Abstract

Females of many vertebrate species have the capacity to store sperm within their reproductive tracts for prolonged periods of time. Termed long-term sperm storage, this phenomenon has many important physiological, ecological, and evolutionary implications, particularly to the study of mating systems, including male reproductive success and post-copulatory sexual selection. Reptiles appear particularly predisposed to long-term sperm storage, with records in most major lineages, with a strong emphasis on turtles and squamates (lizards, snakes, but not the amphisbaenians). Because facultative parthenogenesis is a competing hypothesis to explain the production of offspring after prolonged separation from males, the identification of paternal alleles through genetic analysis is essential. However, few studies in snakes have undertaken this. Here, we report on a wild-collected female Western Diamond-backed Rattlesnake, Crotalus atrox, maintained in isolation from the time of capture in September 1999, that produced two healthy litters approximately one and six years post capture. Genetic analysis of the 2005 litter, identified paternal contribution in all offspring, thus rejecting facultative parthenogenesis. We conclude that the duration of long-term sperm storage was approximately 6 years (71 months), making this the longest period over which a female vertebrate has been shown to store sperm that resulted in the production of healthy offspring.

Highlights

  • The ability for females to store viable spermatozoa in their reproductive tracts, capable of retaining fertilization capacity for months or even years post insemination, has been reported across a variety of vertebrate species [1,2,3,4,5,6,7]; a reproductive phenomenon termed long-term sperm storage (LTSS) [1, 8]

  • Supporting these results demonstrating the presence of paternal alleles in male offspring, COLONY assigned all offspring to one full-sibship when a subset of high minor allele frequency (MAF) loci were analyzed (n = 417)

  • Combining captive history and genomic screening, we unequivocally identify LTSS as the reproductive mechanism underlying the production of offspring from female Ca-149, following a period of approximately 71 months of isolation post capture

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Summary

Introduction

The ability for females to store viable spermatozoa in their reproductive tracts, capable of retaining fertilization capacity for months or even years post insemination, has been reported across a variety of vertebrate species [1,2,3,4,5,6,7]; a reproductive phenomenon termed long-term sperm storage (LTSS) [1, 8]. The non-avian reptiles (i.e., chelonians, crocodilians, Rhynchocephalia, and squamates), LTSS is remarkably prolonged, with duration reports of months to several years

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