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Figure 1. Hermes/Rbpms protein and nanos RNA localize within germinal granules unique to germ plasm. Stage I oocyte. (A) Immunofluorescence (IF) and confocal microscopy showing Hermes/Rbpms protein (green) is ubiquitous in stage I oocyte including the mitochondrial cloud (MC), nucleus (N) and ooplasm; microtubules (red). Enlarged image from (A) showing microtubule organization around MC (B) and Hermes/Rbpms (C); Note how Hermes/Rbpms is enriched within the germ plasm region of the MC; (D) Electron micrograph showing a region of germ plasm within the MC of stage I oocyte. Red arrows indicate two germinal granules; (E) Nanos RNA is localized within germinal granules in stage I oocytes as is Hermes/Rbpms protein (F). Electron microscopy in situ hybridization with antisense Digoxigenin labeled nanos RNA was visualized with nanogold conjugated anti-Dig antibody and silver enhancement. Electron microscopy immunostaining with Hermes/Rbpms polyclonal antibody [49], anti-rabbit nanogold conjugated secondary antibody and silver enhancement. Scale bars are as indicated. GG (germinal granules); M (mitochondria); N (nucleus). |
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Figure 5. Cellular distribution of Hermes/Rbpms, hnRNP I, Vg1RBP/Vera, and Xpat proteins in the stage I/II oocytes. Confocal images showing immunofluorescence (IF) of stage I/II oocyte with (A) anti-Hermes/Rbpms (green) and (A’) anti-Xpat (red) antibodies; (A’’) Superimposed image. White arrows indicate Xpat and Hermes/Rbpms particles are not identical; (B) Superimposed images of hnRNP I (green) and GRP94 (red) proteins. Co-staining with anti-GRP94 reveals the ER enriched within the MC, cortex, and perinuclear region. Note that hnRNP I is excluded from the MC but is present in the nucleus and cytoplasm; (C) Merged images of endogenous Vg1RBP/Vera (red); or (D) hnRNP I (red) with Alexa 488-labeled nanos 3′UTR injected 20 h before fixation (green). Note that while nanos RNA localizes to the MC, both Vg1RB/Vera and hnRNP I are excluded from it. N: nucleus; MC: mitochondrial cloud. Scale bars are as indicated. |