Egg chamber development and eggshell patterning is disrupted in dPPCS1/1. (A) Morphology of dPPCS1/1 ovaries was analyzed and compared with wt using light microscopy. (Aa) Bright field microscopy revealed that at 48 h after eclosion (AE), Wt ovaries are well developed and contain mature eggs (arrowheads). (Ab) At 48 h after eclosion, dPPCS1/1 ovaries were small in size compared to wt. (Ac-e) Ovaries were labeled with rhodamin-phalliodin to detect F-actin and stained with DAPI to visualize DNA. (Ac) At 72 h AE, wt ovaries contain vitellogenic egg chambers, as determined by the increased size of the oocyte compartment (asterisk). (Ad) dPPCS1/1 ovaries remained small in size (1 entire lobe is shown) and no vitellogenic egg chambers were observed. (Ae) At 120 h AE, dPPCS1/1 ovaries contained vitellogenic egg chambers (asterisks) and exhibited features of degenerating egg chambers (arrowheads). The 2 lobes were frequently different in size. (B) Chorion patterning was analysed in dPPCS1/1. (Ba, Bd) Wt embryos have 2 dorsal appendages. (mp = micropyle; p = paddle; s = stalk). (Bb-c, Be-f) Embryos deposited by dPPCS1/1 mothers showed a dumpless phenotype and had a wide range of patterning defects, which were classified in 5 groups: (Bb) embryos with opercula positioned in a different angle in relation to the stalks (bracket, compare with Ba) and 4 appendages; (Bc) abnormal stalks (arrows); (Be) fused appendages (bracket, compare with Bd); and (Bf) embryos without dorsal appendages. Percentages are indicated (n = 142). The remaining 22.8% had either 2 dorsal appendages of different lengths, missing paddles, or a shift of the dorsal appendages posteriorly. Scale bars: 500 μm (Aa-b), 150 μm (Ac-e).