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Hata, T.

Publications and source records attributed to Hata, T..

13 recordsLinked to original sources

Cholinergic interneurons in the dorsal striatum play an important role in the acquisition of duration memory

Although the formation of duration memory is important to optimize the timing of behavior based on previous experiences, the neural mechanism of formation remains unclear. A previous study suggested that muscarinic acetylcholine 1 receptors (M1Rs) in the dorsal striatum of rats are involved in the consolidation of duration memory in interval timing. Therefore, cholinergic interneurons (ChIs) may also be involved in the formation of duration memory in interval timing because ChIs activate M1Rs in the dorsal striatum. In the Exp. 1A, rats underwent a peak interval (PI)-20 s training. During the training, two different trials were randomly presented. One was a food trial in which the initial lever press was reinforced 20 s after the start of the trial (i.e., fixed interval (FI), 20 s). The other was an empty trial in which the reinforcement did not occur for 80 s. After sufficient training, the rats responded at approximately 20 s in the empty trials. They were then subjected to a PI-40 s training (i.e., FI, 40 s) after ChI lesions were present in the dorsal striatum. In this training, the sham-lesioned group responded frequently at approximately 40 s, whereas the ChI-lesioned group responded at approximately 20 s. As the PI-40 s additional learning progressed in the Exp. 1B, the ChI-lesioned group frequently responded at approximately 40 s, similar to that of the sham-lesioned group. In the following PI-20 s re-shift training, the ChI-lesioned group responded similar to the sham-lesioned group. In Exp. 2 of another cohort, the results of a PI-20 s training after the occurrence of ChI lesions were similar to that before the presence of the lesion. Together, these results suggest that ChI lesions delayed only the acquisition of new duration memory, but had no effect on the adjustment of behavior associated with changing the reinforcement schedule of the PI-training and interval timing itself.

animal behavior and cognition↗

Light-exercise-induced dopaminergic and noradrenergic stimulation in the dorsal hippocampus: using a rodent physiological exercise model

Exercise activates the dorsal hippocampus, which triggers the synaptic and cellar plasticity and ultimately promotes memory formation. For decades, these benefits have been explored using demanding and stress-response-inducing exercise at moderate-to-vigorous intensities. In contrast, our translational research with animals and humans has focused on light exercise below the lactate threshold (LT), which almost anyone can safely perform with minimal stress, and found that even light exercise can stimulate hippocampal activity and enhance memory performance. Although the circuit mechanism of this boost remains unclear, arousal promotion even with light exercise implies the involvement of the ascending monoaminergic system, which is essential to modulate hippocampal activity and impact memory. To examine this hypothesis, we employed our physiological exercise model based on the LT of rats that can be applied to human and immunohistochemically assessed the neuronal activation of the dorsal hippocampal sub-regions and brainstem monoaminergic neurons. Also, we monitored the dynamics of monoamine release at the dorsal hippocampus using in vivo microdialysis. We found that even light exercise increased neuronal activity in the dorsal hippocampal sub-regions and induced noradrenaline and dopamine release. Furthermore, we found that tyrosine hydroxylase-positive neurons in the locus coeruleus (LC) and the ventral tegmental area (VTA) were activated even by light exercise and were both positively correlated with the dorsal hippocampal activation. In conclusion, our findings demonstrate that light exercise stimulates hippocampal neurons, possibly through the LC-noradrenergic and/or VTA-dopaminergic neurons. This sheds light on the circuit mechanisms responsible for hippocampal neural activation during exercise, consequently enhancing memory function. Graphical abstractOur previous research with animals and humans has demonstrated that even light exercise can boost neuronal activity in the dorsal hippocampus and improve memory. While the mechanism underlying this remains undetermined, recent studies suggest the involvement of the ascending monoaminergic system. Here, we examined this hypothesis and found a possible contribution of noradrenergic neurons in the locus coeruleus and dopaminergic neurons in the ventral tegmental area to dorsal hippocampal activation during light exercise, implying a circuit mechanism for light-exercise-enhanced memory. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=144 SRC="FIGDIR/small/545490v2_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@1e286e8org.highwire.dtl.DTLVardef@1070708org.highwire.dtl.DTLVardef@5ec2a6org.highwire.dtl.DTLVardef@13d2fdd_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

Inducible expression of the restriction enzyme uncovered genome-wide distribution and dynamic behavior of histones H4K16ac and H2A.Z at DNA double-strand breaks in Arabidopsis

DNA double-strand breaks (DSBs) are among the most serious types of DNA damage and cause mutations and chromosomal rearrangements. In eukaryotes, DSBs are immediately repaired in coordination with chromatin remodeling for the deposition of DSB-related histone modifications and variants. To elucidate the details of DSB-dependent chromatin remodeling throughout the genome, artificial DSBs need to be reproducibly induced at various genomic loci. Recently, in mammals, a comprehensive method for elucidating chromatin remodeling at multiple DSB loci via chemically induced expression of a restriction enzyme was developed. However, this DSB induction system is not suitable for investigating chromatin remodeling during and after DSB repair, and such approach has not been performed in plants. Here, we established a transgenic Arabidopsis plant harboring a restriction enzyme gene Sbf I driven by a heat-inducible promoter. Using this transgenic plant, we performed chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) of histones H4K16ac and H2A.Z and investigated dynamics of these histone marks around the endogenous 623 Sbf I recognition sites. We also precisely quantified DSB efficiency at all cleavage sites using the DNA resequencing data obtained by ChIP-seq procedures. From the results, Sbf I-induced DSBs were detected at 360 loci and induced the transient deposition of H4K16ac and H2A.Z around these regions. Interestingly, we also observed the co-localization of H4K16ac and H2A.Z at some DSB loci. Overall, DSB-dependent chromatin remodeling was found to be a highly conserved between plants and animals. These findings provide new insights into chromatin remodeling that occurs in response to DSBs in Arabidopsis.

molecular biology↗

Paradoxical tumor suppressive role of the musculoaponeurotic fibrosarcoma gene in colorectal cancer

Somatic cell reprogramming using the microRNAs miR200c, miR-302s, and miR-369s leads to increased expression of cyclin-dependent kinase inhibitors in human colorectal cancer (CRC) cells and suppressed tumor growth. Here, we investigated whether these microRNAs inhibit colorectal tumorigenesis in CPC;Apc mice, which are prone to colon and rectal polyps. Repeated administration of microRNAs inhibited polyp formation. Microarray analysis indicated that c-MAF, which reportedly shows oncogene-like behavior in multiple myeloma and T-cell lymphoma, decreased in tumor samples but increased in microRNA-treated normal mucosa. Immunohistochemistry identified downregulation of c-MAF as an early tumorigenesis event in CRC, with low c-MAF expression associated with poor prognosis. Of note, c-MAF expression and p53 protein levels were inversely correlated in CRC samples. c-MAF knockout led to enhanced tumor formation in azoxymethane/dextran sodium sulfate-treated mice, with activation of cancer-promoting genes. c-MAF may play a tumor-suppressive role in CRC development.

molecular biology↗

Intra-dorsal striatal acetylcholine M1 but not dopaminergic D1 or glutamatergic NMDA receptor antagonists inhibit the consolidation of duration memory in interval timing

The striatal beat frequency (SBF) model assumes that striatal medium spiny neurons encode duration via synaptic plasticity. Muscarinic 1 (M1) cholinergic receptors, as well as dopamine and glutamate receptors, are important for neural plasticity in the dorsal striatum. Therefore, we investigated the effect of inhibiting these receptors on the formation of duration memory. After sufficient training in a Peak interval (PI)-20 s procedure, rats were given a single or mixed infusion of a selective antagonist for the dopamine D1 receptor (SCH23390, 0.5 g per side), the NMDA-type glutamate receptor (D-AP5, 3 g), or the M1 receptor (pirenzepine, 10 g) bilaterally in the dorsal striatum, immediately before starting a PI 40 s session (shift session). On the next day, the rats were tested for new duration memory (40 s) in a session in which no lever presses were reinforced (probe session). In the shift session, performance was tie, irrespective of the drug injected. However, in the probe session, the mean peak time (an index of duration memory) of the M1 + NMDA co-blockade group, but not of the D1 + NMDA co-blockade group, was lower than that of the control group (Exp. 1 and 2). In Exp. 3, the effect of the co-blockade of M1 and NMDA receptors was replicated. Moreover, sole blockade of M1 receptors induced the same effect as M1 and NMDA blockade. These results suggest that in the dorsal striatum, the M1 receptor, but not the D1 or NMDA receptors, are involved in the consolidation of duration memory.

animal behavior and cognition↗

Spatiotemporal variations of surface water microplastics near Kyushu, Japan: A quali-quantitative analysis

Microplastics in the ocean are threatening marine ecosystems. Although plastic contaminants are ubiquitous from rivers to polar oceans, their distribution is thought to be heterogeneous, implying that both spatial and temporal variability exist. Here, we elucidate the significant spatial and temporal (seasonal) variations in the quanti-qualitative characteristics of microplastics off the west coast of Kyushu, Japan in the East China Sea. Six surveys across nine stations (n = 54) were conducted over a 14-month period, and a total of 6131 plastic items were identified. The average microplastic abundance (items {middle dot}m-3) and size (mm) {+/-} S.D. were 0.49 {+/-} 0.92 (n = 54), and 1.71 {+/-} 0.93 (n = 6131), respectively. Differences between the highest and lowest abundances were 50-fold among monthly means (1.97 {+/-} 1.49, n = 9; 0.04 {+/-} 0.03, n = 9), and 550-fold across all net tows (5.50; 0.01). With respect to colour, polymer type, and shape, white and transparent (68.5%), polyethylene (80%) fragments (76.0%) were the dominant composition. There were statistically significant differences for each of the analytical microplastic parameters among survey months (p < 0.02). Our results provide baseline data, and lead to a more comprehensive understanding of the spatiotemporal characteristics of microplastic pollution. HighlightsO_LISignificant spatiotemporal variability in microplastic litter was detected based on the quali-quantitative analyses. C_LIO_LIAverage ({+/-} S.D.) microplastic abundance was 0.49 {+/-} 0.92 (items {middle dot}m-3) and size was 1.71 {+/-} 0.93 (mm). C_LIO_LIDifferences between highest and lowest abundances were 50-fold among monthly means, and 550-fold across all net tows. C_LIO_LIWhite and transparent polyethylene fragments were the dominant composition. C_LI

ecology↗

The dorsal hippocampus is required for the formation of long-term duration memories in rats

Interval timing--the perception of durations mainly in seconds or minutes--is a ubiquitous behavior in organisms. Animal studies have suggested that the hippocampus plays an essential role in duration memory; however, the memory processes involved are unclear. To clarify the role of the dorsal hippocampus in the acquisition of long-term duration memories, we adapted the "time-shift paradigm" to a peak-interval procedure. After a sufficient number of training with an initial target duration (20 s), the rats underwent "shift sessions" with a new target duration (40 s) under a muscimol (0.5 g per side) infusion into the bilateral dorsal hippocampus. The memory of the new target duration was then tested in drug-free "probe sessions," including trials in which no lever presses were reinforced. In the probe sessions, the mean response rate distribution of the muscimol group was located leftward to the control group, but these two response rate distributions were superimposed on the standardized time axis, suggesting a scalar property. In the session-by-session analysis, the mean peak time (an index of timing accuracy) of the muscimol group was lower than that of the control group in the probe sessions, but not in the shift sessions. These findings suggest that the dorsal hippocampus is required for the formation of long-term duration memories within the range of interval timing.

neuroscience↗

Cryptic promoter activation occurs by at least two different mechanisms in the Arabidopsis genome

In gene-trap screening of plant genomes, promoterless reporter constructs are often expressed without trapping of annotated gene promoters. The molecular basis of this phenomenon, which has been interpreted as the trapping of cryptic promoters, is poorly understood. In this study, using Arabidopsis gene-trap lines in which a firefly luciferase (LUC) open reading frame (ORF) was expressed from intergenic regions, we found that cryptic promoter activation occurs by at least two different mechanisms: one is the capturing of pre-existing promoter-like chromatin marked by H3K4me3 and H2A.Z, and the other is the entirely new formation of promoter chromatin near the 5 end of the inserted LUC ORF. To discriminate between these, we denoted the former mechanism as "cryptic promoter capturing", and the latter one as "promoter de novo origination". The latter finding raises a question as to how inserted LUC ORF sequence is involved in this phenomenon. To examine this, we performed a model experiment with chimeric LUC genes in transgenic plants. Using Arabidopsis psaH1 promoter-LUC constructs, we found that the functional core promoter region, where transcription start sites (TSS) occur, cannot simply be determined by the upstream nor core promoter sequences; rather, its positioning proximal to the inserted LUC ORF sequence was more critical. This result suggests that the insertion of the LUC ORF sequence alters the local distribution of the TSS in the plant genome. The possible impact of the two types of cryptic promoter activation mechanisms on plant genome evolution and endosymbiotic gene transfer is discussed.

evolutionary biology↗

Plant genome response to incoming coding sequences: stochastic transcriptional activation independent of chromatin configuration

Horizontal gene transfer can occur between phylogenetically distant organisms, such as prokaryotes and eukaryotes. In these cases, how do the translocated genes acquire transcriptional competency in the alien eukaryotic genome? According to the conventional view, specific loci of the eukaryotic genome are thought to provide transcriptional competency to the incoming coding sequences. To examine this possibility, we randomly introduced the promoterless luciferase (LUC)-coding sequences into the genome of Arabidopsis thaliana cultured cells and performed a genome-wide "transgene location vs. expression" scan. We mapped 4,504 promoterless LUC inserts on the A. thaliana chromosomes, and found that about 30% of them were transcribed. Only a small portion of them were explained by the conventional transcriptional fusions with the annotated genes, and the remainder occurred in a quite different manner; (1) they occurred all over the chromosomal regions, (2) independently of the insertion sites relative to the annotated gene loci, inherent transcribed regions, or heterochromatic regions, and (3) with one magnitude lower transcriptional level than the conventional transcriptional fusions. This type of transcriptional activation occurred at about 30% of the inserts, raising a question as to what this 30% means. We tested two hypotheses: the activation occurred at 30% of the entire chromosomal regions, or stochastically at 30% of each insertion event. Our experimental analysis indicates that the latter model could explain this transcriptional activation, a new type of plant genome response to the incoming coding sequences. We discuss the possible mechanisms and evolutionary roles of this phenomenon in the plant genome.

evolutionary biology↗

Kozak sequence acts as a negative regulator for de novo transcription initiation of newborn coding sequences in the plant genome

The manner in which newborn coding sequences and their transcriptional competency emerge during the process of gene evolution remains unclear. Here, we experimentally simulated eukaryotic gene origination processes by mimicking horizontal gene transfer events in the plant genome. We mapped the precise position of the transcription start sites (TSSs) of hundreds of newly introduced promoterless firefly luciferase (LUC) coding sequences in the genome of Arabidopsis thaliana cultured cells. The systematic characterization of the LUC-TSSs revealed that 80% of them occurred under the influence of endogenous promoters, while the remainder underwent de novo activation in the intergenic regions, starting from pyrimidine-purine dinucleotides. These de novo TSSs obeyed unexpected rules; they predominantly occurred ~100 bp upstream of the LUC inserts and did not overlap with Kozak-containing putative open reading frames (ORFs). These features were the output of the immediate responses to the sequence insertions, rather than a bias in the screening of the LUC gene function. Regarding the wild-type genic TSSs, they appeared to have evolved to lack any ORFs in their vicinities. Therefore, the repulsion by the de novo TSSs of Kozak-containing ORFs described above might be the first selection gate for the occurrence and evolution of TSSs in the plant genome. Based on these results, we characterized the de novo type of TSS identified in the plant genome and discuss its significance in genome evolution.

evolutionary biology↗

De novo activated transcription of newborn coding sequences is inheritable in the plant genome

The manner in which inserted foreign coding sequences become transcriptionally activated and fixed in the plant genome is poorly understood. To examine such processes of gene evolution, we performed an artificial evolutionary experiment in Arabidopsis thaliana. As a model of gene-birth events, we introduced a promoterless coding sequence of the firefly luciferase (LUC) gene and established 386 T2-generation transgenic lines. Among them, we determined the individual LUC insertion loci in 76 lines and found that one-third of them were transcribed de novo even in the intergenic or inherently unexpressed regions. In the transcribed lines, transcription-related chromatin marks were detected across the newly activated transcribed regions. These results agreed with our previous findings in A. thaliana cultured cells under a similar experimental scheme. A comparison of the results of the T2-plant and cultured cell experiments revealed that the de novo-activated transcription concomitant with local chromatin remodelling was inheritable. During one-generation inheritance, it seems likely that the transcription activities of the LUC inserts trapped by the endogenous genes/transcripts became stronger, while those of de novo transcription in the intergenic/untranscribed regions became weaker. These findings may offer a clue for the elucidation of the mechanism by which inserted foreign coding sequences become transcriptionally activated and fixed in the plant genome.

evolutionary biology↗

Large DNA virus promoted the endosymbiotic evolution to make a photosynthetic eukaryote

Chloroplasts in photosynthetic eukaryotes originated from a cyanobacterial endosymbiosis far more than 1 billion years ago1-3. Due to this ancientness, it remains unclear how this evolutionary process proceeded. To unveil this mystery, we analysed the whole genome sequence of a photosynthetic rhizarian amoeba4, Paulinella micropora5,6, which has a chloroplast-like organelle that originated from another cyanobacterial endosymbiosis7-10 about 0.1 billion years ago11. Here we show that the predacious amoeba that engulfed cyanobacteria evolved into a photosynthetic organism very quickly in the evolutionary time scale, probably aided by the drastic genome reorganization activated by large DNA virus. In the endosymbiotic evolution of eukaryotic cells, gene transfer from the endosymbiont genome to the host nucleus is essential for the evolving host cell to control the endosymbiont-derived organelle12. In P. micropora, we found that the gene transfer from the free-living and endosymbiotic bacteria to the amoeba nucleus was rapidly activated but both simultaneously ceased within the initiation period of the endosymbiotic evolution, suggesting that the genome reorganization drastically proceeded and completed. During this period, large DNA virus appeared to have infected the amoeba, followed by the rapid amplification and diversification of virus-related genes. These findings led us to re-examine the conventional endosymbiotic evolutionary scenario that exclusively deals with the host and the symbiont, and to extend it by incorporating a third critical player, large DNA virus, which activates the drastic gene transfer and genome reorganization between them. This Paulinella version of the evolutionary hypothesis deserves further testing of its generality in evolutionary systems and could shed light on the unknown roles of large DNA viruses13 in the evolution of terrestrial life.

evolutionary biology↗

Domain organization and conformational change of dynactin p150

Dynactin is a principal regulator of the minus-end directed microtubule motor dynein. The sidearm of dynactin is essential for binding to microtubules and regulation of dynein activity. Although our understanding of the structure of the dynactin backbone (Arp1 rod) has greatly improved recently, structural details of the sidearm part remain elusive. Here, electron microscopy of individual molecules of the dynactin complex revealed that the sidearm was highly flexible and exhibited diverse morphologies. Utilizing mutants for nanogold labeling and deletion analysis, we determined the domain organization of the largest subunit p150 and identified a filamentous structure protruding from the head domain of the sidearm as the coiled-coil 1 (CC1), the dynein-binding domain, in p150. Furthermore, the protrusion formed by CC1 exhibited either a folded or an extended form, suggesting that CC1 works as an extending \"arm\". These findings provide clues to understand how dynactin binds to microtubules and regulates dynein.

biophysics↗