Rapid determination of sperm number in ant queens by flow cytometry

Cournault, Laurent; Aron, Serge

doi:10.1007/s00040-008-1003-8

Cited by 21 publications

(17 citation statements)

References 37 publications

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“…Each egg was placed in an eppendorf tube and ruptured using a fine needle, after which 200 ml of an extraction buffer (CyStain DNA 2 step, PARTEC) was added to extract nuclei and the sample was vortexed for 30 s. Subsequently, 800 ml staining solution (CyStain DNA 2 step, PARTEC) was added, containing the fluorescent stain 4 0 ,6-diamino-2-phenylindole (DAPI) that binds strongly to DNA. The DNA content of each sample was analysed using a PA-I flow cytometer (PARTEC, Partec Gmbh, Mü nster, Germany) with an optical arrangement as used by Cournault & Aron (2008). Nuclei of sperm cells (haploid) and of somatic cells from workers (diploid) were used as standards for calibration.…”

Section: Methodsmentioning

confidence: 99%

Prudent sperm use by leaf-cutter ant queens

et al. 2009

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In many species, females store sperm between copulation and egg fertilization, but the consequences of sperm storage and patterns of sperm use for female life history and reproductive success have not been investigated in great detail. In hymenopteran insect societies (ants, bees, wasps), reproduction is usually monopolized by one or relatively few queens, who mate only during a brief period early in life and store sperm for later use. The queens of some ants are particularly long-lived and have the potential to produce millions of offspring during their life. To do so, queens store many sperm cells, and this sperm must remain viable throughout the years of storage. Queens should also be under strong selection to use stored sperm prudently when fertilizing eggs. We used the leaf-cutter ant Atta colombica to investigate the dynamics of sperm use during egg fertilization. We show that queens are able to fertilize close to 100 per cent of the eggs and that the average sperm use per egg is very low, but increases with queen age. The robustness of stored sperm was found to decrease with years of storage, signifying that senescence affects sperm either directly or indirectly via the declining glandular secretions or deteriorating sperm-storage organs. We evaluate our findings with a heuristic model, which suggests that the average queen has sperm for almost 9 years of normal colony development. We discuss the extent to which leaf-cutter ant queens have been able to optimize their sperm expenditure and infer that our observed averages of sperm number, sperm robustness and sperm use are consistent with sperm depletion being a significant cause of mortality of mature colonies of Atta leaf-cutter ants.

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Section: Methodsmentioning

confidence: 99%

Prudent sperm use by leaf-cutter ant queens

et al. 2009

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“…For this purpose, bumble bees were first killed by deep freezing (at −80°C) and the head was placed each in 1 ml of the DNA-staining solution. Processing, cell cycle analysis and quantification of flow cytometric data were performed according to Cournault and Aron (2008). Both wings were removed for geometric morphometric analyses.…”

Section: Materials and Ploidy Determinationmentioning

confidence: 99%

Discrimination of haploid and diploid males of Bombus terrestris (Hymenoptera; Apidae) based on wing shape

et al. 2015

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-Presence of diploid males in wild bees reflects inbreeding and provides information about the health status of a colony or population. Detection of diploid males, and discrimination from haploid males and workers, has, however, been limited to molecular diagnostics. Here we present a novel method based on differences in wing shape, e.g., venation patterns in wings. The method is easy to apply and results, for Bombus terrestris , in very high discrimination success. Possible applications of the method are discussed.Bombus / decline / diploid males / wing shape / geometric morphometrics

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“…Both spermathecae and seminal vesicles were emptied in a staining solution of DAPI fluorochrome (4, 6-diamidino-2-phenylindole). The number of sperm was then counted by flow cytometry (Ploidy Analyser PAI, Partec), a method that gives reliable and repeatable counts of sperm samples (Aron et al, 2003;Cournault and Aron, 2008).…”

Section: Sperm Countmentioning

confidence: 99%

Multiple mating in the ant Cataglyphis cursor: testing the sperm limitation and the diploid male load hypotheses

et al. 2008

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Abstract. Multiple mating (i.e., polyandry) by queens in social Hymenoptera is expected to weaken social cohesion since it lowers within-colony relatedness, and hence, indirect fitness benefits from kin selection. Yet, there are many species where queens mate multiply. Several hypotheses have been put forward to explain the evolution and maintenance of polyandry. Here, we investigated the sperm limitation and the diploid male load hypotheses in the ant Cataglyphis cursor. Genetic analyses of mother-offspring combinations showed that queens mate with up to 8 males, with an effective mating frequency of 3.79. Significant paternity skew (unequal contribution of the fathers) was detected in 1 out of 5 colonies. The amount of sperm stored in the spermatheca was not correlated with the queen mating frequency, and males carry on average enough sperm in their seminal vesicles to fill one queens spermatheca. Analyses of the nuclear DNA-content of males also revealed that all were haploid. These results suggest that the sperm limitation and the diploid male load hypotheses are unlikely to account for the queen mating frequency reported in this ant. In light of our results and the life-history traits of C. cursor, we discuss alternative hypotheses to account for the adaptive significance of multiple mating by queens in this species.

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Rapid determination of sperm number in ant queens by flow cytometry

Cited by 21 publications

References 37 publications

Prudent sperm use by leaf-cutter ant queens

Prudent sperm use by leaf-cutter ant queens

Discrimination of haploid and diploid males of Bombus terrestris (Hymenoptera; Apidae) based on wing shape

Multiple mating in the ant Cataglyphis cursor: testing the sperm limitation and the diploid male load hypotheses

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