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

Development 135, 311-321 (2008)
Published by The Company of Biologists 2008
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The Sf1-related nuclear hormone receptor Hr39 regulates Drosophila female reproductive tract development and function

Anna K. Allen and Allan C. Spradling*

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


Figure 1
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  		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.

 

Figure 2
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  		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.

 

Figure 3
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  		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.

 

Figure 4
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  		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.

 

Figure 5
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  		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.

 

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