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研究生:方志宇
研究生(外文):Fang, Chih-Yeu
論文名稱:盤谷蟾蜍與黑眶蟾蜍的血液攜氧力與血紅素組成之比較
論文名稱(外文):Comparison of Blood Oxygen Capacity and Hemoglobin Compositions Among Bufo bankorensis and Bufo melanostictus
指導教授:侯平君, 辛致煒
指導教授(外文):Ping-Chun L. Hou, Jyh-Wei Shin
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生物學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:1998
畢業學年度:86
語文別:中文
論文頁數:70
中文關鍵詞:盤谷蟾蜍黑眶蟾蜍氧氣平衡曲線血紅素組成海拔分佈血紅素聚合
外文關鍵詞:Bufo bankorensisBufo melanostictusoxygen equilibrium curvehemoglobin compositionsaltitudinal distributionhemoglobin aggregation
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台灣地區僅有兩種蟾蜍科兩棲動物。其中盤古蟾蜍(Bufo bankorensis
)可由平地一直分佈到3,000公尺山區,而黑眶蟾蜍(Bufo
melanostictus)卻侷限於500公尺以下的低地。在本實驗中,我比較這兩
種蟾蜍的血液攜氧力及血紅素組成,以瞭解是否因血液生理上的不同而導
致蟾蜍的海拔分佈差異。同時檢測血紅素在低氧下聚合的現象是否也存在
這兩種蟾蜍中。我以混合法測量血紅素-氧氣平衡曲線。在25℃時,半飽
和氧分壓(P50)在高、低海拔盤古蟾蜍分別為47.96及41.43 torr;黑眶
蟾蜍則為47.31 torr。25℃時,兩種蟾蜍的血液-氧氣結合的合作性(
cooperativity;Hill n值)在血液趨近飽和時有超過4的現象。在15℃時
,半飽和氧分壓在高、低海拔盤古蟾蜍分別為29.92及25.90 torr;黑眶
蟾蜍則為34.58 torr。15℃下,兩種蟾蜍的Hill n值在任何飽和度下皆沒
有超過4。等電點電泳顯示,盤古蟾蜍的血液由兩種主要血紅素構成,而
黑眶蟾蜍有三種。由Tricine SDS-PAGE得知組成蟾蜍血紅素之
polypeptide至少有3種。HPLC的血紅素分析結果大致與電泳結果相同。在
native電泳中,去氧血紅素在膠體中的相對移動速率(relative
mobility)明顯慢於充氧血紅素,表示血紅素在去氧狀態下有聚合的現象
。除了缺氧狀態下的聚合,蟾蜍的血紅素亦有形成分子間雙硫鍵而結合的
現象。這些結果顯示:盤古蟾蜍的血液-氧氣親和力的確比黑眶蟾蜍來的
好,而氧氣親和力的增加可以幫助盤古蟾蜍適應高海拔低氧環境。在充氧
過程中,蟾蜍血紅素由低氧親和力之聚合物解離為高氧親和力之四合體,
可能是造成血液在接近充氧飽和時,結合氧氣能力躍升的原因。

The Taiwanese toads, Bufo bankorensis, show wide altitudinal
distributionfrom lowland to over 3000 m. Bufo melanostictus, on
the other hand, are restricted to area below 500 m. I compared
blood-oxygen capacity and hemoglobin compositions in these two
species to understand whether the difference in altitudinal
distribution can be attributed to physiological variations. I
also examine the deoxygenated aggregation of hemoglobins in
these two toads. Blood-oxygen equilibrium curves were measured
by mixing technique. At 25 oC, the half saturation pressures
(P50) were 47.96 and 41.43 torr in Bufo bankorensis from high
and low altitudes, respectively, and was 47.31 torr in Bufo
melanostictus. Cooperativity of oxygen binding, evaluated as
Hill's n, exceeded 4 in both species between 75% and 90%
saturation at 25 oC. At 15 oC, P50 were 29.92 and 25.90 torr in
Bufo bankorensis from high and low altitudes, respectively, and
was 34.58 torr in Bufo melanostictus. At 15 oC, Hill's n in both
species within any saturation range never exceeded 4.
Isoelectric focusing showed that Bufo bankorensis has two major
hemoglobins, and Bufo melanostictus has three. Tricine SDS-PAGE
showed that hemoglobins of both toad species were composed of at
least three polypeptides. HPLC analysis also showed similar
results with electrophoresis. In native-PAGE, hemoglobins under
nitrogen treatment had lower relative mobility than in
oxygenated treatment, indicating formation of hemoglobin
aggregations under deoxygenation. Besides deoxygenated
aggregation, hemoglobins of these toads also formed disulfide
bond linkage between tetramers. These results suggest that
blood-oxygen affinity of Bufo bankorensis was higher than Bufo
melanostictus, and may contribute to the adaptation of Bufo
bankorensis to altitudinal hypoxia. Dissociation of hemoglobin
aggregates during oxygenation may enhance oxygen-binding
coopreativity, resulting in rapid loading of oxygen near
saturation.

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