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Dopamine pp 201-214 | Cite as

Genomic Strategies for the Identification of Dopamine Receptor Genes in Zebrafish

  • Wendy Boehmler
  • Jessica Petko
  • Victor A. Canfield
  • Robert LevensonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 964)

Abstract

In this chapter, we describe the identification and cloning of D2-like dopamine receptor (DR) genes in zebrafish, a vertebrate model genetic organism. To identify DR genes, we performed searches of the zebrafish genomic sequence database that yielded contig segments of several D2-like DR genes. From these sequences, we amplified full-length cDNAs encoding three D2, one D3, and three D4 DR receptor subtypes via RT-PCR. The predicted proteins displayed 57–72% amino acid identity when compared to their human DR counterparts. To validate the identity of zebrafish DR genes, each of the genes was mapped by using the T51 radiation hybrid panel. With the exception of drd2b and drd4b, each of the zebrafish DR genes mapped to chromosomal positions that were syntenic with regions of human chromosomes containing orthologs of the zebrafish DR genes. To further validate the identity of the D2-like DR genes in zebrafish, we conducted phylogenetic analysis which supported the predicted identities of the cloned DR receptor cDNAs.

Key words

Cloning D2-like DR RT-PCR Phylogenetic analysis Contig Dopamine 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Wendy Boehmler
    • 1
  • Jessica Petko
    • 2
  • Victor A. Canfield
    • 2
  • Robert Levenson
    • 2
    Email author
  1. 1.Department of Biological SciencesYork College of PennsylvaniaYorkUSA
  2. 2.Department of PharmacologyPenn State College of MedicineHersheyUSA

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