|Alternative Title||A study on the structure and function of hemoglobin in two Phrynocephalus lizards from different altitudes
|Place of Conferral||兰州
|Abstract||高原低氧环境会对动物生活造成独特的生理挑战，红尾沙蜥是迄今为止已知的世居海拔高度最高的爬行动物，但这种动物的高原适应机制尚无报道。本文选择红尾沙蜥（Phrynocephalus erythrurus）和荒漠沙蜥（Phrynocephalus przewalskii）作为研究对象，通过测定两个物种部分血液学参数、全血氧解离曲线及P50<下标!>值、成体α-和β-珠蛋白基因的表达以及β-珠蛋白基因序列，比较分析了这些指标在两个物种间的差异。
|Other Abstract||In the past decades, scientists have found variety of adaptation features and coordination mechanism to high-altitude hypoxia in mammals, birdsand reptiles. As yet, Phrynocephalus erythrurus a native species of Qiangtang plateau is considered to be the highest living reptile in the world. However, the adaptation mechanisms to high altitude in this mysterious species are still unknown.In this study, two species of lizard from the genus Phrynocephalus were collected and used for experiments. Phrynocephalus erythrurus was collected from Tuotuo River (34o<上标!>13'N, 92o<上标!>13'E), Phrynocephalus przewalskii was collected from Minqin (38o<上标!>38'N, 103o<上标!>05'E), Gansu province, China. We mainly studied the structure and function of hemoglobin in both species, including hematological parameters, whole blood oxygen dissociation curves and P50<下标!> values, the expression of α-and β-globin gene in adult and the β-globin gene sequence of the two species.
The results showed that hemoglobin content and red blood cell count in Phrynocephalus erythrurus were significantly higher than Phrynocephalus przewalskii (p<0.001), and the mean corpuscular volume of the two species was not significantly different (p>0.05). Whole blood oxygen dissociation curve of Phrynocephalus erythrurus was on the left of curve of Phrynocephalus przewalskii, and the P50 value of Phrynocephalus erythrurus was obviously lower than Phrynocephalus przewalskii (p<0.001). We used reverse phase high-performance liquid chromatography (RP-HPLC) and electrospray ionization mass spectrometry (ESI-MS) to analysis the expression of hemoglobin gene of the two species, the results showed that the expression of four globins was seen in both Phrynocephalus erythrurus and Phrynocephalus przewalskii and the chromatography map and molecular mass of Hb of the two species were obvious different. We also identified the amino acids substitution sites of the β-chain in two species by the molecular cloning methods. Alignment analysis showed that amino acid sequence of β1<下标!>-globin chains in two species was slightly different, there was only one amino acid alternative (β1<下标!>12Ser in Phrynocephalus erythrurus and β1<下标!>12Thr in Phrynocephalus przewalskii). However, amino acid sequence of β2<下标!>-globin chains was obviously different in two species, there were 24 amino acids alternative with each other.
鲁松松. 不同海拔两种沙蜥血红蛋白结构和功能的研究[D]. 兰州. 兰州大学,2013.
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