Atomic Force Microscopy of Mammalian Urothelial Surface

Kreplak, Laurent; Wang, Huaibin; Aebi, Ueli; Kong, Xiang‐Peng

doi:10.1016/j.jmb.2007.09.040

Cited by 41 publications

(37 citation statements)

References 60 publications

(17 reference statements)

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“…2B), and the border between adjacent cells contained zippered apical membranes (formed by interdigitation of the apical membrane of adjacent cells) (Fig. 2C), which lie just above the tight junctions (47). The angular hinge regions and adjacent plaque regions were visible at higher magnifications (Fig.…”

Section: Temporal Changes In Barrier Morphology In Dmmentioning

confidence: 93%

Impact of diabetes mellitus on bladder uroepithelial cells

Hanna-Mitchell

Ruiz

Daneshgari

et al. 2013

American Journal of Physiology-Regulatory, Integrative and Comp

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Diabetic bladder dysfunction (DBD), a prevalent complication of diabetes mellitus (DM), is characterized by a broad spectrum of symptoms including urinary urgency, frequency, and incontinence. As DBD is commonly diagnosed late, it is important to understand the chronic impact of DM on bladder tissues. While changes in bladder smooth muscle and innervation have been reported in diabetic patients, the impact of DM on the specialized epithelial lining of the urinary bladder, the urothelium (UT), is largely unknown. Quantitative polymerase chain reaction analysis and electron microscopy were used to evaluate UT gene expression and cell morphology 3, 9, and 20 wk following streptozotocin (STZ) induction of DM in female SpragueDawley rats compared with age-matched control tissue. Desquamation of superficial (umbrella) cells was noted at 9 wk DM, indicating a possible breach in barrier function. One causative factor may be metabolic burden due to chronic hyperglycemia, suggested by upregulation of the polyol pathway and glucose transport genes in DM UT. While superficial UT repopulation occurred by 20 wk DM, the phenotype was different, with significant upregulation of receptors associated with UT mechanosensation (transient receptor potential vanilloid subfamily member 1; TRPV1) and UT autocrine/paracrine signaling (acetylcholine receptors AChR-M2 and -M3, purinergic receptors P2X2 and P2X3). Compromised barrier function and alterations in UT mechanosensitivity and cell signaling could contribute to bladder instability, hyperactivity, and altered bladder sensation by modulating activity of afferent nerve endings, which appose the urothelium. Our results show that DM impacts urothelial homeostasis and may contribute to the underlying mechanisms of DBD. streptozotocin; urothelium; barrier; sensory; diabetic bladder dysfunction IN THE UNITED STATES alone it is estimated that approximately 19 million individuals, comprising both men and women, suffer from diabetes mellitus (DM) (13). DM is characterized by defects in the secretion of the hormone insulin by the pancreas and/or insulin signaling (53), which causes a dysregulation of cellular glucose uptake. Glucose is a large hydrophilic molecule that depends on specific glucose transporter (GLUT) proteins, present on the cell membrane, for cellular entry (12). From the GLUT family (13 cloned to date), GLUT4 is regulated by insulin and controls glucose uptake by skeletal muscle [which accounts for ϳ40% of total body mass (70)], cardiac muscle, and adipose tissue (75). Thus Type 1 and 2 diabetics are unable to efficiently transport glucose from the blood into these tissue groups (66) resulting in a state of hyperglycemia. Consequently, glucose inundates cells that express insulinindependent glucose transporters (such as GLUT1). Chronic elevation in cytosolic glucose leads to metabolic abnormalities such as osmotic and oxidative stress, thought to be factors that contribute to tissue injury and dysfunction associated with long-term DM (15, 61).The urinary bladder is one of th...

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Section: Temporal Changes In Barrier Morphology In Dmmentioning

confidence: 93%

Impact of diabetes mellitus on bladder uroepithelial cells

Hanna-Mitchell

Ruiz

Daneshgari

et al. 2013

American Journal of Physiology-Regulatory, Integrative and Comp

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“…The formation of tight junctions creates a tight seal and a high-resistance paracellular barrier between adjacent umbrella cells (7,10). When the epithelial barrier is compromised, leakage of urine components into the underlying bladder layers results, with symptoms of interstitial cystitis (IC) (11,12).…”

mentioning

confidence: 99%

MicroRNA-205 Targets Tight Junction-related Proteins during Urothelial Cellular Differentiation

Chung

Chi

Chen

et al. 2014

Molecular & Cellular Proteomics

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“…DNA, RNA, and protein bind to the membranes in a noncovalent but irreversible manner by a mechanism that is not fully understood. It is believed that a dominant hydrophobic interaction and as well as electrostatic interactions may occur between cellular surface proteins and this membrane (Fang et al 2009;Cretich et al 2010), which allow the freshly dissected tissue samples to be adsorb to the membrane (Kreplak et al 2007). Similarly, the adhesion of cells to the nitrocellulose membrane mainly takes advantage of the nitrocellulose's affinity to membrane proteins.…”

Section: Discussionmentioning

confidence: 99%

“…They have been widely used as a solid-phase immobilization support for DNA, RNA, and proteins in traditional molecular biology assays, such as Southern, Northern, and Western blots, as well as atomic force microscopy (Kreplak et al 2007). These membranes are also used to detect coliforms in water in conjunction with a filtration device (Barry et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Use of nitrocellulose membranes as a scaffold in cell culture

Wang

Deng

et al. 2012

Cytotechnology

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Nitrocellulose membranes, one of the most important and oldest cellulose derivatives, are commonly used for nucleic acid and protein detection in research and diagnostic applications. However, a limited number of studies have explored whether they can act as scaffolds for cell growth. In this study, we investigated this polymeric material for its ability to support the growth of human cells. Eight established cell lines were examined for adherence, growth, spread, and survival on nitrocellulose membranes by optical microscopy after hematoxylin and eosin and/or immunocytochemical staining and by scanning electron microscopy. Apoptosis and leakage of lactate dehydrogenase (LDH) were also assessed. All cells readily adhered to and spread on the surface of nitrocellulose membranes as well as coverslips, and the cells maintained the expression of digestive system-specific genes. No significant change was detected in apoptosis or leakage of LDH from cells grown on nitrocellulose membranes. These results suggested that nitrocellulose membranes have a suitable cytocompatibility towards human cells and that they might be used for tissueengineering scaffolds. Moreover, we demonstrate an additional and underused property of nitrocellulose of specific relevance to microscopic imaging, as it can be rendered virtually transparent, thus the cells growing on such membranes can be observed directly under an optical microscope after staining.

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Atomic Force Microscopy of Mammalian Urothelial Surface

Cited by 41 publications

References 60 publications

Impact of diabetes mellitus on bladder uroepithelial cells

Impact of diabetes mellitus on bladder uroepithelial cells

MicroRNA-205 Targets Tight Junction-related Proteins during Urothelial Cellular Differentiation

Use of nitrocellulose membranes as a scaffold in cell culture

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