Date

2014

Major

Biology (B.S.B.)

Document Type and Release Option

Thesis (restricted to Georgia Southern)

Faculty Mentor

Dr. Christopher Cutler

Abstract

Immunohistochemical staining allowed localization of aquaporin 4 (AQP4) in the basolateral and apical membranes of the renal tubule epithelial cells of the American eel, Anguilla rostrata, acclimated to seawater (SW). In the SW eel, AQP4 is mutually expressed with aquaporin 1Dup-2 (AQP1Dup-2) and aquaporin e (AQPe) with minimal co-localization. Additionally, the conservation of aquaporin genes within the genus A. allowed cloning of aquaporin 3a (AQP3a), a duplicated gene copy of aquaporin 3 (AQP3), in the American eel, using primers designed from aquaporin 3a in the Japanese eel, A. japonica. The gene identified revealed a 933 nucleotide sequence coding for a 295 amino acid (AA) sequence. A 43 nucleotide 5’untranslated region preceded the coding region with a 2 nucleotide 3’ untranslated region flanking the coding region. Alignment of the American eel AQP3a nucleotide sequence against Japanese eel AQP3a revealed 99.04% homology. Alignment of the derived AA sequence against that of the Japanese eel revealed 99.6% nucleotide homology. Alignment of the American eel AQP3a AA sequence to that of American eel AQP3b AA sequence revealed an 82.77% homology with the similar comparative nucleotide alignment revealing a 76.54% homology. Gel electrophoresis of RT-PCR amplified AQP3a cDNA fragments in American eel tissues involved in osmoregulation involvement revealed mRNA expression specific to the SW acclimated-fish kidney.

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