQP4 gene has therefore been hypothesized to be associated with genetic susceptibility to neurodegenerative diseases. This study aimed to investigate whether two specific single nucleotide polymorphisms (SNP) in the AQP4 gene, rs335929, and rs2075575, are associated with the risk and clinical features of PD. MethodsA total of 950 participants, including 475 patients with sporadic PD and 475 independent healthy controls, were included in this case-control study. Two SNPs (rs335929 and rs2075575) of the AQP4 gene were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Sanger sequencing was used to determine whether the genotyping results were accurate. A chi-square (χ2) test was used to compare the frequencies of alleles and genotypes between patients and controls. Logistic regression was used to calculate dominance ratios (OR) and 95% confidence intervals (CI). ResultsThe difference between rs2075575 in the dominant model (GG vs GA + AA: P = 0.019) and the overdominant model (GG + AA vs GA: P = 0.013) was statistically significant. Subgroup analysis showed that the frequency of the rs2075575 A allele was significantly higher in female PD patients than in matched female controls (P = 0.017). rs2075575 A allele was significantly more frequent in LOPD patients than in matched elderly controls (P = 0.033). rs335929 polymorphism was not significantly correlated with PD susceptibility in either the overall or subgroup analysis. Haplotype analysis between the two SNPs did not show an association with PD susceptibility. In addition, we found that the rs2075575 G allele was significantly associated with Rapid Eye Movement Behaviour Disorder (RBD) (P = 0.044), and the rs335929 A allele with memory impairment (P = 0.028) in PD. ConclusionThe AQP4 gene rs2075575 polymorphism may be associated with PD susceptibility, but not the rs335929 polymorphism. rs2075575 is associated with RBD and rs335929 is associated with memory cognition. Regulation of the glymphatic system by interfering with the genetics of AQP4 and thus influencing the pathology of PD may be a direction worth investigating. Studies in larger sample sizes and across ethnicities are essential for further understanding the potential association between AQP genes and PD pathogenesis.

Differential expression of aquaporin genes and the influence of environmental hypertonicity on their expression in juveniles of air-breathing stinging catfish (Heteropneustes fossilis)

Aquaporins (AQPs) are a superfamily of transmembrane channel proteins that are responsible for the transport of water and some other molecules to and from the cell, mainly for osmoregulation under anisotonicity. We investigated here the expression patterns of different AQP isoforms and also during exposure to hypertonicity (300 mOsmol/L) for 48 h in juvenile stages of air-breathing stinging catfish (Heteropneustes fossilis). A total of 8 mRNA transcripts for different isoforms of AQPs and their translated proteins could be detected in the anterior and posterior regions of S1, S2, and S3 stages of juveniles of stinging catfish at variable levels. In general, more expression of mRNAs for different aqp genes was seen in the S2 and S3 juveniles than in the S1 juveniles. Most interestingly, exposure to hypertonicity of S2 juveniles for a period of 48 h led to increased expression of most of the aqp genes both at transcriptional and translational levels, except for aqp3 in the anterior and posterior regions and aqp1 in the anterior region, showing maximum expression at later stages of hypertonic exposure. Thus, it is evident that AQPs play crucial roles in maintaining the water and ionic balances under anisotonic conditions even at the early developmental stages of stinging catfish as a biochemical adaptational strategy to survive and grow in anisotonic environment.

Detection of genomic regions underlying milk production traits in Valle del Belice dairy sheep using regional heritability mapping.

The aim of this study was to identify genomic regions underlying milk production traits in the Valle del Belice dairy sheep using regional heritability mapping (RHM). Repeated measurements for milk yield (MY), fat percentage and yield (F% and FY) and protein percentage and yield (P% and PY), collected over a period of 6 years (2006–2012) on 481 Valle del Belice ewes, were used for the analysis. Animals were genotyped with the Illumina 50k SNP chip. Variance components, heritabilities and repeatabilities within and across lactations were estimated, fitting parity, litter size, season of lambing and fortnights in milk, as fixed; and additive genetic, permanent environment within and across lactations, flock by test‐day interaction and residual as random effects. For the RHM analysis, the model included the same fixed and random effects as before, plus an additional regional genomic additive effect (specific for the region being tested) as random. While the whole genomic additive effect was estimated using the genomic relationship matrix (GRM) constructed from all SNPs, the regional genomic additive effect was estimated from a GRM matrix constructed from the SNPs within each region. Heritability estimates ranged between 0.06 and 0.15, with repeatabilities being between 0.14 and 0.24 across lactations and between 0.23 and 0.39 within lactation for all milk production traits. A substantial effect of flock‐test‐day on milk production traits was also estimated. Significant genomic regions at either genome‐wide (p < .05) or suggestive (i.e., one false positive per genome scan) level were identified on chromosome (OAR) 2, 3 and 20 for F% and on OAR3 for P%, with the regions on OAR3 in common between the two traits. Our results confirmed the role of LALBA and AQP genes, on OAR3, as candidate genes for milk production traits in sheep.

Genomic and expression characterization of aquaporin genes from Siniperca chuatsi

Aquaporins (AQPs) are major intrinsic proteins that form pores in the membranes of biological cells. We first cloned the full-length sequences of aqp0, 1, 3, 4, 7, 8, 9, 10, 11, and 12 genes in Siniperca chuatsi. The 10 S. chuatsi aqp (Sc-aqp) genes included complete open reading frames and exhibited different exon-intron organizations. Sc-aqp1, 3, 8, 9, 10, and 11 were mostly expressed in the gallbladder, gills, gastric cecum, liver, ovaries, and spleen, respectively; Sc-aqp0 and 4 were mostly expressed in larvae at 1 day after hatching and in gastrula; Sc-aqp7 and 12 were mostly expressed in 2K-cell embryos. The expression levels of Sc-aqp1, 3, 7, 8, 9, and 10 after 10 part per thousand (ppt) salt treatment had significantly changed compared with those after 0 ppt salt treatment. Real-time quantitative PCR analysis further showed that in the intestines, the mRNA levels of Sc-aqp1 and 10 significantly decreased by approximately 2.07- and 2.85-fold, respectively, whereas those of Sc-aqp8 and 9 significantly increased by approximately 7.08- and 4.14-fold, respectively. Sc-aqp1, 8, 9, and 10 showed no significant differences in the gills. Sc-aqp3 significantly decreased by approximately 1.51- and 1.67-fold in the gills and intestines, respectively. Sc-aqp7 significantly increased by approximately 4.18- and 7.04-fold in the gills and intestines, respectively. This study was the first to investigate the tissue expression profiles and response to salt stress of aqp genes in S. chuatsi. Moreover, altering diet and suffering from immune stress could cause changes in the expression level of aqps. This study provided valuable reference information for AQPs' roles in osmoregulation in freshwater fish.

Transcriptomics analysis of Paeonia lactiflora Pall. in response to drought stress by high-throughput sequencing

ABSTRACT Drought is an environmental stress that limits agricultural and horticultural production worldwide. Paeonia lactiflora Pall. is a perennial flowering plant with strong environmental adaptability and a good candidate for production in arid areas. Using the Illumina Hiseq platform to study the transcriptome of P. lactiflora leaves under drought stress, 62.27 Gb of clean data was obtained and 87,099 unigenes (N50 = 1,352 bp; mean length = 790.66 bp) were obtained by transcriptome sequencing. After screening 4,198 differentially expressed genes, genes that may be related to the response of P. lactiflora to drought stress were identified: 28 signal transduction genes, 12 protein kinase genes, 14 transcription factor genes, 35 osmotic adjustment genes, 14 antioxidase enzyme genes, 22 photosynthesis genes, 9 AQP genes, 20 HSP genes, and 17 LEA genes. Transcriptomics analysis revealed that transcription factors, such as ERF, WRKY, and NAC transcription factor family members regulate the synthesis of hormones that play a role in signal transduction pathways of peony in response to drought stress. These results enrich currently available herbaceous peony gene sequence resources, fortify the identification of candidate genes and molecular regulatory mechanisms in response to drought stress, allowing genetic engineering to be used to create drought-tolerant plants.

Interference of CsGPA1, the α-submit of G protein, reduces drought tolerance in cucumber seedlings

The G protein α-subunit, GPA1, is an integral component of several signaling pathways in plants, including response to abiotic stress. However, the molecular mechanism behind these processes remains largely unknown in the cucumber plant (Cucumis sativus L.). In order to further understand the role of CsGPA1 in cucumber under drought stress, changes in plant growth, physiological parameters, and gene expression of CsAQPs were all measured under water stress induced by polyethylene glycol (PEG) using wild type (WT) and CsGPA1-interference (RNAi) cucumber seedlings. Our results demonstrated that the RNAi plants had lower drought tolerance, displaying seriously withered leaves, lower relative growth rate, lower root-shoot ratio, and lower root activity under drought stress compared to WT plants. Physiological studies indicated that the suppression of CsGPA1 weakened drought stress tolerance due to higher water loss rate in the leaves, higher levels of hydrogen peroxide (H2O2), increased malondialdehyde (MDA) content, lower free proline content, lower soluble sugar content, lower soluble protein content, and decreased antioxidant enzyme activities. qRT-PCR analysis demonstrated that the interference of CsGPA1 up-regulated the expression of most AQP genes (except for CsPIP2;3 in leaves) and down-regulated the expression of CsPIP1;2, CsPIP1;4, CsPIP2;1, and CsPIP2;4 in roots under drought stress when compared to WT plants. Our results demonstrated that CsGPA1 could function as a positive regulator in drought stress response by decreasing the accumulation of reactive oxygen species (ROS), improving permeable potentials, and reducing water loss in cucumber plants.

Aquaglyceroporin AqpS from Sinorhizobium meliloti conducts both trivalent and pentavalent methylarsenicals

Arsenic is a toxic metalloid that enters cells adventitiously via uptake systems for phosphate transporters, aquaglyceroporins (AQPs) or sugar permeases. However, transport of highly toxic methylarsenite (MAs(III)) and relatively nontoxic methylarsenate (MAs(V)) by bacterial AQPs has not been characterized. MAs(V) has a history of use as an herbicide. Here we used whole genome sequence analysis of AQPs in arsenic resistance (ars) operons. The aqp genes are frequently located next to MAs(III) resistance genes such as arsH, which suggests that they could be involved in MAs(III) uptake. Bacterial AQPs encoded by ars operons can be classified into two subgroups. One subgroup includes AqpS from the plant symbiont Sinorhizobium meliloti 1021. Our data suggests that AqpS has a substrate selectivity filter different from that of other bacterial AQPs. Both Escherichia coli GlpF and AqpS conduct MAs(III) efficiently, but GlpF conducts the MAs(V) anion poorly, so E. coli takes up MAs(V) inefficiently. In contrast, AqpS conducts MAs(V) under physiological conditions. A homology model of AqpS indicates that it has a substrate channel with a selectivity filter containing the nonpolar residue Val177 instead of the charged arginine residue found in other AQPs. While the selectivity filter in most AQPs prevents movement of anions, Val177 is predicted to allow movement of the MAs(V) anion through the channel. We propose that AqpS is a component of an MAs(III) resistance pathway in which MAs(III) enters cells of S. meliloti via AqpS, is oxidized by ArsH to MAs(V), which exits the cells via AqpS.

Inhibition of the water channel aquaporin-1 prevents myocardial remodeling by blocking the transmembrane transport of hydrogen peroxide

Abstract Background Pathological remodeling of the myocardium has long been known to involve oxidant signaling, but so far, strategies using systemic anti-oxidants have generally failed to prevent it. Aquaporins are a family of transmembrane water channels with thirteen isoforms currently known. Some isoforms have been implicated in oxidant signaling. AQP1 is the most abundant aquaporin in cardiovascular tissues but its specific role in cardiac remodeling remains unknown. Purpose We tested the role of AQP1 as a key regulator of oxidant-mediated cardiac remodeling amenable to targeted pharmacological therapy. Methods We used mice with genetic deletion of Aqp1 (and wild-type littermate), as well as primary isolates from the same mice and human iPSC/Engineered Heart Tissue to test the role of AQP1 in pro-hypertrophic signaling. Human cardiac myocyte-specific (PCM1+) expression of AQP's and genes involved in hypertrophic remodeling was studied by RNAseq and bioinformatic GO pathway analysis. Results RNA sequencing from human cardiac myocytes revealed that the archetypal AQP1 is a major isoform. AQP1 expression correlates with the severity of hypertrophic remodeling in patients with aortic stenosis. The AQP1 channel was detected at the plasma membrane of human and mouse cardiac myocytes from hypertrophic hearts, where it colocalizes with the NADPH oxidase-2 (NOX2) and caveolin-3. We show that hydrogen peroxide (H2O2), produced extracellularly, is necessary for the hypertrophic response of isolated cardiac myocytes and that AQP1 facilitates the transmembrane transport of H2O2 through its water pore, resulting in activation of oxidant-sensitive kinases in cardiac myocytes. Structural analysis of the amino acid residues lining the water pore of AQP1 supports its permeation by H2O2. Deletion of Aqp1 or selective blockade of AQP1 intra-subunit pore (with Bacopaside II) inhibits H2O2 transport in mouse and human cells and rescues the myocyte hypertrophy in human induced pluripotent stem cell-derived engineered heart muscle. This protective effect is due to loss of transmembrane transport of H2O2, but not water, through the intra-subunit pore of AQP1. Treatment of mice with clinically-approved Bacopaside extract (CDRI08) inhibitor of AQP1 attenuates cardiac hypertrophy and fibrosis. Conclusion We provide the first demonstration that AQP1 functions as an aqua-peroxiporin in primary rodent and human cardiac parenchymal cells. We show that cardiac hypertrophy is mediated by the transmembrane transport of H2O2 through the AQP1 water channel. Our studies open the way to complement the therapeutic armamentarium with specific blockers of AQP1 for the prevention of adverse remodeling in many cardiovascular diseases leading to heart failure. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): FRS-FNRS, Welbio

Effects of osmotic stress on the expression profiling of aquaporin genes in the roughskin sculpin (Trachidermus fasciatus)

Aquaporins (AQPs) are a family of integral membrane proteins that have been shown to be important for osmoregulation in many vertebrates. To identify potential stress resistance-related aqp genes in salinity adaptation of the roughskin sculpin Trachidermus fasciatus, we investigated the time-course expression dynamics of seven aquaporin genes (aqpl, 4, 7, 8, 10, 11 and 12) in three osmoregulatory tissues (kidney, gill and intestine) and one metabolic tissue (liver). The fish were subjected to two different acute osmotic treatments (seawater-to-freshwater transfer respectively achieved in 1 h and 24 h, namely, E-acute and acute group). The expression profiling of the seven aqp genes were performed using quantitative real-time PCR (qRT-PCR). At the time of all sampling time points (0 h, 12 h, 24 h and 48 h), no expression of aqp4 was found in the gill, liver and intestine; no expression of aqp7 was found in the gill and liver. Significant differences of aqp expression were determined in the four target tissues, and the mRNA levels were largely variable among gene members and tissues. Similar patterns of the time-course expression were detected in most of the aqp genes in T. fasciatus between the two acute groups, except that only one gene (aqpl2) in the kidney and three genes (aqp7, aqp8 and aqp10) in the intestine revealed different expression patterns. These results suggest that the expression response of aqp genes was similar under osmotic changes with different rates.

Aquaporin 5 promotes tumor migration and angiogenesis in non-small cell lung cancer cell line H1299.

Non-small cell lung cancer (NSCLC) constitutes the majority of all lung-cancer cases. Aquaporin 5 (AQP5) may be involved in NSCLC by promoting lung-cancer initiation and progression. The present study aimed to determine the role of AQP5 in migration and angiogenesis using NSCLC cells and HUVECs. AQPs 1, 3, 4, 5, 8 and 9 were screened in the NSCLC cell line H1299, and the present results showed that AQP5 mRNA was upregulated compared with the other AQP genes. At the protein level, AQP5 was significantly increased in H1299 cells compared with 16HBE cells. AQP5 knockdown in H1299 cells significantly decreased cell migration compared with untransfected cells, as demonstrated by both Transwell and wound closure assays. The present study further investigated H1299 ability to promote HUVEC vascularisation. The supernatants of both transfected and untransfected H1299 cells were used as conditioned medium for HUVECs, and tube formation was measured. The supernatant of AQP5-downregulated cells exhibited significantly low tube formation potential compared with untransfected cells. Similarly, vascular endothelial growth factor was significantly increased in control cells (si-NC) compared with cells transfected with small interfering RNA targeting AQP5. The present study found that AQP5 downregulation significantly decreased the phosphorylation level of epidermal growth factor receptor and the activity of the ERK1/2 pathway. In summary, the present study suggested that AQP5 influenced migration and angiogenesis in NSCLCs in vitro and may potentially exhibit similar in vivo effects.

Expression patterns of different isoforms of aquaporins in isoproterenolinduced myocardial infarction model in rat treated with Terminalia arjuna Bark extract

Aquaporins (AQPs) are found to express in myocardium and their involvement in myocardial water balance have been figured out quite well as they are primarily associated with the transportation of water and other solutes like glycerol and urea. In the present study, we investigated the levels of expression of AQP sub-types in normal myocardium and their changes in the process of edema formation and cardiac dysfunction following myocardial infarction by isoproterenol as well as after treatment with Terminalia arjuna methanolic extract. The qPCR analysis demonstrated an up-regulation of certain aquaporin genes like aqp1, aqp4, aqp5 and aqp7 in the myocardial infarcted group of rats induced by isoproterenol. Again, the treatment with methanolic extract of T. arjuna was found to down-regulate these aqp genes, thereby suggesting a possible co-relation between these two. In addition, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were found to increase in the infarcted group of rats, followed by lowering of these two enzyme levels in the treated group. Histopathological study quite clearly demonstrated that the myocardium from infarcted group displayed muscle separation and infiltration of lymphocytes, whereas treatment of myocardium from infarcted group with T. arjuna extract revealed a decrease in the degree of muscle separation and inflammatory response. Thus, our findings suggest the possible role of certain AQPs in isoproterenol induced myocardial infarction and also the remedial effects of T. arjuna in reverting the condition back to normal by lowering their expression levels. The enhanced expression levels of the aforementioned AQPs are quite indicative of the fact that these can serve as prognostic markers for myocardial infarction.

Genome-wide identification, structure characterization, and expression pattern profiling of aquaporin gene family in cucumber

BackgroundAquaporin (AQP) proteins comprise a group of membrane intrinsic proteins (MIPs) that are responsible for transporting water and other small molecules, which is crucial for plant survival under stress conditions including salt stress. Despite the vital role of AQPs, little is known about them in cucumber (Cucumis sativus L.).ResultsIn this study, we identified 39 aquaporin-encoding genes in cucumber that were separated by phylogenetic analysis into five sub-families (PIP, TIP, NIP, SIP, and XIP). Their substrate specificity was then assessed based on key amino acid residues such as the aromatic/Arginine (ar/R) selectivity filter, Froger’s positions, and specificity-determining positions. The putative cis-regulatory motifs available in the promoter region of each AQP gene were analyzed and results revealed that their promoter regions contain many abiotic related cis-regulatory elements. Furthermore, analysis of previously released RNA-seq data revealed tissue- and treatment-specific expression patterns of cucumber AQP genes (CsAQPs). Three aquaporins (CsTIP1;1, CsPIP2;4, and CsPIP1;2) were the most transcript abundance genes, with CsTIP1;1 showing the highest expression levels among all aquaporins. Subcellular localization analysis in Nicotiana benthamiana epidermal cells revealed the diverse and broad array of sub-cellular localizations of CsAQPs. We then performed RNA-seq to identify the expression pattern of CsAQPs under salt stress and found a general decreased expression level of root CsAQPs. Moreover, qRT-PCR revealed rapid changes in the expression levels of CsAQPs in response to diverse abiotic stresses including salt, polyethylene glycol (PEG)-6000, heat, and chilling stresses. Additionally, transient expression of AQPs in N. benthamiana increased leaf water loss rate, suggesting their potential roles in the regulation of plant water status under stress conditions.ConclusionsOur results indicated that CsAQPs play important roles in response to salt stress. The genome-wide identification and primary function characterization of cucumber aquaporins provides insight to elucidate the complexity of the AQP gene family and their biological functions in cucumber.

Combining genome-wide and transcriptome-wide analyses reveal the evolutionary conservation and functional diversity of aquaporins in cotton

BackgroundAquaporins (AQPs) are integral membrane proteins from a larger family of major intrinsic proteins (MIPs) and function in a huge variety of processes such as water transport, plant growth and stress response. The availability of the whole-genome data of different cotton species allows us to study systematic evolution and function of cotton AQPs on a genome-wide level.ResultsHere, a total of 53, 58, 113 and 111 AQP genes were identified in G. arboreum, G. raimondii, G. hirsutum and G. barbadense, respectively. A comprehensive analysis of cotton AQPs, involved in exon/intron structure, functional domains, phylogenetic relationships and gene duplications, divided these AQPs into five subfamilies (PIP, NIP, SIP, TIP and XIP). Comparative genome analysis among 30 species from algae to angiosperm as well as common tandem duplication events in 24 well-studied plants further revealed the evolutionary conservation of AQP family in the organism kingdom. Combining transcriptome analysis and Quantitative Real-time PCR (qRT-PCR) verification, most AQPs exhibited tissue-specific expression patterns both in G. raimondii and G. hirsutum. Meanwhile, a bias of time to peak expression of several AQPs was also detected after treating G. davidsonii and G. hirsutum with 200 mM NaCl. It is interesting that both PIP1;4 h/i/j and PIP2;2a/e showed the highly conserved tandem structure, but differentially contributed to tissue development and stress response in different cotton species.ConclusionsThese results demonstrated that cotton AQPs were structural conservation while experienced the functional differentiation during the process of evolution and domestication. This study will further broaden our insights into the evolution and functional elucidation of AQP gene family in cotton.

Genome-wide comparison reveals divergence of cassava and rubber aquaporin family genes after the recent whole-genome duplication

BackgroundAquaporins (AQPs) are a class of integral membrane proteins that facilitate the passive transport of water and other small solutes across biological membranes. Despite their importance, little information is available in cassava (Manihot esculenta), a perennial shrub of the Euphorbiaceae family that serves the sixth major staple crop in the world.ResultsThis study presents a genome-wide analysis of the AQP gene family in cassava. The family of 42 members in this species could be divided into five subfamilies based on phylogenetic analysis, i.e., 14 plasma membrane intrinsic proteins (PIPs), 13 tonoplast intrinsic proteins (TIPs), nine NOD26-like intrinsic proteins (NIPs), four X intrinsic proteins (XIPs), and two small basic intrinsic proteins (SIPs). Best-reciprocal-hit-based sequence comparison and synteny analysis revealed 34 orthologous groups (OGs) present in the Euphorbiaceae ancestor, and nearly one-to-one or two-to-one orthologous relationships were observed between cassava with rubber/physic nut, reflecting the occurrence of one so-called ρ recent whole-genome duplication (WGD) in the last common ancestor of cassava and rubber. In contrast to a predominant role of the ρ WGD on family expansion in rubber, cassava AQP duplicates were derived from the WGD as well as local duplication. Species-specific gene loss was also observed in cassava, which includes the entire NIP4 group and/or six OGs. Comparison of conserved motifs and gene expression profiles revealed divergence of paralogs in cassava as observed in rubber.ConclusionsOur findings will not only improve our knowledge on family evolution in Euphorbiaceae, but also provide valuable information for further functional analysis of AQP genes in cassava and rubber.

Comparative Analysis of the aquaporin Gene Family in 12 Fish Species.

Simple SummaryAquaporins (Aqps) are a group of membrane proteins. In this study, 166 Aqp genes were identified in 12 fish species. Gene organization, motif distribution, recombination, and selection pressure were performed to investigate their evolutionary characteristics. In addition, expression profiles of Aqps were also examined under pathogens infection and organophosphorus pesticide stress. This study will provide a useful reference for further functional study.Aquaporins (Aqps) are a class of water channel proteins that play key roles in many physiological functions and cellular processes. Here, we analyzed 166 putative Aqp genes in 12 fish species and divided them into four groups. Gene organization and motif distribution analyses suggested potentially conserved functions in each group. Several recombination events were identified in some members, which accelerate their divergence in evolution. Furthermore, a few positive selection sites were identified, and mutations at these sites could alter the stability of Aqp proteins. In addition, expression profiles of some Aqp genes under pathogen infection and organophosphorus pesticide stress were also investigated. The result implied that several Aqp genes may affect different immune responses and osmoregulation. This study provides a comparative analysis of the fish Aqp gene family to facilitate further functional analyses.

Cytoprotective effect of aquaporin 1 against lipopolysaccharide-induced apoptosis and inflammation of renal epithelial HK-2 cells.

Sepsis is the most common underlying disease of disseminated intravascular coagulation. Acute kidney injury is a common and serious complications of sepsis. In the present study, a lipopolysaccharide (LPS)-induced human proximal tubule cell line (HK-2 cells) was selected as an in vitro model of septic acute kidney injury. The aim of the present study was to investigate whether aquaporin 1 (AQP-1) has a cytoprotective role in LPS-induced HK-2 cells. HK-2 cells were treated with 0–16 µg/ml LPS for 0–24 h to establish the in vitro model of sepsis. The results demonstrated that AQP-1 levels were the lowest of the eight AQP genes expressed in LPS-induced HK-2 cells. Prior to LPS treatment, HK-2 cells were transfected with pcDNA-AQP-1 or small interfering-AQP-1 and cell counting kint-8 and flow cytometry assays were performed to assess cell viability and apoptosis rate, respectively. Changes in the expression of proinflammatory cytokines and chemokines, as well as important factors in the p38, extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK) pathways, were assessed using reverse transcription-quantitative polymerase chain reaction, western blotting and ELISA, respectively. LPS treatment reduced viability, increased apoptosis and upregulated the expression of proinflammatory cytokines and chemokines in HK-2 cells. AQP-1 overexpression significantly reversed the effects of LPS and downregulated the expression of tumor necrosis factor-α, interleukin (IL)-8, IL-1β and monocyte chemoattractant protein-1. The p38, ERK1/2 and JNK pathways were activated by LPS; however, the p38 and ERK1/2 pathways were blocked in AQP-1-overexpressing cells. AQP-1 overexpression was demonstrated to confer a survival advantage to LPS-injured HK-2 cells by controlling cell viability, apoptosis and inflammation, possibly via modulation of the p38 and ERK1/2 pathways. The results of the present study suggest that AQP-1 may be an effective treatment for acute kidney injury caused by sepsis.