A Method for the Determination of Genetic Sex in the Fathead Minnow, Pimephales promelas, To Support Testing of Endocrine-Active Chemicals

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• 作者：Allen W. Olmstead ; Daniel L. Villeneuve ; Gerald T. Ankley ; Jenna E. Cavallin ; Annelie Lindberg-Livingston ; Leah C. Wehmas ; Sigmund J. Degitz
• 刊名：Environmental Science & Technology
• 出版年：2011
• 出版时间：April 1, 2011
• 年：2011
• 卷：45
• 期：7
• 页码：3090-3095
• 全文大小：769K
• 年卷期：v.45,no.7(April 1, 2011)
• ISSN：1520-5851

文摘

Certain endocrine-active toxicants have been reported to completely sex reverse both male and female individuals in amphibian, avian, fish, invertebrate, and reptile species, resulting in a phenotype indistinguishable from unaffected individuals. Detection of low-level sex reversal often requires large numbers of organisms to achieve the necessary statistical power, especially in those species with predominantly genetic sex determination and cryptic/homomorphic sex chromosomes. Here we describe a method for determining the genetic sex in the commonly used ecotoxicological model, the fathead minnow (Pimephales promelas). Analysis of amplified fragment length polymorphisms (AFLP) in a spawn of minnows resulted in detection of 10 sex-linked AFLPs, which were isolated and sequenced. No recombination events were observed with any sex-linked AFLP in the animals examined (n = 112). A polymerase chain reaction (PCR) method was then developed that determined the presence of one of these sex-linked polymorphisms for utilization in routine toxicological testing. Analyses of additional spawns from our in-house culture indicate that fathead minnows utilize a XY sex determination strategy and confirm that these markers can be used to genotype sex; however, this method is currently limited to use in laboratory studies in which breeders possess a defined genetic makeup. The genotyping method described herein can be incorporated into endocrine toxicity assays that examine the effects of chemicals on gonad differentiation.