WormMine: LifeStage WBls:0000035

• Definition: The third stage larva. At 25 Centigrade, it ranges 32.5-40 hours after fertilization, 18.5-26 hours after hatch.
• Primary Identifier: WBls:0000035
• Public Name: L3 larva Ce

7 Anatomy Terms

Definition	Name	Synonym	Primary Identifier
A chain of very large cuboidal cells forming a wide central lumen in which food arrives from the posterior pharynx, is digested, and from which waste products proceed to the rectum. Intestinal rings form in groups of two and four cells surrounding the common lumen; thus the epithelium is only one cell deep at any point, with neighboring cells firmly secured to their neighbors by apical adherens junctions. These cells have very large nuclei and many large vacuoles, yolk granules, and other inclusions; the latter increase in number and electron density as the animal ages.	intestine	gut	WBbt:0005772
type of cells that make up muscle layers in the pharynx.	pharyngeal muscle cell		WBbt:0005451
Major cell type of nervous tissue, specialized for transmission of information in the form of patterns of impulses.	neuron	neurone	WBbt:0003679
H-shaped cell associated with the excretory system, largest cell in C. elegans.	excretory cell	excretory canal cell	WBbt:0005812
anchor cell, induces vulva, part of hermaphrodite gonad.	anchor cell	AC	WBbt:0004522
organ producing either sperm or ova.	gonad		WBbt:0005175
Neuron class of one pharyngeal interneuron, posterior sensory.	I6 neuron	I6	WBbt:0004739

2 Contained In

Remark	Definition	Other Name	Public Name	Primary Identifier
	A developmental life stage of the nematode Caenorhabditis elegans that occurs from egg hatching until adulthood.		larva Ce	WBls:0000023
	The third stage larva of nematodes.		L3 larval stage	WBls:0000108

129 Expression Clusters

Regulated By Treatment	Description	Algorithm	Primary Identifier
	miRNA with decreased expression in N2 L3 larva comparing to in N2 L2 larva.	DEGseq and GFOLD were used to analyze miRNAs expression. Authors chose miRNAs which had more than two-fold difference in expression (P < 0.001, q < 0.01 of Storey) from DEGseq, and miRNAs which had more than two-fold difference in expression (GFOLD score > 0 for up-regulation and GFOLD score < 0 for down-regulation) from GFOLD outcomes. Then authors obtained the intersection of up-regulated miRNAs and down-regulated miRNAs for each stage from the chosen miRNAs, respectively.	WBPaper00045316:miRNA_N2_L3_vs_L2_downregulated_adult
	Genes with expression level regulated by genotype (N2 vs CB4856) and age at L3 larva and Late reproduction stage (96 hours at 24 centigrade).	For model 2, authors used 100 permutations to estimate the FDR threshold. Per permutation, genotypes and ages were independently randomly distributed, keeping the among-gene structure intact. Then for each spot (23,232) on the array, model 2 was tested. The obtained P-values were used to estimate a threshold for each of the explanatory factors. Authors also used a genome-wide threshold of -log10 P-value = 2, which resembles an FDR of 0.072 and 0.060 for marker and the interaction age-marker for the developing worms and FDR of 0.050 and 0.065 for marker and age-marker for the aging worms. For the physiological age effect, authors used a log10 P-value = 8 in developing worms (0.012 FDR) and -log10 P-value = 6 (0.032 FDR).	WBPaper00040858:eQTL_age_regulated_developing
	Genes with expression level regulated by genotype (N2 vs CB4856) at L3 larva and Late reproduction stage (96 hours at 24 centigrade).	For model 2, authors used 100 permutations to estimate the FDR threshold. Per permutation, genotypes and ages were independently randomly distributed, keeping the among-gene structure intact. Then for each spot (23,232) on the array, model 2 was tested. The obtained P-values were used to estimate a threshold for each of the explanatory factors. Authors also used a genome-wide threshold of -log10 P-value = 2, which resembles an FDR of 0.072 and 0.060 for marker and the interaction age-marker for the developing worms and FDR of 0.050 and 0.065 for marker and age-marker for the aging worms. For the physiological age effect, authors used a log10 P-value = 8 in developing worms (0.012 FDR) and -log10 P-value = 6 (0.032 FDR).	WBPaper00040858:eQTL_regulated_developing
	Transcripts that showed significantly increased expression in rrf-3(pk1426) comparing to in N2 at L3 larva stage.	DESeq2v 1.18.1, fold change > 1.5, adjusted p-value < 0.01.	WBPaper00056169:rrf-3(pk1426)_upregulated_L3
	Expression Pattern Group C, enriched for genes involved in metabolic processes.	The significance (P 0.0001) of the relative age (time) was used to determine if a gene was differentially expressed between the three age (time) groups. The effect of this factor explaining gene expression differences was used to determine if the expression went up or down during the two age/time periods (t1 - t2 and t2 -t3). Authors used a permutation approach to determine the thresholds for the different mapping strategies. For each of the used models for eQTL mapping, authors used 23,000 permutations. For each permutation, authors randomly picked a spot; each spot could only be picked once. The gene expression and relative lifespan values were than randomly distributed over the RILs (and time points) and used for mapping. In this way, authors obtained a threshold for each of the explaining factors. For the single time points, authors used a FDR of 0.01 to adjust for multiple testing. The genome-wide threshold for this FDR is -log10 P = 3.8 for each of the three time points. For the combined models (t1 to t2 and t2 to t3), authors used a genome-wide threshold of -log10 P = 4, which resembles an FDR of 0.006, 0.001, and 0.006 for marker, age, and the interaction between marker and age, respectively. To determine the threshold for the single gene examples, authors used 1000 permutations as in the genome-wide threshold. The difference is that they use the gene under study in all of the permutations. The P-values for the gene specific thresholds were determined at FDR = 0.05.	WBPaper00036286:Pattern_C
	Genes that showed oscillating mRNA expression level throughout the 16 hour time courses from L3 larva to young adult.	The following three lines of R code were used to perform the classification: increasing <-2*amplitude-PC1 < -1.7; oscillating <-!increasing & (amplitude > 0.55); flat <-!increasing & !oscillating; Note that the amplitude of a sinusoidal wave corresponds to only half the fold change between trough and peak.	WBPaper00044736:oscillating_dev_expression
	Genes that showed expression levels higher than the corresponding reference sample (L3/L4 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:dopaminergic-neurons_L3-L4-larva_expressed
	Genes that showed expression levels higher than the corresponding reference sample (L3/L4 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:hypodermis_L3-L4-larva_expressed
	Genes that showed expression levels higher than the corresponding reference sample (L3/L4 all cell reference).	A Mann-Whitney U test with an empirical background model and FDR correction for multiple testing was used to detect expressed transcripts (Benjamini and Hochberg 1995). Genes and TARs with an FDR <= 0.05 were reported as expressed above background. Authors detected differentially expressed transcripts using a method based on linear models. Genes and TARs were called differentially expressed if the FDR was <= 0.05 and the fold change (FC) >= 2.0. To more strictly correct for potential false-positives resulting from multiple sample comparisons, authors divided individual FDR estimates by the number of samplesor sample comparisons, respectively. This resulted in an adjusted FDR of 1.3 * 0.0001 for expression above background and of 7.4 * 0.0001 for differential expression. Authors called genes selectively enriched in a given tissue if they met the following requirements: (1) enriched expression in a given tissue (FDR <= 0.05 and FC >= 2.0), (2) fold change versus reference among the upper 40% of the positive FC range observed for this gene across all tissues, and (3) fold-change entropy among the lower 40% of the distribution observed for all genes.	WBPaper00037950:PVD-OLL-neurons_L3-L4-larva_expressed
	Transcripts that showed significantly increased expression in dpy-21(e428) comparing to in N2 during L3 stage.	DESeq v1.6.3. Fold change > 1.5.	WBPaper00050370:dpy-21(e428)_L3_upregulated
	Transcripts that showed significantly increased expression in blmp-1(tm548) comparing to in N2.	DESeq2, fold change > 2, p-value < 0.05.	WBPaper00062347:blmp-1(tm548)_upregulated
	Genes that showed decreased expression in nhr-8(hd117) comparing to N2.	Expression data were read in the statistical programming environment R and processed for background correction and quantile normalization and summarized for genes that differed in expression >= 1.5-fold (N2 versus nhr-8) and a p value <= 0.01 using the Limma package.	WBPaper00044030:nhr-8(hd117)_downregulated
	Transcripts that showed significantly decreased expression in animals carrying hsp-16.2p-LIN-29 (induced at early L3 larva stage), comparing to animals carrying empty vectors.	DESeq2 v.1.12.3, FDR < 0.05, fold change > 1.7.	WBPaper00059191:hsp-16.2p-LIN-29_downregulated
	Transcripts that showed significantly decreased expression in rrf-3(pk1426) comparing to in N2 at L3 larva stage.	DESeq2v 1.18.1, fold change > 1.5, adjusted p-value < 0.01.	WBPaper00056169:rrf-3(pk1426)_downregulated_L3
	Transcripts that showed significantly increased expression in ints-5(RNAi) comparing to in N2.	DESeq2 and edgeR.	WBPaper00056284:ints-5(RNAi)_upregulated
	Transcripts that showed significantly increased expression in lin-22(ot269) comparing to in N2 at L3 larva.	Differences in gene expression were then calculated using the negative binomial test in the DESeq package (FDR = 0.1).	WBPaper00053295:lin-22(ot269)_upregulated
	Expression Pattern Group D, enriched for genes involved in catabolic processes.	The significance (P 0.0001) of the relative age (time) was used to determine if a gene was differentially expressed between the three age (time) groups. The effect of this factor explaining gene expression differences was used to determine if the expression went up or down during the two age/time periods (t1 - t2 and t2 -t3). Authors used a permutation approach to determine the thresholds for the different mapping strategies. For each of the used models for eQTL mapping, authors used 23,000 permutations. For each permutation, authors randomly picked a spot; each spot could only be picked once. The gene expression and relative lifespan values were than randomly distributed over the RILs (and time points) and used for mapping. In this way, authors obtained a threshold for each of the explaining factors. For the single time points, authors used a FDR of 0.01 to adjust for multiple testing. The genome-wide threshold for this FDR is -log10 P = 3.8 for each of the three time points. For the combined models (t1 to t2 and t2 to t3), authors used a genome-wide threshold of -log10 P = 4, which resembles an FDR of 0.006, 0.001, and 0.006 for marker, age, and the interaction between marker and age, respectively. To determine the threshold for the single gene examples, authors used 1000 permutations as in the genome-wide threshold. The difference is that they use the gene under study in all of the permutations. The P-values for the gene specific thresholds were determined at FDR = 0.05.	WBPaper00036286:Pattern_D
Exposure to Au-NP.	Genes with differeiential expression after exposed to Au-NP.	Hierarchical clustering was performed in Partek to confirm that the samples match to the treatment groups. Analysis of variance (ANOVA) was used to partition the variance due to treatment from technical and biological noise. The list of differentially expressed genes was generated by identifying the genes showing fold change of more than 1.5 and less than -1.5 at p < 0.05 with and without multiple sample correction, False Discovery Rate (FDR). False discovery correction according to Benjamini and Hochberg produced a list of 37 significant transcripts. FDR was not applied when selecting differential expressed genes, because this approach can increase the type II error and result in elimination of the genes responsive to the treatment.	WBPaper00040821:Au-NP_regulated
	Genes with expression level regulated by genotype (N2 vs CB4856) at L3 larva stage	Authors permuted transcript values and used a genome-wide threshold of log10 P-value = 2, which resembles a false discovery rate (FDR) of 0.0129.	WBPaper00040858:eQTL_regulated_juvenile
	Genes that showed significantly decreased experssion after 22.5 hours of treatment in 200nM delta7-dafachronic acid comparing with in ethanol vehicle control.	To identify the differentially expressed genes, we applied Significance Analysis of Microarrays (SAM) analysis using the R package samr [46]. Genes with median false discovery <5% and fold changes >2.0 were considered differentially expressed.	WBPaper00046548:dafachronic-acid_downregulated
	Transcripts that showed significantly increased expression in ints-1(RNAi) animals comparing to control RNAi animals, at late L3 larva stage.	DESeq2, v1.12.0, fold change > 2, FDR < 0.05	WBPaper00057068:ints-4(RNAi)_upregulated
	Transcripts that showed significantly increased expression in lin-22(icb38) comparing to in N2 at L3 larva.	Differences in gene expression were then calculated using the negative binomial test in the DESeq package (FDR = 0.1).	WBPaper00053295:lin-22(icb38)_upregulated
	Transcripts that showed significantly increased expression in lin-22(ot267) comparing to in N2 at L3 larva.	Differences in gene expression were then calculated using the negative binomial test in the DESeq package (FDR = 0.1).	WBPaper00053295:lin-22(ot267)_upregulated
	Genes with expression 1.5X depleted in PVD and OLL neurons.	Data sets were normalized by RMA and transcripts showing relative PVD enrichment (>=1.5X) vs. the reference sample were identified by SAM analysis (False Discovery Rate, FDR < 1%)	WBPaper00036375:depleted_in_PVD_OLL
	Genes enriched in AMsh glia.	Two different statistical methods were used for differential gene expression analysis, DESeq and voom. For DEseq analysis, DESeq2 was applied to normalize count matrix and to perform differential gene expression on the counts using negative binomial distribution; for voom analysis, edgeR was applied to normalize count matrix, and voom was applied for gene differentiation analysis. Significant genes from both analyses were combined. To identify transcripts enriched in AMsh glia compared to other cells (control AMsh versus control non-AMsh), authors used a fold change of 3.5 and an adjusted p value threshold of <0.05.	WBPaper00049489:AMsh-glia_enriched
	Genes eniched in hermaphrodites by comparing tra-2(ar221ts);xol-1(y9) XX phenotype males (at 25C) with either N2 or xol-1(y9) XX hermaphrodites during L3 larva stage	Student t-test. An mRNA was considered to have sex-enriched expression if it showed at least a two-fold enrichment within a 99% confidence interval (P < 0.01).	WBPaper00026629:L3_male_enriched
	Significantly up-regulated genes (>= 1.5-fold change, q value < 0.05) in blmp-1(tm548) mutant versus N2 L3 larvae as obtained from RNA sequencing analysis (n = 3).	Differential gene expression was performed with cuffdiff - which is part of the cufflinks package (v2.0.2) Genes with a FPKM value >0.5 in at least one of the N2 and blmp1 samples were considered expressed. Those with a q-value < 0.05 and fold change > +/- 1.5 were classified as differentially expressed.	WBPaper00045017:blmp-1(tm548)_upregulated
	Transcripts that showed significantly increased expression in sir-2.1(ok434) comparing to in N2 at L3 larva stage.	DESeq2, fold change > 2, FDR < 0.05.	WBPaper00062335:sir-2.1(ok434)_upregulated_L3
	Genes that showed flat mRNA expression level throughout the 16 hour time courses from L3 larva to young adult.	The following three lines of R code were used to perform the classification: increasing <-2*amplitude-PC1 < -1.7; oscillating <-!increasing & (amplitude > 0.55); flat <-!increasing & !oscillating; Note that the amplitude of a sinusoidal wave corresponds to only half the fold change between trough and peak.	WBPaper00044736:flat_dev_expression
	Expression Pattern Group B, enriched for genes involved in embryonic development. These patterns have in common that they all have genes of which the expression goes up after the juvenile stage. The expression of the genes in these patterns remains high or even goes up after reproduction.	The significance (P 0.0001) of the relative age (time) was used to determine if a gene was differentially expressed between the three age (time) groups. The effect of this factor explaining gene expression differences was used to determine if the expression went up or down during the two age/time periods (t1 - t2 and t2 -t3). Authors used a permutation approach to determine the thresholds for the different mapping strategies. For each of the used models for eQTL mapping, authors used 23,000 permutations. For each permutation, authors randomly picked a spot; each spot could only be picked once. The gene expression and relative lifespan values were than randomly distributed over the RILs (and time points) and used for mapping. In this way, authors obtained a threshold for each of the explaining factors. For the single time points, authors used a FDR of 0.01 to adjust for multiple testing. The genome-wide threshold for this FDR is -log10 P = 3.8 for each of the three time points. For the combined models (t1 to t2 and t2 to t3), authors used a genome-wide threshold of -log10 P = 4, which resembles an FDR of 0.006, 0.001, and 0.006 for marker, age, and the interaction between marker and age, respectively. To determine the threshold for the single gene examples, authors used 1000 permutations as in the genome-wide threshold. The difference is that they use the gene under study in all of the permutations. The P-values for the gene specific thresholds were determined at FDR = 0.05.	WBPaper00036286:Pattern_B

2354 Expression Patterns

• Primary Identifier: Expr8361
• Remark: Picture: Figure 1.
• Pattern: GFP expression initiated in the early gastrula. Robust expression of Prncs-1::GFP was observed in the midgut (E cell lineage) starting at the 28-cell stage and continuing into adulthood. By the comma stage, fluorescence was also visible in the embryo periphery in cells that give rise to hypodermis. In L1 larva and subsequent stages, strong expression of GFP was seen in hypodermal cells, including Hyp 7 syncytium and head and tail hypodermis. The expression pattern was identical in hermaphrodites and males, but adult hermaphrodites displayed fluorescence in vulval epithelium. Expression was absent in seam cells, nervous system, and pharynx. The Prncs-1::GFP reporter showed increased expression during starvation. Although fluorescence intensity was enhanced under starved conditions, the spatial expression pattern was unchanged. Expression of the Prncs-1::GFP transgene was also enhanced in males. An ~2.5-fold increase in rncs-1 expression in total RNA prepared from wild-type, well fed males, compared with hermaphrodites.

• Primary Identifier: Expr4899
• Remark: Picture: Figure 2.
• Pattern: Levels of the drh-3 transcripts in animals at adult stages were approximately threefold higher than that in larval nematodes.

• Primary Identifier: Expr4890
• Remark: Picture: Fig 5.
• Pattern: Several globin genes (C06E4.7, C09H10.8, C36E8.2, C52A11.2, F52A8.4, R01E6.6, R13A1.8, R90.5, and W01C9.5) are similarly upregulated in L3 and dauers relative to young adults, although some reach significance in dauers only. Many genes exhibited more than 2- fold upregulation but didn't reach statistical significance because strong upregulation was only seen in 2 biological replicates, A significant downregulation in L3 stage relative to young adults was observed for C26C6.7, T22C1.2 and ZK637.13. A similar trend was seen in dauers. C26C6.7 was the only globin which exressed at a significantly higher level in dauers relative to L3. Quantitative real-time RT-PCR experiments were done to compare the relative bundance of all 33 globins in wild type adults. Results demonstrate T22C1.2 and ZK637.13 are expressed at substantially higher levels. The difference with the other globins ranges within 1-3 orders of magnitude.

• Primary Identifier: Expr4892
• Remark: Picture: Figure 4A, B, D.
• Pattern: INA-1::GFP was present in the DTCs at the L2 stage prior to migration and maintained throughout L4. In wild-type N2 adults, INA-1::GFP was down-regulated with the cessation of migration (2% GFP-positive, n = 49).

• Primary Identifier: Expr4881
• Remark: Picture: Fig. 2A, B.
• Pattern: The Venus expression began during the early L2 stage when DTCs start to migrate and was maintained specifically in DTCs until the adult stage.

• Primary Identifier: Expr4882
• Remark: Picture: Fig. 2C.
• Pattern: Although the signal was very faint, expression of the full-length mig-24::venus translational fusion construct in hermaphrodites, which fully rescues the mig-24 phenotype, was detected only in DTC nuclei. Expression of mig-24::venus was observed in males, where the signal was detected specifically in MLCs from L2 through L4 stages. Thus, MIG-24 is expressed specifically in gonadal leader cells both in hermaphrodites and males.
• Subcellular Localization: Expressed in nuclei.

• Primary Identifier: Expr4883
• Remark: Picture: Fig. 4A.
• Pattern: Low GFP signals were detected exclusively in the intestinal cells of late embryos, L1L4, and adult hermaphrodites.

• Primary Identifier: Expr4874
• Remark: Picture: Figure 4D, 4E. nfyb-1 and nfyc-1 displayed identical expression patterns.
• Pattern: nfyb-1 was expressed in many cells in the developing embryo. At the larval stages, the expression level of nfyb-1 was reduced in most somatic cells except in some head neurons and in the developing hermaphrodite vulva and male tail.
• Subcellular Localization: NFYB-1 was localized in both the nucleus and the cytoplasm.

• Primary Identifier: Expr4875
• Remark: Picture: Figure 4D, 4E. nfyb-1 and nfyc-1 displayed identical expression patterns.
• Pattern: nfyc-1 was expressed in many cells in the developing embryo. At the larval stages, the expression level of nfyc-1 was reduced in most somatic cells except in some head neurons and in the developing hermaphrodite vulva and male tail.
• Subcellular Localization: NFYC-1 was localized in both the nucleus and the cytoplasm.

• Primary Identifier: Expr4876
• Remark: Picture: Figure 4H, 4I.
• Pattern: The expression of nfya-2 was restricted to few tissues, including the spermatheca, some neurons in the head and other body regions. Notably, it was highly expressed in intestine cells at all developmental stages.
• Subcellular Localization: NFYA-2 localized to the nucleus.

• Primary Identifier: Expr4873
• Remark: Picture: Figure 4A, B, C.
• Pattern: NFYA-1 was localized to the nucleus and was ubiquitously expressed in all nuclei at all developmental stages. In larvae and adult animals, strong expression of nfya-1 was observed in the head ganglia neurons and also in the developing hermaphrodite vulva and mail tail, while its expression was lower in most somatic cells.
• Subcellular Localization: NFYA-1 was localized to the nucleus.

• Primary Identifier: Expr4838
• Remark: Picture: Figure 8 C and D.
• Pattern: The major site of agrin expression was around the pharynx and the staining was particularly enriched in the anterior part. The posterior bulb was labeled more weakly correlating with the fainter GFP reporter expression in the posterior part. Polyclonal antiserum staining resulted in the same staining pattern in wild type worms of different developmental stages. Young larvae (L1) generally showed stronger agrin staining compared to young adults. In addition to the pharynx staining in the wild type worms, the polyclonal antiserum stained the gut lumen both in the wild type worms as well as in the agrin mutants, but not when preimmune serum was used. The staining of the lumen of the gut represents an unrelated cross-reactivity of the antiserum, possibly corresponding to the background bands detected on the western blots.
• Subcellular Localization: Agrin was detected in the basal lamina around the pharynx procorpus and anterior bulb. Posterior bulb staining was weaker possibly due to poor antibody penetration.

• Primary Identifier: Expr4836
• Remark: Picture: Figs. 4A-D.
• Pattern: In hermaphrodites, the expression of bro-1 was restricted to seam cells. Its expression was first detected at bean-stage embryos and persisted throughout the developmental stages. In the male tail, bro-1 was also expressed in the ray precursor cells.
• Subcellular Localization: GFP::BRO-1 was localized to both the cytoplasm and the nucleus.

• Primary Identifier: Expr4837
• Remark: Picture: Figure 5.
• Pattern: Fluorescence started to be visible in two cells of young embryos at around the 64 AB cell stage. Towards the end of gastrulation expression was visible in about 40 cells throughout the embryo including neuronal precursors, ventral hypodermal cells, and pharyngeal precursor cells. At the 1 to 2 fold stages fluorescence was observed in IL1 neurons (the identity was determined post-embryonically), the nine buccal epidermal cells, and additional cells in the head, most likely arcade cells. Transient expression was also observed in embryonic motoneurons (no longer visible in 3 fold stage embryos) and in a few apoptotic cells in the head. Based on their position they could be the sister cells of some of the IL1 neurons, which are known to undergo programmed cell death at this developmental stage. At the 3 fold stage expression was restricted to the buccal epidermal cells, most of the arcade cells (3 anterior and the DL and DR posterior arcade cells), and the six IL1 neurons. The two lateral IL1 neurons expressed the marker only weakly also in the L1 larval stage (but not later during development), whereas the dorsal and ventral IL1 neurons expressed GFP strongly throughout all larval stages and in the adults. Starting from the L1 larval stage expression could also be observed in the posterior cells of the gut. Starting from the L2 stage, when gonad development and migration begins, fluorescence became also visible in the distal tip cells of the gonad.

• Primary Identifier: Expr4829
• Remark: Picture: Fig. 6A, 6B. Reporter gene fusion type not specified.
• Pattern: Exclusively expressed throughout the nervous system in C. elegans. F25B3.3::gfp is a postmitotic pan-neuronal marker, i.e. its onset of expression is observed after the terminal division of neurons (around 450 minutes of embryonic development).

• Primary Identifier: Expr4821
• Remark: Picture: Fig. 10, A and B.
• Pattern: The hex-1 promoter was particularly active in coelomocytes as well as in neurons of the pharyngeal region and nerve cord, as compared with the head and tail pattern observed in strain BC14144 (see Expr6695). Expressed throughout the life-cycle.

• Primary Identifier: Expr4822
• Remark: Picture: Fig. 10, C.
• Pattern: The hex-2::gfp construct appeared to be active in the hypodermal cells, vulval toroids, and various adult head and tail neurons, Expressed throughout the life-cycle.

• Primary Identifier: Expr4823
• Remark: Picture: Fig. 10 H.
• Pattern: The hex-3 construct was expressed in gut granules. Expressed throughout the life-cycle.

• Primary Identifier: Expr4825
• Remark: Picture: Fig. 10, J and K.
• Pattern: Expression of hex-5 was restricted to certain cells at the 3-fold stage but was also present in the vulval, head (muscle), and tail regions in larval and adult worms. Expressed throughout the life-cycle.

• Primary Identifier: Expr4818
• Remark: Picture: Fig. 1A, 1B, 1C.
• Pattern: aqp-8 localized exclusively to the excretory system of the worm. Expression of aqp-8 also appears to be localized to an additional cell. The aqp-8p::GFP-PEST-expressing worms displayed an identical spatial pattern to the worms carrying the usual aqp-8::GFP construct, but due to the short half-life of the GFP-PEST construct, authors were able to determine that aqp-8 is transcribed only in the interval between the first larval stage and early adulthood. The relative levels of expression in the excretory cell and the excretory gland cell appeared to be similar to each other. Expression patterns derived from extrachromosomal arrays may be confounded by somatic loss of the transgene (leading to mosaically expressing transgenes). Therefore, the expression pattern of aqp-8 was confirmed by generating a genome-integrated aqp-8p::GFP transgenic line to prevent the sporadic loss of the transgene in somatic tissue.

• Primary Identifier: Expr4813
• Remark: Picture: Figure 7, C and D.
• Pattern: Expression was observed throughout development, starting at midembryogenesis. VHA-5 was also detected at the lumen of the vulva and rectum. In addition, authors found VHA-5 expressed in the sheath cells associated with head and tail sensory organs. Three-dimensional reconstructions showed that VHA-5 and RDY-2 formed a sixfold symmetrical pattern, which includes a larger spot that presumably corresponds to the amphid.
• Subcellular Localization: In the amphid sheath cell, VHA-5 was found in the most distal part of the cell lining the sheath pocket, which can be equated to its apical side.

• Primary Identifier: Expr4690
• Remark: Reporter gene fusion type not specified.
• Pattern: It was observed that the fusion gene expression continued from embryonic and post-embryonic stages. At embryonic stage, several cells were stained in post-gastrulating embryos whereas in postembryonic stages staining of cells was seen from L1 to young adults.

• Primary Identifier: Expr4693
• Remark: nsy-5 = T16H5.1.
• Pattern: A GFP reporter transgene with 5.8 kb of the nsy-5 promoter was expressed exclusively in sensory neurons and interneurons in the head and tail. The neurons that expressed nsy-5::GFP included AWC, ASH, AFD, ASI, ADL, ASK, BAG, AWB, and ADF (head sensory neurons); ADA, AIZ, RIC, AIY, and AIM (head interneurons); PHA and PHB (tail sensory neurons); and PVC and PVQ (tail interneurons). Expression began about halfway through embryogenesis, was strongest in late embryogenesis and the L1 larval stage, and faded thereafter. Adults maintained weak expression in several neurons, including ASH but not AWC.

• Primary Identifier: Expr4687
• Pattern: Embryonic expression of pgp-2::gfp was first seen in the daughters of the E blastomere (E2 stage), which generate the intestine. Intestinal expression persisted through embryogenesis and into adulthood. Rarely, weak expression of pgp-2::gfp was detected in embryonic and adult hypodermal cells. Pharyngeal or AWA expression of the pgp-2::gfp reporter were never detected.

• Primary Identifier: Expr4689
• Pattern: It was found that the amount of transcript of tbg-1 varies significantly in different stages during the development. In embryos the expression of the gene was high; it has extremely low level of gene expression during L1 larval stage, increased from L2 to L4 stages and showed the maximum expression in young adult stage. In gravid adult stage, the expression was more than that of embryos.

• Primary Identifier: Expr4683
• Pattern: hlh-29/hlh-28 mRNA is present at all developmental stages and does not vary significantly during later larval stages. Embryos and early L1-stage larvae produce significantly more hlh-29/hlh-28 RNA than later larvae. In separate assays from three independent cDNA samples, L1-stage larvae produced an average of 3 1/2 times more hlh-29 RNA than did L4 stage larvae and adults.

• Primary Identifier: Expr4684
• Pattern: GFP expression was detected at most developmental stages, with the spatial expression depending on the developmental stage of the animal. Neuronal expression of hlh-29 was detected in larvae and adults in both amphid and phasmid sockets, in the ALA and PVT neurons, in the chemosensory and mechanosensory neurons, ASI, ASK, PHA, and PQR, and in neurons of the anterior pharyngeal bulb. Weaker expression was also detected in the ASG chemosensory neurons in some transgenic lines. L1 animals show strong expression of hlh-29 in intestinal cells, and weaker expression in the rectal glands and the pharyngeal muscle cell PM1. By L3 stage, intestinal expression of the hlh-29::GFP is limited to the posterior intestinal cells, and PM1 expression is no longer detected. Expression is also detected in the ventral posterior coelomocytes in the later L3-stage larvae, and in the spermatheca and vulval muscles of L4 and adult animals.

• Primary Identifier: Expr4791
• Remark: Reporter gene fusion type not specified.
• Pattern: The glt-7::gfp fusion shows strong expression in the excretory canal cell from embryonic to larval stages. Strikingly, there was a complete disappearance of the glt-7::gfp signal in most adult animals.

• Primary Identifier: Expr4793
• Pattern: For all arrays examined, the reporter gene was expressed exclusively in hypodermal cells. In males, expression of dpy-5::gfp generally resembles that of hermaphrodites. Fluorescence is observed within the head and tail hypodermal cells, the P cells and hyp7, and is absent or of low abundance within the seam cells. This is more easily seen in a higher magnification of the tail region in which GFP is notably absent in the V5-derived seam cell, yet abundant in its sister set cell and the V6- and T-derived specialized hypodermal cells that envelope the sensory rays used in copulation. In summary, expression of the dpy-5::gfp reporter gene in the hypodermal cells begins in L1, and continues throughout larval development, ending in adulthood. In seam cells, expression is variable, suggesting that it may be regulated differently than in the hypodermal cells.

• Primary Identifier: Expr4796
• Remark: Reporter gene fusion type not specified.
• Pattern: cnx-1 was expressed ubiquitously in every blastomere of the embryos up to the gastrulation stage but expression became gradually restricted to the head and tail regions at the comma stage during embryogenesis. During post-embryonic development, cnx-1 was expressed prominently in the H-shaped excretory cell, in the neurons of head and tail, in the dorsal and ventral nerve cords, and in the spermatheca. cnx-1 expression was also observed in the spicules of the male tail. The two head neurons expressing cnx-1 are ASK and ADL, and two tail neurons are PHA and PHB. Therefore, cnx-1 is expressed in head neurons including ASK and ASI chemosensory neurons and tail neurons including PHA and PHB.

2 Followed By

Remark	Definition	Other Name	Public Name	Primary Identifier
	The fourth stage larva. At 25 Centigrade, it ranges 40-49.5 hours after fertilization, 26-35.5 hours after hatch.		L4 larva Ce	WBls:0000038
	The stage when an animal shifts from L3 larva to L4 larva. It includes the synthesis of new cuticle, cease of phrayngeal pumping during a lethargus stage, and the shed off of old cuticle.		L3-L4 molt Ce	WBls:0000037

2 Preceded By

Remark	Definition	Other Name	Public Name	Primary Identifier
	The second stage larva. At 25 Centigrade, it ranges 25.5-32.5 hours after fertilization, 11.5-18.5 hours after hatch.		L2 larva Ce	WBls:0000027
	The stage when an animal shifts from L2 larva to L3 larva. It includes the synthesis of new cuticle, cease of phrayngeal pumping during a lethargus stage, and the shed off of old cuticle.		L2-L3 molt Ce	WBls:0000029

0 Sub Stages