Synthesis of two Drosophila male accessory gland proteins and their fate after transfer to the female during mating

Abstract Drosophila males, like males of most other insects, transfer a group of specific proteins to the females during mating. These proteins are produced primarily in the accessory gland and are likely to influence the female's reproduction. The results of studies of DNA sequence polymorphism and divergence in two genes coding for male accessory gland proteins of Drosophila are reported here. The Mst26Aa and Mst26Ab transcription units are tandemly arranged in a approximately 1.6-kb segment in Drosophila sechellia, Drosophila mauritiana and Drosophila simulans as they were reported to be in Drosophila melanogaster. The DNA sequences of 10 alleles from D. melanogaster and one allele each from the three sibling species reveals a high degree of amino acid replacement variation. A substantial part of the variation is due to insertion/deletion differences. Possible functional significance of these amino acid sequence changes is discussed. Statistical analyses based on the neutral theory of molecular evolution show that the distribution of polymorphism over the 1.6-kb region is inconsistent with the pattern of divergence between the species. The amount of 4-cutter restriction map polymorphism in a larger sample of 75 alleles from the same D. melanogaster population is similar to that obtained from the DNA sequence of the 10 alleles (a pairwise average of 0.007 difference per site). The 6-cutter restriction map survey of a 18-kb region containing the Mst26A genes indicates that polymorphism in the region flanking these genes maybe higher. The failure of polymorphisms and divergence in the Mst26A region to conform to the expectations of a simple mutation-drift-equilibrium model indicates that selection in or near this region has played a role in the history of these genes.

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Molecular Population Genetics of Male Accessory Gland Proteins in Drosophila

Genetics ◽

10.1093/genetics/156.4.1879 ◽

2000 ◽

Vol 156 (4) ◽

pp. 1879-1888 ◽

Cited By ~ 5

Author(s):

David J Begun ◽

Penn Whitley ◽

Bridget L Todd ◽

Heidi M Waldrip-Dail ◽

Andrew G Clark

Keyword(s):

Population Genetics ◽

Accessory Gland ◽

High Rate ◽

Sequence Evolution ◽

Male Accessory Gland ◽

Accessory Gland Proteins ◽

Molecular Sequence ◽

Molecular Population Genetics ◽

Genes Encoding ◽

Interspecific Comparisons

Abstract Drosophila seminal proteins have an unusually high rate of molecular sequence evolution, suggesting either a high rate of neutral substitution or rapid adaptive evolution. To further quantify patterns of polymorphism and divergence in genes encoding seminal proteins, also called accessory gland proteins (Acp’s), we conducted a sequencing survey of 10 Acp genes in samples of Drosophila melanogaster and D. simulans (Acp29AB, Acp32CD, Acp33A, Acp36DE, Acp53Ea, Acp62F, Acp63F, Acp76A, Acp95EF, and Acp98AB). Mean heterozygosity at replacement sites in D. simulans was 0.0074 for Acp genes and 0.0013 for a set of 19 non-Acp genes, and mean melanogaster-simulans divergence at replacement sites was 0.0497 for Acp genes and 0.0107 at non-Acp genes. The elevated divergence of Acp genes is thus accompanied by elevated within-species polymorphism. In addition to the already-reported departures of Acp26A, Acp29AB, and Acp70A from neutrality, our data reject neutrality at Acp29AB and Acp36DE in the direction of excess replacements in interspecific comparisons.

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Different Forces Drive the Evolution of the Acp26Aa and Acp26Ab Accessory Gland Genes in the Drosophila melanogaster Species Complex

Genetics ◽

10.1093/genetics/150.3.1079 ◽

1998 ◽

Vol 150 (3) ◽

pp. 1079-1089 ◽

Cited By ~ 15

Author(s):

Montserrat Aguadé

Keyword(s):

North Carolina ◽

Drosophila Melanogaster ◽

Species Complex ◽

Amino Acid Replacement ◽

Accessory Gland ◽

Male Accessory Gland ◽

Accessory Gland Proteins ◽

Length Polymorphisms ◽

African Populations ◽

Nonsynonymous Variation

Abstract The Acp26Aa and Acp26Ab genes that code for male accessory gland proteins are tandemly arranged in the species of the Drosophila melanogaster complex. An ∼1.6-kb region encompassing both genes has been sequenced in 10, 24, and 18 lines from Spain, Ivory Coast, and Malawi, respectively; the previously studied 10 lines from North Carolina have also been included in the analyses. A total of 110 nucleotide and 4 length polymorphisms were detected. Silent variation for the whole Acp26A region was slightly higher in African than in non-African populations, while for both genes nonsynonymous variation was similar in all populations studied. Based on Fst estimates no major genetic differentiation was detected between East and West Africa, while in general non-African populations were strongly differentiated from both African populations. Comparison of polymorphism and divergence at synonymous and nonsynonymous sites revealed that directional selection acting on amino acid replacement changes has driven the evolution of the Acp26Aa protein in the last 2.5 myr.

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Male accessory gland proteins induce female monogamy in anopheline mosquitoes

Medical and Veterinary Entomology ◽

10.1111/j.1365-2915.2009.00849.x ◽

2010 ◽

Vol 24 (1) ◽

pp. 91-94 ◽

Cited By ~ 31

Author(s):

B. SHUTT ◽

L. STABLES ◽

F. ABOAGYE-ANTWI ◽

J. MORAN ◽

F. TRIPET

Keyword(s):

Accessory Gland ◽

Male Accessory Gland ◽

Accessory Gland Proteins ◽

Anopheline Mosquitoes

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Post-mating Gene Expression Profiles of FemaleDrosophila melanogasterin Response to Time and to Four Male Accessory Gland Proteins

Genetics ◽

10.1534/genetics.108.086934 ◽

2008 ◽

Vol 179 (3) ◽

pp. 1395-1408 ◽

Cited By ~ 89

Author(s):

Lisa A. McGraw ◽

Andrew G. Clark ◽

Mariana F. Wolfner

Keyword(s):

Gene Expression ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Accessory Gland ◽

Male Accessory Gland ◽

Accessory Gland Proteins

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Fates and targets of male accessory gland proteins in mated female Drosophila melanogaster

Insect Biochemistry and Molecular Biology ◽

10.1016/j.ibmb.2005.05.001 ◽

2005 ◽

Vol 35 (9) ◽

pp. 1059-1071 ◽

Cited By ~ 94

Author(s):

K. Ravi Ram ◽

S. Ji ◽

M.F. Wolfner

Keyword(s):

Drosophila Melanogaster ◽

Accessory Gland ◽

Male Accessory Gland ◽

Accessory Gland Proteins ◽

Mated Female

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New Genes for Male Accessory Gland Proteins in Drosophila melanogaster

Insect Biochemistry and Molecular Biology ◽

10.1016/s0965-1748(97)00056-8 ◽

1997 ◽

Vol 27 (10) ◽

pp. 825-834 ◽

Cited By ~ 74

Author(s):

Mariana F. Wolfner ◽

Heidi A. Harada ◽

Michael J. Bertram ◽

Thomas J. Stelick ◽

Kevin W. Kraus ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Accessory Gland ◽

Male Accessory Gland ◽

Accessory Gland Proteins ◽

New Genes

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FACS-based isolation and RNA extraction of Secondary Cells from the Drosophila male Accessory Gland

10.1101/630335 ◽

2019 ◽

Author(s):

Clément Immarigeon ◽

François Karch ◽

Robert K. Maeda

Keyword(s):

Cell Population ◽

Seminal Fluid ◽

Rna Extraction ◽

Cell Types ◽

Accessory Gland ◽

Specific Cell ◽

Male Accessory Gland ◽

Accessory Gland Proteins ◽

Accessory Glands ◽

Study Gene Expression

ABSTRACTTo appreciate the function of an organ, it is often critical to understand the role of rare cell populations. Unfortunately, this rarity often makes it difficult to obtain material for study. This is the case for the Drosophila male accessory gland, the functional homolog of mammalian prostate and seminal vesicle. In Drosophila, this gland is made up of two morphologically distinct cell types: the polygonally-shaped main cells, which compose 96% of the organ, and the larger, vacuole-containing secondary cells (SCs), which represent the remaining 4% of cells (~40 cells per lobe). Both cell types are known to produce accessory gland proteins (Acps), which are important components of the seminal fluid and are responsible for triggering multiple physiological and behavioral processes in females, collectively called the post-mating response (PMR). While a few genes are known to be specific to the SCs, the relative rarity of SCs has hindered the study of their whole transcriptome. Here, a method allowing for the isolation of SCs is presented, enabling the extraction and sequencing of RNAs from this rare cell population. The protocol consists of dissection, protease digestion and mechanical dissociation of the glands to obtain individual cells. Then, the cells are sorted by FACS, and living GFP-expressing SC singulets are isolated for RNA extraction. This procedure is able to provide SC-specific RNAs from ~40 males per condition in the course of one day. Given the speed and low number of flies required, this method enables the use of downstream RT-qPCR and/or RNA sequencing to the study gene expression in the SCs from different genetic backgrounds, ages, mating statuses or environmental conditions.SUMMARYHere, we describe the dissociation and sorting of a specific cell population from the Drosophila male accessory glands (Secondary cells), followed by RNA extraction for sequencing and RT-qPCR. The dissociation consists of dissection, proteases digestion and mechanical dispersion, followed by FACS purification of GFP-expressing cells.

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