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Experiment details for pgat

Butler AM et al. (2018) Assay

A novel role for sox7 in Xenopus early primordial germ cell development: mining the PGC transcriptome.

Gene Clone Species Stages Anatomy
pgat.L laevis NF stage 11.5 to NF stage 33 and 34 primordial germ cell

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  Fig. 3. WISH analysis of PGC transcripts. Expression of selected PGC-enriched mRNAs at post-MBT st. 11.5 (gastrula), 16 (neurula) and 33/34 (tailbud). pgat expression marks germ plasm in PGCs. Transcripts detected in PGCs at neurula are shown at higher magnification in insets. WISH analysis was performed on ≥21 total embryos from at least three adult female frogs. Insets show magnifications of the boxed areas. A, anterior; b, brain; ba, brachial arches; cg, cranial ganglia; D, dorsal; gl, cement gland; isr, intersegmental region; L, left; n, notochord; np, nasal placodes; nt, neural tube; ov, otic vesicle; P, posterior; pn, pronephros; R, right; s, somites; V, ventral; vbi, ventral blood islands; Vg, vegetal view. Scale bars: 200 μm.

Gene Clone Species Stages Anatomy
pgat.L laevis NF stage 11.5 germ plasm
pgat.L laevis NF stage 33 and 34 primordial germ cell

  Fig. 4. PGC number increased after PGC-directed sox7 knockdown or overexpression in early development. (A-D) The number of PGCs per embryo was determined in control, sox7-MO, sox7-FL, and sox7-MO rescue embryos at gastrula (st. 11.5) (A,B) and tailbud (st. 33-34) (C,D) stages. Representative images are shown (B,D). In box and whisker plots, middle bar is the median, top of the box is the third quartile, bottom of the box is the first quartile, and the whiskers represent the minimum and maximum number of PGCs observed in one embryo of the set analyzed (i.e. the range). ctrl n=23, sox7-MO n=24, sox7-FL n=23, sox7-MO rescue n=25. *P<0.05, **P<0.005 (compared with uninjected control); # P<0.05 (compared with sox7-MO). pgat expression by WISH was used to identify PGCs. Analysis shown for one of at least three independent experiments. ctrl, control; FL, full-length, flagtagged; MO, morpholino. Scale bars: 500 μm (B); 1 mm (D).

Gene Clone Species Stages Anatomy
pgat.L laevis NF stage 11.5 germ plasm
pgat.L laevis NF stage 33 and 34 primordial germ cell

  Figure S2. PGC-directed Injections. A) Live 16-32-cell embryo, vegetal pole up. Germ plasm is detected as dark regions in vegetal blastomeres (*). Typical injection sites are shown. B&C) Lineage tracer (red, outlined in black) showing gastrula (st. 11.5, B) and tailbud (st. 33-34, C) progeny injected as in A). xpat (pgat) WISH (purple, arrows) indicates PGCs. Note: quality of targeting of PGCs.

Gene Clone Species Stages Anatomy
pgat.L laevis NF stage 11.5 germ plasm
pgat.L laevis NF stage 33 and 34 primordial germ cell

  Figure S5. PGC number increased after PGC-directed pou5f3.3 (oct60) knockdown in early development. The number of PGCs per embryo was determined in control, pou5f3.3-MO, pou5f3.3-FL, and pou5f3.3-MO rescue embryos at gastrula (st. 11.5) (A&B) and tailbud (st. 33- 34) (C&D) stages. Representative images are shown (B&D). ctrl n=23, pou5f3.3-MO n=24, pou5f3.3-FL n=25, pou5f3.3-MO rescue n=24. ** statistically significant compared to uninjected control (p<0.005). pgat (xpat) expression by WISH was used to identify PGCs. Analysis shown is representative of 5 out of 8 independent experiments. ctrl = control; MO = morpholino; FL = full length, flag-tagged. Scale bar = 500um (B), or 1mm (D).