Figure 5.
Oys and Nes are required for individualization and are expressed in the germline. (A) oysΔ6 nesΔ52 mutant males (0–5 d old) have an average of 2 normal individualization structures per testis (n = 20), while precise excision controls (revertant, “rev”) have more than 8 (n = 31). Males homozygous for either oysΔ67 or nesΔ52 and heterozygous for the other also have more than 8 normal individualization structures per testis on average (n ≥ 24 each). (B–E) oys nes mutant testes have scattered ICs (B, arrowheads, phalloidin, green, close-up in D) and waste bags devoid of actin cones (E). The average number of abnormal ICs in the oys nes mutant (4.9, n = 22) is significantly increased compared to the revertant control (0.36, n = 11), p <0.001 (C). (F–G) RNA in situ hybridization to endogenous oys (F) or nes (G) in w1118 testes shows expression in the germline beginning in premeiotic spermatocytes (blue). Endogenous transcripts are not detected in the germinal proliferation center (black arrowheads). (H-I) Phase contrast microscopy of testis squashes reveals no cytokinesis defects in either oysΔ67nesΔ52 homozygotes (H) or sibling heterozygotes (I, “bal” = SM6-TM6B balancer). Each phase dark Nebenkern has one associated phase light nucleus.