Figure 5

H. robusta. Whole mount in situ hybridizations stained with NBT/BCIP. The staining in the pharynx is background due the influx of RNA probe into the mouth cavity. A. Live image of a H. robusta TX individual to show position of pigmented eyes. B. Expression of gq-protein in extraocular PRCs in H. robusta CA. C. Negative control. D. Expression of H. robusta TX opsin B in extraocular PRCs. E’E”. Vertical section of LSM stacks with reflection of the NBT/BCIP staining (red) and nuclei stained by DAPI (blue) to show that the expression is restricted to the cell body and present neither in the microvilli nor in the nucleus. F. Higher magnification of boxed area from D. Darker and lighter colours of the staining in adjacent phaosomal PRCs are caused by the focus levels. The phaosomal cavities are visible as lighter areas. G. Z-projection of cLSM stack showing one PRC. Note the dark phaosomal cavity without any signal. Reflection of the NBT/BCIP staining (red (blue). H. TEM micrograph of a similar single phaosomal PRC. The phaosomal cavity completely filled with sensory microvilli corresponding to the dark central space in G. - A-D, F Bright field images. E+G confocal laser scanning microscope images. arrows: extraocular phaosomal PRC; asterisks: pigmented eyes; psc pigmented supporting cell; mo mouth opening; phc phaosomal cavity; n nucleus; smv sensory microvilli.