Aquaporin Isoforms Research Articles

The Role of Aquaporin in Stress Physiology: A Review

The concept of water movement across cell membranes eventually lead to identification of water channels, known as ‘Aquaporins’. The transport function exerted by aquaporins (AQPs) is carried out efficiently by 13 distinct isoforms of AQPs in mammals. AQPs help in regulation of physiological functions of various organ systems and have a functional role in stress associated with various pathological conditions. This review summarises the involvement of AQPs in transport of ROS and oxidative stress regulation in various organ systems. AQP1, AQP4, AQP7 and AQP9 are the major AQPs in the cardiac tissues of various animals, playing vital role in the patho-physiology of congestive heart failure, myocardial oedema, myocardial ischemia and polymicrobial sepsis. In digestive system, AQP1, AQP3, AQP4, AQP7, AQP8, AQP10 and AQP11 are localised in gastro-intestinal tract, modulating patho-physiology of pseudomembranous colitis, bile induced diarrhoea, irritable bowel syndrome and intestinal ulcers. AQPs in enteric tissues are engaged in metabolism of H2O2, providing a protective mechanism against oxidative stress. AQP1, AQP2, AQP3, AQP4 and AQP11 are major renal AQPs involved in ureteral obstruction, nephrogenic diabetes insipidus and kidney injury. In respiratory system, AQP1, AQP2, AQP4 and AQP5 are expressed in various pathological circumstances like lung injury and pulmonary inflammation due to oxidative stress. Thus, the evidences regarding the role of AQPs in oxidative stress offer a foundation for comprehending the adaptive mechanisms to oxidative injury at cellular level, which helps in strategizing breeding programmes and serves as biomarkers for efficient diagnosis.

Structural Basis for the Interaction between the Ezrin FERM-Domain and Human Aquaporins.

The Ezrin/Radixin/Moesin (ERM) family of proteins act as cross-linkers between the plasma membrane and the actin cytoskeleton. This mechanism plays an essential role in processes related to membrane remodeling and organization, such as cell polarization, morphogenesis and adhesion, as well as in membrane protein trafficking and signaling pathways. For several human aquaporin (AQP) isoforms, an interaction between the ezrin band Four-point-one, Ezrin, Radixin, Moesin (FERM)-domain and the AQP C-terminus has been demonstrated, and this is believed to be important for AQP localization in the plasma membrane. Here, we investigate the structural basis for the interaction between ezrin and two human AQPs: AQP2 and AQP5. Using microscale thermophoresis, we show that full-length AQP2 and AQP5 as well as peptides corresponding to their C-termini interact with the ezrin FERM-domain with affinities in the low micromolar range. Modelling of the AQP2 and AQP5 FERM complexes using ColabFold reveals a common mode of binding in which the proximal and distal parts of the AQP C-termini bind simultaneously to distinct binding sites of FERM. While the interaction at each site closely resembles other FERM-complexes, the concurrent interaction with both sites has only been observed in the complex between moesin and its C-terminus which causes auto-inhibition. The proposed interaction between AQP2/AQP5 and FERM thus represents a novel binding mode for extrinsic ERM-interacting partners.

Advancing Sustainable Malting Practices: Aquaporins as Potential Breeding Targets for Improved Water Uptake during Controlled Germination of Barley (Hordeum vulgare L.).

The conversion of raw barley (Hordeum vulgare L.) to malt requires a process of controlled germination, where the grain is submerged in water to raise the moisture content to >40%. The transmembrane proteins, aquaporins, influence water uptake during the initial stage of controlled germination, yet little is known of their involvement in malting. With the current focus on sustainability, understanding the mechanisms of water uptake and usage during the initial stages of malting has become vital in improving efficient malting practices. In this study, we used quantitative proteomics analysis of two malting barley genotypes demonstrating differing water-uptake phenotypes in the initial stages of malting. Our study quantified 19 transmembrane proteins from nine families, including seven distinct aquaporin isoforms, including the plasma intrinsic proteins (PIPs) PIP1;1, PIP2;1, and PIP2;4 and the tonoplast intrinsic proteins (TIPs) TIP1;1, TIP2;3, TIP3;1, and TIP3;2. Our findings suggest that the presence of TIP1;1, TIP3;1, and TIP3;2 in the mature barley grain proteome is essential for facilitating water uptake, influencing cell turgor and the formation of large central lytic vacuoles aiding storage reserve hydrolysis and endosperm modification efficiency. This study proposes that TIP3s mediate water uptake in malting barley grain, offering potential breeding targets for improving sustainable malting practices.

The differential expressions of aquaporins underline the diverse strategies of cucumber and tomato against salinity and zinc stress.

Salinity and excess zinc are two main problems that have limited agriculture in recent years. Aquaporins are crucial in regulating the passage of water and solutes through cells and may be essential for mitigating abiotic stresses. In the present study, the adaptive response to moderate salinity (60 mM NaCl) and excess Zn (1 mM ZnSO4 ) were compared alone and in combination in Cucumis sativus L. and Solanum lycopersicum L. Water relations, gas exchange and the differential expression of all aquaporins were analysed. The results showed that cucumber plants under salinity maintained the internal movement of water through osmotic adjustment and the overexpression of specific PIPs aquaporins, following a "conservation strategy". As tomato has a high tolerance to salinity, the physiological parameters and the expression of most aquaporins remained unchanged. ZnSO4 was shown to be stressful for both plant species. While cucumber upregulated 7 aquaporin isoforms, the expression of aquaporins increased in a generalized manner in tomato. Despite the differences, water relations and transpiration were adjusted in both plants, allowing the RWC in the shoot to be maintained. The aquaporin regulation in cucumber plants facing NaCl+ZnSO4 stress was similar in the two treatments containing NaCl, evidencing the predominance of salt in stress. However, in tomato, the induced expression of specific isoforms to deal with the combined stress differed from independent stresses. The results clarify the key role of aquaporin regulation in facing abiotic stresses and their possible use as markers of tolerance to salinity and heavy metals in plants.

Nitric oxide modulates the expression of aquaporin isoforms (PIP2 and TIP1) on oil body membranes in sunflower (Helianthus annuus L.) seedling cotyledons in response to salt stress

Nitric oxide modulates the expression of aquaporin isoforms (PIP2 and TIP1) on oil body membranes in sunflower (Helianthus annuus L.) seedling cotyledons in response to salt stress

Red LED Light Improves Pepper (Capsicum annuum L.) Seed Radicle Emergence and Growth through the Modulation of Aquaporins, Hormone Homeostasis, and Metabolite Remobilization

Red LED light (R LED) is an efficient tool to improve seed germination and plant growth under controlled environments since it is more readily absorbed by photoreceptors' phytochromes compared to other wavelengths of the spectrum. In this work, the effect of R LED on the radicle emergence and growth (Phase III of germination) of pepper seeds was evaluated. Thus, the impact of R LED on water transport through different intrinsic membrane proteins, via aquaporin (AQP) isoforms, was determined. In addition, the remobilization of distinct metabolites such as amino acids, sugars, organic acids, and hormones was analysed. R LED induced a higher germination speed index, regulated by an increased water uptake. PIP2;3 and PIP2;5 aquaporin isoforms were highly expressed and could contribute to a faster and more effective hydration of embryo tissues, leading to a reduction of the germination time. By contrast, TIP1;7, TIP1;8, TIP3;1 and TIP3;2 gene expressions were reduced in R LED-treated seeds, pointing to a lower need for protein remobilization. NIP4;5 and XIP1;1 were also involved in radicle growth but their role needs to be elucidated. In addition, R LED induced changes in amino acids and organic acids as well as sugars. Therefore, an advanced metabolome oriented to a higher energetic metabolism was observed, conditioning better seed germination performance together with a rapid water flux.

Role of aquaporins in corneal healing post chemical injury

Aquaporins (AQPs) are transmembrane water channel proteins that regulate the movement of water through the plasma membrane in various tissues including cornea. The cornea is avascular and has specialized microcirculatory mechanisms for homeostasis. AQPs regulate corneal hydration and transparency for normal vision. Currently, there are 13 known isoforms of AQPs that can be subclassified as orthodox AQPs, aquaglyceroporins (AQGPs), or supraquaporins (SAQPs)/unorthodox AQPs. AQPs are implicated in keratocyte function, inflammation, edema, angiogenesis, microvessel proliferation, and the wound-healing process in the cornea. AQPs play an important role in wound healing by facilitating the movement of corneal stromal keratocytes by squeezing through tight stromal matrix and narrow extracellular spaces to the wound site. Deficiency of AQPs can cause reduced concentration of hepatocyte growth factor (HGF) leading to reduced epithelial proliferation, reduced/impaired keratocyte migration, reduced number of keratocytes in the injury site, delayed and abnormal wound healing process. Dysregulated AQPs cause dysfunction in osmolar homeostasis as well as wound healing mechanisms. The cornea is a transparent avascular tissue that constitutes the anterior aspect of the outer covering of the eye and aids in two-thirds of visual light refraction. Being the outermost layer of the eye, the cornea is prone to injury. Of the 13 AQP isoforms, AQP1 is expressed in the stromal keratocytes and endothelial cells, and AQP3 and AQP5 are expressed in epithelial cells in the human cornea. AQPs can facilitate wound healing through aid in cellular migration, proliferation, migration, extracellular matrix (ECM) remodeling and autophagy mechanism. Corneal wound healing post-chemical injury requires an integrative and coordinated activity of the epithelium, stromal keratocytes, endothelium, ECM, and a battery of cytokines and growth factors to restore corneal transparency. If the chemical injury is mild, the cornea will heal with normal clarity, but severe injuries can lead to partial and/or permanent loss of corneal functions. Currently, the role of AQPs in corneal wound healing is poorly understood in the context of chemical injury. This review discusses the current literature and the role of AQPs in corneal homeostasis, wound repair, and potential therapeutic target for acute and chronic corneal injuries.

A high-throughput yeast approach to characterize aquaporin permeabilities: Profiling the Arabidopsis PIP aquaporin sub-family.

Engineering membrane transporters to achieve desired functionality is reliant on availability of experimental data informing structure-function relationships and intelligent design. Plant aquaporin (AQP) isoforms are capable of transporting diverse substrates such as signaling molecules, nutrients, metalloids, and gases, as well as water. AQPs can act as multifunctional channels and their transport function is reliant on many factors, with few studies having assessed transport function of specific isoforms for multiple substrates. High-throughput yeast assays were developed to screen for transport function of plant AQPs, providing a platform for fast data generation and cataloguing of substrate transport profiles. We applied our high-throughput growth-based yeast assays to screen all 13 Arabidopsis PIPs (AtPIPs) for transport of water and several neutral solutes: hydrogen peroxide (H2O2), boric acid (BA), and urea. Sodium (Na+) transport was assessed using elemental analysis techniques. All AtPIPs facilitated water and H2O2 transport, although their growth phenotypes varied, and none were candidates for urea transport. For BA and Na+ transport, AtPIP2;2 and AtPIP2;7 were the top candidates, with yeast expressing these isoforms having the most pronounced toxicity response to BA exposure and accumulating the highest amounts of Na+. Linking putative AtPIP isoform substrate transport profiles with phylogenetics and gene expression data, enabled us to align possible substrate preferences with known and hypothesized biological roles of AtPIPs. This testing framework enables efficient cataloguing of putative transport functionality of diverse AQPs at a scale that can help accelerate our understanding of AQP biology through big data approaches (e.g. association studies). The principles of the individual assays could be further adapted to test additional substrates. Data generated from this framework could inform future testing of AQP physiological roles, and address knowledge gaps in structure-function relationships to improve engineering efforts.

Changes in root hydraulic conductivity in wheat (Triticum aestivum L.) in response to salt stress and day/night can best be explained through altered activity of aquaporins.

Salt stress reduces plant water flow during day and night. It is not known to which extent root hydraulic properties change in parallel. To test this idea, hydroponically grown wheat plants were grown at four levels of salt stress (50, 100, 150 and 200 mM NaCl) for 5-8d before harvest (d14-18) and subjected to a range of analyses to determine diurnal changes in hydraulic conductivity (Lp) at cell, root and plant level. Cell pressure probe analyses showed that the Lp of cortex cells was differentially affected by salt stress during day and night, and that the response to salt stress differed between the main axis of roots and lateral roots. The Aquaporin (AQP) inhibitor H2 O2 reduced Lp to a common, across treatments, level as observed in salt-stressed plants during the night. Analyses of transpiring plants and exuding root systems provided values of root Lp which were in the same range as values modeled based on cell-Lp. The results can best be explained through a change in root Lp in response to salt stress and day/night, which results from an altered activity of AQPs. qPCR gene expression analyses point to possible candidate AQP isoforms.

Effect of drought on aquaporin expression in grafted and ungrafted grapevine cultivars

Drought stress severely affects growth, development and productivity in most agricultural crops. Since ancient times, rootstocks have been used to enable crop cultivation in unsuitable soil conditions. In the present study, three factors were evaluated: 1) cultivar: Vitis vinifera L. cv. ‘Horozkarası’ (drought-tolerant) and cv. ‘Kabarcık’ (drought-sensitive) were used; 2) rootstock: each cultivar was self-rooted and grafted onto ‘Rupestris du Lot’ rootstock; 3) drought stress: half of each cultivar/rootstock combination underwent drought stress and the other half was irrigated at field capacity for seven days. In order to estimate the responses of the cultivars, relative water content, proline content and aquaporin isoform expression levels (VvPIP2;1, VvPIP2;2, VvTIP1;1, and VvTIP2;1) were quantified. The results revealed that drought stress caused more reduction in relative water content (RWC) in ‘Kabarcık’ cultivar (drought-sensitive) than in ‘Horozkarası’ cultivar (drought-tolerant). Proline content increased in both cultivars in response to drought stress but to a relatively greater extent in the grafted ‘Kabarcık’ cultivar. Considering expression levels of genes, VvPIP2;1, VvPIP2;2, and VvTIP2;1 were downregulated whilst VvTIP1;1 was upregulated in the leaf. Both ‘Horozkarası’ and ‘Kabarcık’ cultivars showed similar trends in terms of their responses to drought stress. Grafting significantly increased the proline content in both cultivars exposed to drought stress. The rootstock conferred better drought protection to ‘Kabarcık’ cultivar than to ‘Horozkarası’ cultivar.

Expression patterns of maize PIP aquaporins in middle or upper leaves correlate with their different physiological responses to drought and mycorrhiza.

Here we report the effect of Rhizophagus irregularis on maize leaf expression of six plasma membrane aquaporin isoforms from PIP1 and PIP2 subfamilies under severe drought development and recovery. The novelty of our study is the finding that leaf-specific mycorrhizal regulation of aquaporins is dependent on the position of the leaf on the shoot and changes in parallel with the rate of photosynthesis and the stomatal response to drought. The transcripts were isolated from the upper third (L3) or ear (L5) leaf, which differed greatly in physiological response to stress within each symbiotic variant. Aquaporins expression in upper L3 leaves appeared to be largely not sensitive to drought, regardless of symbiotic status. In contrast, L5 leaf of non-mycorrhizal plants, showed strong down-regulation of all PIPs. Mycorrhiza, however, protected L5 leaf from such limitation, which under maximal stress was manifested by 6-fold and circa 4-fold higher transcripts level for PIP1s and PIP2s, respectively. Distinct expression patterns of L3 and L5 leaves corresponded to differences in key parameters of leaf homeostasis - stomatal conductance, photosynthetic rates, and accumulation of ABA and SA as phytohormonal indicators of drought stress. In result symbiotic plants showed faster restoration of photosynthetic capability, regardless of leaf position, which we recognize as the hallmark of better stress tolerance. In summary, arbuscular mycorrhiza alleviates short-term drought effects on maize by preventing the down-regulation of plasma membrane aquaporins within middle leaves, thereby affecting stomatal conductance.

Genome-wide identification of Fagus sylvatica aquaporins and their comparative spring and summer expression profiles

Key messageA total of 45 aquaporins was identified in Fagus sylvatica, 35 of which were differentially expressed in spring and summer in the leaves, phloem and xylem of 11-year-old trees.European beech (Fagus sylvatica) has been widely studied in terms of its water relations and local adaptation. However, to date, the underlying basis conferring adaptation to differences in water availability are unknown. Therefore, we examined the expression of aquaporins in trees of four different beech provenances representing the southern and northern range margins, as well as core populations, grown in a common garden. We sampled their xylem, phloem and leaf tissue, when leaves had fully expanded, and in late summer. A total of 45 aquaporin isoforms were identified in the beech genome, of which 35 were detected across all sampled tissues. In our phylogenetic analysis, beech aquaporins clustered into the five subfamilies found in other woody species. Members of the plasma membrane intrinsic protein subfamily generally displayed the highest levels of expression, followed by tonoplast intrinsic proteins. Isoforms of the remaining subfamilies, Noduline-26-like intrinsic proteins, small basic intrinsic proteins and uncharacterised intrinsic proteins, were expressed at very low to moderate levels. The expression of most isoforms was stable or declined from spring to summer. Leaves followed a different expression profile from that of vascular tissues, whereas both phloem and xylem were found to express the same FsMIPs. Tissue-specific aquaporin expression was very similar amongst the four beech provenances, indicating that there is no inherent difference in the capability of these provenances to regulate aquaporin activity. The general decrease in FsMIP expression toward the end of the growing period indicates that aquaporins are involved in tree water relations and growth.

Genome-wide identification and biological relevance of broccoli aquaporins.

Broccoli (Brassica oleracea var. italica) is an important crop worldwide, and its regular consumption is associated with health benefits due to the presence of various bioactive compounds. An optimal water balance and homeostasis are needed for plant growth; in this sense, aquaporins play a crucial role. As a result of a genome-wide search, a total of 65 aquaporin genes were identified in broccoli. The aquaporins were classified according to their phylogenetic relationships with other Brassicas species and Arabidopsis thaliana, and evolutionary events of gene duplication were also assessed, highlighting the tendency of NIPs (Nodulin-26-like Intrinsic Proteins) to duplicate. Also, the chromosomal localization, gene duplication, the study of the conserved motifs, and the tertiary structure were determined in broccoli. Functional predictive analyses were also carried out, which, together with the expression analyses in different broccoli plant tissues, allowed the prediction of the biological role of each aquaporin isoform. BoiPIP1-2a and BoiPIP1-2b showed higher expression in all the plant tissues when compared with other aquaporins. BoiTIP1-2b also showed high expression levels and was associated with nitrogen compounds transport such as urea. However, NIPs, through their differential expression and the tandem duplications of the isoforms, were revealed as the putative main actors in the response of broccoli plants to abiotic stress responses. The results of this work pointed to the physiological significance of each aquaporin isoform of broccoli, opening a new field of knowledge and constituting the first step of further in vivo analyses.

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.

Changes in the expression of certain osmosensitive channel and transporter genes in primary hepatocytes of air-breathing catfish, Clarias magur: A strategy to adapt under osmotic stress

Environmental salinity/osmolarity is an important abiotic ecological factor that affects several metabolic activities of aquatic organisms. Air-breathing magur catfish (Clarias magur) frequently encounter a wide range of external osmolarity changes in their natural habitats as well as internal osmolarity due to the absorption of amino acids and other metabolites through the intestine. The major objective of the present investigation was to demonstrate the influences of anisotonicity exposure on the expression of certain isoforms of aquaporin (AQP) channels (aqp7, 8, and 11), taurine transporter (TauT, slc6a6), urea transporter (UT1, slc14a1) and Na+-K+-ATPase (NKA, atp1a1, and atp1b1) transporter genes, and their translated protein products in the primary hepatocytes of magur catfish. These genes are known to play major roles in cellular volume regulation and water homeostasis under osmotic stress. Exposure of primary hepatocytes to hypertonic medium (+40 mOsmol/L) for a period of 48 h resulted in a significant increase in the expression of aqp7, aqp8, aqp11, slc6a6, slc14a1, atp1a1, and atp1b1 genes, as evidenced by a rise in the levels of both mRNAs of all the above-mentioned genes, and also their translated protein products compared to isotonic controls. However, exposure of primary hepatocytes to hypotonic medium (_40 mOsmol/L) for the same period led to a significant decrease in mRNA levels for aqp7, aqp8, aqp11, slc6a6, and slc14a1 genes, and also their translated protein products compared to isotonic controls as a consequence of downregulation of corresponding genes. Interestingly, NKA enzyme activity in primary hepatocytes was found to increase significantly after exposure to both hypertonic and hypotonic media. Hypertonicity induced the genes for α- and β-subunits of this enzyme both at transcriptional and translational levels. In contrast, hypotonicity induced only the atp1a1 gene and the expression of corresponding translated protein without causing any changes in the expression of the atp1b1 gene. Thus, it may be contemplated that osmotic stress, which results due to anisotonicity of the external environment or in the body fluids, leads to differential expression of different osmosensitive channels such as AQPs, and transporters such as TauT, UT1, UT2, and NKA in hepatic cells of magur catfish and work in coordination to defend the osmotically-induced changes of cellular volume. These regulatory responses of channels and transporters could be one of the vital molecular mechanisms that may have evolved in this catfish to defend against the osmotically-induced stresses.

Role of aquaporin 9 in hyperglycaemia-induced testicular leydig cell apoptosis

Hyperglycaemia is an early symptom of diabetes mellitus (DM) and hyperglycaemia-induced apoptosis is the most widely accepted cause for diabetes induced male infertility/subfertility. Aquaporin (AQP) isoforms (AQP0-12), are water channels involved in maintaining water homeostasis and regulate fluid absorption and/or excretion; thus, play a key role in reproductive function. Further, AQPs regulate diabetic complications, apoptosis induced by hyperglycaemia/high glucose (HG) and also involved in insulin regulation. Herein, we investigated AQP 9 role in hyperglycaemia-induced testicular Leydig cell apoptosis. In-vivo and in-vitro role of AQP 9 on hyperglycaemia induced Leydig cell apoptosis was analysed in diabetic rat testis and LC-540 rat Leydig cells by Real Time-PCR, western blotting and siRNA knock-down experiments. In addition, HG effect on viability and morphological changes due to apoptosis in cells were assessed by MTT and fluorescence microscopy. In-vivo and in-vitro AQP 9 expressions were increased in diabetic testicular Leydig cells. Hyperglycaemia caused apoptosis in testicular Leydig cells by increasing Cytochrome c, Bax and decreasing Bcl-2. Furthermore, AQP 9 knock-down resulted in downregulation of pro-apoptotic proteins and in contrast, upregulated anti-apoptotic protein expressions; thereby decreased apoptosis in testicular Leydig cells under hyperglycemia. Our findings collectively suggest, AQP 9 role in hyperglycaemia-induced apoptosis in testicular Leydig cells.

Aquaporin-mediated transport: Insights into metalloid trafficking.

Metalloids in plants have diverse physiological effects. From being essential to beneficial to toxic, they have significant effects on many physiological processes, influencing crop yield and quality. Aquaporins are a group of membrane channels that have several physiological substrates along with water. Metalloids have emerged as one of their important substrates and they are found to have a substantial role in regulating plant metalloid homeostasis. The present review comprehensively details the multiple isoforms of aquaporins having specificity for metalloids and being responsible for their influx, distribution or efflux. In addition, it also highlights the usage of aquaporin-mediated transport as a selection marker in toxic screens and as tracer elements for closely related metalloids. Therefore, aquaporins, with their imperative contribution to the regulation of plant growth, development and physiological processes, need more research to unravel the metalloid trafficking mechanisms and their future applications.

Differences in the localization of AQP1 and expression patterns of AQP isoforms in rat and mouse sciatic nerve and changes in rat AQPs expression after nerve crush injury.

In the peripheral nervous system aquaporins (AQPs) have been reported in both peripheral neurons and glial cells. Previously we described the precise localization of AQP1 in the rat sciatic nerve, which is present in both Remak and myelin Schwann cells, and is enriched in the Schmidt-Lanterman incisures. In this work, we found that AQP1 in mouse is only present in Remak cells, showing a different localization between these species. However, after nerve crush injury the level of AQP1 mRNA expression remains constant at all times studied in rat and mouse. We then performed RT-PCR of nine AQP (AQP1–9) isoforms from rat and mouse sciatic nerve, we found that in rat only five AQPs are present (AQP1, AQP4, AQP5, AQP7 and AQP9), whereas in mouse all AQPs except AQP8 are expressed. Then, we studied the expression by RT-PCR of AQPs in rat after nerve crush injury, showing that AQP1, AQP4 and AQP7 expression remain constant at all times studied, while AQP2, AQP5 and AQP9 are upregulated after injury. Therefore, these two closely related rodents show different AQP1 localization and have different AQPs expression patterns in the sciatic nerve, possibly due to a difference in the regulation of these AQPs. The expression of AQP1 in Remak cells supports the involvement of AQP1 in pain perception. Also, in rat the upregulation of AQP2, AQP5 and AQP7 after nerve injury suggests a possible role for these AQPs in promoting regeneration following injury.

Molecular characterization and structural dynamics of Aquaporin1 from walking catfish in lipid bilayers

Aquaporin's (AQPs) are the major superfamily of small integral membrane proteins that facilitates transportation of water, urea, ammonia, glycerol and ions across biological cell membranes. Despite of recent advancements made in understanding the biology of Aquaporin's, only few isoforms of aquaporin 1 (AQP1) some of the teleost fish species have been characterized at molecular scale. In this study, we made an attempt to elucidate the molecular mechanism of water transportation in AQP1 from walking catfish (Clarias batrachus), a model species capable of breathing in air and inhabits in challenging environments. Using state-of-the-art computational modelling and all-atoms molecular dynamics simulation, we explored the structural dynamics of full-length aquaporin 1 from walking catfish (CbAQP1) in lipid mimetic bilayers. Unlike AQP1 of human and bovine, structural ensembles of CbAQP1 from MD revealed discrete positioning of pore lining residues at the intracellular end. Snapshots from MD simulation displayed differential dynamics of aromatic/arginine (ar/R) filter and extracellular loop C bridging transmembrane (TM) helix H3 and H4. Distinct conformation of large extracellular loops, loop bridging TM2 domain and HB helix along with positioning of selectivity filter lining residues controls the permeability of water across the bilayer. Moreover, the identified unique and conserved lipid binding sites with 100% lipid occupancy signifies lipid mediated structural dynamics of CbAQP1. All-together, this is the first ever report on structural-dynamics of aquaporin 1 in walking catfish which will be useful to understand the molecular basis of transportation of water and other small molecules under varying degree of hyperosmotic environment.

Nutritional and Physiological Regulation of Water Transport in the Conceptus.

Water transport during pregnancy is essential for maintaining normal growth and development of conceptuses (embryo/fetus and associated membranes). Aquaporins (AQPs) are a family of small integral plasma membrane proteins that primarily transport water across the plasma membrane. At least 11 isoforms of AQPs (AQPs 1-9, 11, and 12) are differentially expressed in the mammalian placenta (amnion, allantois, and chorion), and organs (kidney, lung, brain, heart, and skin) of embryos/fetuses during prenatal development. Available evidence suggests that the presence of AQPs in the conceptus mediates water movement across the placenta to support the placentation, thehomeostasis of amniotic and allantoic fluid volumes, as well as embryonicand fetal survival,growth and development. Abundances of AQPs in the conceptus can be modulated by nutritional status and physiological factors affecting the pregnant female. Here, we summarize the effects of maternal dietary factors (such as intakes of protein, arginine, lipids, all-trans retinoic acid, copper, zinc, and mercury) on the expression of AQPs in the conceptus. We also discuss the physiological changes in hormones(e.g., progesterone and estrogen), oxygensupply, nitric oxide,pH, and osmotic pressure associated with the regulation of fluid exchange between mother and fetus. These findings may help to improve the survival, growth, and development of embryo/fetus in livestock species and other mammals (including humans).