Schistocerca gregaria. Anterior is down in (A), and to the left in (B) and (C). (A) Adult female locust. (B) Germarium (g) in an adult female ovary. The panoistic ovaries of S. gregaria are made up of approximately 50 ovarioles per ovary, and do not have nurse cells. Oocytes proliferate in the germarium and move posterior in the ovariole as they develop. The oocytes are surrounded by a monolayer of somatic follicle cells. Vasa protein (red) is located in the cytoplasm of early oocytes, with some concentration around the nucleus (arrowheads). Most somatic cells are undergoing mitosis (blue) in the germarium and during the early stages of oogenesis. Cells in mitosis are labeled with an antibody against phosphorylated Histone 3 (blue). (C) Early stages of oogenesis. Vasa protein (red) continues to concentrate in juxtanuclear aggregates (arrowhead). In (B) and (C) nuclear counter stain (YO-PRO-3 iodide) is in green. Scale bars: (B) 80 um, (C) 40 um.
Thermobia domestica. Anterior is up in (A), and to the left in (B) and (C). (A) Adult female firebrat. (B) Germarium (g) and early stages of oogenesis in an adult female ovary. The panoistic ovaries of T. domestica are organised in a similar manner to those of S. gregaria, but with only five ovarioles per ovary. Vasa protein (red) is concentrated in asymmetric aggregates next to the nuclei of early oocytes (arrowheads). (C) At later stages of oogenesis, Vasa protein (red) continues to concentrate in asymmetric juxtanuclear aggregates (arrowhead). High concentrations of Vasa are also seen in the cortical ooplasm. In (B) and (C) nuclear counter stain (YO-PRO-3 iodide) is in green. Scale bars: (B) and (C) 40 um.
Oncopeltus fasciatus. Anterior is up in all panels. (A) Adult female milkweed bug with freshly laid eggs (top right). (B) Germarium (g) and early stages of oogenesis in an adult female ovary. The ovaries of O. fasciatus contain seven ovarioles per ovary, and like those of Drosophila melanogaster, are meroistic (containing nurse cells). However, unlike D. melanogaster, nurse cells are shared between all oocytes of an ovariole. All nurse cells are located in the germarium, liked by cytoplasmic bridges to a pool of common cytoplasm called the trophic core. Tubes called trophic cords (arrowheads) originate in trophic core, thus linking each oocyte to the nurse cells. Vasa (red) protein is detected in cells of the germarium and within the trophic cords (arrowheads). The actin cytoskeleton (blue) is concentrated along the sides of the trophic core and around the oocyte cortex. (C) At later stages of oogenesis, Vasa protein (red) is distributed in the ooplasm but excluded from the oocyte nucleus (on). The actin cytoskeleton (blue) has become further enriched at the cortical ooplasm. (D) As oogenesis progresses, Vasa protein (red) disappears from the central ooplasm, but remains at the oocyte cortex, with highest levels at the posterior pole. In (B) through (D), Vasa protein is in red, F-actin (marked with Alexa 647-conjugated phalloidin) is in blue, and nuclear counter stain (YO-PRO-3 iodide) is in green. Scale bars: (B) 80 um, (C) 40 um, (D) 80 um.
Artemia franciscana. Anterior is up in all panels. (A) Adult female carrying eggs in brood pouch (arrow). (B) 8 day nauplius larva stained with cross-reacting Vasa antibody. Vasa is detected specifically in germ cells (arrowheads), which lie in the segmental mesoderm lateral to the gut (g). (C) Close up of (B). Vasa protein is cytoplasmic in germ cells (arrowheads).
Parhyale hawaiensis. Anterior is to the right in all panels. (A) Adult female carrying embryos in brood pouch (arrowheads). Natural purplish colour of developing oocytes can be seen in the ovary (arrows). (B) Late germ band stage embryo stained with cross-reacting Vasa antibody (pink). Vasa is detected in a group of cells among the endomesodermal precursors, posterior to the head lobes (hl). These cells are the likely descendants of the a micromere of the 8 cell stage, which has been shown to be the unique germ cell precursor by cell lineage studies. p: posterior. (C) Anterior end of an adult ovary with mature oocytes. The paired ovaries of P. hawaiensis are organised with a strand of primordial germ cells (PGCs) along the dorsal midline (towards top of panel). There are no nurse cells. Maturing oocytes move laterally in the ovary, so that the most mature oocytes are located farthest away from the PGCs (towards bottom of panel). The oocytes are surrounded by a monolayer of somatic follicle cells. Vasa protein (red) is concentrated in a perinuclear ring around the oocyte nucleus. (D) Orthogonal sections through an adult testis. The testes of P. hawaiensis have a compartmental organization, with PGCs lying along the dorsal midline (bottom of panel). Maturing spermatocytes move laterally in the testis, so that the most advanced spermatocytes are farthest away from the PGCs (towards top of panel). Spermatids and later spermatogenic stages are located in a long tube extending posterior from the testis (not shown). Green lines indicate planes of orthogonal sectioning. Vasa protein (red) is concentrated in primordial germ cells (PGCs) in the medial "pgc zone" (pgc). Primary spermatocytes (region 1) and secondary spermatocytes (region 2) are located laterally to the PGCs, and do not have high levels of Vasa protein. Orthogonal section panels (bottom, right) indicate intensity of Vasa (red) and nuclear (blue) staining. Spikes of red next to high levels of blue indicate perinuclear distribution of Vasa protein. Nuclei are counterstained with TO-PRO-3 iodide (blue) in (B), (C) and (D).
Schistocerca gregaria. Anterior is down in (A), and to the left in (B) and (C). (A) Adult female locust. (B) Germarium (g) in an adult female ovary. The panoistic ovaries of S. gregaria are made up of approximately 50 ovarioles per ovary, and do not have nurse cells. Oocytes proliferate in the germarium and move posterior in the ovariole as they develop. The oocytes are surrounded by a monolayer of somatic follicle cells. Vasa protein (red) is located in the cytoplasm of early oocytes, with some concentration around the nucleus (arrowheads). Most somatic cells are undergoing mitosis (blue) in the germarium and during the early stages of oogenesis. Cells in mitosis are labeled with an antibody against phosphorylated Histone 3 (blue). (C) Early stages of oogenesis. Vasa protein (red) continues to concentrate in juxtanuclear aggregates (arrowhead). In (B) and (C) nuclear counter stain (YO-PRO-3 iodide) is in green. Scale bars: (B) 80 um, (C) 40 um.
Thermobia domestica. Anterior is up in (A), and to the left in (B) and (C). (A) Adult female firebrat. (B) Germarium (g) and early stages of oogenesis in an adult female ovary. The panoistic ovaries of T. domestica are organised in a similar manner to those of S. gregaria, but with only five ovarioles per ovary. Vasa protein (red) is concentrated in asymmetric aggregates next to the nuclei of early oocytes (arrowheads). (C) At later stages of oogenesis, Vasa protein (red) continues to concentrate in asymmetric juxtanuclear aggregates (arrowhead). High concentrations of Vasa are also seen in the cortical ooplasm. In (B) and (C) nuclear counter stain (YO-PRO-3 iodide) is in green. Scale bars: (B) and (C) 40 um.
Oncopeltus fasciatus. Anterior is up in all panels. (A) Adult female milkweed bug with freshly laid eggs (top right). (B) Germarium (g) and early stages of oogenesis in an adult female ovary. The ovaries of O. fasciatus contain seven ovarioles per ovary, and like those of Drosophila melanogaster, are meroistic (containing nurse cells). However, unlike D. melanogaster, nurse cells are shared between all oocytes of an ovariole. All nurse cells are located in the germarium, liked by cytoplasmic bridges to a pool of common cytoplasm called the trophic core. Tubes called trophic cords (arrowheads) originate in trophic core, thus linking each oocyte to the nurse cells. Vasa (red) protein is detected in cells of the germarium and within the trophic cords (arrowheads). The actin cytoskeleton (blue) is concentrated along the sides of the trophic core and around the oocyte cortex. (C) At later stages of oogenesis, Vasa protein (red) is distributed in the ooplasm but excluded from the oocyte nucleus (on). The actin cytoskeleton (blue) has become further enriched at the cortical ooplasm. (D) As oogenesis progresses, Vasa protein (red) disappears from the central ooplasm, but remains at the oocyte cortex, with highest levels at the posterior pole. In (B) through (D), Vasa protein is in red, F-actin (marked with Alexa 647-conjugated phalloidin) is in blue, and nuclear counter stain (YO-PRO-3 iodide) is in green. Scale bars: (B) 80 um, (C) 40 um, (D) 80 um.
Artemia franciscana. Anterior is up in all panels. (A) Adult female carrying eggs in brood pouch (arrow). (B) 8 day nauplius larva stained with cross-reacting Vasa antibody. Vasa is detected specifically in germ cells (arrowheads), which lie in the segmental mesoderm lateral to the gut (g). (C) Close up of (B). Vasa protein is cytoplasmic in germ cells (arrowheads).
Parhyale hawaiensis. Anterior is to the right in all panels. (A) Adult female carrying embryos in brood pouch (arrowheads). Natural purplish colour of developing oocytes can be seen in the ovary (arrows). (B) Late germ band stage embryo stained with cross-reacting Vasa antibody (pink). Vasa is detected in a group of cells among the endomesodermal precursors, posterior to the head lobes (hl). These cells are the likely descendants of the a micromere of the 8 cell stage, which has been shown to be the unique germ cell precursor by cell lineage studies. p: posterior. (C) Anterior end of an adult ovary with mature oocytes. The paired ovaries of P. hawaiensis are organised with a strand of primordial germ cells (PGCs) along the dorsal midline (towards top of panel). There are no nurse cells. Maturing oocytes move laterally in the ovary, so that the most mature oocytes are located farthest away from the PGCs (towards bottom of panel). The oocytes are surrounded by a monolayer of somatic follicle cells. Vasa protein (red) is concentrated in a perinuclear ring around the oocyte nucleus. (D) Orthogonal sections through an adult testis. The testes of P. hawaiensis have a compartmental organization, with PGCs lying along the dorsal midline (bottom of panel). Maturing spermatocytes move laterally in the testis, so that the most advanced spermatocytes are farthest away from the PGCs (towards top of panel). Spermatids and later spermatogenic stages are located in a long tube extending posterior from the testis (not shown). Green lines indicate planes of orthogonal sectioning. Vasa protein (red) is concentrated in primordial germ cells (PGCs) in the medial "pgc zone" (pgc). Primary spermatocytes (region 1) and secondary spermatocytes (region 2) are located laterally to the PGCs, and do not have high levels of Vasa protein. Orthogonal section panels (bottom, right) indicate intensity of Vasa (red) and nuclear (blue) staining. Spikes of red next to high levels of blue indicate perinuclear distribution of Vasa protein. Nuclei are counterstained with TO-PRO-3 iodide (blue) in (B), (C) and (D).