Male x Female Protein Interactions Mediating Reproductive Success in the Drosophila Mating Plug
Project Number5F32HD111231-02
Former Number1F32HD111231-01
Contact PI/Project LeaderCARLISLE, JOLIE ANNA
Awardee OrganizationCORNELL UNIVERSITY
Description
Abstract Text
PROJECT SUMMARY
In both mammals and invertebrates, initiation of post-mating responses in the female post-copulation is known
to contribute to reproductive success. Seminal fluid proteins (Sfps) have been shown to initiate many of these
responses. However, male x female interactions are underexplored as contributors to infertility. Investigating
the mechanisms and evolution of male x female interactions is critical for understanding the complexities of
reproduction. I will use D. melanogaster as a model system for investigating post-copulatory male x female
interactions via characterization of male and female contributions to the Drosophila mating plug’s (MP)
formation and ejection. In Drosophila, the MP forms in the uterus of the female via coagulation of male-
ejaculated Sfps and some female reproductive tract proteins. Genetic disruption of the MP impacts
reproductive outcomes by affecting sperm retention; however, little is known about the male and female
proteins (and their interactions) that modulate mating plug ejection (MPE) rates either directly by contributing to
MP composition/degradation or indirectly by regulating female ejection behavior. In Aims 1 & 2 I will
respectively use female or male phenotypic variation in the Drosophila Genomic Reference Panel to
perform a GWAS on MPE timing. I will then functionally investigate how top gene candidates mediate MPE
timing. I have concluded a GWAS on female MPE timing and have so far identified 4 neuronal genes that
regulate female MPE (Aim 1). Using highly tissue- and cell type- specific knockdown of gene candidate
expression via RNA interference, I will more deeply characterize neuronal regulation of female MPE timing.
After also identifying and characterizing male genes regulating MPE timing (Aim2) I will perform a 6x6 grid
cross of males and females with disrupted function of MPE genes to investigate the presence of
complex non-additive male x female MPE interactions. Finally, I will investigate the molecular evolution and
function of male and female genes contributing to MP composition (Aim 3). I discovered that many male and
female MP genes are closely paralogous to each other and evolving under positive selection,
potentially suggestive of evolution under sexual conflict. After fully characterizing the evolution and
coevolution of these male and female MP paralogs, I will investigate their sex-specific functions in MPE
and MP formation. Observation of opposing functions for paralogous male and female MP genes would point
to sexual conflict driven evolution caused by opposing sex-specific optimal mating strategies. Observation of
complementary functions would indicate similar evolutionary pressures acting on male and female paralogs to
cooperatively ensure optimal reproductive outcomes.
Public Health Relevance Statement
PROJECT NARRATIVE
Male x Female interactions are necessary for reproduction, however, an understanding of the
intersexual interactions mediating reproductive phenotypes is still in its nascent stages. By
harnessing genetic variation from within and between Drosophila species and the
genetic and genomic resources available in D. melanogaster, I will disentangle the
contributions from both male and female flies to the mechanisms of mating plug
formation and ejection and characterize the potential coevolution of male and female MP
interaction partners. My results will lead to an understanding of the intersexual interactions
that mediate reproduction and how they are shaped evolutionarily.
Eunice Kennedy Shriver National Institute of Child Health and Human Development
CFDA Code
865
DUNS Number
872612445
UEI
G56PUALJ3KT5
CCV3WG2JG248
D4H1NV4APKP3
ELS2M3C6V2S5
EQA8NBEN9WD5
FFAZGE9NH3M8
K6JRCJJXFET1
M8FBSLHASMT3
P4LRVQT1H4K5
PJUVN8AT5416
RT1JPM9UMGM5
ZBMGUAZYFGC4
ZMP8BDLJTUW9
Project Start Date
01-January-2024
Project End Date
31-December-2026
Budget Start Date
01-January-2025
Budget End Date
31-December-2025
Project Funding Information for 2025
Total Funding
$74,284
Direct Costs
$74,284
Indirect Costs
Year
Funding IC
FY Total Cost by IC
2025
Eunice Kennedy Shriver National Institute of Child Health and Human Development
$74,284
Year
Funding IC
FY Total Cost by IC
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