| Abstrakt: |
Background: Cortical folding is related to the functional organization of the brain. The TMF-1 regulated protein (TRNP1) regulates the expansion and folding of the mammalian cerebral cortex, a process that may have been accelerated by the domestication of dogs. The objectives of this study were to sequence the TRNP1 gene in dogs and related canid species, provide evidence of its expression in dog brain and compare the genetic variation within dogs and across the Canidae. The gene was located in silico to dog chromosome 2. The sequence was experimentally confirmed by amplifying and sequencing the TRNP1 exonic and promoter regions in 72 canids (36 purebred dogs, 20 Gy wolves and wolf-dog hybrids, 10 coyotes, 5 red foxes and 1 Gy fox). Results: A partial TRNP1 transcript was isolated from several regions in the dog brain. Thirty genetic polymorphisms were found in the Canis sp. with 17 common to both dogs and wolves, and only one unique to dogs. Seven polymorphisms were observed only in coyotes. An additional 9 variants were seen in red foxes. Dogs were the least genetically diverse. Several polymorphisms in the promoter and 3'untranslated region were predicted to alter TRNP1 function by interfering with the binding of transcriptional repressors and miRNAs expressed in neural precursors. A c.259_264 deletion variant that encodes a polyalanine expansion was polymorphic in all species studied except for dogs. A stretch of 15 nucleotides that is found in other mammalian sequences (corresponding to 5 amino acids located between Pro58 and Ala59 in the putative dog protein) was absent from the TRNP1 sequences of all 5 canid species sequenced. Both of these aforementioned coding sequence variations were predicted to affect the formation of alpha helices in the disordered region of the TRNP1 protein. Conclusions: Potentially functionally important polymorphisms in the TRNP1 gene are found within and across various Canis species as well as the red fox, and unique differences in protein structure have evolved and been conserved in the Canidae compared to all other mammalian species. Plain English Summary: The folded shape of mammalian brains allows the cerebral cortex and other structures to attain a large surface area to fit into the skull. Over the past several thousand years, the domestication of dogs has led to increased folding as well as a great variety of skull size and shape amongst various breeds. Cortical folding, also known as gyrification, is regulated by numerous genes, including one that encodes the TMF-1 regulated protein (TRNP1). The TRNP1 protein in the brain affects the development of specialized cells in the brain that are involved in gyrification. It is not known whether a functionally distinct TRNP1 protein in dogs is responsible for the increased folding. Variations in the DNA sequence of the gene that encodes TRNP1 may be responsible for these dramatic changes in brain structure in dogs. This study sought to discover the differences in the TRNP1 DNA sequence in seventy-two canids, represented by thirty-six dogs of various breeds, twenty gray wolves and wolf-dog hybrids, ten coyotes, five red foxes and one gray fox. After finding evidence of the expression of this gene in dog brain, we located thirty genetic changes or variants in the canids, with seventeen common to both dogs and wolves, and only one unique to dogs. Another seven of these genetic variants were observed only in coyotes. An additional nine variants were seen in red foxes. Dogs were the least genetically diverse species, an expected result of the inbreeding that characterizes domestication. Several of these changes may affect the function of the TRNP1 gene by affecting the binding of other biomolecules to regions in the DNA which regulate this gene. This study also found two other changes, one only found in dogs, and the other one only found in canids (compared to all other mammalian TRNP1 proteins) may change the length and three-dimensional structure and hence the function of the TRNP1 protein. This study concluded that numerous, potentially functionally significant dissimilarities in the TRNP1 gene exist between dogs and their wild relatives, as well as between canid and all other mammalian species. [ABSTRACT FROM AUTHOR] |
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