|
Picture: Fig. 3G,H. |
|
Expr4830
|
|
DAB-1 expression was sdetected in a punctate pattern localised at, or close to, the membranes of oocytes; a pattern of immunoreactivity not present in dab-1 mutants. |
|
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). |
|
Expr4759
|
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. |
|
|
|
Expr4271
|
Anti-MIG-22 stained DTCs and the proximal arms of the gonad in a punctate pattern. Strong signals were detected in oocytes in adult gonads. MIG-22 was also detected in the developing vulval epithelium and seam cells. All these signals were significantly lower in mig-22(tk24) animals. |
|
|
|
|
Expr4577
|
|
WGA-G and MBK-2 colocalized in the cortex of oocytes in the proximal germline and in the cortex of newly fertilized embryos. Furthermore, cortical WGA-G and MBK-2 punctae were completely coincident at anaphase of meiosis I. MBK-2 localizes to centrosomes and chromosomes by first mitosis, at which point, WGA-G punctae are no longer detected. |
|
|
|
Expr4524
|
|
In wild-type oocytes, RAB-7 appeared to be mostly membrane associated. |
|
Picture: Figure 2. |
|
Expr7892
|
|
Affinity-purified polyclonal antibodies raised against DEPS-1 stain both P granules and nuclei in germ lines, and developing embryos of all stages. Several findings suggest that the P-granule stain is due to DEPS-1, but that the nuclear stain is due to cross-reactive material. First, in deps-1 mutants, the P-granule stain and a DEPS-1-sized band (~69 kDa) on western blots are absent, while nuclear stain and a band of ~120 kDa persist. Second, a DEPS-1::GFP fusion protein expressed in the C. elegans germ line is cytoplasmic and concentrated on P granules in adult germ lines and late stage embryos; nuclear GFP is not observed. |
|
Picture: Fig 2, 3, 4. The controls of the in situ hybridiza tion reactions showed negative results. |
|
Expr8634
|
A different expression pattern of CeNF-Y genes was observed in infertile worms (12 days after L1 stage). The positivity in the gonads disappeared, while CeNF-YB and CeNF-YC expression decreased in the pharingeal secretory cells and the neurons. Apart from the gonads and developing embryos, the CeNF-Y mRNAs were detected in other tissue from C. elegans. In 6-day-old worms, CeNF-YB and CeNF-YC, but not CeNF-YA1 nor CeNF-YA2, were expressed in the cytoplasm of the secretory cells located in the pharyngeal terminal bulb wall, in the neuron cell bodies of the small ganglia surrounding the pharynx and in the neurons running anteriorly to the sensory organs in the animal's head. NF-YC was also found in the excretory cell. Subunits were not expressed in any other organs, such as the intestine, the hypodermis and body wall muscle surrounding the pseudocoelomic space. The in situ hybridization procedure revealed the expression of CeNF-Y gene subunits in adult, fertile, hermaphrodite (6 days after L1 stage) C. elegans. High mRNA expression of all the subunits was evident in various parts of the gonads with differing degrees of intensity. In particular, positivity increased in oocytes from the distal to the proximal arm of the gonad in function of the stage of maturation. The cytoplasm of the syncytial ovaries and immature oocytes at the beginning of the proximal arm of the gonad were weakly positive, while oocytes, which increased in size along the oviduct toward the spermatheca, were strongly positive. Fertilized oocytes present in the uterus were strongly positive. In the late embryo stage, the expression of CeNF-Y mRNAs did not appear to be uniformly distributed, and presented an asymmetrical organization with anterior-posterior polarity. |
cytoplasm |
|
|
|
Expr13242
|
We did not detect GFP fluorescence in sydn-1(ju1415) knock-in animals, probably reflecting the low endogenous expression levels of SYDN-1. By immunostaining with FLAG antibody, we detected weak signals in multiple tissues, including oocytes, germ cells, neurons and epidermis. |
|
|
|
|
Expr10590
|
cyp-31A3 expression is confined to the gonads, oocytes and can be detected in eggs. |
|
|
|
We employed a transgene-based assay that tests the post-transcriptional control activity of 3' UTRs in vivo. In this assay, the GFP::H2B reporter fusion is placed upstream of the selected 3' UTR such that the entire fusion-GFP::H2B-3' UTR-is produced as a single transcript. Expression in the germline is achieved by using pie-1 promoter, which has been shown earlier to drive transgene expression in the germline. |
Expr9719
|
The transgene containing T01C3.3 3' UTR expressed the GFP::H2B reporter strongly in oocytes, but not in mitotic or early pachytene regions. |
|
|
|
|
Expr14451
|
ALLO-1 and GFP-IKKE-1 co-localized in embryos and oocytes. We did not observe obvious anti-ALLO-1 antibody staining in spermatozoa. |
|
|
|
|
Expr14452
|
ALLO-1 and GFP-IKKE-1 co-localized in embryos and oocytes. |
|
|
The cellular expression pattern for nlp-3 fragments was indistinguishable from the cellular expression pattern for nlp-3 subcloned constructs. Transgenic lines were generated by coinjection of the nlp::gfp construct (5070 ng/ul) and lin-15 rescue construct (pJM24,100 ng/ul) into lin-15(n765ts) animals. At least two independent transgenic lines were analyzed for each nlp gene; 5-10 animals were scored per line. 1,1'-Dioctadecyl-3,3,3',3 -tetramethylindodicarbocyanine perchlorate (Molecular Probes) was used to stain amphid and phasmid neurons to facilitate identification. |
|
Expr1688
|
Expressed in ADF, ASE, ASH, AWB, ASJ, BAG, HSN, I1, I2, I3, I4, MI, M3, NSMR, 3 head neurons, VNC, oocytes, I6, M2, pm1VL, intestine. |
|
|
Similar expression patterns and subcellular localizations were observed by both antibody staining and RME-8::GFP reporter expression, except that RME-8::GFP was not expressed in the germline, most likely because of a general silencing of transgenes in the C. elegans germline. These observations suggest that this RME-8::GFP fusion is an accurate reporter of endogenous RME-8 expression and subcellular localization. |
|
Expr2399
|
RME-8 expression in other cell types was examined by whole mount immunostaining. RME-8 is expressed in almost all cells. RME-8 was highly expressed in the lateral hypodermis, muscle, and the developing germ cells in larvae. RME-8 staining was extremely abundant in the developing uterus of L4 worms. RME-8 expression was examined in dissected adult ovaries by immunofluorescence. RME-8 was highly expressed in oocytes. |
Immunofluorescence: RME-8 staining was concentrated in the cortex of oocytes beneath the plasma membrane. At high magnification, RME-8 staining was localized to punctate structures that were 0.20.4 um in diameter. RME-8 staining in the rme-8(b1023) missense mutants was diminished and mostly diffuse in the oocyte cytoplasm with little localization to cortical punctate structures . Similar mislocalization was also observed in other cell types in the mutant worms. Similar expression patterns of RME-8 in somatic cells were observed with the RME-8::GFP reporter. In most cells, RME-8::GFP-labeled small puncta with sizes similar to the puncta stained with RME-8 antibodies. In live coelomocytes, RME-8::GFP labeled the limiting membranes of larger vesicles, with diameters ranging from 0.5 to 4.0 um. By serial thin-section confocal microscopy, these vesicles appeared to be round regardless of their sizes, but after fixation, these RME-8::GFP-labeled vesicles appeared smaller, as if they had collapsed. Whole mount immunostaining: in all the cells examined, RME-8 staining was localized to cytoplasmic punctate structures similar to those seen in the oocytes. |
|
|
|
Expr2719
|
TBB-2 was expressed throughout the female gonad, from the distal syncytial arm to maturing oocytes. TBB-2 expression was not germline and embryo specific, since the protein was also expressed in a glp-1 mutant, which lack germ line and fertilized embryos. TBB-2 was detected post-embryonically in larval and adult neuronal tissues. |
TBB-2 was expressed in every subset of spindle microtubules recognized by the generic anti-alpha-tubulin antibody, including spindle midzone and astral microtubules. TBB-2 expression persisted in mitotic spindles and asters throughout embryogenesis, beyond the end of the temperature-sensitive period of mei-1(ct46gf). |
|
|
|
Expr11704
|
Expression of the transcriptional fusion Ptop-1::gfp was not detected in early embryonic cells due to maternal germline silencing of the multi-copy transgenic gene. The GFP expression, however, was observed in most cells at the late embryonic stage and decreased with larval development. In the larval stages, GFP expression was predominantly present in the neuronal system, including sensory neurons (ILso, URX, RIC, IL1/IL2, AIY/AIM and RIG/RIF), motor neurons (VC4, VC5, HSN, PVD and PVM) of hermaphrodites and tail neurons (SPD and SPV/SPC) of males. The expression of the transcriptional fusion Ptop-1::gfp was strong in DTCs during the L3-L4 stage when gonad migration proceeds. |
|
|
|
|
Expr16363
|
DAF-2::mNeonGreen was detected in neurons, XXX cells, vulval cells, germ cells, and oocytes. In the last three types of cells, DAF-2::mNeonGreen had clear plasma membrane (PM) localization as expected for a cell surface receptor. In the neurons and XXX cells, a pair of specialized hypodermal cells of neural endocrine function, strong DAF-2::mNeonGreen signals were seen in the cell bodies and along processes. The DAF-2::mNeonGreen-expressing neurons included all of the ciliated sensory neurons marked by the osm-6 promoter-driven mCherry protein. |
|
|
|
|
Expr12193
|
The nkcc-1 reporter transgene was expressed primarily in the body wall muscles and neurons, including a subset of head and tail neurons, and the ventral cord neurons. It was also expressed in the vulval muscles and the posterior intestine. Unlaid eggs inside the adults showed high expression. |
|
|
|
|
Expr16365
|
We found that DAF-16::GFP was expressed ubiquitously in most or all somatic tissues, such as neurons, intestine, BWM, and hypodermis, and also in the germ cells and oocytes. Germline expression of DAF-16::GFP was not detected by earlier transgene reporters.Temporally, ubiquitous expression of DAF-16::GFP from the endogenous locus was detected from the embryonic bean stage to adulthood. |
|
|
|
|
Expr16552
|
UNC-85::wrmScarlet fusion protein is ubiquitously expressed. |
|
|
In a negative control, embryos incubated only with the secondary anti-rabbit antibody showed very weak background staining. |
|
Expr2879
|
RFP-1-specific staining is observed in nuclei, but not in nucleoli, of C. elegans embryos. In L1 larvae, specific RFP-1 staining was observed in the nuclei of pharyngeal, intestinal, and nerve ring cells, but not in nucleoli. The nuclear staining of RFP-1 in most tissues was observed in L2, L3, and L4 animals. In adult animals, RFP-1 was localized to the nuclei of oocytes and germ cells in the syncytial gonad. Again, the nucleoli did not stain for RFP-1. Competition with RFP-1 abolished this signal. RFP-1 nuclear staining was also observed in pharyngeal and intestinal cell nuclei in the head region of the adult animal. |
Expressed in nuclei but not in nucleoli. |
|
|
Transgenic worms that expressed RME-6 with either N- or C-terminal GFP tags, each under the control of the rme-6 promoter. Expression of these RME-6 fusion proteins fully rescues the coelomocyte endocytosis defect of rme-6(b1014), indicating that both GFP fusions are functional and are very likely to be correctly localized. |
Expr3238
|
GFP-RME-6 was expressed in many tissues, including coelomocytes, oocytes, hypodermis, intestinal and pharyngeal cells. |
In intestinal and pharyngeal cells, GFP-RME-6 was clearly concentrated on the plasma membrane. In coelomocytes, GFP-RME-6 was also localized on or very close to the plasma membrane. Most GFP-RME-6 did not overlap with mRFP1-EEA-1 or mRFP1-RAB-5. In contrast, strong colocalization of GFP-RME-6 was found with mRFP1-CHC-1 and mRFP1-alpha-adaptin on or near the plasma membrane of coelomocytes. mRFP1-CHC-1 and mRFP1-alpha-adaptin as well as GFP-RME-6 were distributed unevenly around the plasma membrane, with regions of high and low labelling, reminiscent of the uneven distribution of endocytic pit structures that can be seen in coleomocytes by electron microscopy. Similar colocalization was observed with mRFP-RME-6 and GFP-CHC-1. These results indicate that most RME-6 is associated with clathrin-coated pits and that only a minor fraction of the RME-6 in the cells is associated with early endosomes. Similar subcellular localization of GFP-RME-6 was observed in oocytes and the hypodermis. To analyse RME-6 distribution in oocytes, strains that express GFP-RME-6 in the rme-6(b1014) background under pie-1 promoter control were established . Expression of GFP-RME-6 in oocytes completely rescued the yolk endocytosis defect of the rme-6(b1014)-null mutant. GFP-RME-6 was concentrated on small puncta, which were on or close to the plasma membrane of oocytes. Strong colocalization of this GFP-RME-6 with mRFP1CHC-1 surface puncta was observed, indicating that RME-6 labels coated pits or vesicles. On the other hand, GFP-RME-6 in oocytes showed little spatial overlap with endogenous EEA-1, as assayed using a previously published anti-EEA-1 antibody. Clear colocalization of GFP-RME-6 and mRFP-CHC-1 was also evident in hypodermal cells. RME-6 and coated pit marker colocalization was not perfect and that the ratio of intensities of GFP-RME-6 and mRFP1-CHC-1 varied from puncta to puncta, implying that GFP-RME-6 may associate with clathrin-coated pits at a specific stage in their formation. |
|
|
|
Expr2291
|
SQV-4 Abs stained the cytoplasm of many cells, including (but not limited to) oocytes and vulval cells, as well as uterine, seam, pharyngeal, and spermathecal cells. During the early L4 stage, 10 of the 22 vulval nuclei were in cells with dramatically increased expression of SQV-4. These 10 nuclei are the six inner nuclei of the P5.p and P7.p descendants and the four outer nuclei of the P6.p descendants. During the mid-late L4 stage and thereafter, cells containing the inner four nuclei of the P6.p descendants also increased SQV-4 expression, bringing the total of vulval nuclei that highly expressed SQV-4 to 14. Thus, the 22 vulval nuclei define three classes based on the levels and timing of SQV-4 expression. |
cytoplasm |
|
Picture: Fig. 4, A and C. |
|
Expr7879
|
This antibody stains the two to three most proximal oocytes only in the presence of sperm. The MAPK-YT antibody only recognizes mpk-1 MAP kinase gene products because no staining is observed in gonads from mpk-1(ga117) homozygotes, a likely protein null. |
|
|
Picture: Figure 3M. |
|
Expr7864
|
|
DAB-1 protein localizes to the oocyte cytoplasm and is enriched at the oocyte cell cortex between oocytes. DAB-1 protein localization is altered in the absence of sperm, no longer exhibiting cortical enrichment between oocytes. |
|
Picture: Fig. 1. The OMA-1a, OMA-1b and OMA-2a signals in oocytes were completely abolished by absorption using their corresponding antigenic peptides. Furthermore, RNAi treatment of oma-1 and oma-2 abolished the fluorescence signals from anti-OMA-1b, anti-OMA-1a and anti-OMA-2a antibodies. |
|
Expr7830
|
Anti-OMA antibodies strongly and exclusively stained wild-type developing oocytes. |
|
|
Picture: Fig. 1. The OMA-1a, OMA-1b and OMA-2a signals in oocytes were completely abolished by absorption using their corresponding antigenic peptides. Furthermore, RNAi treatment of oma-1 and oma-2 abolished the fluorescence signals from anti-OMA-1b, anti-OMA-1a and anti-OMA-2a antibodies. |
|
Expr7832
|
Anti-OMA antibodies strongly and exclusively stained wild-type developing oocytes. |
|
|
Picture: Fig. 1. The OMA-1a, OMA-1b and OMA-2a signals in oocytes were completely abolished by absorption using their corresponding antigenic peptides. Furthermore, RNAi treatment of oma-1 and oma-2 abolished the fluorescence signals from anti-OMA-1b, anti-OMA-1a and anti-OMA-2a antibodies. |
|
Expr7831
|
Anti-OMA antibodies strongly and exclusively stained wild-type developing oocytes. |
|
|
|
|
Expr12265
|
The localization of tetraThymosin-beta is shown in oocytes (in dissected gonads of adult worms), in embryos and in whole adult organisms. TetraThymosin-beta was maternally expressed and in the adult gonad diffusely present in the cytoplasm of the oocytes, with strong staining at the cell cortex where also actin is present. In the distal arm of the gonad, tetraThymosin-beta localized to the inside edges of the membrane cubicles surrounding germ cell nuclei. Here, tetraThymosin-beta did not localize to the lateral side of the membrane cubicles where actin is arranged in a 'honeycomb' structure (Strome, 1986). In two-cell stage embryos, tetraThymosin-beta was enriched in the cell-cell contact where actin was also concentrated. The tetraThymosin-beta signal was clearly apparent until the four-cell stage and became weaker later on. At the comma stage (290 min after the first cell division), tetraThymosin-beta staining was observed at the developing nerve ring that is the largest axonal bundle in the nematode body and that positions around the isthmus of the pharynx. TetraThymosin-beta was evident in this region even before a clear actin signal was apparent. In larvae and adults, immunofluorescence yielded a diffuse but specific staining of the entire worm body with enrichment at the intestinal tract and spermatheca. At these stages, in contrast to what is observed in developing embryo, no prominent signal was observed in the head region where the nerve ring is located. In young adults is expressed in theposterior bulb of the pharynx. These immunostaining patterns were reproducible in three or more experiments and in 20 or more worms or embryos. |
|
|
This information was extracted from published material (Archana Sharma-Oates, Andrew Mounsey and Ian A. Hope). |
|
Expr660
|
9h after fertilization: strong staining in intestinal and hypodermal nuclei; Weak neuronal staining. Early L1: staining in nuclei of most postembryonic blast cells. Strong staining in nuclei of hypodermic blast cells H1, H2, V1-V6, T and all intestinal (E) cells. Weak staining nuclei of neuroblasts Q1 and Q2, mesoblast M cells and P cells. 9h after fertilization: strong staining in intestinal and hypodermal nuclei; Weak neuronal staining. Early L1: staining in nuclei of most postembryonic blast cells. Strong staining in nuclei of hypodermic blast cells H1, H2, V1-V6, T and all intestinal (E) cells. weak staining nuclei of neuroblasts Q1 and Q2, mesoblast M cells and P cells. Adult: staining observed in the mature oocyte nuclei of hermaphrodites, at meiotic prophase I when the chromosomes are condensed. (Possible artifact, detected in lin-14 loss-of-function mutant strains n536n540, n355n726). In embryo, first observed in embryo at 7h after fertilization (half way through embryogenesis). Strong staining in intestinal and hypodermal nuclei. L3: Pn.p stains weakly before division (staining fades by L4). Occasional weak staining of hypodermal, intestinal and neuronal nuclei and cytoplasm at L2 and L3. Late L1: staining of all nuclei except for neuronal nuclei is weaker. More neuron of the nerve ring and posterior ganglion stain than in earlier stages. Intestinal and hypodermal cell lineages stain strongest at mid to late L1 (Fade entirely by L2) similarly with many of the neuronal cells. Mid L1: staining in nuclei of hypodermic blast cells H1, H2, V1-V6 and T. The nuclei of intestinal (E) cells also stain. Weak staining in nuclei of P cells (staining fades before migration into ventral cord). Strong staining in nuclei of embryo-derived nuclei in hypodermal syncytial cell hyp7, ABarpppapa, ABplaapppp, Cpaaaa, Cpaapa, Cpaapp, Cpapaa, terminally differentiated nuclei from embryonic body muscle also stain for lin-14. Staining observed in nuclei of neuronal cells BDU, ALM, and CAN. All embryonic generated ventral cord neurons and some neurons of the nerve ring and posterior ganglion stain for lin-14. |
lin-14 is localized to the nuclei. |
