Sayaka Hori

Nara Women's University · Faculty Division of Natural Sciences

The transcription factor atonal contributes to patterning and cell fate determination in specialized epithelial cells in various animals, but its function in hypodermis is unknown. Here, we analyzed the atonal homolog lin-32 in C. elegans to clarify whether atonal acts in hypodermal development. The lin-32 null mutants exhibited bulges and cavities...

The central neural network optimizes avoidance behavior depending on the nociceptive stimulation intensity and is essential for survival. How the property of hub neurons that enables the selection of behaviors is genetically defined is not well understood. We show that the transcription factor unc-130, a human FOXD3/4 ortholog, is required to optim...

Optimization of the types and timing of avoidance behaviors depending on the intensity of a noxious stimulus is essential for survival; however, processing in the central nervous system and its developmental basis are largely unknown. Here, we report that Caenorhabditis elegans preferentially selects one of three different types of avoidance behavi...

A list of all screened transcription factors. A list of all screened transcription factors in the RNAi screening. Percent change was calculated by normalizing to the rates of avoidance behaviors of the animals during 2-sec stimulation that were treated with negative control RNAi (empty L4440 vector). Note that we did not distinguish the type of avo...

Summary of cell differentiation defects of avoidance circuits in lin-32 and fax-1 mutants. The rates of normal cells and analyzed neuron numbers are shown. The deficiencies were defined as the following: a change in the number of marker-positive cells and/or reduced intensity. lin-32 mutants showed differentiation defects in the AIB neurons, while...

Results of the control experiments. (A) Avoidance behaviors of ASH::ChR2(H134R); lite-1 animals without food with the 2-sec stimulation. n = 7,7,6,10,9,9. (B) The lite-1 mutants as a negative control with the 2-sec stimulation. n = 4,7,7,4,7,7. (C) Wild type animals adjust probability of omega turns depending on the glycerol drop concentration. n =...

AIB marker-positive cells in the lin-32 mutants show functional defects. lin-32 animals expressing AIB marker Pinx-1::Venus ("lin-32 AIB marker +") show defects that are comparable to those of the AIB marker-negative siblings ("lin-32 AIB marker -"). *p < 0.05, ANOVA followed by the Tukey's post hoc tests. n = 6,3,8,4. n = plate (cohort) of approxi...

S1_Dataset file contains the all relevant datasets which were analyzed for graphs or summary statistics in the research described in this paper. (XLSX)

Normal differentiation of ASH sensory neurons, AIA, AVA and RIM interneurons, and DA/VA motor neurons in the lin-32 and fax-1 mutants. (A-C) The normal expression pattern of Psra-6::GFP, which is expressed in the ASH sensory neurons and faintly expressed in the ASI neurons of all animal types (arrowheads). (D-F) Normal dye-filling patterns, which s...

Cells in the AIB sister lineage normally differentiate in the lin-32 mutants. (A) Schematic of the ABpl/raapap cell lineage. AIB shares a common precursor cell with AWA, ASG and ASI sensory neurons. The sister cells of ASI are removed via programmed cell death. (B) Schematic of the wild type AIB, AWA, ASG, and ASI morphologies (green). (C-H) ASI, A...

Calcium concentration changes in the muscles of Pmyo-3::G-CaMP2 transgenic animals. Calcium concentration changes in the muscles of Pmyo-3::G-CaMP2 transgenic animals during the three types of avoidance behaviors in the drop tests. (MOV)

Eighteen candidate transcription factors are involved in avoidance behavior. A list of candidate transcription factors in the screenings. We performed three screenings: a combination of optogenetics and enhanced neuronal RNAi methods ("RNAi & ChR2"); optogenetics or the high osmolarity ring assays using each mutant of the candidate genes ("Mutant &...

Avoidance behaviors of the ASH::ChR2(H134R); lite-1 animals. Avoidance behaviors of the ASH::ChR2(H134R); lite-1 animals are shown. They exhibited omega turns under 100% light irradiation and short reversals under 25% irradiation. (MOV)

AIB excitation correlates omega turn via gap junction. (A) lite-1 mutants expressing ChR2(H134R) in AIB neurons (AIB::ChR2(H134R); lite-1) with ATR exhibit omega turns even if without food condition. n = 50 each. n means the number of individuals (animals). (B) Behavioral frequencies during 2- or 5-sec 100% light stimulations in ASH::ChR2-expressin...

Avoidance behaviors of the ASH::ChR2(H134R); lin-32 lite-1 animals. Avoidance behaviors of the ASH::ChR2(H134R); lin-32 lite-1 animals are shown. They exhibited reversals under 100% light irradiation and no avoidance under 25% light irradiation. (MOV)

Calcium concentration changes in the muscles of lin-32; Pmyo-3::G-CaMP2 transgenic animals. Calcium concentration changes in the muscles of lin-32; Pmyo-3::G-CaMP2 transgenic animals during the three types of avoidance behaviors in the drop tests. (MOV)

Balancer chromosomes are critical tools for genetic research. In C. elegans, reciprocal translocations that lead to aneuploidy have been widely used to maintain lethal and sterile mutations in stable stocks. Here, we generated a set of aneuploidy-free and structurally defined crossover suppressors that contain two overlapping inversions using the C...

We report tm6429 and tm6475 as novel deletion alleles of the gene Y73E7A.1 that is a homologue of mammalian Coiled-coil domain containing 124 (Ccdc124)1. The Ccdc124 is a conserved gene from invertebrates to human. In human cell lines, Ccdc124 is a component of the centrosome during interphase and at the G2/M transition. During cell division, Ccdc1...

We report tm5468, tm5625 and tm5626 as novel deletion alleles of the gene Y48E1C.1 that is the only ortholog of human calmodulin-lysine N-methyltransferase (CAMKMT)1. CAMKMT encodes an evolutionarily conserved enzyme class I protein methyltransferase that acts in the formation of trimethyllysine in calmodulin for calcium-dependent signaling2. CAMKM...

We report tm4476 and tm4561 as novel alleles of the gene C38D4.9 that is an ortholog of human METTL5 (methyltransferase like 5)1. The alleles were isolated from the comprehensive screening of gene deletions generated by TMP/UV2. In the screening, both the alleles were detected by nested PCR using the following primer sets, 5’-CCGCCTATATCATGGCGCTT-3...

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Balancer chromosomes are convenient tools used to maintain lethal mutations in heterozygotes. We established a method for engineering new balancers in C. elegans by using the CRISPR/Cas9 system in a non-homologous end-joining mutant. Our studies will make it easier for researchers to maintain lethal mutations and should provide a path for the devel...

C. elegans strains used in this study. DOI: http://dx.doi.org/10.7554/eLife.14599.019

Conditional knockout (cKO) based on site-specific recombination (SSR) technology is a powerful approach for estimating gene functions in a spatially and temporally specific manner in many model animals. In Caenorhabditis elegans (C. elegans), spatial- and temporal-specific gene functions have been largely determined by mosaic analyses, rescue exper...

Marine subsurface sediments on the Pacific margin harbor diverse microbial communities even at depths of several hundreds meters below the seafloor (mbsf) or more. Previous PCR-based molecular analysis showed the presence of diverse reductive dehalogenase gene (rdhA) homologs in marine subsurface sediment, suggesting that anaerobic respiration of o...

The mushroom bodies (a higher center) of the honeybee (Apis mellifera L) brain were considered to comprise three types of intrinsic neurons, including large- and small-type Kenyon cells that have distinct gene expression profiles. Although previous neural activity mapping using the immediate early gene kakusei suggested that small-type Kenyon cells...

In situ hybridization of mKast in the nurse bee and forager MBs. Nurse bee MB sections (right panels) and forager MB sections (left panels) hybridized with antisense probes (upper panels) or sense probes (lower panels, control experiments). Red arrowheads indicate mKast expression. (Upper right panel) Schematic drawing of lKCs (green), sKCs (blue)...

Quantitative RT-PCR analysis of the mKast expression level in the nurse bee and forager brains. The amounts of the mKast transcript normalized with that of the EF-1alpha transcript are indicated. Relative expression levels of mKast in the brain regions that mainly contained the MBs did not differ significantly between the nurse bees and foragers. (...

Sensory neurons adopt distinct morphologies and functional modalities to mediate responses to specific stimuli. Transcription factors and their downstream effectors orchestrate this outcome but are incompletely defined. Here, we show that different classes of mechanosensory neurons in C. elegans are distinguished by the combined action of the trans...

Bacteria form unique ecosystems by coexisting with large organisms. Here we present the first evidence of active flora surrounding xenophyophorea revealed through clone analyses of environmental ribosomal RNA gene sequences. The flora included eight phyla in the xenophyophorean cells with agglutinated test. The major operational taxonomic units wer...

Table S1. Comparison of single- or low-copy integration methods.

Figure S2. Heat-shock induction of Venus mRNA in integrant strains. Relative expression of venus mRNA determined by quantitative RT-PCR. Total RNA was extracted from a 100-μl pellet of worms collected from 0-min, 30-min, and 1-h culture at 20°C after heat-shock at 32°C for 1 h. The expression of mRNA (normalized to act-2) is presented as a ratio to...

Figure S1. A positive-negative selection scheme. The tm246(vps-45) mutants exhibited a temperature-sensitive phenotype. Only tm246 mutants carrying vps-45 rescue transgenes survived at 20°C (positive selection). The tm234(ben-1) mutants were resistant to benzimidazole. tm234 mutants carrying ben-1 rescue transgenes are sensitive and unable to survi...

Table S2. PCR primers.

Transgenic strains of Caenorhabditis elegans are typically generated by injecting DNA into the germline to form multi-copy extrachromosomal arrays. These transgenes are semi-stable and their expression is silenced in the germline. Mos1 transposon or microparticle bombardment methods have been developed to create single- or low-copy chromosomal inte...

To identify candidate microRNAs involved in post-transcriptional regulation of brain (region)-selective gene expression in the adult honeybee brain, we isolated eight microRNAs: seven known microRNAs, ame-mir-2-1, −8, 13a, −34, −276, −317, −1000, and one novel one, named mir-hbd, that has significant sequence similarity with the Drosophila dme-mir-...

In situ hybridization of Amfutsch in the queen brains. In situ hybridization using DIG-labeled RNA antisense (B, D–I) and sense (C) Amfutsch probes and the queen brain sections. (A) Schematic representation of the signals detected in the left-brain hemisphere of the queen brain. Black circles and black check marks indicate the stronger and intermed...

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In situ hybridization of Amtau in the nurse bee brains. In situ hybridization using DIG-labeled RNA antisense (B, D–I) and sense (C) Amtau probes with nurse bee brain sections. (A) Schematic representation of signals detected in the left-brain hemisphere of the forager brain. Black circles indicate stronger signals. (D–I) Magnified views of parts o...

In situ hybridization of Amfutsch in the forager brains. In situ hybridization using DIG-labeled RNA antisense (B, D–I) and sense (C) Amfutsch probes with forager brain sections. (A) Schematic representation of the signals detected in the left-brain hemisphere of the forager brain. Black circles indicate the stronger signals. (D–I) Magnified views...

In situ hybridization of Amfutsch in the drone brains. In situ hybridization using DIG-labeled RNA antisense (B, D–I) and sense (C) Amfutsch probes with drone brain sections. (A) Schematic representation of the signals detected in the left-brain hemisphere of the drone brain. Black circles and black check marks indicate the stronger and intermediat...

Expression analysis of Amfutsch in the developing pupal brain. In situ hybridization using DIG-labeled RNA Amfutsch antisense probes with developing pupal brain sections (Stage P2, P4, and P5). (A) Schematic representation of signals detected in the left hemisphere of the developing pupal brain. Grey regions indicate the part of the brain cortex wi...

Amplification of the cDNA fragment that contained both Clone #2 and the predicted exon region of AmMESK2. (A) The predicted gene structure of AmMESK2 (GB18470, middle line) is indicated below the Linkage group 6.13 (upperline), where the putative exons of AmMESK2 are indicated with vertical solid boxes. Numbers above the Linkage group indicate nucl...

In situ hybridization with the exon probe of AmMESK2 in the nurse bee brains. In situ hybridization using DIG-labeled RNA antisense (B, D–H) and sense (C) AmMESK2 probes with nurse bee brain sections. (A) Schematic representation of signals detected in the left-brain hemisphere of the nurse bee brain. Black circles indicate stronger signals. (D–H)...

In situ hybridization with the exon probe of AmMESK2 in the queen brains. In situ hybridization using DIG-labeled RNA antisense (B, D–H) and sense (C) AmMESK2 probes with queen brain sections. (A) Schematic representation of signals detected in the left-brain hemisphere of the queen brain. Black circles indicate stronger signals. (D–H) Magnified vi...

Expression analysis of Amtau in the developing pupal brain. In situ hybridization using DIG-labeled RNA antisense Amtau probes with developing worker brain sections. (A) Results of the in situ hybridization using a section from the right hemisphere of the developing pupal brain. (B) A magnified view of the right pupal MB, indicated by the box in pa...

In situ hybridization with the intron probe of AmMESK2 in the nurse bee brains. In situ hybridization using DIG-labeled RNA antisense (B, D–H) and sense (C) AmMESK2 probes with nurse bee brain sections. (A) Schematic representation of signals detected in the left-brain hemisphere of the nurse bee brain. Black circles indicate stronger signals. (D–H...

Comparison of the amounts of Amfutsch-, Amtau-, and AmMESK2-transcripts in the whole brains of queens, nurse bees, and foragers. Real-time RT-PCR was performed to compare the amounts of Amfutsch-, Amtau-, and AmMESK2-transcripts in the whole brains of nurse bees and foragers. N: nurse bee, F: forager. The amount of Amfutsch-, Amtau-, and AmMESK2-tr...

The importance of visual sense in Hymenopteran social behavior is suggested by the existence of a Hymenopteran insect-specific neural circuit related to visual processing and the fact that worker honeybee brain changes morphologically according to its foraging experience. To analyze molecular and neural bases that underlie the visual abilities of t...

We previously studied a conditioning paradigm to associate the proboscis extension reflex (PER) with monochromatic light (conditioned stimulus; CS) in harnessed honeybees. Here, we established a novel conditioning paradigm to associate the PER with a motion cue generated using graphics interchange format (GIF) animations with a speed of 12 mm/s spe...

We studied associative visual learning in harnessed honeybees trained with monochromatic lights associated with a reward of sucrose solution delivered to the antennae and proboscis, to elicit the proboscis extension reflex (PER). We demonstrated five properties of visual learning under these conditions. First, antennae deprivation significantly inc...

SII-T1 is a tissue-specific member of the transcription elongation factor S-II that is expressed specifically in male germ cells. In the present study, we have identified a protein named GRIP1tau interacting with SII-T1 by yeast two-hybrid screening. GRIP1tau is a novel isoform of glutamate receptor-interacting protein 1 (GRIP1) that associates wit...