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Georgia Journal of Science

Article Title

PARTIAL CHARACTERIZATION OF THE PITX1 GENE OF EMPETRICHTHYINE FISHES (GOODEIDAE).

Abstract

Vertebrate pelvic appendages are complex structures comprising many different tissues. Nevertheless, taxa including sticklebacks (Gasterosteidae, Teleostei) and manatees (Sirenia, Mammalia) have independently lost these structures as a result of defects in a common, cis-acting, upstream regulatory element (pel) of the Pituitary homeobox transcription factor 1 (PITX1) gene. Here we began investigating whether independent pelvic loss in the Empetrichthyinae (Goodeidae, Teleostei) was caused by similar PITX1 regulatory defects. Toward this end we desired to sequence PITX1 from Crenichthys baileyi and the goodeine Goodea atripinnis (a related clade with pelvic fins) using standard molecular protocols. Multiple, overlapping sets of primers were developed within the four exons of this nuclear gene using comparative bioinformatics approaches with publicly available sequence data for Oryzias latipes and Fundulus heteroclitus. Most primer combinations did not successfully amplify products of the predicted size. Partial sequences were successfully determined for exon 2, intron 2 and exon 3. Comparisons of exons 2 (89 bp) and 3 (148 bp) of Crenichthys and Goodea revealed 96 and 97% sequence identity, respectively. These same exons shared 84 and 92% identity with Oryzias, and 96 and 96% identity with Fundulus. These coding regions yielded no significant hits on other genes. We have successfully sequenced part of the PITX1 gene of goodeids with and lacking pelvic fins.

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