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First published online 12 December 2007
doi: 10.1242/dev.015156

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The Sf1-related nuclear hormone receptor Hr39 regulates Drosophila female reproductive tract development and function

Howard Hughes Medical Institute, Department of Embryology, Carnegie Institution, 3520 San Martin Drive, Baltimore, MD 21218, USA.

View larger version (52K): [in this window] [in a new window]   Fig. 1. Hr39 mutants alter spermathecal and parovarial development. (A) The female lower reproductive tract (RT). (B) Hr39 transcript structure showing the four different isoforms with insertion mutations indicted above. (C-G) Nomarski images of RT. All RT images possess a single seminal receptacle (asterisk). (C) Wild-type RT with two spermathecae and two parovaria (one out of focus). (D) Hr3903508 RT with no spermatheca or parovarium. (E,F) Magnifications of parovaria (circled); (E) wild type, (F) Hr3904443. (G) Hr3904443 RT with three spermathecae and two parovaria. (H) Percentage of homozygous females of each genotype possessing 0, 1, 2 or 3 spermathecae. Arrows, spermathecae; arrowheads, parovaria; asterisks, seminal receptacles. Scale bar: 100 µm in C,D,G.

View larger version (97K): [in this window] [in a new window]   Fig. 2. Hr39 expression. (A) Hr39 expression levels in mutants using RT-PCR. As an internal control, RpS17 was amplified. (B) Comparison of Hr39 expression among strains using real-time quantitative RT-PCR. Standard deviation of each triplicate (±s.d.) is shown. (C-E,L-N) Green indicates Hr39-lacZ detected via anti-β-gal antibody. Magenta indicates DNA. (F-K) Hr39 in situ hybridization (green) (C) Larval ring gland, Hr3903508/+. Blue indicates anti-Fas2. Scale bar: 75 µm. (D) Female genital disc, Hr3903508/+. Scale bar: 50 µm. (E) Female larval gonad, Hr3907154/+. Scale bar: 50 µm. Wild type (F-J); Hr3904443 (K,L). (F) Reproductive tract; (G) parovaria; (H) seminal receptacle; (I) spermathecae-associated fat body; (J-L) spermathecae. (M) Hr3903508/+ ovariole. Scale bar: 100 µm. (N) Hr3903508/+ testis. Scale bar: 100 µm. SR, seminal receptacle; SP, spermatheca; P, parovarium; O, oviduct; FB, fat body.

View larger version (54K): [in this window] [in a new window]   Fig. 3. An Hr39 cDNA rescues Hr39 mutations and expands spermathecal number. Nomarski images of adult female reproductive tracts of different genotypes. (A) yw; hsp70GAL4::UAS-Hr39(7-3) with three spermathecae (SP). (B) Hr39ly92; hsp70GAL4::UAS-Hr39(7-5) with one SP. (C) Hr3907154; hsp70GAL4::UAS-Hr39(7-5) with two abnormal SP. (D) Hr3903508; hsp70GAL4::UAS-Hr39(7-5) with two SP. (E) Hr3903508; hsp70GAL4::UAS-Hr39(7-3) with three SP. (F) Hr3903508; hsp70GAL4::UAS-Hr39(7-3) with four SP. Red arrows indicate SP.

View larger version (67K): [in this window] [in a new window]   Fig. 4. Hr39 controls spermathecal secretion. Electron microscopy images of wild-type and Hr39 mutant spermathecae (SP) and parovaria (P). (A-D) Wild-type SP. (A) Gland cells (red outline) surround the lumen (L). N, nucleus; EA, end apparatus. Scale bar: 10 µm. (B) SP lumen with sperm (S) surrounded by extracellular material (M). Scale bar: 2 µm. (C) EA in active state showing secretion. Scale bar: 2 µm. (D) EA (white outline) showing little secretion among the microvilli. Scale bar: 2 µm. (E) Wild-type parovaria with little secretion. Scale bar: 10 µm. (F) Hr3907154 SP containing less secretory product than wild type (C). Scale bar: 2 µm. Red arrows indicate SP secretion; L, lumen; M, luminal material; S, sperm.

View larger version (91K): [in this window] [in a new window]   Fig. 5. Hr3904443 spermathecae lack secretory material and parovaria contain extra secretory material. (A-C) Hr3904443 spermathecae (SP). (A) Sperm remain in lumen (L) but cell death (asterisk) is increased; gland cell, red outline. Scale bar: 10 µm. (B) Lumen with sperm (S) surrounded by extracellular material (M). Scale bar: 2 µm. (C) Secretion appears absent from end apparatus (EA). Scale bar: 1 µm. (D) Hr3904443 parovaria. Secretion (arrows) appears to fill the end apparati (EA). Scale bar: 10 µm. N, nucleus.