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. 2017 Mar;23(3):284–296. doi: 10.1261/rna.059527.116

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© 2017 Lin et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 4. — Tailor and Dis3L2 are required for fertility and gametogenesis, particularly in males. We tested the phenotypic characteristics of Tailor, Dis3l2, and double mutant animals. We analyzed two trans-heterozygous combinations of Tailor, homozygous and hemizygous combinations of Dis3l2, and double mutants. (A) Quantification of individual female fertility. Tailor and Dis3L2 mutants are subfertile, but the double mutants are nearly fully sterile. (B) Quantification of individual male fertility. Tailor mutants are subfertile, while Dis3L2 single or double mutants are nearly completely sterile. (C–J) Immunostaining of 5-d-old testes, using DAPI to label DNA (in gray), anti-myosin VI (in green), phalloidin to label F-actin (in red), and anti-activated-caspase-3 (in blue). (C) Control w[1118] testis. The 64 haploid meiotic products remain together in a cyst and their cytoplasmic contents are extruded synchronously and form a cystic bulge (CB), they detach from the distal sperm tails to form a wastebag (WB). (D) Tailor trans-heterozygous mutant exhibits cystic bulges, but these exhibit mildly scattered actin cones. (E) Dis3L2 hemizygous mutant completely lacks cystic bulges; all actin cones are separate and only reach the middle portion of the testis. (F) Tailor, Dis3l2 double mutant is similar to the Dis3l2 single mutant. (G–J) Close-ups of individualization complexes in which actin cones are normally well organized within a cystic bulge (G, control) and mildly scattered in Tailor mutants (H). Dis3L2 single (I) or Tailor, Dis3L2 double (J) mutants lack cystic bulges and present only scattered actin cones in the testis basal region. (K) Quantification of ICs and wastebags in Tailor and Dis3L2 single and double mutants. (L–S) Seminal vesicles stained for DAPI; the sperm bear needle-shaped nuclei. Compared to control (L,M), Tailor mutant has a reduced seminal vesicle (N,O), while Dis3L2 (P,Q) mutant, and Tailor, Dis3L2 (R,S) double mutants bear empty seminal vesicles lacking sperm.