In this paper, we propose a generative model, Temporal Generative Adversarial Nets (TGAN), which can learn a semantic representation of unlabeled videos, and is capable of generating videos. Unlike existing Generative Adversarial Nets (GAN)-based methods that generate videos with a single generator consisting of 3D deconvolutional layers, our model exploits two different types of generators: a temporal generator and an image generator. The temporal generator takes a single latent variable as input and outputs a set of latent variables, each of which corresponds to an image frame in a video. The image generator transforms a set of such latent variables into a video. To deal with instability in training of GAN with such advanced networks, we adopt a recently proposed model, Wasserstein GAN, and propose a novel method to train it stably in an end-to-end manner. The experimental results demonstrate the effectiveness of our methods.

Primary cilia are displayed during the G0/G1 phase of many cell types. Cilia are reabsorbed as cells prepare to re-enter the cell cycle, but the causal and molecular link between these two cellular events remains unclear. We show that phospho(T94)Tctex-1 is recruited to ciliary transition zones prior to S-phase entry and plays a pivotal role in both ciliary disassembly and cell cycle progression. Tctex-1’s role in S-phase entry, however, is dispensable in non-ciliated cells. Exogenously added phosphomimic Tctex-1 T94E accelerates cilium disassembly and S-phase entry. These results support a model in which the cilia act as a brake to prevent cell cycle progression. Mechanistic studies show the involvement of actin dynamics in Tctex-1 regulated cilium resorption. Phospho(T94)Tctex-1 is also selectively enriched at the ciliary transition zones of cortical neural progenitors, and plays a key role in controlling G1 length, cell cycle entry, and fate determination of these cells during corticogenesis.

Gangliosides are known as modulators of transmembrane signaling by regulating various receptor functions. We have found that insulin resistance induced by tumor necrosis factor-␣ (TNF-␣) in 3T3-L1 adipocytes was accompanied by increased GM3 ganglioside expression caused by elevating GM3 synthase activity and its mRNA. We also demonstrated that TNF-␣ simultaneously produced insulin resistance by uncoupling insulin receptor activity toward insulin receptor substrate-1 (IRS-1) and suppressing insulin-sensitive glucose transport. Pharmacological depletion of GM3 in adipocytes by an inhibitor of glucosylceramide synthase prevented the TNF-␣-induced defect in insulin-dependent tyrosine phosphorylation of IRS-1 and also counteracted the TNF-␣-induced serine phosphorylation of IRS-1. Moreover, when the adipocytes were incubated with exogenous GM3, suppression of tyrosine phosphorylation of insulin receptor and IRS-1 and glucose uptake in response to insulin stimulation was observed, demonstrating that GM3 itself is able to mimic the effects of TNF on insulin signaling. We used the obese Zucker fa/fa rat and ob/ob mouse, which are known to overproduce TNF-␣ mRNA in adipose tissues, as typical models of insulin resistance. We found that the levels of GM3 synthase mRNA in adipose tissues of these animals were significantly higher than in their lean counterparts. Taken together, the increased synthesis of cellular GM3 by TNF may participate in the pathological conditions of insulin resistance in type 2 diabetes.

Depolarization of the sarcolemma of smooth muscle cells activates voltage-gated Ca2+ channels, influx of Ca2+ and activation of cross-bridge cycling by phosphorylation of myosin catalysed by Ca2+/calmodulin-dependent myosin light-chain kinase (MLCK). Agonist stimulation of smooth muscle contraction often involves other kinases in addition to MLCK. In the present study, we address the hypothesis that membrane depolarization-induced contraction of rat caudal arterial smooth muscle may involve activation of Rho-associated kinase (ROK). Addition of 60 mM K+ to de-endothelialized muscle strips in the presence of prazosin and propranolol induced a contraction that peaked rapidly and then declined to a steady level of force corresponding to approx. 30% of the peak contraction. This contractile response was abolished by the Ca2+-channel blocker nicardipine or the removal of extracellular Ca2+. An MLCK inhibitor (ML-9) inhibited both the phasic and tonic components of K+-induced contraction. On the other hand, the ROK inhibitors Y-27632 and HA-1077 abolished the tonic component of K+-induced contraction, and slightly reduced the phasic component. Phosphorylation levels of the 20-kDa light chain of myosin increased rapidly in response to 60 mM K+ and subsequently declined to a steady-state level significantly greater than the resting level. Y-27632 abolished the sustained and reduced the phasic elevation of the phosphorylation of the 20-kDa light chain of myosin, without affecting the K+-induced elevation of cytosolic free Ca2+ concentration. These results indicate that ROK activation plays an important role in the sustained phase of K+-induced contraction of rat caudal arterial smooth muscle, but has little involvement in the phasic component of K+-induced contraction. Furthermore, these results are consistent with inhibition of myosin light-chain phosphatase by ROK, which would account for the sustained elevation of myosin phosphorylation and tension in response to membrane depolarization.

Clostridium botulinum neurotoxins (BoNTs) act on nerve endings to block acetylcholine release. Their potency is due to their enzymatic activity and selective high affinity binding to neurons. Although there are many pieces of data available on the receptor for BoNT, little attempt has been made to characterize the receptors for BoNT/C and BoNT/D. For this purpose, we prepared the recombinant carboxyl-terminal domain of the heavy chain (H C ) and then examined its binding capability to rat brain synaptosomes treated with enzymes and heating. Synaptosomes treated with proteinase K or heating retained binding capability to both H C /C and H C /D, suggesting that a proteinaceous substance does not constitute the receptor component. We next performed a thin layer chromatography overlay assay of H C with a lipid extract of synaptosomes. Under physiological or higher ionic strengths, H C /C bound to gangliosides GD1b and GT1b. These data are in accord with results showing that neuraminidase and endoglycoceramidase treatment decreased H C /C binding to synaptosomes. On the other hand, H C /D interacted with phosphatidylethanolamine but not with any ganglioside. Using cerebellar granule cells obtained from GM3 synthase knock-out mice, we found that BoNT/C did not elicit a toxic effect but that BoNT/D still inhibited glutamate release to the same extent as in granule cells from wild type mice. These observations suggested that BoNT/C recognized GD1b and GT1b as functional receptors, whereas BoNT/D induced toxicity in a ganglioside-independent manner, possibly through binding to phosphatidylethanolamine. Our results provide novel insights into the receptor for clostridial neurotoxin.

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The Crk-associated substrate (Cas) is a unique docking protein that possesses a repetitive stretch of tyrosine-containing motifs and an Src homology 3 (SH3) domain. Embryonic fibroblasts lacking Cas demonstrated resistance to Src-induced transformation along with impaired actin bundling and cell motility, indicating critical roles of Cas in actin cytoskeleton organization, cell migration, and oncogenesis. To gain further insight into roles of each domain of Cas in these processes, a compensation assay was performed by expressing a series of Cas mutants in Cas-deficient fibroblasts. The results showed that motifs containing YDxP were indispensable for actin cytoskeleton organization and cell migration, suggesting that CrkII-mediated signaling regulates these biological processes. The C-terminal Src-binding domain played essential roles in cell migration and membrane localization of Cas, although it was dispensable in the organization of actin stress fibers. Furthermore, the Src-binding domain was also a prerequisite for Src transformation possibly, because of its crucial role in the phosphorylation of Cas during transformation. Overall, differential uses of the Cas domains in individual biological processes were demonstrated. Cas1 docking protein was initially identified and cloned as a major phosphotyrosine-containing protein in cells transformed by v-src and v-crk oncogenes (1, 2). It has a structure with a number of protein-protein interaction domains, including an N-terminal Src homology 3 (SH3) domain, a substrate domain (SD) that consists of a cluster of YxxP motifs (one YLVP, four YQxPs, nine YDxPs, and one YAVP), and a C-terminal Srcbinding domain containing motifs YDYV (amino acids 762-765) and RPLPSPP (2). The substrate domain offers docking sites for several molecules including adaptor proteins Crk, Nck, and an inositol 5Ј-phosphatase, SHIP2 (SH2-containing inositol 5Ј-phosphatase), through their SH2 domains in a phosphorylation dependent manner (3-5). Motifs RPLPSPP and YDYV in the SB domain serve as direct binding sites for SH3 and SH2