Threespin Stickleback Fish

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Pitx1 is a gene with in a larger family of similar genes, referred to as homeobox genes that direct the formation of many body structures during early embryonic development. Pitx1 in particular, encodes for a protein of some 283 amino acids (varying slightly in different species) that acts as a transcription factor and is in charge of regulating the expression of other genes involved in the differentiation and function of certain developmental regions. The scientists in this study have been interested in the role of this Pitx1 protein and the differentiation and function of the development in particular of the hind limb regions in the threespin stickleback fish. Through their previously conducted studies these scientists concluded that the …show more content…

In order to test the hypothesis and ensure that other genes in the larger chromosome region were not the real cause of pelvic loss the scientists generated F1 hybrids between both pelvic-completed and pelvic-reduced sticklebacks as well as two pelvic-complete populations. These F1 hybrids allowed for combination of the alleles of Pitx1 from pelvic-complete (FRIL, LITC) and pelvic-reduced (PAXB) populations. Brain and pelvic tissues were isolated to compare the expression of either the LITC or PAXB allele to the FRIL allele. The results supported the initial hypothesis as expression of the PAXB Pitx1 allele was greatly reduced in the pelvis but not the head of F1 hybrids, indicating a tissue-specific, cis-regulatory change in the Pitx1 locus. If indeed other genes in the larger chromosome region were the real cause of pelvic reduction then there would have been a secondary or trans-acting reduction seen in the expressions of Pitx1 in both …show more content…

EGFP is a powerful reporter molecule for monitoring gene expression, protein localization and protein-point interaction. Through the use of EGFP, the scientists were able to see what areas of embryonic development this intergenic region of the Pitx1 was correlated with. Therefore, EGFP will be visibly expressed in areas within the intergenic region of the Pitx1 gene as well as within any of the developing embryonic regions that it is correlated with. Seeing that the only region where EGFP was consistently expressed was in the hindfin area, the scientists concluded that the non-coding region upstream of Pitx1 contained a tissue-specific enhancer for hindfin expression. Again, supporting their hypothesis that indeed Pitx1, and no other genes in the larger chromosome region, is the real cause of pelvic loss in sticklebacks. This region, which the scientists termed as “Pel” was then cloned and injected into unfertilized eggs of pelvic-reduced fish. The scientists did this in order to see if the region is a major determinant of pelvic formation in sticklebacks, which it ended up

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