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The specificity of binding of anti-Ce-GnRHR antibody to GnRHR was demonstrated by the lack of staining throughout the worm (including the germline, pharyngeal muscle and intestinal cells) when the antibody was preincubated with its antigen (the C-terminus amino acids 386 to 401). |
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Expr4759
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To better localize cellular staining in worms, authors permeabilized worms and performed whole-mount fluorescent immunohistochemistry. These experiments indicated that Ce-GnRHR was localized to the nucleus of maturing oocytes and intestinal cells, to sperm, pharyngeal muscles, but not other cells such as hypodermal cells. Similar staining of these structures was evident for both the anti-human-GnRHR1 and anti-Ce-GnRHR antibodies. Ce-GnRHR staining clearly illustrates an increase in the size of the oocyte nuclei as they mature along the gonadal arm. Upon fertilization, Ce-GnRHR staining becomes diffuse throughout the developing egg, although staining appears to increase during egg development. Only uniform background autofluorescence was apparent in worms treated with secondary antibody alone. Ce-GnRHR also was detected along the myofilament lattice of the pharyngeal muscles. staining along the three parallel muscles that comprise the pharyngeal musculature beginning at the tip of the head and extending to the pharyngeal bulb, staining of all 8 pharyngeal muscle (pm1 to 8) domains, but not in the gaps between contractile zones of adjacent pharyngeal muscle domains. staining of myofilaments that run radially towards the lumen and which are most obvious in the bulbs. These results indicate Ce-GnRHR is present on the pharyngeal musculature. |
Ce-GnRHR was localized to the nucleus of maturing oocytes and intestinal cells, to sperm, pharyngeal muscle. |
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Expr4210
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High levels in sperm, strong around 50% of mature sperm DNA. |
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Expr4211
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High levels on sperm DNA during and after meiotic divisions. |
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Expr4212
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High levels on sperm DNA during and after meiotic divisions. |
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Expr4209
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High levels in sperm, strong around meiotic and mature sperm DNA. |
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Expr16432
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We detected DMD-9::GFP expression in 10 head neurons of hermaphrodites at the L4 stage. High dmd-9 expression was detected in the BAG, AFD, AWCon, AWCoff, ASEL, and AWB neurons, with lower expression in ASER, and the ASGs. We also observed weak and inconsistent expression in other head neurons and transient weak expression in the tail of hermaphrodites and males at the L4 stage. No overt differences were observed in DMD-9::GFP expression in neurons of hermaphrodites and males. We also detected DMD-9::GFP expression in non-neuronal tissues. Transient DMD-9::GFP expression was observed in the uterine cells at specific sub-stages of the L4 stage. DMD-9 is not expressed from L4.0 to L4.3, with expression first detected in the early L4.4 stage, with the highest level during L4.4. before gradual loss by the L4.9 stage (Mok et al. 2015). DMD-9::GFP was not detected in adult uterine cells. We also detected DMD-9::GFP expression in sperm of hermaphrodites and males. In addition, we found that in a small proportion of L4 animals DMD-9::GFP shows asymmetric expression in the ASEs and AWCs neurons. |
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Expr9883
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Immunolocalization of GSP-3/4 in wild-type males revealed their expression is limited to sperm-producing germlines. |
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Expr13145
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We raised antibodies to a C-terminal region of GSP-3/4 and confirmed the previously reported localization pattern in the sperm. |
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Expr15065
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In hermaphrodites, ENRI-3::GFP was detected in the maturing germline of L4 larvae, with its expression peaking during spermatogenesis and in mature sperm in young adult hermaphrodites. ENRI-3 localization was concentrated in distinctive perinuclear puncta around germline nuclei, and co-localizes with the P granule marker PGL-3. The strongest expression of ENRI-3 was detected in the germline cytoplasm of male animals. ENRI-3 could also be detected in embryos and early larval stages, but its expression was limited to the precursor cells of the germline (Z2 and Z3). |
ENRI-3 localization was concentrated in distinctive perinuclear puncta around germline nuclei, and co-localizes with the P granule marker PGL-3. |
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Expr3701
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Using two independent antibodies elicited to two different FRK-1 peptides, FRK-1 was initially present both in nuclei and at cell-cell contact points of all cells in early embryos. Expression later became restricted to epithelial cells, body wall muscle and the germline, including mature sperm. Staining was eliminated in frk-1(RNAi) embryos, confirming specificity of the antibody. Epidermal expression of FRK-1, which is apparently sufficient for morphogenesis, requires ELT-1, a transcription factor required to generate all epidermal cells: FRK-1 was seen only in body wall muscle cells in late elt-1(zu180) mutant embryos. |
Localization of FRK-1 to nuclei correlates with phases of active cell division: nuclear localization was prominent in early embryos and the adult germline. The protein remained in the nucleus and at the cell surface until shortly before embryonic enclosure, at which stage most cells became mitotically inactive and nuclear FRK-1 became undetectable. In elongated embryos, FRK-1 was present in the cytoplasm and at the membrane of epidermal cells; cell surface staining was especially prominent in the seam cells. While nuclear function for FRK-1 in morphogenesis cannot be excluded, the protein was not detectable in nuclei during this phase of development. Later, during larval development, FRK-1 reappeared in the nuclei of seam cells and in the nuclei of the developing germline, correlated with their active post-embryonic division. In the adult soma, FRK-1 stably localized to apical junctions of the intestine and somatic cells of the gonad. |
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Expr10766
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SHN-1::GFP expressed from embryo to adult. SHN- 1::GFP is expressed in neurons including nerve cords, pharynx, pharyngeal-intestinal valve, intestine, vulva, rectal epithelial cells, tail ganglia and male tail. SHN-1 protein is expressed from the embryonic stage to adulthood and tissue localization overlapped with SHN-1::GFP expression patterns in the pharynx, pharyngeal-intestinal valve, intestine, nerve cords, and tail region in addition to sperm. |
Immunogold electron microscopy in wild-type hermaphrodites and males showed signals in the vesicle-like structures of intestinal and pharyngeal cells, and in the tail region, and most clearly seen in the male germline in developing sperm and in the presumptive pseudopodia, structures for motility of mature sperm. |
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Expr2412
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CRT-1 was detected in the cytoplasm, presumably from the endoplasmic reticulum network, of the early embryo (~100-cell stage) and a strong intestinal staining was observed in both larval and adult worms. Signals were also observed in the terminal bulb region of the pharynx and in excretory cells in the head. Faint but specific staining was also observed in head neurons and body-wall muscle cells. |
CRT-1 was detected in the cytoplasm, presumably from the endoplasmic reticulum network. |
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Expr15987
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Like TSP-12, TSP-14 is also widely expressed in multiple somatic tissues throughout hermaphrodite development, including the nose, pharynx, hypodermis, and developing vulva, as well as in the germline, in particular in the sperm. |
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See Expr593, and Expr595 for expression patterns for the same locus. This information was extracted from published material (Archana Sharma-Oates, Andrew Mounsey and Ian A. Hope). |
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Expr594
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ADM-1 localized to sperm in the gonads of adult males and inside spermathecae of hermaphrodite. Embryos from 2 cells - beginning of morphogenesis shows punctate staining between cells. Embryos from comma - 2-fold stages of elongation show staining localized between ventral hypodermal cells and in the head. Embryos from 2-fold to 3-fold show strong staining in luminal domains of the pharynx and buccal cavity. After hatching in different larval stages, there are punctate hypodermal staining, bp2 of ADM-1 staining in the head, two large cells stain close to sheath cells associated with chemosensory organs. Two smaller cells in tail correspond to the sheath cells of the phasmids. Staining in the area where sheath cells surround the sensory dendrites and in processes of the sheath cells. The monoclonal antibody stains in the sperm between embryonic cells in the pharyngeal apical domain in sheath cells surrounding sensory neurons, in vulva and in hypodermis. Expression is maintained from sperm throughout all of the embryonic and larval stages and in adults. |
Staining was punctate in structures inside and surrounding the sperm but excluding their nuclei. |
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Picture: Fig. 5. To test if EGRH-1 antibody staining was specific, authors stained egrh-1(tm1736) mutants, which are predicted to encode a truncated protein that would be recognized by our antibody, and egrh-1(tm1736) with egrh-1 expression further reduced by RNAi. Staining was decreased in both cases, indicating that this antibody stain is specific for EGRH-1. |
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Expr9107
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To understand EGRH-1 function in regulating oocyte development, authors examined egrh-1::gfp expression and endogenous EGRH-1 expression using an antibody targeting a unique region at the EGRH-1 N-terminus in dissected adult hermaphrodites. Using both techniques, EGRH-1 expression was observed in the sheath cells and distal tip cells of the somatic gonad, as well as in the intestine and sperm. egrh-1::gfp was expressed in embryos through adult stages in most somatic tissues, including the gut, pharynx, body wall muscle, and nervous system. |
EGRH-1 was localized to nuclei in all tissues except sperm, where it appeared perinuclear. Importantly, no egrh-1::gfp expression or EGRH-1 antibody staining was observed in oocytes. |
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Expr14682
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We first show the localization of UNC-59 at the cleavage furrow (previously shown with antibody staining, (Nguyen et al. 2000)) during a time lapse of cell divisions in early 2- to 4-cell stages of embryogenesis and throughout embryogenesis (cleavage rings in older embryos. Septins are also important for gonad morphogenesis and distal tip cell (DTC) migration (Nguyen et al. 2000) where UNC-59 protein is detected throughout gonad development in the rachis (previously shown with endogenously tagged unc-59::mKate, (Priti et al. 2018)) and DTCs. We highlight UNC-59/Septin localization in the DTC (previously shown with a transgene, (Finger et al. 2003)) at the L2 and L3 stages where it is organized into bundles (DeMay et al. 2011) and ring structures. The two bilateral sex myoblast cells express UNC-59 during their posterior to anterior migration in the L2 and early L3 stage and continue to express UNC-59 in these cells as they differentiate into vulval muscles in the late L3 to early L4 stages. Lastly, we show UNC-59/Septin expression and localization in tissue not previously reported: in the pharynx (cells of the buccal cavity, anterior procorpus, and terminal bulb); in the seam cells, both in bundles and at the cleavage furrows, beginning in the L1 stage and continuing throughout development and into the adult; and in sperm surrounding an embryo that has exited the spermatheca. |
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Expr16272
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GSKL-1-FLAG and GSKL-2-Ollas immunofluorescence was also detected specifically in the hermaphrodite sperm. |
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Expr2454
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kvs-1 expression was detected in more than 10 cells in the head, including the amphid neurons ADL, ASK, ASH, ADF, ASE, AWC, and ASG, in ventral cord neurons, in the motoneuron PDA in the anal depressor muscle and in sperm. |
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This information was extracted from published material (Archana Sharma-Oates, Andrew Mounsey and Ian A. Hope). |
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Expr668
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Postembryonic expression is observed in the rectum epithelium. A major site of EGL-5 expression is in the rectal epithelium. At hatching, the rectal expression is in K, F, B, U and Y. In addition expression is seen in (Y differentiates into) PDA motor neuron, (K divides to rise to) part of dorsal rectal epithelium and a cell that becomes DVB motor neuron. In males male-specific neurons show expression. In males Ab staining is observed in B.a and B.p as well as Y.p and Y.p in L1 and early L2. It appears that most/all B, Y, U, F, K descendants express EGL-5. Ventral neuroblast P12, staining is first seen in P12.a and P12.p in 12-h worms. Staining is maintained in P12 descendants in 15-h until adulthood. Both sexes' mechanosensory neurons, expression is seen in PLM neurons throughout larval development. In addition two cells express EGL-5, one in anterior region of each lumbar ganglion, likely to be PVC interneurons. Both sexes' muscles cells, expression is detected in 4-6 left/right pairs of posterior body-wall muscle cells in L1 larvae at earliest examined time 10-12 h. At L2 staining is detected in 12 left/right pairs of nuclei. Staining is strongest in the most posterior nuclei and tapers of towards the anterior. Staining in posterior body wall muscle cells remains throughout larval development and into adulthood in both sexes. In L3 males, sex-specific muscle lineages and sex-specific muscles stain strongly. These muscles include the diagonal muscles, muscles of spicule, gubernaculum and other sex muscles. Staining in these muscles persist until adulthood. HSN neurons, expression from L1 onwards through to adulthood. Male gonad, first detected in the male gonad in late L1 in a group of 6 cells at the anterior end. It appears that expression is clustered in a region that consists of both somatic cells and germ cells. Later at the beginning of the late mitotic period, staining nuclei lose their clustered arrangement. By 34 h, staining is seen in several dividing cells that form the primordium of the seminal vesicles as well as in two large nuclei in the valve region. In the nuclei of diving cells staining surrounds a condensed chromatin. This pattern persists until the end of the late mitotic period (35-37 h posthatching) when staining is also detected in sperm cells. No staining was observed in cells of the vas deferens. Lateral hypodermis, expression is seen in male seam from mid-L2. Staining first appears in V6.ppp at 20-22 h postembryonic development. Staining persists in V6.pppa and V6.pppp but at a lower level. Intensity of staining increases in R5 and R6 and to lesser degree in R4. Identification of staining cells in ray sublineages was not possible due to intense fluorescence of B-lineage cells lying in the same region. However it was possible to observe expression of a reporter gene in R4, R5, and R6 and also in cells of the R5 and R6 sublineages. |
Expressed in the nuclei. |
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See Expr593, and Expr594 for expression patterns for the same locus. This information was extracted from published material (Archana Sharma-Oates, Andrew Mounsey and Ian A. Hope). |
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Expr595
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Earliest staining against antipeptide antibodies was in embryo 430 mins after first cleavage that had elongated about 1.5-fold. Staining persists throughout development in the sheath cells of amphids and phasmids. At L2, the 16 sheath cells of anterior sensilla localized at the tip of the head area are detected by antibody against extracellular domains of ADM-1. Antibody against peptides bp2 and bp3 show syncytial hypodermis, vulva and mature sperm. |
Staining in sperm, between embryonic cells, in the sheath cells and in the hypodermis appear to be associated with intracellular membranes and the plasma membrane. |
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Protein_Description: ICB4 antigen. |
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Expr2784
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Antigen expressed in chemosensory neruons (inner labial 2, cephalic companion cell, and possibly ray neruon B), intestine and sperm. |
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Expr9272
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such-1 is expressed throughout the developing germline as well as in the meiotic embryo and throughout embryogenesis. Limited expression is observed in the soma of the adult, including some head neurons and vulval precursor cells. In hermaphrodites, such-1-driven transgene expression is prominent in mature sperm stored in the spermatheca. Male transgenics were generated and was found that such-1 transgene is expressed throughout the male germline. |
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Expr9271
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gfi-3 is expressed throughout the germline, meiotically, and in all embryonic stages. The gfi-3 transgene is observed in the soma of the L1-L4 larval stages and is also expressed in the gut cells of the adult animal. In hermaphrodites, gfi-3-driven transgene expression is prominent in mature sperm stored in the spermatheca. Male transgenics were generated and was found that gfi-3 transgene is expressed throughout the male germline. |
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Reporter gene fusion type not specified. Similar patterns of HSP16 distribution were observed with two other polyclonal antibodies, one with greater specificity for HSP16-41 and another which cross-reacts with all four stress-inducible HSP16s. Thus the staining pattern described here reflects the distribution of the HSP16 family in general. Non-stressed animals showed little or no detectable signal. |
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Expr1117
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HSP16-2 is ubiquitously expressed throughout most somatic tissues of larvae after heat-shock, and the intensity gradually decreased with age. HSP16 labelling is prominent in spermathecae and in the vulval region in L4 and adult hermaphrodites. The spermathecal labelling seems to consist of cytoplasmic localization of HSP16-2 in sperm, and perhaps also in the spermathecal cage cells. The prominent localization of HSP16 to intestinal cells was previously noted in transgenic animals expressing a lacZHSP16 gene fusion. Vulval expression of HSP16 was also seen in transgenic reporter experiments. In males, HSP16-2 was distributed more generally, though it was slightly concentrated in the lower testis in the region corresponding to spermatids, and in tail structures. The latter expression pattern was also seen with transgenic reporter constructs. |
Counterstaining with DAPI revealed HSP16 localized in discrete regions within the cytoplasm of intestinal cells, characterized by their large nuclei. Concentrations of HSP16-2 occur along the luminal border of these cells. |
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Temporal description |
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Expr11524
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By antibody staining, MCM-4 was found to be expressed in dividing cells during all stages of development in wild-type animals. Embryos showed the highest levels of MCM-4 expression, in agreement with the fact that more than half of the somatic cells are formed during embryogenesis. Even dauer larvae that had been arrested in cell division for 2 weeks still contained detectable MCM-4 protein levels. These results suggest that a pool of MCM-4 is retained during prolonged periods of quiescence, so that MCM-4 might function in the re-initiation of DNA synthesis when conditions improve. Immunostaining of wild-type animals for MCM-4 showed strong nuclear staining in the gonad, embryos and postembryonic lineages. MCM-4 was detectable in sperm and accumulated during oocyte maturation in the nucleus but did not show overlap with the condensed chromosomes in diakinesis of meiotic prophase. MCM-4 was not chromatin-associated during MeiosisI of the fertilized oocyte, and the first polar body did not contain MCM-4. This finding is consistent with the absence of S phase between Meiosis I and -II. The second polar body and maternal pronucleus received some MCM-4. Subsequently, embryonic cells in interphase showed strong nuclear staining. In prophase, MCM-4 localization did not overlap with the condensing chromosomes. Upon nuclear envelope degradation, MCM-4 became diffusely localized throughout the cell and clearly did not co-localize with the metaphase-aligned chromosomes. MCM-4 remained cytoplasmic at the onset of anaphase; however, chromatin association became apparent in late anaphase. These data show that chromosome association of MCM-4 is tightly controlled, consistent with origin licensing taking place at the end of mitosis and disappearing during S phase. Similar observations were made during larval divisions. Matching the MCM-4::mCherry reporter, endogenous MCM-4 expression was detectable prior to and during mitosis. Staining of synchronized L1 animals revealed the timing of MCM-4 expression, which in general preceded mitosis by 1-2 h. After 5 h of L1 development at 20 C, MCM-4 immunostaining was predominantly detected in the epithelial seam cells, Q neuroblast daughters and gonad primordium. The somatic gonad precursor cells Z1 and Z4 showed nuclear staining, while the mitotically arrested germline precursor cells Z2 and Z3 showed diffuse cytoplasmic staining. At 6 hours of L1 development, the mesoblast (M) also stained strongly as well as the most anterior ventral cord precursors cells (W, P1 and P2). Subsequently at 7 h, additional P cells showed nuclear MCM-4 expression, which became apparent prior to migration of the nucleus into the ventral nerve cord. At 8 h of L1 development, the intestinal nuclei showed MCM-4 expression, which preceded nuclear division by at least 4 h. At subsequent time points, daughter cells that continued division, such as the Pn.a and M descendants, retained strong nuclear staining. L2 animals stained at 16 h of larval development showed strong MCM-4 expression in the gonad, the H1.a, H2.p, V1-6.p and T.ap seam cells and, weakly, the intestinal nuclei (data not shown). Importantly, MCM-4 staining did not overlap with DNA in prophase and metaphase, while in late anaphase co-localization with the chromosomes was clearly detectable. Similar to our observations with the MCM-4::mCherry reporter, we could not detect any asymmetry in MCM-4 segregation. Thus, even if only one daughter cell continued cell division, both daughters received a similar amount of MCM-4in mitosis. Furthermore, the MCM-4protein became undetectable during quiescence, i.e. the P3.p-P8.p daughter cells that resume DNA replication in the L3 stage did not show detectable expression in the L2 stage. Altogether, our reporter gene and antibody staining analysis show that MCM-4 is dynamically expressed and localized during larval development as well as during different phases of the cell cycle. Expression of MCM-4::mCherry was specifically induced in all postembryonic blast cell lineages well before mitotic entry, at the expected time of S-phase induction. The fusion protein localized to the cell nucleus until degradation of the nuclear envelope in prometaphase, at which point MCM-4 became diffusely localized through the cell. This diffuse localization indicates that MCM-4 is not chromatin-associated in mitosis. MCM-4::mCherry did not disappear upon completion of mitosis but was segregated to both daughter cells. Even cells that permanently withdrew from cell division, such as the motor neurons of the ventral nerve cord, initially retained MCM-4::mCherry expression. However, this expression subsequently disappeared in differentiated cells as well as in cells that temporarily arrested cell division, such as the Pn.p vulval precursor cells in the ventral cord. These experiments indicate that mcm-4 is transcriptionally activated at approximately the time of G1/S transition and that MCM-4 protein is segregated to both daughter cells in mitosis. |
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TSP-14A isoform expression. |
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Expr16022
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We found that TSP-14A and TSP-14B exhibit distinct expression and localization patterns. First, only TSP-14A, but not TSP-14B, is detectable in the germline and sperm cells, as well as at the tip of the anterior sensory cilia, while TSP-14B is detectable in the pharynx. Both TSP-14A and TSP-14B are found in hypodermal cellsand the developing vulva. However, the two isoforms exhibit different subcellular localization patterns. In hypodermal cells, TSP-14A is primarily localized in intracellular vesicles, while TSP-14B is mainly localized on the cell surface. In the developing vulva, TSP-14A is localized on the apical side, while TSP-14B is localized on the basolateral membrane. Similar localization patterns hold true in the pharynx. |
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Expr11861
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Expression of snf-10 was visible in the male and hermaphrodite germline in cells undergoing differentiation into spermatocytes as well as at later stages of spermatogenesis. No fluorescence was visible in somatic tissues,in the mitotic or meiotic-prophase regions of the germline, or in germ-line cells developing into oocytes. |
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Expr12966
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ets-4 showed pan-germline expression. Reporter expression was observed in the distal region of the germline, in mitotic progenitor cells, as well as in the syncytial region, in the germline bend, in oocytes, and in embryos. ets-4 reporter expression remained strong in sperm (n=12/23), suggesting that at least some 3'UTRs can direct retention of sperm specific expression. |
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Expr12971
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set-2 3'UTR showed faint GFP expression in the distal end of the germline followed by an increased expression in the syncytial region, which then decreased around the recellularization region and oocytes. As with ets-4, the GFP expression remained strong in sperm (n=11/15), providing another example of a 3'UTR that can direct expression of a reporter in male gametes (see also Expr12966). |
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Expr15070
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mScarlet::PEZO-1, and PEZO-1::mScarlet were widely expressed from embryonic stages through adulthood. Notably, PEZO-1 is strongly expressed in several tubular tissues, including the pharyngeal-intestinal and spermathecal-uterine valves. Under higher magnification, we observed PEZO-1 on the plasma membranes of oocytes and embryonic cells during a variety of embryonic stages, suggesting PEZO-1 is a transmembrane protein. PEZO-1 is expressed in multiple reproductive tissues, including the germline, somatic oviduct, and spermatheca. Higher magnification imaging of the spermatheca revealed expression of PEZO-1 on sperm membranes as well. Live imaging and detailed analysis of PEZO-1 expression patterns during reproduction revealed that GFP::PEZO-1 is expressed in sheath cells, sperm, both spermathecal valves and the spermathecal bag cells. |
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