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First published online October 5, 2007
Journal of Experimental Biology 210, 3601-3606 (2007)
Published by The Company of Biologists 2007
doi: 10.1242/jeb.009035

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Male accessory glands of Drosophila melanogaster make a secreted angiotensin I-converting enzyme (ANCE), suggesting a role for the peptide-processing enzyme in seminal fluid

Caroline M. Rylett^1,*, Michael J. Walker^1,*, Gareth J. Howell², Alan D. Shirras³ and R. Elwyn Isaac^1,

¹ Institute of Integrative and Comparative Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK
² Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK
³ Department of Biological Sciences, Lancaster University, Lancaster, LA1 4YQ, UK

View larger version (26K): [in this window] [in a new window]   Fig. 1. Western blot analysis shows the presence of ANCE, but not ACER in male accessory glands (AGs). Western blots of male AGs using anti-ANCE antibodies (lanes 1 and 2) and anti-ACER antibodies (lanes 3 and 4). Lane 1, protein from 10 AGs; lane 2, 0.5 ng of recombinant ANCE; lane 3, protein from 10 AGs; lane 4, recombinant ACER, 2 ng.

View larger version (54K): [in this window] [in a new window]   Fig. 2. (A) Immunohistochemical staining of ANCE in the secondary cells of the distal region of the male accessory gland using ANCE-specific primary antiserum and the ABC detection kit. (B) Control experiment using pre-immune serum.

View larger version (35K): [in this window] [in a new window]   Fig. 3. ANCE is localised to the giant vesicles of the secondary cells of the male accessory gland (AG). (A) Immunofluorescence staining of ANCE in the secondary cells of the distal region of the male AG using ANCE-specific primary antiserum and FITC-conjugated secondary antibodies. Giant vesicles are numbered clockwise and the two nuclei (N) are labelled. (B) Three-dimensional representation of the vesicles (red) in the secondary cell (green) of the AG. Serial confocal sections were collected through the sample, with an optical depth of 1 µm. One of these sections is shown. (C) Electron micrograph of a giant vesicle of an AG secondary cell showing the dense core material (star) and filaments (f). Scale bar, 2 µm.

View larger version (95K): [in this window] [in a new window]   Fig. 4. Ance is transcribed in the secondary cells of the male accessory gland (AG). In situ hybridisation using digoxigenin-labelled Ance antisense riboprobe (A,C) and sense strand control (B,D) reveals higher levels of mRNA in the large secondary cells (arrow points to one of several secondary cells in the field of view). C and D are higher magnification images of secondary cells and AG lumen. Powerpoint was used to manipulate all the images equally to increase contrast. Scale bars, 10 µm.

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