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First published online 23 April 2008
doi: 10.1242/dev.021949

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Post-meiotic transcription in Drosophila testes

Carine Barreau, Elizabeth Benson, Elin Gudmannsdottir, Fay Newton and Helen White-Cooper^*,

Department of Zoology, University of Oxford, South Parks Road, Oxford OX1 3PS, UK.

View larger version (72K): [in this window] [in a new window]   Fig. 1. RNA in situ hybridisation reveals comet and cup transcript patterns. (A) Drosophila testis viewed by phase contrast. Asterisk, apical tip; bracket, spermatocytes; arrowheads, early spermatids; arrows, elongating spermatid bundles. (B-D) Control genes. (E,F) Typical cup and comet localisation. Scale bar: 100 µm for to A-F. (G-J) Typical cup localisation at spermatid distal ends. Scale bar: 25 µm. (K) Diagram of a spermatid bundle showing the location of comet and cup transcripts relative to spermatid nuclei, and cyst cells (grey).

View larger version (74K): [in this window] [in a new window]   Fig. 2. Q-RT-PCR confirms post-meiotic transcription. (A) Individual cyst Q-RT-PCR assay schematic. (B) Cysts used for RNA extraction, from primary spermatocytes (I1, I2) and round spermatids (R1, R2; scale bars: 25 µm), and from elongating spermatids (0.19-1.89 mm; scale bars: 100 µm). (C,D) Control Q-RT-PCRs. (E-H) Peak of expression in spermatids confirms post-meiotic transcription of comet and cup genes. hale (E) and schuy (F) were expressed in more bundles than were d-cup (G) and sunz (H).

View larger version (34K): [in this window] [in a new window]   Fig. 3. Comet and cup transcription precedes the histone-protamine switch. Bars above graphs and colours within graphs indicate H2A-mRFP (histone, red) and Mst35Ba-EGFP (protamine, green; overlap orange/yellow) detection. (A-C) Control Q-RT-PCRs. (D-J) Comet and cup Q-RT-PCR expression profiles show that expression initiates in cysts negative for protamine. (K) Nuclear fluorescence in elongating cysts.

View larger version (61K): [in this window] [in a new window]   Fig. 4. soti is required for normal individualisation complex progression. Confocal maximum brightness projections of the entire cyst depth. DNA is labelled with propidium iodide (A-D, magenta); actin is labelled with FITC-phalloidin (A'-D', green). Scale bar: 10 µm. (A-B'') In wild-type testes, investment cones assembled above spermatid nuclei (A-A'') and moved along spermatid bundles as a complex (B-B''). (C-D'') In soti mutant testes, investment cones assembled normally, but complexes dissociated during individualisation.

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