An understanding of how synaptic vesicles are recruited to and maintained at presynaptic compartments is required to discern the molecular mechanisms underlying presynaptic assembly and plasticity. We have previously demonstrated that cadherin-beta-catenin complexes cluster synaptic vesicles at presynaptic sites. Here we show that scribble interacts with the cadherin-beta-catenin complex to coordinate vesicle localization. Scribble and beta-catenin are colocalized at synapses and can be coimmunoprecipitated from neuronal lysates, indicating an interaction between scribble and beta-catenin at the synapse. Using an RNA interference approach, we demonstrate that scribble is important for the clustering of synaptic vesicles at synapses. Indeed, in scribble knockdown cells, there is a diffuse distribution of synaptic vesicles along the axon, and a deficit in vesicle recycling. Despite this, synapse number and the distribution of the presynaptic active zone protein, bassoon, remain unchanged. These effects largely phenocopy those observed after ablation of beta-catenin. In addition, we show that loss of beta-catenin disrupts scribble localization in primary neurons but that the localization of beta-catenin is not dependent on scribble. Our data supports a model by which scribble functions downstream of beta-catenin to cluster synaptic vesicles at developing synapses.
The cuticle is a protective layer that coats the primary aerial surfaces of land plants and mediates plant interactions with the environment. It is synthesized by epidermal cells and is composed of a cutin polyester matrix that is embedded and covered with cuticular waxes. Recently, we have discovered a novel regulatory mechanism of cuticular wax biosynthesis that involves the ECERIFERUM7 (CER7) ribonuclease, a core subunit of the exosome. We hypothesized that at the onset of wax production, the CER7 ribonuclease degrades an mRNA specifying a repressor of CER3, a wax biosynthetic gene whose protein product is required for wax formation via the decarbonylation pathway. In the absence of this repressor, CER3 is expressed, leading to wax production. To identify the putative repressor of CER3 and to unravel the mechanism of CER7-mediated regulation of wax production, we performed a screen for suppressors of the cer7 mutant. Our screen resulted in the isolation of components of the RNA-silencing machinery, RNA-DEPENDENT RNA POLYMERASE1 and SUPPRESSOR OF GENE SILENCING3, implicating RNA silencing in the control of cuticular wax deposition during inflorescence stem development in Arabidopsis (Arabidopsis thaliana).
Cadherins and neuroligins (NLs) represent two families of cell adhesion proteins that are essential for the establishment of synaptic connections in vitro; however, it remains unclear whether these proteins act in concert to regulate synapse density. Using a combination of overexpression and knockdown analyses in primary hippocampal neurons, we demonstrate that NL1 and N-cadherin promote the formation of glutamatergic synapses through a common functional pathway. Analysis of the spatial relationship between N-cadherin and NL1 indicates that in 14-day in vitro cultures, almost half of glutamatergic synapses are associated with both proteins, whereas only a subset of these synapses are associated with N-cadherin or NL1 alone. This suggests that NL1 and N-cadherin are spatially distributed in a manner that enables cooperation at synapses. In young cultures, N-cadherin clustering and its association with synaptic markers precede the clustering of NL1. Overexpression of N-cadherin at this time point enhances NL1 clustering and increases synapse density. Although N-cadherin is not sufficient to enhance NL1 clustering and synapse density in more mature cultures, knockdown of N-cadherin at later time points significantly attenuates the density of NL1 clusters and synapses. N-cadherin overexpression can partially rescue synapse loss in NL1 knockdown cells, possibly due to the ability of N-cadherin to recruit NL2 to glutamatergic synapses in these cells. We demonstrate that cadherins and NLs can act in concert to regulate synapse formation.Synapse formation begins with the recognition of appropriate targets and formation of incipient contacts and is followed by the recruitment of pre-and postsynaptic proteins to exquisitely localized microdomains at points of cell-cell contact (1, 2). Transsynaptic cell adhesion complexes have come to the forefront as key players in the formation and maturation of synaptic connections (3, 4). Among these adhesion complexes, the homophilic cadherin complex and the heterophilic neurexin-neuroligin complex have been extensively studied and are known to exert key roles in synapse development (5-7).The distributions of cadherins and neuroligins (NLs) 2 at synaptic compartments have previously been analyzed; however, their spatial distribution with respect to one another is still unclear. This information is essential for understanding the functional interplay between these two adhesion systems. Cadherins and their associated catenins have been observed in both pre-and postsynaptic compartments in many neuronal populations in the CNS (8, 9). Previous reports demonstrate that in the brain, two of these, N-and E-cadherin, are localized to synaptic complexes in mutually exclusive distributions (8). As synapses mature, N-cadherin is excluded from inhibitory synapses, becoming primarily localized at glutamatergic synapses (10).Evidence that cadherins play an important role in establishing synaptic junctions includes observations that cadherins rapidly accumulate at points of cell-cell contact prior ...
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