RESUMO
Heterothermy (hibernation and daily torpor) is a key strategy that animals use to survive in harsh conditions and is widely employed by bats, which are found in diverse habitats and climates. Bats comprise more than 20% of all mammals and although heterothermy occurs in divergent lineages of bats, suggesting it might be an ancestral condition, its evolutionary history is complicated by complex phylogeographic patterns. Here, we use Leptin, which regulates lipid metabolism and is crucial for thermogenesis of hibernators, as molecular marker and combine physiological, molecular and biochemical analyses to explore the possible evolutionary history of heterothermy in bat. The two tropical fruit bats examined here were homeothermic; in contrast, the two tropical insectivorous bats were clearly heterothermic. Molecular evolutionary analyses of the Leptin gene revealed positive selection in the ancestors of all bats, which was maintained or further enhanced the lineages comprising mostly heterothermic species. In contrast, we found evidence of relaxed selection in homeothermic species. Biochemical assays of bat Leptin on the activity on adipocyte degradation revealed that Leptin in heterothermic bats was more lipolytic than in homeothermic bats. This shows that evolutionary sequence changes in this protein are indeed functional and support the interpretation of our physiological results and the molecular evolutionary analyses. Our combined data strongly support the hypothesis that heterothermy is the ancestral state of bats and that this involved adaptive changes in Leptin. Subsequent loss of heterothermy in some tropical lineages of bats likely was associated with range and dietary shifts.
Assuntos
Adaptação Fisiológica/fisiologia , Temperatura Corporal/genética , Temperatura Corporal/fisiologia , Quirópteros/genética , Quirópteros/fisiologia , Evolução Molecular , Leptina/metabolismo , Adipócitos/citologia , Adipócitos/metabolismo , Animais , Contagem de Células , Sobrevivência Celular/genética , Quirópteros/metabolismo , Escherichia coli/genética , Hibernação/genética , Hidroliases/metabolismo , Leptina/genética , Lipólise/genética , Filogenia , Seleção Genética , Clima TropicalRESUMO
Leptin, a 16-kDa hormone produced by mature adipocytes, has been shown to regulate the hibernation of mammals. In this study, the leptin gene sequences of both hibernating (Rhinolophus ferrumequinum) and non-hibernating (Rousettus leschenaultii) bats were determined, and the leptin proteins from these two different species of bats were expressed in Escherichia coli for the first time. Results showed that the amino acid sequence of the leptin protein from hibernating bats had a lower degree of identity than that from non-hibernating bats to those of several non-hibernating mammals. The leptin protein of hibernating bats had a stronger growth inhibitory effect on 3T3-L1 cells than that of non-hibernating bats. Structural modeling revealed that the structures of the receptor binding site III, which is critical for signal transduction, of the two bat leptins were very different. Similar to the human leptin, the leptin protein of non-hibernating bats was predicted to have a random loop, whereas that of hibernating bats had a helical structure in this region. This observation provided a clue as to the differential effects of the two different leptins on 3T3-L1 cells.
Assuntos
Quirópteros/metabolismo , Hibernação/fisiologia , Leptina/química , Leptina/farmacologia , Células 3T3-L1 , Sequência de Aminoácidos , Animais , Western Blotting , Sobrevivência Celular/efeitos dos fármacos , Eletroforese em Gel de Poliacrilamida , Leptina/genética , Leptina/metabolismo , Camundongos , Dados de Sequência Molecular , Estrutura Secundária de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/farmacologia , Homologia de Sequência de AminoácidosAssuntos
Quirópteros/genética , Proteínas de Ligação a RNA/genética , Sequência de Aminoácidos , Animais , Clonagem Molecular , Humanos , Masculino , Mamíferos/genética , Dados de Sequência Molecular , Isoformas de Proteínas/genética , Proteínas de Ligação a RNA/fisiologia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Espermatogênese/genéticaRESUMO
The homeobox (Hox) genes Hoxd12 and Hoxd13 control digit patterning and limb formation in tetrapods. Both show strong expression in the limb bud during embryonic development, are highly conserved across vertebrates, and show mutations that are associated with carpal, metacarpal, and phalangeal deformities. The most dramatic evolutionary reorganization of the mammalian limb has occurred in cetaceans (whales, dolphins, and porpoises), in which the hind limbs have been lost and the forelimbs have evolved into paddle-shaped flippers. We reconstructed the phylogeny of digit patterning in mammals and inferred that digit number has changed twice in the evolution of the cetacean forelimb. First, the divergence of the early cetaceans from their even-toed relatives coincided with the reacquisition of the pentadactyl forelimb, whereas the ancestors of tetradactyl baleen whales (Mysticeti) later lost a digit again. To test whether the evolution of the cetacean forelimb is associated with positive selection or relaxation of Hoxd12 and Hoxd13, we sequenced these genes in a wide range of mammals. In Hoxd12, we found evidence of Darwinian selection associated with both episodes of cetacean forelimb reorganization. In Hoxd13, we found a novel expansion of a polyalanine tract in cetaceans compared with other mammals (17/18 residues vs. 14/15 residues, respectively), lengthening of which has previously been shown to be linked to synpolydactyly in humans and mice. Both genes also show much greater sequence variation among cetaceans than across other mammalian lineages. Our results strongly implicate 5'HoxD genes in the modulation of digit number, web forming, and the high morphological diversity of the cetacean manus.
Assuntos
Cetáceos/genética , Evolução Molecular , Membro Anterior/crescimento & desenvolvimento , Genes Homeobox/genética , Morfogênese/genética , Animais , Variação Genética , Seleção GenéticaRESUMO
Bone morphogenetic protein 2 (BMP2) plays an important role in skeletogenesis, osteoblastic differentiation and limb patterning. Its protein coding region consists of the signal peptide, the pro-domain (that regulates post-translational control of synthesis) and the mature domain (that carries out gene function). This gene has been considered previously to be conserved. By re-analyzing the coding region of BMP2 in 31 species of vertebrates, we found that the mature domain region is indeed conserved in mammals, but not among non-mammalian taxa. Moreover, compared to the mature domain, the signal peptide and pro-domain have experienced dramatic variation in all vertebrates. Six amino acid sites in the pro-domain were identified to be under diversifying Darwinian selection in mammals. These results indicate that the signal peptide and pro-domain of BMP2 may be involved in skeletal poly-morphology during mammal evolution and the mature domain may also contribute to this function in non-mammals. This supports the hypothesis that morphological variations in mammals result mainly from a change in post-translational control of synthesis, whereas in non-mammals they result mainly from gene functional change.
