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Amp-Activated Protein Kinase Knockdown In Labyrinthine Trophoblast Cells Results In Altered Morphology And Function, Erica Ashton Kayleigh Carey

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The placenta is a transient organ that develops upon initiation of pregnancy and is essential for fetal development and survival. The rodent placenta consists of three layers with predominantly analogous cell types in the human placenta. The labyrinth layer, which lies closest to the fetus, facilitates nutrient and waste exchange between mother and baby. Abnormalities of the placenta may occur as a result of cellular stress and have been associated with the pregnancy-associated disorders, intrauterine growth restriction and placental insufficiency. AMP-activated protein kinase (AMPK) is a master sensor of cellular stress and changes in AMPK expression or activation could be …

Novel Approaches For The Eradication Of Hiv Latently Infected Cells, Sally Al Ali

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The development of a suitable experimental cell model to study HIV latency in primary cells could have a massive effect on the current approaches to eradicate virus in latently infected cells. The main proposal of this paper is to develop an in vitro HIV cell model that represents HIV latency in vivo, then to create a more effective viral vector in order to target HIV reservoirs. For this goal, a directed evolution method is suggested to be used in order to mutate the AAV cap gene to generate a recombinant AAV vector that is capable of infecting primary resting CD4+ …

Co2/Ph Signaling Of Respiratory Control Neurons In The Bullfrog, Lithobates Catesbeianus: Development Of A Comparative Model, Joseph M. Santin

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The locus coeruleus (LC) in the brainstem senses alterations in CO₂/pH and influences ventilatory adjustments that restore blood gas values to starting levels in bullfrogs (Lithobates catesbeianus). We hypothesized that neurons of the bullfrog LC are sensitive to changes in CO₂/pH and that chemosensitive responses are intrinsic to individual neurons. In addition, we hypothesized putative respiratory control neurons of the bullfrog LC would be stimulated by hypercapnic acidosis within physiological ranges of PCO₂/pH. 84% of LC neurons depolarized and increased firing rates during exposure to hypercapnic acidosis (HA). A pH dose response …

The Phylogenetics Of Tachinidae (Insecta: Diptera) With An Emphasis On Subfamily Structure, Daniel J. Davis

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The parasitoid flies of the Tachinidae family are an important and diverse (>10,000 species) lineage of insects. However, tachinids are not well studied partially due to their confusing classification and taxonomy. DNA sequences were obtained from twenty tribal representatives of Tachinidae, along with eight outgroups in order to phylogenetically reconstruct the superfamilial, subfamilial and tribal relationships of Tachinidae. Seven gene regions of six genes (18S, 28S, COI, CAD, Ef1a, and TPI) were sequenced for each taxon (6214 bp total). Both maximum likelihood and Bayesian methods were used to infer phylogenies. The Sarcophagidae and Oestridae were usually reconstructed as monophyletic. …

Hpai H5n1: A Global Pandemic Concern, With Implications For Pandemic Preperation And Public Health Policy, Lauren M. Koontz

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Research on highly pathogenic avian influenza A (HPAI) H5N1 has gained much attention in recent years due to its devastating impact within the bird population. While transmission to humans is rare, infections are fatal in more than half of the cases. One of the main concerns among the scientific community is the ability for this virus to mutate in a way much like seasonal influeanza that would allow it to be transmitted efficiently among the human population. Two researchers were able to separately create mutant isolates of H5N1 influenza A virus that could be transmitted via the respiratory route between …

Fabrication Of Responsive Polymer Brushes For Patterned Cell Growth And Detachment, Ashley B. Sutherland

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Patterned polymer brushes are used in biological applications to enhance cell function and to achieve selective cell growth. These patterned surfaces have a variety of applications, including cell sheet harvesting and tissue engineering. This work describes creation of chemical, topological, and responsive patterns on glass by sequential surface-initiated polymerization of poly(N-isopropylacrylamide)-co-(2-hydroxyethyl methacrylate) (pNIPAM-co-HEMA) and subsequent use of UV lithography to pattern the surface with pNIPAM. The cell adhesion peptide, RGD, is then immobilized on the surface of the copolymer pHEMA via DSC-coupling to spatially enhance cell adhesion properties. By culturing cells at 37 degrees celsius, cells selectively grow on DSC-activated …

Type I Interferon Activation Of Natural Killer (Nk) Cells By Cytomegalovirus (Cmv) And Their Interaction With Dendritic (Dc) And Nkt Cells, Howaida A. Oulad Abdelati

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In our current study the roles of natural killer (NK) cells in regulation of the acute phase of murine cytomegalovirus (MCMV) infection were demonstrated. NK cells utilize perforin and gamma interferon to kill MCMV infected cells. Activation of NK cells is controlled by the balance between inhibitory and activation receptors and modulated by cytokines produced from the infected cells. Type I interferons are produced mainly by plasmacytoid dendritic cells (PDCs). Type I interferons have a substantial role in enhancing NK cells cytotoxic activity and NK cells proliferation and maintain cell survival through STAT mediated signaling pathway. IL-12 produced by dendritic …

The Aryl Hydrocarbon Receptor Regulates An Essential Transcriptional Element In The Immunoglobulin Heavy Chain Gene, Michael J. Wourms

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2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental contaminant that inhibits immunoglobulin (Ig) expression and Ig heavy (IgH) chain gene transcription. Transcription of the IgH gene involves several regulatory elements including the 3'lgh regulatory region (3'lghRR) which is composed of four enhancers (hs3A, hs1,2, hs4, and hs3B). Dioxin responsive elements (DRE) in the hs4 and hs1,2 enhancers of the 3lghRR that bind the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that regulates dioxin sensitive genes suggest that the 3'lghRR may be a transcriptional target of TCDD. The current study utilized an IgA secreting mouse …

Role Of Bax, Ibpa, Ibpb And Csph Genes In Protecting Cft073 (Uropathogenic Escherichia Coli) Against Salt And Urea Stress, Pavani Beesetty

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Uropathogenic Escherichia coli (UPEC) are the primary cause of 80 to 90% of uncomplicated urinary tract infections. After entering the urinary tract, uropathogenic E.coli has to tolerate high levels of salt and urea in urine to cause a successful infection. The osmotic stress imposed by urea is different from that by NaCl, as urea can freely move across the cell membrane and is a protein denaturant. Hence, microarray experiments were performed to observe the differential expression of genes in CFT073 (UPEC) due to the presence of 0.3 M NaCl and 0.6 M urea in K medium individually. Based on the …

Enhancing Herbicide Efficacy On Reed Canary Grass (Phalaris Arundinacea) By Testing A Plant Growth Hormone, Application Times, And Herbicide Type, Denise Lynn Fong

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Phalaris arundinacea, also known as reed canary grass (RCG), is a non-native invasive grass that thrives in floodplains. RCG plants displace native flora which reduces diversity in otherwise species rich wetlands. This is a widespread problem throughout many parts of the USA. RCG can grow by its seeds or its rhizomes. Its seeds are capable of surviving long periods in soil. RCG can potentially be controlled by multi-year treatments in early spring and early fall with broad spectrum or grass specific herbicides. The goal of this study was to optimize methods to control RCG, in order to increase species …

Long-Term Effects Of Deer Browsing On Northern Wisconsin Forest Plant Communities, Danielle Rae Begley

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The effects of excessive white-tailed deer (Odocoileus virginianus) browsing on forest understory plant communities are well-documented. However, these studies are usually short-term, and fail to focus on phylogenetic diversity and additional variables that may explain species composition. This study examined ecological and phylogenetic diversity, vegetation structure, and light (% insolation) in short (7 year) and long-term (22 year) exclosures (and paired controls) to identify if competitive exclusion occurs in long-term exclosures. A deer browsing susceptibility index (DBS) was also developed to identify species reliant on exclosures for persistence. Statistical analysis using 2-way ANOVAs showed increased percent cover, vegetation height, and …

Tolfenamic Acid Induces Cell Apoptosis And Inhibits Collagen Accumulation In Keloid Fibroblasts, Dan Yi

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Keloid scar is a fibroproliferative disorder which results from the accumulation of collagen through the activation of TGF-B/Smad signaling. Tolfenamic acid (TA) has been found to inhibit collagen synthesis in animal and induce cancer cell apoptosis. In this study, we investigated the effect of DMSO on cell apoptosis, TA on collagen deposition, and TA on collagen type I and smad2 protein expression in human normal fibroblasts (NFs) and human keloid fibroblasts (KFs). In the dose- and exposure time- response assays, we found that 1% DMSO for 48h treatment did not obviously induce NF and KF apoptosis and that 0.55x10-3 M …

Histone Deacetylation As A Mechanism Of Ypel3 Down-Regulation In Er-A Positive Breast Cancer Cell Lines, Remah Ali

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YPEL3 is a growth inhibitory gene that was established by our laboratory to trigger senescence in both primary and tumor cells. We recently reported YPEL3 to be directly repressed by estrogen in estrogen receptor-a positive (ER-a +ve) breast cancer cell lines. Here, we set out to determine whether the repression of YPEL3 involves histone deacetylation and, if so, elucidate the molecular mechanism of this activity. MCF-7, T-47D, and ZR-75.1 (all ER-a +ve) breast cancer cell lines were treated with varying doses of the global histone deacetylase inhibitors (HDACIs), trichostatin A (TSA) or suberoyl anilide hydroxamic acid (SAHA), for 24, 48 …

Can Antibiotics From Recently Discovered Marine Actinobacteria Slow The Tide Of Antibiotic Resistance?, Lorraine Susan Tangeman

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Actinobacteria, one phylum of gram positive bacteria, are found throughout all the environments on earth. Actinobacteria have long been studied for the benefits they provide, both to their environment and to humans, and have a great capacity for adaptation and evolution. They decompose organic matter, replenishing nutrients into the soil, and as such are important members of the food chain. Humans benefit from the exploitation of Actinobacterial metabolites as antimicrobial drugs. These antimicrobials have been effectively utilized for decades in the fight against infectious disease. Despite the success of this drug arsenal we are now in the midst of an …

Likely Successors Of Ash Species In Response To The Emerald Ash Borer In Ohio Forests, Brian Michael Good

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Invasive species have the capability to alter landscapes and change the composition of a forest in a very short time. The recent invasive pest, Agrilus planipennis, emerald ash borer, was unintentionally introduced to the United States via ship route to Michigan. The pest attacks and kills all five native ash species in Ohio. This study focused on an area in west central Ohio not yet affected by the borer. Ash centered plots were used to record all species and sizes (diameter at breast height) within a 5m radius of a central ash tree. Plots ranged in topography and all five …

Effects Of Ang 1-7 And Endothelial Microvesicles On Ang Ii-Induced Dysfunction And Apoptosis In Cerebral Endothelial Cells, Xiang Xiao

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Angiotensin II (Ang II) induces endothelial dysfunction and is implicated in the pathogenesis of vascular diseases. Angiotensin 1-7 (Ang 1-7) has been reported to counteract many deleterious effects of Ang II. Endothelial microvesicles (EMVs) are small membrane vesicles released from endothelial cells (ECs) undergoing stress and apoptosis. But their functions are largely unknown. In this study, we investigated the effects of Ang 1-7 and EMVs on apoptosis and dysfunction of human brain microvascular endothelial cells (HbmECs). Reactive oxygen species (ROS) and nitric oxide (NO) production, and Nox2, p-Akt/Akt, p-eNOS/eNOS expression were analyzed. We found that Ang II dose-dependently induced HbmEC …

Conversion Of The U937 Monocyte Into "Macrophage-Like" Populations Exhibiting M1 Or M2 Characteristics, Bradley M. Sharp

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The U937 cell line is an oncogenic human monocyte cell line. These monocytes have the potential of differentiating into either macrophages or dendritic cells (Lawrence et al., 2011). This differentiation pattern depends on the characteristics of the tissue microenvironment (Kigerl et al., 2009). PMA (Phorbol 12-Myristate 13-Acetate) is a phorbol ester capable of transforming monocytic cells toward the macrophage pathway. Upon treatment with PMA, U937 cells under-go a series of morphological and functional changes. Traditionally monocytic cell lines are used as a model of macrophage function, because current human macrophage cell lines require a T-cell conditioned growth medium and contact …

Insulin Treatment Attenuates Renal Adam17 And Ace2 Shedding In Akita Diabetic Mice, Esam Said Busah Salem

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Diabetic patients have a 40%-50% lifetime chance of developing chronic kidney disease, which remains one of the leading causes of morbidity and mortality. Alterations within renin angiotensin system balance contribute to the pathogenesis of diabetic kidney disease. Angiotensin converting enzyme 2 (ACE2) has an endogenous renoprotective role due to its ability to form angiotensin (1-7) (Ang 1-7) by degrading angiotensin II (Ang II). We have shown previously that hyperglycemia increases urinary ACE2 and albumin excretion in db/db diabetic mice. The protease, disintegrin and metalloprotease (ADAM) 17, is involved in the shedding of several transmembrane proteins, including ACE2 in vitro. Tissue …

Physical Exercise Training But Not Metformin Attenuates Albuminuria And Shedding Of Ace2 In Type 2 Diabetic Db/Db Mice, Hari Krishna Somineni

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Angiotensin II (Ang II), a potent vasoconstrictor cleaved from Ang I, is responsible for renal damage in diabetes. Angiotensin converting enzyme 2 (ACE2) is highly expressed in the kidney and has been shown to be renoprotective by degrading Ang II to Ang-(1-7). A Disintegrin and Metalloproteinases (ADAMs) were recently identified as an ectodomain sheddases of transmembrane proteins. ADAM17 mediated shedding of renal ACE2 could contribute to the pathogenesis of diabetic nephropathy. In our previous study, rosiglitazone treatment normalized hyperglycemia and improved renal injury by preventing ACE2 shedding. The aim of this study is to test the hypothesis that improved glucose …

Regulation Of The Stability And Cellular Localization Of The Coxsackievirus And Adenovirus Receptor (Car), Ran Yan

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In polarized epithelia, the seven exon isoform of the Coxsackievirus and adenovirus receptor (CAR^Ex7) is a basolateral cell-cell adhesion protein that is inaccessible for viral infection. In contrast, the eight-exon CAR isoform (CAR^Ex8) localizes at the apical surface and mediates adenovirus (AdV) infection. A PDZ-domain containing protein, MAGI-1, interacts with both isoforms of CAR. I hypothesized that each CAR isoform interacts with specific MAGI-1 PDZ domain(s). Co-immunoprecipitation, FRET and binding assays showed that CAR^Ex7 and CAR^Ex8 both interact with MAGI-1-PDZ3 with high affinity. CAR^Ex8 also interacts with MAGI-1-PDZ1. Whereas the CAR^Ex7-PDZ3 …

Bioaccumulation And Neuroinflammation Of Gold Nanoparticles In The Central Nervous System, Fahimeh Fallahi

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Gold nanoparticles (GNPs) possess unique physicochemical properties that may facilitate entry into the central nervous system (CNS) where they may act therapeutically. There is little information on biodistribution or inflammatory effects of GNPs in specific brain regions. Brain Localization and neuroinflammatory response to citrate-capped spherical GNP (10 nm) was determined 24 hours after intravenous (IV) injection in male C57Bl mice. A known inflammogen, lipopolysaccharide (LPS, 2 mg kg-1, SC), was tested as a positive control supplement. Aggregation of GNPs was measured using various Phosphate-Buffered Saline (PBS) concentrations (10, 1, 0.1, 0.01 X) to determine the optimal buffer concentration to maintain …

K-Cl Cotransport: Role Of Kcc3 In Cellular Potassium (K) Homeostasis In Kcc3- Transfected Hek-293 Cells, Nagendra Babu Ravilla

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K-Cl cotransport (KCC) mediated by four protein isoforms, KCC1 to KCC4, plays a significant role in cell volume regulation, and in K and Cl homeostasis. In this study, we demonstrate the importance of KCC3 and its two threonine (T) phosphorylation sites T991 and T1048 in cellular K homeostasis in isogenic human embryonic kidney (HEK-293) cells transfected with KCC3 wild type (WT) or constitutively active double mutant KCC3 (AA) expressed under tetracycline or doxycycline control. In both WT and AA cells, under baseline conditions, the Na-K-2Cl cotransport (NKCC) (55 %- 60 %) was the major contributor of Rb influx followed by …

Hypoxia Inducible Factor 1 Alpha (Hif-1a): A Major Regulator Of Placental Development, Renee E. Albers

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Hypoxia-inducible factor-1 alpha (HIF-1a) is a critical component of the cellular oxygen-sensing machinery and is essential for placental formation and embryonic survival. In this study, we promoted prolonged expression of HIF-1a, by using a form that is insensitive to oxygen, denoted as CA-HIF-1a. In order to have continual placental specific expression of the CA-HIF-1a, lentiviral infection was performed on embryos at the blastocyst stage of development and transferred into pseudo-pregnant mothers. Placental analysis was performed via in situ hybridization on embryonic day 14.5 (E14.5) to determine the effects of CA-HIF-1a prolonged expression. Data indicate that prolonged activity of CA-HIF-1a restricted …

Chronic Swim Stress Attenuates Endocrine Response And Improves Cardiac Function In Mice Lacking The Angiotensin Ii Type 1a Receptor, Najat Khalifa Almahroug

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Stress is a stimulus that causes an often abrupt but always large change in autonomic activity and hormone secretion. Angiotensin II (AngII) is one of the main neurohormonal mediators that are stimulated by stress. AngII mediates most of its relevant biological effects via AngII type1 receptor (AT1R) activation. Stress increases tissue and circulating levels of Ang II which contributes to stimulation of adrenal AT1R, mediating stressinduced hormone release. Ang II through the activation of AT1R not only elevates blood pressure but also has a patho-physiological relevance in cardiac remodeling. The goal of this study was to assess the influence of …

Establishment Of A Quiscence Herpes Simplex Type 1 Infection In L929 Fibroblasts And Neuro-2a Cells By A Nucleoside Analogue Acyclovir, Noura Shaklawoon

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Herpes simplex type 1 (HSV-1) quiescent infection was established in L929 cells, murine fibroblasts, and in Neuro-2A (N2A) cells, mouse neuroblastoma, by treating them with the nucleoside analogue acyclovir (ACV) 24 hours before infection. Subsequent release of virus from the non-productive state was accomplished by treating the cells with the histone deacetylase inhibitor trichostatin A (TSA). Treatment of both L929 and N2A cell lines with ACV 24 hours before infection induced protection from HSV-1 cytopathic effect. A quiescent state was confirmed by absence of virus plaques when supernatant fluids from ACV-treated HSV-1 infected cultures were titered on Vero cell monolayers. …

Effects Of Red Diamondback Rattlesnake Venom On Keloid Dermal Fibroblasts In Vitro, Mackenzie Shelby Newman

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Keloid scarring is an inflammatory healing response to physical injury such as incision or piercing in the dermis. It is characterized by aberrant extracellular matrix production, the overaccumulation of mature collagen, and excessive fibroblast proliferation and migration beyond the borders of the original wound site. This results in swelling, depigmentation, itchiness, and pain akin to a benign tumor. Although there are myriad treatments for the condition, most are invasive and exhibit a high recurrence rate. Previous studies have shown that rattlesnake venom stimulates apoptosis in the skin via multiple specific mechanisms, largely composed of extracellular matrix and its receptors' interactions. …

The Effect Of Hsv-1 Infection On Differentiated And Polarized U937 Cells, Allolo Dreiwish Aldreiwish

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Herpes simplex virus type 1 (HSV-1) challenges the host immune system through several mechanisms (Frey, et al., 2009). In vitro, U937 cells (human macrophage-like precursor cell line) are not susceptible to HSV-1 infection when they are not differentiating (Lopez-Guerrero and Alonso, 1997). Differentiation of these cells resistance can abrogates their resistance to HSV-1 (Tenney and Morahan, 1991). In this study, we examined the effect of HSV-1 infection on differentiated and polarized U937 cells. U937 cells are differentiated to M0 cells.

Then, M0 cells are polarized to distinct phenotypes, M1 or M2. M1 are proinflammatory macrophages while M2 are anti-inflammatory cells. …

"Characterization Of Red Diamondback Rattlesnake Venom Proteins On Cell Death And Function", Venus Ebrahimian

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Snake venom is composed of many different toxins and protein components such as metalloproteases and phospholipase A2. Together, these proteins cause an up regulation in cell death pathways and disrupt the overall homeostasis of a living cell. The Red Diamondback Rattlesnake is abundantly available and well-known specie and has been used as a source for development of new drugs. It has been mainly used for the purposes of making anti-venom but it has also been therapeutically used for surgical purposes to act as an anti-aggregate. Integrins derived from RDB venom has been used for cancer cell treatment as well. Specifically …

Locating Chelerythrine, An Alkaloid, Within A Cytosolic Environment By Maldi-Tof Mass Spectrometry, Bradley Allen Williams

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The quaternary benzo-phenanthridine alkaloid (QBA) chelerythrine (CET) is a well-known inhibitor of PKC and cancer growth (Herbert et al., 1990). In human lens epithelial cells (HLECs), CET almost completely inhibits the Na+/K+ pump ATPase (NKA) without changes in phosphorylation (Lauf et al. 2013). Protein alignment studies revealed B-lymphocyte type 2 protein (Bcl-2) BH1- like motifs within the a1 subunit of the NKA to which CET, a drug mimicking Bcl-2 protein BH3-like motifs, might bind thus disrupting NKA function (Lauf et al, 2013). It is unknown by which mechanism CET crosses the plasma membrane to reach its intracellular targets, especially the …

N-Glycosylation, Localization And Trafficking Of Endogenous Nkcc1 In Cos7 Cells, Richa Singh

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The Na+K+2Cl^- co-transporter-1 (NKCC1) effects the electroneutral movement of Na+, K+ and 2Cl^- ions across plasma membranes. NKCC1 is considered a widely distributed, highly N-glycosylated ion transporter involved in the regulation of the intracellular chloride concentration ([Cl^-]i), cell volume and salt secretion. NKCC1 locates in the plasma membrane of all cells studied so far, particularly on the basolateral side of most polarized cells. However, the majority of immunoreactive NKCC1 distributes in intracellular compartments of unknown nature whereas a small proportion of the transporter appears to be located in the plasma membrane. Here, we identified and characterized …