Expression patterns of relaxin family peptides in Danio rerio
LE3 .A278 2009
2009
Wilson, Brian
Acadia University
Bachelor of Science
Honours
Biology
The family of relaxin peptides is a diverse group of hormones implicated in a number of physiological processes in mammals. This study investigated the functional nature of five of the six known teleostean relaxin family genes in zebrafish (Danio rerio), namely telrln3a & 3b, telinsl3, and telinsl5a & 5b. While telrln3a, telinsl3 and telinsl5a are orthologous to mammalian rln3, insl3 and insl5 genes respectively, telrln3b and telinsl5b are considered paralogous to telrln3a and telinsl5a. The level of relaxin gene expression within and across tissues was assessed using SYBR Green Real Time (quantitative) PCR (qPCR). Additionally, protein assays obtained from five zebrafish tissues were screened for the different forms of teleostean relaxin family peptides using 2D-PAGE. The protein spots suspected of containing the desired peptides were sent for spectroscopic MALDI analysis. The template cDNA used in the qPCR reactions was obtained via reverse transcription of total RNA extracted from brain, eye, gill, heart, gut and gonads of both male and female zebrafish. The relative expression of each of the five relaxin family genes was normalized by comparison to the level of expression of the housekeeping gene b2m. Our qPCR results indicate that telinsl5b was barely detectable in any tissue, telrln3a was moderately expressed in the brain, but was also detected in gonads, telrln3b exhibited moderately low expression in the brain, and finally that telinsl3 exhibited the highest expression of all the relaxin genes, and was found predominantly in ovaries and testes. Although 2D-PAGE showed separation of proteins in the range of isoelectric points and molecular weights predicted for teleost relaxin-like peptides, mass spectrometry analysis did not identify any of the expected molecules due to technical failure.
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https://scholar.acadiau.ca/islandora/object/theses:550
