Urothelial barrier function is maintained by apical membrane plaques and intercellular tight junctions (TJ). Little is known about the composition and regulation of TJ expression in human urothelium. In this study, we have characterised the expression of TJ components in situ and their regulation in an in vitro model of differentiating normal human urothelial (NHU) cells. In normal ureteric urothelium in situ, there was a differentiation-associated profile of claudins 3, 4, 5, 7, ZO1 and occludin proteins. Proliferating NHU cells in vitro expressed predominantly claudin 1 protein and transcripts for claudins 1-5 and 7. Following induction of differentiation by pharmacological activation of PPARγand blockade of EGFR, there was de novo expression of claudin 3 mRNA and protein and downregulation of claudin 2 transcription. There was also a massive increase in expression of claudin 4 and 5 proteins which was due to inhibition of proteasomal degradation of claudin 4 and consequential stabilisation of the claudin 5 heterodimerisation partner. NHU cell differentiation was accompanied by relocalisation of TJ proteins to intercellular junctions. The differentiation-associated development of TJ formation in vitro reflected the stage-related TJ expression seen in situ. This was distinct from changes in TJ composition of NHU cells mediated by increasing the calcium concentration of the medium. Our results imply a role for PPARγ and EGFR signalling pathways in regulating TJ formation in NHU cells and support the hypothesis that TJ development is an integral part of the urothelial differentiation programme.The maintenance of epithelial barrier function requires that the transepithelial passage of water and solutes be tightly regulated. Ion channels and membrane pumps located in the apical and basolateral membrane compartments control transcellular ion transport, whereas tight junctions (TJ), located at the superior aspect of the intercellular junctional complex, control paracellular diffusion (Schneeberger and Lynch, 2004). TJ are composed of cytoplasmic plaque proteins, such as the zonular occludens (ZO) proteins that link the TJ to the cytoskeleton, and integral transmembrane proteins, such as occludin, junctional adhesion molecule (JAM) and claudins that define the properties of the paracellular pore (Tsukita and Furuse, 2002; GonzalezMariscal et al., 2003). The TJ not only limits paracellular movement, but maintain polarity by restricting the movement of proteins and lipids between apical and basolateral membrane compartments. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptThe claudins, which represent a multigene family of some 24 members ranging from 20,000 to 27,000 Mr, are considered to represent the primary seal-forming fibrils of the TJ. Claudins are tetra-spanning proteins, comprising two extracellular loops and short amino and carboxy termini (Schneeberger and Lynch, 2004). The expression of different claudin proteins and the pairing of claudins to form homotypic or heterotypic f...

LKT and IE contributed equally assessed by the acquisition of a transitional cell morphology, a switch from a cytokeratin (CK)13 lo /CK14hi to a CK13 hi /CK14 lo phenotype, expression of claudin 3, 4 and 5 proteins, and induction of uroplakin gene transcription. RESULTSTwo of 12 SUI cell lines showed early senescent changes in culture and were not characterized further; one of seven IC, one of five IDO and a further three SUI cell lines had some evidence of senescence at passage 3. Of the seven IC-derived cell lines, four showed a near normal range of differentiationassociated responses, but the remainder showed little or no response. Most IDO cell lines (four of five) showed a normal differentiation response, but at least three of the 10 SUI cell lines showed some compromise of differentiation potential. CONCLUSIONThis study supports the existence of a subset of patients with IC in whom a failure of urothelial cytodifferentiation might contribute to the disease, and provides a novel platform for investigating the cell biology of urothelium from SUI and other benign dysfunctional conditions.

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Objective: Simulation is now firmly established in modern surgical training and is applicable not only to acquiring surgical skills but also to non-surgical skills and professionalism. A 5-day intensive Urology Simulation Boot Camp was run to teach emergency procedural skills, clinical reasoning, and communication skills using clinical scenario simulations, endoscopic and laparoscopic trainers. This paper reports the educational value of this first urology boot camp. Subjects and methods: Sixteen urology UK trainees completed pre-course questionnaires on their operative experience and confidence level in common urological procedures. The course included seven modules covering basic scrotal procedures, laparoscopic skills, ureteroscopy, transurethral resection of the prostate and bladder tumour, green light laser prostatectomy, familiarisation with common endoscopic equipment, bladder washout to remove clots, bladder botox injection, setting up urodynamics. Emergency urological conditions were managed using scenarios on SimMan®. The main focus of the course was hands-on training using animal models, bench-top models and virtual reality simulators. Post-course assessment and feedback on the course structure and utility of knowledge gained together with a global outcome score was collected. Results: Overall all the sections of feedback received score of over 4.5/5, with the hands-on training on simulators getting the best score 4.8/5. When trainees were asked “The training has equipped me with enhanced knowledge, understanding and skills,” the average score was 4.9/5.0. The vast majority of participants felt they would recommend the boot camp to future junior trainees. Conclusion: This first UK Urology Simulation Boot Camp has demonstrated feasibility and effectiveness in enhancing trainee’s experience. Given these positive feedbacks there is a good reason to expect that future courses will improve the overall skills of a new urology trainee

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