Delineating the roles of males and females in sperm competition

Disentangling the relative roles of males, females and their interactive effects on competitive fertilization success remains a challenge in sperm competition. In this study, we apply a novel experimental framework to an ideally suited externally fertilizing model system in order to delineate these roles. We focus on the chinook salmon, Oncorhynchus tshawytscha, a species in which ovarian fluid (OF) has been implicated as a potential arbiter of cryptic female choice for genetically compatible mates. We evaluated this predicted sexually selected function of OF using a series of factorial competitive fertilization trials. Our design involved a series of 10 factorial crosses, each involving two ‘focal’ rival males whose sperm competed against those from a single ‘standardized’ (non-focal) rival for a genetically uniform set of eggs in the presence of OF from two focal females. This design enabled us to attribute variation in competitive fertilization success among focal males, females (OF) and their interacting effects, while controlling for variation attributable to differences in the sperm competitive ability of rival males, and male-by-female genotypic interactions. Using this experimental framework, we found that variation in sperm competitiveness could be attributed exclusively to differences in the sperm competitive ability of focal males, a conclusion supported by subsequent analyses revealing that variation in sperm swimming velocity predicts paternity success. Together, these findings provide evidence that variation in paternity success can be attributed to intrinsic differences in the sperm competitive ability of rival males, and reveal that sperm swimming velocity is a key target of sexual selection

Data from: Delineating the roles of males and females in sperm competition

Disentangling the relative roles of males, females and their interactive effects on competitive fertilization success remains a challenge in sperm competition. In this study, we apply a novel experimental framework to an ideally suited externally fertilizing model system in order to delineate these roles. We focus on the chinook salmon, Oncorhynchus tshawytscha, a species in which ovarian fluid (OF) has been implicated as a potential arbiter of cryptic female choice for genetically compatible mates. We evaluated this predicted sexually selected function of OF using a series of factorial competitive fertilization trials. Our design involved a series of 10 factorial crosses, each involving two ‘focal’ rival males whose sperm competed against those from a single ‘standardized’ (non-focal) rival for a genetically uniform set of eggs in the presence of OF from two focal females. This design enabled us to attribute variation in competitive fertilization success among focal males, females (OF) and their interacting effects, while controlling for variation attributable to differences in the sperm competitive ability of rival males, and male-by-female genotypic interactions. Using this experimental framework, we found that variation in sperm competitiveness could be attributed exclusively to differences in the sperm competitive ability of focal males, a conclusion supported by subsequent analyses revealing that variation in sperm swimming velocity predicts paternity success. Together, these findings provide evidence that variation in paternity success can be attributed to intrinsic differences in the sperm competitive ability of rival males, and reveal that sperm swimming velocity is a key target of sexual selection

Sperm traits in Chinook salmon depend upon activation medium: implications for studies of sperm competition in fishes

Sperm traits of externally fertilizing fish species are typically measured in fresh (or salt) water, even though the spawning environment of their ova contains ovarian fluid. In this study, we measured sperm traits of Chinook salmon ( Oncorhynchus tshawytscha (Walbaum in Artedi, 1792)) in both fresh water and dilute ovarian fluid at 10 and 20 s postactivation, using a computer-assisted sperm analysis system. Spermatozoa swam faster, and had both higher percent motility and a straighter path trajectory for a longer period of forward motility when activated in ovarian fluid compared with activation in fresh water. Comparing sperm activity of 10 males in water versus ovarian fluid, we found a weak but significant correlation for sperm swimming speed at 10 s postactivation (r = 0.34, p = 0.01), but not for any other sperm traits measured. Most important, across males, mean sperm swimming speed in water accounted for &lt;10% of the observed variation in mean sperm swimming speed in ovarian fluid. Thus, we argue that sperm traits measured in fresh water are not particularly relevant to those same traits during normal spawning in this species. We suggest that sperm performance measured in fresh water should be used with caution when comparing the potential for individual males to fertilize ova, especially in studies of sperm competition in externally fertilizing species.</jats:p

Data for Evans et al. 2013

Contains raw data used to test for interacting sperm-ovarian fluid interactions on sperm motility (sheet 1) and for partitioning variance in sperm competitiveness (sheet 2

Proteomic Analysis of Chinook Salmon (<i>Oncorhynchus tshawytscha</i>) Ovarian Fluid

<div><p>The ovarian, or coelomic, fluid that is released with the egg mass of many fishes is increasingly found to play an important role in several biological processes crucial for reproductive success. These include maintenance of oocyte fertility and developmental competence, prolonging of sperm motility, and enhancing sperm swimming speed. Here we examined if and how the proteome of chinook salmon (<i>Oncorhynchus tshawytscha</i>) ovarian fluid varied among females and then sought to examine the composition of this fluid. Ovarian fluid in chinook salmon was analyzed using 1D SDS PAGE and LC-MS/MS tryptic digest screened against Mascot and Sequest databases. We found marked differences in the number and concentrations of proteins in salmon ovarian fluid across different females. A total of 174 proteins were identified in ovarian fluid, 47 of which were represented by six or more peptides, belonging to one of six Gene Ontology pathways. The <i>response to chemical stimulus</i> and <i>response to hypoxia</i> pathways were best represented, accounting for 26 of the 174 proteins. The current data set provides a resource that furthers our understanding of those factors that influence successful egg production and fertilisation in salmonids and other species.</p></div

Predominant biological pathways identified in chinook salmon ovarian fluid. Gene Ontology annotations (biological process) based on the AgBase v2.00 database.

<p>Predominant biological pathways identified in chinook salmon ovarian fluid. Gene Ontology annotations (biological process) based on the AgBase v2.00 database.</p

One-dimensional SDS-PAGE profiles of chinook salmon ovarian fluid.

<p>A) Comparison of profiles for a representative subset (n = 11) of the 25 ovarian fluids sampled. Lane 1 contains the standard (260 - 3.5 kDa; Invitrogen). Lanes 2–12 contain ovarian fluid samples. Stained with Simply Blue (Invitrogen). 1D-SDS PAGE run conditions were 160 V, 140 mA, 35 min. <b>B</b>) Pooled sample of ovarian fluid from 20 females, detailing fractions and bands that were excised for LC-MS/MS. Stained with conventional Coomassie. 1D-SDS PAGE run conditions were 160 V, 140 mA, 35 min.</p

Ovarian Fluid Mediates the Temporal Decline in Sperm Viability in a Fish with Sperm Storage

A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF) influences the decline in sperm viability (the proportion of live sperm in the ejaculate) over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness) exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection