When autoplay is enabled, a suggested video will automatically play next. Zxuan playing 1 on 1 against Babyshark with Kagura?!! (Bahasa Red = Babyshark. Mengejutkan, Zxuan Main Solo Ranked.. (Permainannya Sulit Untuk Ditiru Oleh User Fanny yg Manapun). ==================================== In the Video:- Zxuan's truth Mobile Legends zxuan zxuan mobile legends zxuan ml zxuan is a guy or girl where is zxuan Old zxuan vs New zxuan Zxuan vs Xsanz. xSanz apology to Zxuan and Syno! Accused of VPN even after returning to his Original Server?. THAT FRIDAY NIGHT STREAM BUT ITS NOW SATURDAY Mobile Legends. This feature is not available right now. Please try again later. These are just my #Theories. Don't Take Them Too Seriously. If You have any questions, Comment down and let me know. #Zxuan. Rating is available when the video has been rented. You're viewing YouTube in English (US). You can change this preference below. #MobileLegendsBangBang =================================== Music:- Diviners feat. Contacreast - Tropic Love [NCS Release]. Evangelio de Hoy Viernes 17 de Mayo de 2019 l Padre Carlos Yepes. OLD VIDEO: Z4PNU VS ZXUAN WHO WILL WIN???. tz·zχυαи 小敢 Fanny Gameplay and Build #14 Mobile Legends. BEASTMODE! - Top Global Fanny by Z4pnu - Gameplay, Build & Tips. Top 7 Heroes who Master with Flicker Tricks Part 2 - Mobile Legends: Bang bang. Z4pnu & YuriGaming Play Together in Deathbattle (DEATHBATTLE - NEW MODE). Truth behind Zxuan's Fall - Mobile Legends: Bang Bang. The identity of many of the proteins within the active zone remains unknown, especially those responsible for keeping vesicles clustered nearby. Xuan, Manning et al. therefore introduced mutations at random into the genome of the roundworm C. elegans, and searched for mutant worms with altered patterns of vesicles. Worms with mutations in a previously unknown gene, which they named clarinet, showed abnormal distribution of vesicles within the presynaptic compartment. They also released fewer vesicles compared to non-mutant worms. Problems with synapses contribute to many brain disorders, including autism and intellectual disability. Xuan, Manning et al. hope that an increased understanding of how synapses form, and how they work, will provide insights into these and other conditions. To determine whether CLA-1L regulates synaptic vesicle clustering cell-autonomously in NSM, we manipulated its expression in specific neurons using CRISPR-based strategies. Briefly, if CLA-1L acts cell-autonomously in NSM, cell-specific knockouts of CLA-1 should result in a cell-specific synaptic vesicle mutant phenotype, even in the context of all other cells expressing wild type CLA-1L. Conversely, in the context of all other cells lacking CLA-1L, cell-specific expression of wild type CLA-1L should result in cell-specific rescue of the synaptic vesicle phenotype. Like I said, it's better to remember me and smile, at least I was once a part of your life. ( A ) Schematic diagram of the nematode head and the NSM neuron (in red inside blue-boxed region). ( B ) Cytosolic mCherry (cyto::mCh) and the synaptic vesicle marker CAT-1::GFP expressed cell specifically in NSM. Scale bar = 5 μm. ( C–F ) Synaptic vesicle markers in NSM: CAT-1::GFP ( C–D ) or RAB-3::mCherry ( E–F ) in ventral neurite in wild type (WT; C and E ) and cla-1(ok560) ( D and F ). Note how cla-1 mutants exhibit diffuse ( D, F ) rather than the wild type punctate ( C, E ) fluorescence patterns. Scale bar = 5 μm. ( G ) Schematics of the genomic region of cla-1 and the structure of three main isoforms of the CLA-1 protein. The locations of loxP sites and the genetic lesions of the cla-1 alleles examined in this study are indicated. In addition to the common C-terminus, CLA-1L contains a large N-terminal repetitive region (see Figure 1—figure supplement 1G ). ( H ) Synaptic enrichment (ΔF/F) of CAT-1::GFP in NSM is greatly reduced in all cla-1 mutants compared to wild type (WT). n = number of animals. ( I ) The PDZ sequence of CLA-1 was aligned to RIM, Piccolo and Fife from C. elegans (CeUNC-10), Drosophila (DmRIM, DmFife), mouse (MmRIM1/2, MmPCLO) and human (HsRIM1/2, HsPCLO) by neighbor joining with 100 bootstrap replicates. PDZ domains of Dishevelled family proteins were used as an outgroup (grey). Based on a phylogenetic analysis using the PDZ domain sequences, we found that the cla-1 PDZ domain is most similar to that of RIM, but constitutes a distinct clade ( Figure 1I ). This result, along with the lack of sequence homology between the rest of the CLA-1 protein (other than the C2 domains) and any known active zone proteins, suggests that cla-1 encodes a novel active zone protein. Its role in synaptic vesicle clustering suggested that it may be functionally homologous to Piccolo, Bassoon and Fife, and hence was given the name Clarinet (CLA-1) to reflect its large size. To achieve cell-specific knockouts of CLA-1L, we created transgenic strains with loxP sites inserted within the introns flanking exon 3 and exon 13 of cla-1L ( Figure 1H and Figure 2—figure supplement 1B ). Insertion of loxP sites did not affect synaptic vesicle clustering in NSM, as predicted ( Figure 2E ). However, cell-specific expression of Cre in NSM, which leads to NSM-specific deletion of CLA-1L, resulted in the cla-1L mutant phenotype in NSM. Namely, we observed a diffuse distribution of synaptic vesicle proteins in NSM ( Figure 2F and G ). These findings indicate that CLA-1L is required in NSM for synaptic vesicle clustering and are consistent with it acting cell-autonomously in NSM. Active zone proteins cluster synaptic vesicles at presynaptic terminals and coordinate their release. In forward genetic screens, we isolated a novel Caenorhabditis elegans active zone gene, clarinet ( cla-1 ). cla-1 mutants exhibit defects in synaptic vesicle clustering, active zone structure and synapse number. As a result, they have reduced spontaneous vesicle release and increased synaptic depression. cla-1 mutants show defects in vesicle distribution near the presynaptic dense projection, with fewer undocked vesicles contacting the dense projection and more docked vesicles at the plasma membrane. cla-1 encodes three isoforms containing common C-terminal PDZ and C2 domains with homology to vertebrate active zone proteins Piccolo and RIM. The C-termini of all isoforms localize to the active zone. Specific loss of the ~9000 amino acid long isoform results in vesicle clustering defects and increased synaptic depression. Our data indicate that specific isoforms of clarinet serve distinct functions, regulating synapse development, vesicle clustering and release. Upgrade your browser for an improved Twitter mobile experience. Download Chrome for Android. The coordinated and precise release of synaptic vesicles from presynaptic compartments underlies neuronal communication and brain function. This is achieved through the concerted action of conserved proteins that make up the cytomatrix at the active zone, a protein dense region within the presynaptic bouton that is surrounded by synaptic vesicles. Active zone proteins regulate neurotransmission by recruiting synaptic vesicles to the plasma membrane, positioning calcium channels adjacent to the site of exocytosis, and priming synaptic vesicles for calcium-dependent release. In vertebrates, the main active zone proteins that coordinate synaptic vesicle release are Liprin-α, RIM, RIM-BP, Elks and Munc-13 ( Südhof, 2012; Ackermann et al., 2015 ). cla-1 mutants display disrupted synaptic vesicle clustering in NSM neurons. Further experiments revealed that the clarinet gene encodes three different proteins with varying sizes, all found at the active zone. Using microscopy and electrode recordings, as well as a genetic technique called CRISPR, Xuan, Manning et al. showed that the three forms of clarinet have different roles. The shorter ones contribute to the development of synapses. They help ensure that the active zone forms correctly and that neurons have an appropriate number of synaptic connections. The longest form of clarinet is responsible for clustering vesicles, which allows cells to continue releasing vesicles during bursts of repeated neuronal firing. To probe the subcellular localization of CLA-1L, we inserted GFP at the N-terminus of the endogenous cla-1 locus via CRISPR ( Figure 2—figure supplement 1A; Dickinson et al., 2015 ). Using this strain, we determined that GFP::CLA-1L (homozygous endogenous) localizes to synapses at the developmental period in which the embryonic nervous system begins to form (three-fold stage: Figure 1—figure supplement 2D,E ). CLA-1L localized in a pattern reminiscent of synaptic vesicle marker RAB-3. When we expressed mCherry::rab-3 cDNA under the NSM-specific promoter in the CRISPR strain, CLA-1L colocalized with RAB-3 in NSM ( Figure 2A–C ), indicating that CLA1L localizes to synapses, at or near synaptic vesicle clusters. Structure, homology and expression pattern of CLA-1 isoforms. cla-1 is predicted to encode six isoforms of different lengths ( Figure 1G ). Based on the length of the proteins, we classified them into three categories: CLA-1L (long) including CLA-1a and b; CLA-1M (medium) including CLA-1c and d; CLA-1S (short) including CLA-1e and f ( Figure 1G ). Distinct alleles affect different isoforms. cla-1(ok560) results in a deletion of the promoter and part of the coding region of cla-1L, and will be referred to henceforth as cla-1(L). cla-1(wy1048), an allele we generated using CRISPR, eliminates most of cla-1S and M, including the PDZ and C2 domains. Because these domains are shared by all isoforms, this deletion is likely a null and the allele will henceforth be referred to as cla-1(S/M/L). Importantly, in cla-1(L) deletion mutants, the shorter isoforms are still expressed, as evidenced by RT-PCR to the C-terminal PDZ domain ( Figure 1—figure supplement 1H ). Synaptic vesicle clustering was examined in five alleles affecting different isoforms ( Figure 1G ), and all alleles examined showed defects in synaptic vesicle clustering in NSM ( Figure 1H ). Since the long-isoform-specific allele cla-1(L) exhibited as dramatic a defect as the null allele, we hypothesize that CLA-1L may thus be specifically required for proper clustering of vesicles at the synapse. To determine the expression pattern of CLA-1 isoforms, we created GFP reporters under the cla-1 promoters (2 kb fragments upstream of the L, M and S isoforms). We found that each isoform is expressed broadly within the nervous system, as evidenced by a high degree of colocalization with an mCherry reporter under the pan-neuronal rab-3 promoter ( Figure 1—figure supplement 2A–C ). CLA-1S was expressed broadly throughout the nervous system, while CLA-1M and L were expressed in a subset of neurons. Open annotations. The current annotation count on this page is being calculated. To examine whether cell-specific expression of CLA-1L is sufficient to mediate synaptic vesicle clustering in cla-1L null mutant animals, we created a conditional cla-1L -expressing strain. We inserted a GFP followed by a transcriptional terminator before the start codon of cla-1L ( Figure 2—figure supplement 1C ). This construct drives GFP expression off the endogenous CLA-1L promoter, preventing the expression of the endogenous CLA-1L gene. In these animals, synap Remember me and smile, for it's better to forget than to remember me and feel sad. It has been a wonderful journey on Mobile Legends, entertaining many and receiving relentless support. Remember me, and I'll remember each and everyone of you forever. Goodbye! pic.twitter.com/7aBE62ax7w. Clarinet (CLA-1), a novel active zone protein required for synaptic vesicle clustering and release. We performed forward genetic screens in C. elegans for proteins required for synaptic vesicle clustering, and identified clarinet ( cla-1 ). CLA-1 is required for normal synapse number and cla-1 null mutants exhibit reduced spontaneous synaptic vesicle release. Cla-1 mutants have a smaller dense projection and display defects in the clustering of the active zone protein SYD-2/Liprin-α. They exhibit a dramatic reduction in the number of synaptic vesicles contacting the dense projection, and increased synaptic depression. The cla-1 gene encodes three main isoforms (CLA-1L, CLA-1M and CLA-1S) containing PDZ and C2 domains with sequence homology to vertebrate Piccolo and RIM. While all three isoforms share a C-terminal region that localizes to the active zone, their genetic requirement in synapse function and development differ: the N-terminus of CLA-1L is specifically required for synaptic vesicle clustering and proper synaptic function during repeated stimulations, whereas the shorter isoforms or C-terminus are required for active zone assembly and proper synapse number. Together our findings indicate that cla-1 encodes novel active zone proteins that are required for proper synapse development, active zone structure and synaptic vesicle clustering, and thus play a role in synaptic function during prolonged activation.