LE3 .A278 2006

2006

Wilson, Brian

Acadia University

Bachelor of Science

Honours

Biology

Biology

In recent years, relaxin research in mammalian systems has shown that there are multiple functional forms of relaxin with distinct physiological roles. However, evidence of relaxin’s presence in teleost fish has been limited to comparative genomics. These studies have shown that there are multiple relaxin-like gene sequences in teleosts with uncharacterized expression and function. To explore the expression of relaxin in teleosts, zebrafish (Danio rerio) were used as a model species because of the recent publication of its assembled genome. Searching of the NCBI and Ensembl databases yielded five relaxin-like sequences in the zebrafish genome. These were used to develop novel, nondegenerate primers for use in PCR. Zebrafish were raised at Acadia University to maturity and DNA was extracted for gene amplification through PCR. Attempts to isolate relaxin from genomic DNA were unsuccessful. RNA was then extracted from tissue specific regions (brain, liver/intestine, ovary), and used to generate cDNA. These analyses generated PCR products of the expected 300 bp size for three of the five relaxin sequences, however sequencing showed that product was zebrafish rRNA. The five relaxin sequences identified in the zebrafish genome were compared to the sequences in humans, mice, and Takifugu rubripes using a Minimum Evolution tree, which showed that there are multiple forms of relaxin in zebrafish. This suggests that the broad teleost “Relaxin-3” nomenclature needs to be subdivided. Though expression of relaxin in zebrafish was not definitively shown, bioinformatic evidence suggests it is present in multiple forms.

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https://scholar.acadiau.ca/islandora/object/theses:498