Asynchronous Embryo Transfer Followed by Comparative Transcriptomic Analysis of Conceptus Membranes and Endometrium Identifies Processes Important to the Establishment of Equine Pregnancy.

Preimplantation horse conceptuses require nutrients and signals from histotroph, the composition of which is regulated by luteal progesterone and conceptus-secreted factors. To distinguish progesterone and conceptus effects we shortened the period of endometrial progesterone-priming by asynchronous embryo transfer. Day 8 embryos were transferred to synchronous (day 8) or asynchronous (day 3) recipients, and RNA sequencing was performed on endometrium and conceptuses recovered 6 and 11 days later (embryo days 14 and 19). Asynchrony resulted in many more differentially expressed genes (DEGs) in conceptus membranes (3473) than endometrium (715). Gene ontology analysis identified upregulation in biological processes related to organogenesis and preventing apoptosis in synchronous conceptuses on day 14, and in cell adhesion and migration on day 19. Asynchrony also resulted in large numbers of DEGs related to 'extracellular exosome'. In endometrium, genes involved in immunity, the inflammatory response, and apoptosis regulation were upregulated during synchronous pregnancy and, again, many genes related to extracellular exosome were differentially expressed. Interestingly, only 14 genes were differentially expressed in endometrium recovered 6 days after synchronous versus 11 days after asynchronous transfer (day 14 recipient in both). Among these, KNG1 and IGFBP3 were consistently upregulated in synchronous endometrium. Furthermore bradykinin, an active peptide cleaved from KNG1, stimulated prostaglandin release by cultured trophectoderm cells. The horse conceptus thus responds to a negatively asynchronous uterus by extensively adjusting its transcriptome, whereas the endometrial transcriptome is modified only subtly by a more advanced conceptus.

Bovine pericardium membrane as new tool for mesenchymal stem cells commitment.

Acellular matrices are widespread biomaterials used in surgical practice as tissue reinforcement and anatomical support to favor tissue regeneration. It is clear that a fundamental role in the regeneration of tissue is played by cell-material interaction. In this work, the interaction between a bovine pericardium membrane and human adult stem cells was investigated by microscopy analysis and gene expression analysis. Parallel cell cultures were prepared on the pericardium membrane or tissue culture plate. They were incubated in basal growth medium or in adipogenic differentiation medium to perform experiments on the seventh and the 14th day of culture. Results demonstrated that the membrane allows cell viability, adhesion, and proliferation of human stem cells. During adipogenic commitment on the membrane, the accumulation of cytoplasmatic lipid droplets and the expression of adipogenic gene PPARG, CEBPA, GLUT4, FABP4, and ADIPOQ were detected. Concurrently, a downregulation of mesenchymal stem cell gene CD29, CD90, and CD105 was detected. In basal medium, the adipogenic gene expression was upregulated, whereas the mesenchymal markers were indifferently expressed. These findings suggest that the bovine pericardium membrane is a biocompatible matrix and that their rough surface allows cell adhesion, spreading, and proliferation. The surface morphology activates mechanochemical signals that stimulate the adipogenic commitment of stem cells in basal medium and potentiate their commitment in adipogenic differentiation medium.

Form-function relationship in artificial lateral lines.

We examined the form-function relationship of laboratory-constructed artificial lateral line canals. These biomimetic flow sensors consisted of a transparent silicone bar located inside a fluid filled canal equipped with canal pores. The silicone bar guided the light from a LED towards a position- sensitive photodiode. Fluid motion inside the canal deflected the silicone bar which was detected by the photodiode. We found that the resonance frequency of the silicone bar determined the resonance frequency of the artificial lateral line (frequency at which the sensor was most sensitive). The thickness and length of the silicone bar influenced both, the resonance frequency and the sensitivity (across all tested frequencies) of the artificial lateral line sensor. Sensitivity was also influenced by the length and diameter of the artificial lateral line canals. The distance between canal pores determined the spatial resolution of the sensor. The functionality of the sensor in detecting oscillatory fluid motions remained when the canal pores were covered with flexible membranes. Tension, diameter and thickness of the membranes altered the temporal filter properties of the artificial lateral line neuromast. The density and viscosity of the fluid inside the artificial lateral line canals also influenced the sensitivity and temporal filter properties of the artificial lateral line. The acquired knowledge will allow us to optimize artificial lateral line systems for specific technical applications.

Rehabilitation of distal radioulnar joint instability.

Distal radioulnar joint (DRUJ) instabilities are common and often combined with other injuries of the interosseous membrane and/or the proximal radioulnar joint. Once they are diagnosed and the treatment is chosen, physiotherapists have limited choices due to the lack of validated protocols. The benefits of proprioception and neuromuscular rehabilitation have been brought to light for the shoulder, knee and ankle joints, among others. However, no program has been described for the DRUJ. The purpose of this article is to study the muscular elements responsible for active DRUJ stability, and to propose a proprioceptive rehabilitation program suited to this condition.

Meeting report - Cellular dynamics: membrane-cytoskeleton interface.

The first ever 'Cellular Dynamics' meeting on the membrane-cytoskeleton interface took place in Southbridge, MA on May 21-24, 2017 and was co-organized by Michael Way, Elizabeth Chen, Margaret Gardel and Jennifer Lippincott-Schwarz. Investigators from around the world studying a broad range of related topics shared their insights into the function and regulation of the cytoskeleton and membrane compartments. This provided great opportunities to learn about key questions in various cellular processes, from the basic organization and operation of the cell to higher-order interactions in adhesion, migration, metastasis, division and immune cell interactions in different model organisms. This unique and diverse mix of research interests created a stimulating and educational meeting that will hopefully continue to be a successful meeting for years to come.

Membrane mediated development of the vertebrate blood-gas-barrier.

During embryonic lung development, establishment of the gas-exchanging units is guided by epithelial tubes lined by columnar cells. Ultimately, a thin blood-gas barrier (BGB) is established and forms the interface for efficient gas exchange. This thin BGB is achieved through processes, which entail lowering of tight junctions, stretching, and thinning in mammals. In birds the processes are termed peremerecytosis, if they involve cell squeezing and constriction, or secarecytosis, if they entail cutting cells to size. In peremerecytosis, cells constrict at a point below the protruding apical part, resulting in fusion of the opposing membranes and discharge of the aposome, or the cell may be squeezed by the more endowed cognate neighbors. Secarecytosis may entail formation of double membranes below the aposome, subsequent unzipping and discharge of the aposome, or vesicles form below the aposome, fuse in a bilateral manner, and release the aposome. These processes occur within limited developmental windows, and are mediated through cell membranes that appear to be of intracellular in origin. In addition, basement membranes (BM) play pivotal roles in differentiation of the epithelial and endothelial layers of the BGB. Laminins found in the BM are particularly important in the signaling pathways that result in formation of squamous pneumocytes and pulmonary capillaries, the two major components of the BGB. Some information exists on the contribution by BM to BGB formation, but little is known regarding the molecules that drive peremerecytosis, or even the origins and composition of the double and vesicular membranes involved in secarecytosis.

Membrane-mediated regulation of vascular identity.

Vascular diseases span diverse pathology, but frequently arise from aberrant signaling attributed to specific membrane-associated molecules, particularly the Eph-ephrin family. Originally recognized as markers of embryonic vessel identity, Eph receptors and their membrane-associated ligands, ephrins, are now known to have a range of vital functions in vascular physiology. Interactions of Ephs with ephrins at cell-to-cell interfaces promote a variety of cellular responses such as repulsion, adhesion, attraction, and migration, and frequently occur during organ development, including vessel formation. Elaborate coordination of Eph- and ephrin-related signaling among different cell populations is required for proper formation of the embryonic vessel network. There is growing evidence supporting the idea that Eph and ephrin proteins also have postnatal interactions with a number of other membrane-associated signal transduction pathways, coordinating translation of environmental signals into cells. This article provides an overview of membrane-bound signaling mechanisms that define vascular identity in both the embryo and the adult, focusing on Eph- and ephrin-related signaling. We also discuss the role and clinical significance of this signaling system in normal organ development, neoplasms, and vascular pathologies.

Structure, regulation, and functional diversity of microvilli on the apical domain of epithelial cells.

Microvilli are actin-based structures found on the apical aspect of many epithelial cells. In this review, we discuss different types of microvilli, as well as comparisons with actin-based sensory stereocilia and filopodia. Much is known about the actin-bundling proteins of these structures; we summarize recent studies that focus on the components of the microvillar membrane. We pay special attention to mechanisms of membrane microfilament attachment by the ezrin/radixin/moesin family and regulation of this protein family. We also discuss the NHERF family of scaffolding proteins that are found in microvilli and their role in microvilli regulation. Microvilli on cultured cells are not static structures, and their dynamics and those of their components are discussed. Finally, we mention diseases related to microvilli and outline questions that our current knowledge will allow the field to address in the near future.

Quantification of Lysosomal Membrane Permeabilization by Cytosolic Cathepsin and ß-N-Acetyl-Glucosaminidase Activity Measurements.

Programmed cell death involving lysosomal membrane permeabilization (LMP) is an alternative cell death pathway induced under various cellular conditions and by numerous cytotoxic stimuli. The method presented here to quantify LMP takes advantage of the detergent digitonin, which creates pores in cellular membranes by replacing cholesterol. The difference in cholesterol content between the plasma membrane (high) and lysosomal membrane (low) allows titration of digitonin to a concentration that permeabilizes the plasma membrane but leaves lysosomal membranes intact. The extent of LMP is determined by measuring the cytosolic activity of lysosomal hydrolases (e.g., cysteine cathepsins) and/or ß-N-acetyl-glucosaminidase in the digitonin-extracted cytoplasm and comparing it to the total cellular enzyme activity. Digitonin extraction of the cytosol can be combined with precipitation of protein and/or western blot analysis for detection of lysosomal proteins (e.g., cathepsins).

Methods for Probing Lysosomal Membrane Permeabilization.

Cell death triggered by lysosomal membrane permeabilization (LMP) is gaining increased interest as target for cancer therapy, but the death pathway also plays an important role in normal physiology (e.g., during involution of the mammary gland). LMP-induced cell death is triggered by release of hydrolases including cysteine cathepsin proteases from the lysosomal lumen into the cytosol. Limited release of proteases to the cytoplasm induces apoptosis or apoptosis-like cell death, whereas massive LMP results in rapid cellular necrosis. Here we introduce three complementary methods for quantifying and visualizing LMP: (i) monitoring LMP by immunocytochemistry, (ii) visualizing LMP by fluorescent dextran release, and (iii) quantification of LMP by activity measurements of lysosomal enzymes in digitonin-extracted cytosol.

Extracellular Membrane Vesicles as Vehicles for Brain Cell-to-Cell Interactions in Physiological as well as Pathological Conditions.

Extracellular vesicles are involved in a great variety of physiological events occurring in the nervous system, such as cross talk among neurons and glial cells in synapse development and function, integrated neuronal plasticity, neuronal-glial metabolic exchanges, and synthesis and dynamic renewal of myelin. Many of these EV-mediated processes depend on the exchange of proteins, mRNAs, and noncoding RNAs, including miRNAs, which occurs among glial and neuronal cells. In addition, production and exchange of EVs can be modified under pathological conditions, such as brain cancer and neurodegeneration. Like other cancer cells, brain tumours can use EVs to secrete factors, which allow escaping from immune surveillance, and to transfer molecules into the surrounding cells, thus transforming their phenotype. Moreover, EVs can function as a way to discard material dangerous to cancer cells, such as differentiation-inducing proteins, and even drugs. Intriguingly, EVs seem to be also involved in spreading through the brain of aggregated proteins, such as prions and aggregated tau protein. Finally, EVs can carry useful biomarkers for the early diagnosis of diseases. Herein we summarize possible roles of EVs in brain physiological functions and discuss their involvement in the horizontal spreading, from cell to cell, of both cancer and neurodegenerative pathologies.

Anatomy and biomechanics of the forearm interosseous membrane.

PURPOSE: To examine the anatomy and function of the forearm interosseous membrane by exploring the anatomical insertions of the central band (CB) on the radius and the ulna and by quantifying the length of the intact ligament and replacement grafts located at the original CB attachment sites and alternative locations. METHODS: Eight fresh cadaver forearms were supinated and pronated and the wrist was extended and flexed while the motion between the distal radius and ulna were recorded. The length of the CB was computed for the intact CB as well for several alternative graft orientations and positions. RESULTS: The maximum length of the CB did not significantly change among different wrist motions. However, with the wrist in a static neutral position, the CB length was significantly shorter in forearm supination than in neutral. During active forearm rotation when CB replacement grafts were positioned distal or proximal to the original CB site, yet still parallel to it, each had a similar trend to be longer in neutral than in supination. If a graft was more transversely oriented, the computed CB length would be 1.6 mm shorter in supination than in neutral. CONCLUSIONS: These results support tensioning a CB graft with the forearm in supination if the goal is to maximize graft tension and to maintain the native 22° angle for a CB graft between the radius and ulna. The results also suggest that the CB graft can probably be located slightly distal or slightly proximal to its original attachment sites. CLINICAL RELEVANCE: Reconstruction of the interosseous membrane has been hampered by a lack of understanding of its length changes with forearm or wrist motion. These results provide a starting point in helping clinicians understand how to more precisely reconstruct this ligament in an anatomical manner.

Confined mobility in biomembranes modeled by early stage Brownian motion.

An equation of motion, derived from the fractal analysis of the Brownian particle trajectory, makes it possible to calculate the time dependence of the mean square displacement for early times, before the Einstein formula becomes valid. The diffusion coefficient increases with the distance travelled which can be restricted by the geometrical conditions. The corresponding diffusion coefficient cannot increase further to achieve a value characteristic for unrestricted environment. Explicit formula is derived for confined diffusivity related to the unrestricted one as dependent on the maximum particle mean square displacement possible normalized by the square of its mean free path. The model describes the lipid and protein diffusion in tubular membranes with different radii, originally fitted by the modified Saffman-Delbrück equation, and the lateral mobility of synthetic model peptides for which the diffusion coefficient is inversely proportional to the radius of the diffusing object and to the thickness of the membrane.

Diacylglycerol activates the light-dependent channel TRP in the photosensitive microvilli of Drosophila melanogaster photoreceptors.

Drosophila light-dependent channels, TRP and TRPL, reside in the light-sensitive microvilli of the photoreceptor's rhabdomere. Phospholipase C mediates TRP/TRPL opening, but the gating process remains unknown. Controversial evidence has suggested diacylglycerol (DAG), polyunsaturated fatty acids (PUFAs, a DAG metabolite), phosphatidylinositol bisphosphate (PIP2), and H(+) as possible channel activators. We tested each of them directly in inside-out TRP-expressing patches excised from the rhabdomere, making use of mutants and pharmacology. When patches were excised in darkness TRP remained closed, while when excised under illumination it stayed constitutively active. TRP was opened by DAG and silenced by ATP, suggesting DAG-kinase (DGK) involvement. The ATP effect was abolished by inhibiting DGK and in the rdgA mutant, lacking functional DGK, implicating DGK. DAG activated TRP even in the presence of a DAG-lipase inhibitor, inconsistent with a requirement of PUFAs in opening TRP. PIP2 had no effect and acidification, pH 6.4, activated TRP irreversibly, unlike the endogenous activator. Complementary liquid-chromatography/mass-spectrometry determinations of DAG and PUFAs in membranes enriched in rhabdomere obtained from light- and dark-adapted eyes showed light-dependent increment in six DAG species and no changes in PUFAs. The results strongly support DAG as the endogenous TRP agonist, as some of its vertebrate TRPC homologs of the same channel family.

Silencing of TBC1D15 promotes RhoA activation and membrane blebbing.

Membrane blebs are round-shaped dynamic membrane protrusions that occur under many physiological conditions. Membrane bleb production is primarily controlled by actin cytoskeletal rearrangements mediated by RhoA. Tre2-Bub2-Cdc16 (TBC) domain-containing proteins are negative regulators of the Rab family of small GTPases and contain a highly conserved TBC domain. In this report, we show that the expression of TBC1D15 is associated with the activity of RhoA and the production of membrane blebs. Depletion of TBC1D15 induced activation of RhoA and membrane blebbing, which was abolished by the addition of an inhibitor for RhoA signaling. In addition, we show that TBC1D15 is required for the accumulation of RhoA at the equatorial cortex for the ingression of the cytokinetic furrow during cytokinesis. Our results demonstrate a novel role for TBC1D15 in the regulation of RhoA during membrane blebbing and cytokinesis.

Avaliação radiográfica e histológica comparativa entre uma nova membrana eletrofiada de PCL/poli(rotaxona) e uma membrana de colágeno suíno na regeneração óssea guiada de defeito crítico em calvária de ratos Wistar / Radiographic and histological evaluation between a new electrospinning PCL/polyrotaxane membrane and a porcine collagen membranes in a guided bone regeneration model in Wistar rat's critical sized calvaria defects

Dentre as diversas possibilidades de reconstrução de tecidos ósseos atróficos, a regeneração óssea guiada é uma das mais promissoras. Neste contexto, muitas membranas reabsorvíveis tem sido desenvolvidas e precisam ser testadas como parte de sua caracterização. O objetivo do presente estudo foi avaliar radiográfica e histologicamente em um estudo in vivo, se uma nova membrana polimérica eletrofiada de PCL/poli(rotaxona) demonstra comportamento semelhante a uma membrana de colágeno, comercialmente consagrada, quanto à promoção de regeneração óssea guiada. Foi realizado defeito crítico de 8mm de diâmetro na calvária de 60 ratos Wistar machos. Em dois grupos iguais (n=20) os defeitos foram recobertos aleatoriamente por uma membrana de colágeno suíno ou por uma membrana polimérica mista de policaprolactona (PCL) e poli(rotaxona). Em um terceiro grupo (n=20) os defeitos não foram recobertos e permaneceram apenas com o coágulo. Os animais sofreram eutanásia em 7, 14, 21 e 42 dias pós operatórios. Espécimes da região foram radiografadas e preparadas para análise histológica. Radiograficamente, os defeitos recobertos pela membrana de colágeno suíno apresentaram diminuição mais significativa da área radiográfica dos defeitos de acordo com a progressão dos períodos pós-operatórios do que nos outros grupos. A histomorfologia do reparo mostrou agrupamentos mais expressivos de células gigantes no grupo PCL/poli(rotaxona) sugerindo resposta à corpo estranho. Na histomorfometria, a neoformação óssea foi significativamente mais intensa e com osso neoformado mais maduro no grupo Colágeno. Concluímos que para um modelo de regeneração óssea guiada, a membrana de PCL/poli(rotaxona) não superou a membrana de colágeno.

The need to rebuild lost bone tissue, shows up as one of the great challenges of modern dentistry. Among several possibilities, guided bone regeneration is one of the most established techniques. In this context, many resorbable membranes have been developed and need to be tested as part of their characterization. The aim of this study was to evaluate by an in vivo model, if a new electrospinning PCL/polyrotaxane polymer membrane promotes similar guided bone regeneration when compared to a collagen membrane. An 8mm diameter critical defect was made in 60 male Wistar rats calvaria. In two equal groups (n = 20) the defects were randomly covered with a porcine collagen or a PCL/polyrotaxane membranes. In a third group (n = 20) the defects remained uncovered and just the blood clot occupied the defect. The animals were euthanized at 7, 14, 21 and 42 days post-operative. Specimens were x-rayed and prepared for histological analysis. Radiographically, the defects covered by porcine collagen membrane, showed more significant reduction in defect area, according to postoperative period evolution. Histomorphology showed intense giant cells presence in the PCL/polyrotaxane group, suggesting a foreign body response. The histomorphometric analysis showed new and mature bone formation more intense in collagen group. Under the limits of this study, the collagen membrane performance in guided tissue regeneration was far superior to the PCL/polyrotaxane membrane.

Extramembrane control of ion channel peptide assemblies, using alamethicin as an example.

Ion channels allow the influx and efflux of specific ions through a plasma membrane. Many ion channels can sense, for example, the membrane potential (the voltage gaps between the inside and the outside of the membrane), specific ligands such as neurotransmitters, and mechanical tension within the membrane. They modulate cell function in response to these stimuli. Researchers have focused on developing peptide- and non-peptide-based model systems to elucidate ion-channel protein functions and to create artificial sensing systems. In this Account, we employed a typical peptide that forms ion channels,alamethicin, as a model to evaluate our methodologies for controlling the assembly states of channel-forming molecules in membranes. As alamethicin self-assembles in membranes, it prompts channel formation, but number of peptide molecules in these channels is not constant. Using planar-lipid bilayer methods, we monitored the association states of alamethicin in real time. Many ligand-gated, natural-ion channel proteins have large extramembrane domains. As these proteins interact with specific ligands, those conformational alterations in the extramembrane domains are transmitted to the transmembrane, pore-forming domains to open and close the channels. We hypothesized that if we conjugated suitable extramembrane segments to alamethicin, ligand binding to the extramembrane segments could alter the structure of the extramembrane domains and influence the association states or association numbers of alamethicin in the membranes. We could then assess those changes by using single-channel current recording. We found that we could modulate channel assembly and eventual ion flux with attached leucine-zipper extramembrane peptide segments. Using conformationally switchable leucine-zipper extramembrane segments that respond to Fe(3+), we fabricated an artificial Fe(3+)-sensitive ion channel; a decrease in the helical content of the extramembrane segment led to an increase in the channel current. When we added a calmodulin C-terminus segment, we formed a channel that was sensitive to Ca(2+). This result demonstrated that we could prepare artificial channels that were sensitive to specific ligands by adding appropriate extramembrane segments from natural protein motifs that respond to external stimuli. In conclusion, our research points to the possibility of creating tailored sensor or signal transduction systems through the conjugation of a conformationally switchable extramembrane peptide/protein segment to a suitable transmembrane peptide segment.

Cells, tissues, organs and systems.

This article, which forms part of the life sciences series, examines cellular organisation in the formation of body tissues, membranes, organs and systems. Four main types of body tissue are described: epithelial, connective, muscle and nervous tissue. Each class of tissue is described in terms of its structure and function. Where appropriate, subgroups within the classifications are identified. Different membranes in the body are considered and the organisation of tissues and membranes into more complex structures, such as organs and body systems, is outlined.

Morphological features and clinical significance of epidural membrane in the cervical spine.

STUDY DESIGN: A prospective clinical study. OBJECTIVE: To elucidate the histomorphological features and clinical significance of the epidural membrane (EM) in the cervical spine based on operative and histological findings. SUMMARY OF BACKGROUND DATA: The anatomical features of the EM have been mostly discussed on the basis of cadaver studies in the whole spine. However, the histomorphological features and clinical significance of the EM in the cervical spine based on operative findings have never been reported. METHODS: Eighty-seven patients with cervical spondylotic myelopathy who had undergone an expansive open-door laminoplasty under microscopy were evaluated with a more than 2-year follow-up period. The most damaged spinal segment was determined in each patient from the preoperative neurological and image findings along with the remaining symptoms at follow-up. The morphological features of the EM were observed and recorded in each patient during decompression. For histology, specimens of common and remarkable types of the EM obtained from 16 patients were examined. RESULTS: The age at surgery averaged 64.5 years; there were 58 men and 29 women. With regard to the most damaged spinal segment, there were 14 cases at the C3-C4 level, 37 at the C4-C5 level, 32 at the C5-C6 level, and 4 at the C6-C7 level. The EM was an adipo-fibro-vascular tissue with various histomorphologies, blending with the periradicular sheath. Some EMs showed notable findings: obstructing dural tube expansion (13 cases, 14.9%), compressing a nerve root or disturbing its mobility (4 cases, 4.6%), and the combined type (1 case, 1.1%). All of them were located at approximately the most damaged spinal segment. In addition, some EMs had interesting histological features, such as harboring many small arteries, calcified debris, and metaplastic bone fragments. CONCLUSION: The EM can develop into remarkable structures with spondylosis and aging in patients with cervical spondylotic myelopathy, affecting surgical outcomes as well as successful decompression procedures. A sound understanding of the histomorphological features of the EM is required to obtain satisfactory surgical outcomes in the limited field afforded by minimally invasive surgery.

Differential regulation of adherens junction dynamics during apical-basal polarization.

Adherens junctions (AJs) in epithelial cells are constantly turning over to modulate adhesion properties under various physiological and developmental contexts, but how such AJ dynamics are regulated during the apical-basal polarization of primary epithelia remains unclear. Here, we used new and genetically validated GFP markers of Drosophila E-cadherin (DE-cadherin, hereafter referred to as DE-Cad) and ß-catenin (Armadillo, Arm) to quantitatively assay the in vivo dynamics of biosynthetic turnover and membrane redistribution by fluorescence recovery after photobleaching (FRAP) assays. Our data showed that membrane DE-Cad and Arm in AJs of polarizing epithelial cells had much faster biosynthetic turnover than in polarized cells. Fast biosynthetic turnover of membrane DE-Cad is independent of actin- and dynamin-based trafficking, but is microtubule-dependent. Furthermore, Arm in AJs of polarizing cells showed a faster and diffusion-based membrane redistribution that was both quantitatively and qualitatively different from the slower and exchange-based DE-Cad membrane distribution, indicating that the association of Arm with DE-Cad is more dynamic in polarizing cells, and only becomes stable in polarized epithelial cells. Consistently, biochemical assays showed that the binding of Arm to DE-Cad is weaker in polarizing cells than in polarized cells. Our data revealed that the molecular interaction between DE-Cad and Arm is modulated during apical-basal polarization, suggesting a new mechanism that might be crucial for establishing apical-basal polarity through regulating the AJ dynamics.