The blood-testis barrier: the junctional permeability, the proteins and the lipids

The elucidation of how individual components of the Sertoli cell junctional complexes form and are dismantled to allow not only individual cells but whole syncytia of germinal cells to migrate from the basal to the lumenal compartment of the seminiferous epithelium without causing a permeability leak in the blood-testis barrier is amongst the most enigmatic yet, challenging and timely questions in testicular physiology. The intriguing key event in this process is how the barrier modulates its permeability during the periods of formation and dismantling of individual Sertoli cell junctions. The purpose of this review is therefore to first provide a reliable account on the normal formation, maintenance and dismantling process of the Sertoli cells junctions, then to assess the influence of the expression of their individual proteins, of the cytoskeleton associated with the junctions, and of the lipid content in the seminiferous tubules on the regulation of the their permeability barrier function. To help focus on the formation and dismantling of the Sertoli cell junctions, several considerations are based on data gleaned not only from rodents but from seasonal breeders as well because these animal models are characterized by exhaustive periods of junction assembly during development and the onset of the seasonal re-initiation of spermatogenesis as well as by an extensive junction dismantling period at the beginning of testicular regression, something unavailable in normal physiological conditions in continual breeders. Thus, the modulation of the permeability barrier function of the Sertoli cell junctions is analyzed in the physiological context of the blood-epidydimis barrier and in particular of the blood-testis barrier rather than in the context of a detailed account of the molecular composition and signalisation pathways of cell junctions. Moreover, the considerations discussed in this review are based on measurements performed on seminiferous tubule-enriched fractions gleaned at regular time intervals during development and the annual reproductive cycle.     

Role of Neuronal Cell Adhesion Molecules and N-Cadherin in Passive Avoidance Training of Rats

A comparative study of the role of specific adhesion proteins, NCAM (neuronal cell adhesion molecules) and N-cadherin, was carried out on rats subjected to passive avoidance training procedure. It was shown that antibodies against the Ca²⁺-dependent adhesion protein N-cadherin injected into the rat somatosensory cortical zone 6 h after passive avoidance training had been completed did not evoke a loss of the habit by experimental animals. At the same time, an absolute amnestic effect with respect to this reflex developed after injection of antibodies against NCAM. After injection of antibodies against the above-mentioned proteins into the dorsal part of the hippocampus, the avoidance habit also disappeared in the case of treatment with antibodies against NCAM and was kept under the influence of antibodies against N-cadherin. The data obtained testify that NCAM and N-cadherin play dissimilar roles in the formation of a memory trace in the course of training.     

Radial Glial Fibers Promote Neuronal Migration and Functional Recovery after Neonatal Brain Injury

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N-cadherin-based adhesion enhances Aβ release and decreases Aβ42/40 ratio

In neurons, Presenilin 1(PS1)/γ-secretase is located at the synapses, bound to N-cadherin. We have previously reported that N-cadherin-mediated cell–cell contact promotes cell-surface expression of PS1/γ-secretase. We postulated that N-cadherin-mediated trafficking of PS1 might impact synaptic PS1-amyloid precursor protein interactions and Aβ generation. In the present report, we evaluate the effect of N-cadherin-based contacts on Aβ production. We demonstrate that stable expression of N-cadherin in Chinese hamster ovary cells, expressing the Swedish mutant of human amyloid precursor protein leads to enhanced secretion of Aβ in the medium. Moreover, N-cadherin expression decreased Aβ_42/40 ratio. The effect of N-cadherin expression on Aβ production was accompanied by the enhanced accessibility of PS1/γ-secretase to amyloid precursor protein as well as a conformational change of PS1, as demonstrated by the fluorescence lifetime imaging technique. These results indicate that N-cadherin-mediated synaptic adhesion may modulate Aβ secretion as well as the Aβ_42/40 ratio via PS1/N-cadherin interactions.     

Increased expression of VE-cadherin correlates temporally with differentiation of a restrictive endothelial barrier during normal angiogenesis in vivo

The purpose of this study was to evaluate temporal expression of VE- and N-cadherins within the angiogenic chick chorioallantoic membrane (CAM). Whether their relative patterns of expression changed in conjunction with abrupt differentiation of the restrictive CAM endothelial barrier between days 4.5 and 5.0 of the 21 days gestation was evaluated. Immunoblots against VE-cadherin depicted an increase of VE-cadherin expression between days 4.5 and 5.0, but no change in expression was detected between days 5.0 and 6.0. N-cadherin expression, on the other hand, remained uniform from day 4.5 to day 6.0. Immunogold-labeled anti-VE-cadherin was found exclusively on the CAM endothelium, and principally along the lateral inter-endothelial junctions. Hence, VE-cadherin expression by the angiogenic endothelium was similar to that of adult endothelium. That VE-cadherin expression by the CAM endothelium was increased between days 4.5 and 5.0 serves to suggest a temporal correlation with the ontogeny of restrictive barrier function in angiogenic endothelium in vivo.     

Induction of apoptosis in cultured endothelial cells by a cadherin antagonist peptide: involvement of fibroblast growth factor receptor-mediated signalling

Cadherins are a family of transmembrane glycoproteins mediating calcium-dependent, homophilic cell-cell adhesion. In addition, these molecules are involved in signaling events, regulating such processes as cell motility, proliferation, and apoptosis. Members of the cadherin subfamily, called either classical or type I cadherins, contain a highly conserved sequence at their homophilic binding site consisting of the three amino acids--histidine-alanine-valine (HAV). Previous studies have shown that peptides containing the HAV motif inhibit cadherin-dependent events such as cell aggregation, compaction, and neurite outgrowth. We report here that a cyclic peptide, N-Ac-CHAVC-NH2 can perturb cadherin-mediated endothelial cell interactions, resulting in a progressive apoptotic cell death. This effect depends on cell density, as it is only observed when dense cultures are treated with the peptide. Adherens junction (AJ)-associated cadherin and catenins are differentially affected by the N-Ac-CHAVC-NH2 treatment, as judged by double immunofluorescence labeling followed by immunofluorescence-ratio imaging. However, cell-cell adhesions are largely retained during the first few hours after addition of the peptide. It was also observed that following treatment, actin filaments partially lose their plasma membrane anchorage at AJs and translocate towards the cell center. Interestingly, addition of basic fibroblast growth factor to confluent, peptide-treated, endothelial cell cultures, completely blocks apoptosis and the inhibitory peptide reduce the phosphorylation of the FGF receptor target protein FRS2, suggesting that the peptide exerts its effect by inhibiting cadherin-mediated activation of fibroblast growth factor receptor signaling. We propose that cadherin-mediated signaling is essential for maintaining viability of confluent endothelial cells, and that its perturbation by N-Ac-CHAVC-NH2 drives these cells to apoptosis.     

Soluble N-cadherin stimulates fibroblast growth factor receptor dependent neurite outgrowth and N-cadherin and the fibroblast growth factor receptor co-cluster in cells

A chimeric molecule consisting of the extracellular domain of the adhesion molecule, N-cadherin, fused to the Fc region of human IgG (NCAD-Fc) supports calcium-dependent cell adhesion and promotes neurite outgrowth following affinity-capture to a tissue culture substrate. When presented to cerebellar neurons as a soluble molecule, the NCAD-Fc stimulated neurite outgrowth in a manner equivalent to that seen for N-cadherin expressed as a cell surface glycoprotein. Neurons expressing a dominant-negative version of the fibroblast growth factor (FGF) receptor did not respond to soluble NCAD-Fc. In cells transfected with full-length N-cadherin and the FGF receptor, antibody-clustering of N-cadherin resulted in a co-clustering of the FGF receptor to discrete patches in the cell membrane. The data demonstrate that the ability of N-cadherin to stimulate neurite outgrowth can be dissociated from its ability to function as a substrate associated adhesion molecule. The N-cadherin and the FGF receptor co-clustering in cells provides a basis for the neurite outgrowth response stimulated by N-cadherin being dependent on FGF receptor function.