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【medical-news】防治疟疾的研究新进展——移植

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http://www.sciencedaily.com/releases/2010/02/100215174119.htm

Scientists Transplant Nose of Mosquito, Advance Fight Against Malaria

ScienceDaily (Feb. 16, 2010) — Scientists at Vanderbilt and Yale universities have successfully transplanted most of the "nose" of the mosquito that spreads malaria into frog eggs and fruit flies and are employing these surrogates to combat the spread of the deadly and debilitating disease that afflicts 500 million people.

The research is described in two complimentary papers, one published this week in the early online edition of the Proceedings of the National Academy of Sciences and the other which appeared online Feb. 3 in the journal Nature.

The mosquito's "nose" is centered in its antennae, which are filled with nerve cells covered with special "odorant receptors" that react to different chemical compounds. The insect ORs are comparable to analogous receptors in the human nose and taste buds on the tongue.

"We've successfully expressed about 80 percent of the Anopheles mosquito's odorant receptors in frog's eggs and in the fruit fly antennae," says Laurence Zwiebel, professor of biological sciences at Vanderbilt, whose lab performed the frog egg transplantation. The fruit-fly (Drosophila melanogaster) work was done in the laboratory of John Carlson, Eugene Higgins Professor of Molecular, Cellular and Developmental Biology at Yale.

Both accomplishments are part of a five-year project supported by the Grand Challenges in Global Health Initiative funded by the Foundation for NIH through a grant from the Bill & Melinda Gates Foundation with the goal of producing novel ways to inhibit the spread of malaria. Scientists from the Wageningen University in the Netherlands, the African Insect Science for Food and Health Institute in Kenya, Ifakara Health Institute in Tanzania and the Medical Research Council Laboratories in the Gambia are also participating in the project.

Previously, scientists have used frog eggs to study the olfactory receptors of moths, honeybees and fruit flies. DNA that encodes insect receptors are injected into a frog egg and given sufficient time to produce and localize proteins. As a result, the surface of the egg is covered with the mosquito odorant receptors. An engineered egg is placed in a voltage clamp system and an odorant is dissolved in the buffer solution in which the egg is floating. If the mosquito receptors react to the compound, the electrical properties of the egg change in a measurable fashion.

"The frog egg system is relatively rapid, highly sensitive and allows us to do very precise measurements of odorant response," says Guirong Wang, a senior research associate in the Zwiebel lab who was the lead author on the PNAS study and carried out several thousand egg/odorant recordings. "However, we call this a medium throughput system because, while it is relatively quick to set up, we have to make the odorant solutions by hand, which goes relatively slowly."

By comparison, Yale's Drosophila system is a somewhat lower throughput system because it takes about three months to engineer a fruit fly with a mosquito odorant receptor in its antennae. The system, originally developed in the Carlson lab, uses mutant flies that are missing an odor receptor. Allison Carey, a graduate student in the Yale lab, systematically inserted mosquito genes into fruit flies one at a time so that a mosquito odorant receptor was expressed in place of the missing receptor. Although the method is slightly slower than the frog egg approach, it has some distinct advantages: Most notably it responds to volatilized odorants so it works with compounds that don't dissolve readily in water. It is also effective in detecting chemicals that inhibit receptors rather than exciting them.

"Both teams used the same set of 72 Anopheles odorant receptors and tested them using the same panel of 110 odorants," says Wang. The Vanderbilt team got responses from 37 of the odorant receptors in the frog eggs while testing 6,300 odorant-receptor combinations. "The results of the two systems were quite similar. There were only a few small differences."

Both studies found that most mosquito receptors are "generalists" that react to a number of different odors while a few are "specialists" that respond to a single or small number of odors. In some cases, the researchers found that a single odorant triggers several receptors while in other cases receptors are specifically tuned to unique compounds. In particular, they found 27 Anopheles receptors that respond strongly to compounds in human sweat.

"We're now screening for compounds that interact with these receptors. We call those that do BDOCs (behaviorally disruptive olfactory compounds)," Zwiebel says. "Compounds that excite some of these receptors could help lure mosquitoes into traps or repel them away from people while others that block receptor activity may help mask people. Ultimately we are looking for cocktails of multiple compounds that demonstrate activity in the field."

The project has already developed and patented a blend of BDOCs that is more attractive to mosquitoes than humans and has also identified several repellant BDOCs. It is currently in product development discussions with several private sector companies.

Researcher Guirong Wang sitting at the workstation where he tested the response of mosquito odorant receptors insert into frog eggs to a panel of 110 different odorants. (Credit: Steve Green, Vanderbilt University)

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Scientists Transplant Nose of Mosquito, Advance Fight Against Malaria
疟疾防治新进展---移植蚊子气味感受器

ScienceDaily (Feb. 16, 2010) — Scientists at Vanderbilt and Yale universities have successfully transplanted most of the "nose" of the mosquito that spreads malaria into frog eggs and fruit flies and are employing these surrogates to combat the spread of the deadly and debilitating disease that afflicts 500 million people.
每日科学（2010年2月16日）—范德比尔特（Vanderbilt）大学和耶鲁（Yale）大学的科学家们成功将大多数传播疟疾的蚊子的“鼻子”植入青蛙卵和果蝇体内，并利用这些替代品来控制严重威胁五亿人群的致死性疾病-疟疾的扩散。

The research is described in two complimentary papers, one published this week in the early online edition of the Proceedings of the National Academy of Sciences and the other which appeared online Feb. 3 in the journal Nature.
该研究发表于两种杂志的增刊上，一个是本周网络版的《美国国家科学院院刊》，另一个是《自然》2月3号的网络版。

The mosquito's "nose" is centered in its antennae, which are filled with nerve cells covered with special "odorant receptors" that react to different chemical compounds. The insect ORs are comparable to analogous receptors in the human nose and taste buds on the tongue.
蚊子的“鼻子”位于两只触角的正当中，里面布满了神经细胞，这些神经细胞表明布满了特殊的“气味感受器”，能识别不同的化合物。昆虫的气味感受器（ORs）相当于人类鼻中的感受器和舌头上的味蕾。

"We've successfully expressed about 80 percent of the Anopheles mosquito's odorant receptors in frog's eggs and in the fruit fly antennae," says Laurence Zwiebel, professor of biological sciences at Vanderbilt, whose lab performed the frog egg transplantation. The fruit-fly (Drosophila melanogaster) work was done in the laboratory of John Carlson, Eugene Higgins Professor of Molecular, Cellular and Developmental Biology at Yale.
“我们已经成功在青蛙卵和果蝇的触角中表达出80%的按蚊气味感受器，” 范德比尔特大学的生物科学教授Laurence Zwiebel说。他的实验室负责向青蛙卵中的移植。而在果蝇（黑腹果蝇）身上进行的移植则由耶鲁大学的分子细胞发展生物学教授John Carlson的实验室进行。

Both accomplishments are part of a five-year project supported by the Grand Challenges in Global Health Initiative funded by the Foundation for NIH through a grant from the Bill & Melinda Gates Foundation with the goal of producing novel ways to inhibit the spread of malaria. Scientists from the Wageningen University in the Netherlands, the African Insect Science for Food and Health Institute in Kenya, Ifakara Health Institute in Tanzania and the Medical Research Council Laboratories in the Gambia are also participating in the project.
这两部分实验都是美国国立卫生研究院（NIH）基金会资助的全球卫生创新大挑战活动支持的一项为期五年的项目的一部分，而NIH的资金则来自以发现抑制疟疾扩散的新方法为目标的比尔及***达-盖茨基金会的捐款。荷兰瓦格宁根大学（Wageningen University）、肯尼亚非洲食品卫生昆虫科学研究院、坦桑尼亚伊法卡拉卫生研究院和冈比亚医学研究委员会实验室的科学家们也参与了这一项目。

Previously, scientists have used frog eggs to study the olfactory receptors of moths, honeybees and fruit flies. DNA that encodes insect receptors are injected into a frog egg and given sufficient time to produce and localize proteins. As a result, the surface of the egg is covered with the mosquito odorant receptors. An engineered egg is placed in a voltage clamp system and an odorant is dissolved in the buffer solution in which the egg is floating. If the mosquito receptors react to the compound, the electrical properties of the egg change in a measurable fashion.
在此之前，科学家们已经利用青蛙卵研究了飞蛾、蜜蜂和果蝇的嗅觉感受器。将为昆虫感受器编码的DNA注入青蛙卵并给以足够的时间以生产蛋白并使其定位。结果青蛙卵的表面被布满了蚊子的嗅觉感受器。一个经过基因改造的青蛙卵被放入电压钳系统，然后一种有气味的物质被溶入有青蛙卵的缓冲液中。如果蚊子的嗅觉感受器对这种物质有反应，那么青蛙卵的电学特性将会改变并可被检测出。

"The frog egg system is relatively rapid, highly sensitive and allows us to do very precise measurements of odorant response," says Guirong Wang, a senior research associate in the Zwiebel lab who was the lead author on the PNAS study and carried out several thousand egg/odorant recordings. "However, we call this a medium throughput system because, while it is relatively quick to set up, we have to make the odorant solutions by hand, which goes relatively slowly."
“青蛙卵系统反应相对比较迅速并且敏感性高，并且还可以让我们很准确的测量气味反应，” Zwiebel实验室的高级研究员Guirong Wang说。他是这项PNAS研究的主要作者并且作了上千条的卵/气味记录。“但是，我们只把它称为有中等生产能力的系统，因为尽管它建立起来较快，但我们不得不手工制备气味溶液，而这就相对较慢。”

By comparison, Yale's Drosophila system is a somewhat lower throughput system because it takes about three months to engineer a fruit fly with a mosquito odorant receptor in its antennae. The system, originally developed in the Carlson lab, uses mutant flies that are missing an odor receptor. Allison Carey, a graduate student in the Yale lab, systematically inserted mosquito genes into fruit flies one at a time so that a mosquito odorant receptor was expressed in place of the missing receptor. Although the method is slightly slower than the frog egg approach, it has some distinct advantages: Most notably it responds to volatilized odorants so it works with compounds that don't dissolve readily in water. It is also effective in detecting chemicals that inhibit receptors rather than exciting them.
通过比较发现，耶鲁大学的果蝇系统是个比较低效的系统，因为它需要大约三个月的时间才能将蚊子的气味感受器基因转入果蝇的触角内。最初由卡尔森实验室开发的这一系统使用缺失了一个嗅觉感受器的变异果蝇。耶鲁实验室的研究生Allison Carey一次只能将蚊子基因系统性的植入一只果蝇体内，以让蚊子气味感受器能在缺失的感受器的位置进行表达。尽管这一方法与青蛙卵方法相比有些慢，但它仍有一些优点：最突出的一点是它可以与挥发性气味剂发生反应，因而它可以与不易溶入水的气味化合物一起进行研究。它在探测抑制感受器而非激动感受器的化合物方面也非常有效。

"Both teams used the same set of 72 Anopheles odorant receptors and tested them using the same panel of 110 odorants," says Wang. The Vanderbilt team got responses from 37 of the odorant receptors in the frog eggs while testing 6,300 odorant-receptor combinations. "The results of the two systems were quite similar. There were only a few small differences."
“两个团队均使用同样的按蚊的72种嗅觉感受器和同样的110种气味剂来进行实验，”Wang说。范德尔比特大学研究团队在检测了6300个气味-受体结合后在青蛙卵的37个嗅觉感受器上发现有反应。“两个系统的研究结果非常相似，仅有很小的差异。”

Both studies found that most mosquito receptors are "generalists" that react to a number of different odors while a few are "specialists" that respond to a single or small number of odors. In some cases, the researchers found that a single odorant triggers several receptors while in other cases receptors are specifically tuned to unique compounds. In particular, they found 27 Anopheles receptors that respond strongly to compounds in human sweat.
两组研究均发现大多数蚊子嗅觉感受器可以对许多不同的气味产生反应而少数的感受器仅对一种或数种气味起反应。研究者们发现在某些情况下一种气味可能触发数种感受器，而在另外的情况下，感受器只对特定的气味起反应。尤为特别的是，他们发现27种按蚊感受器会对人体汗液中的化合物产生强烈反应。

"We're now screening for compounds that interact with these receptors. We call those that do BDOCs (behaviorally disruptive olfactory compounds)," Zwiebel says. "Compounds that excite some of these receptors could help lure mosquitoes into traps or repel them away from people while others that block receptor activity may help mask people. Ultimately we are looking for cocktails of multiple compounds that demonstrate activity in the field."
“我们目前正在筛选那些可以与这些感受器互相作用的化合物。我们把它们称为行为中断嗅觉化合物（BDOCs）。” Zwiebel说。“可以激动某些感受器的化合物可以帮助诱捕或驱赶蚊子，而那些阻断感受器活性的化合物可以帮助保护人们免受蚊子干扰。最终我们希望能得到一种含多种化合物并具有驱蚊效果的混合体。”

The project has already developed and patented a blend of BDOCs that is more attractive to mosquitoes than humans and has also identified several repellant BDOCs. It is currently in product development discussions with several private sector companies.
这一研究项目已经研发了一种可以吸引蚊子的BDOCs混合物，并且已经申请了专利，他们也确认了几种具有驱蚊效果的BDOCs。目前正在与几家私营公司就产品开发进行讨论。

编译：共1515字
疟疾防治新进展---移植蚊子气味感受器

每日科学（2010年2月16日）—范德比尔特（Vanderbilt）大学和耶鲁（Yale）大学的科学家们成功将大多数传播疟疾的蚊子的“鼻子”植入青蛙卵和果蝇体内，并利用这些替代品来控制严重威胁五亿人群的致死性疾病-疟疾的扩散。该研究发表于两种杂志的增刊上，一个是本周网络版的《美国国家科学院院刊》，另一个是《自然》2月3号的网络版。蚊子的“鼻子”位于两只触角的正当中，里面布满了神经细胞，这些神经细胞表明布满了特殊的“气味感受器”，能识别不同的化合物。昆虫的气味感受器（ORs）相当于人类鼻中的感受器和舌头上的味蕾。

“我们已经成功在青蛙卵和果蝇的触角中表达出80%的按蚊气味感受器，” 范德比尔特大学的生物科学教授Laurence Zwiebel说。他的实验室负责向青蛙卵中的移植。而在果蝇（黑腹果蝇）身上进行的移植则由耶鲁大学的分子细胞发展生物学教授John Carlson的实验室进行。这两部分实验都是美国国立卫生研究院（NIH）基金会资助的全球卫生创新大挑战活动支持的一项为期五年的项目的一部分，而NIH的资金则来自以发现抑制疟疾扩散的新方法为目标的比尔及***达-盖茨基金会的捐款。荷兰瓦格宁根大学（Wageningen University）、肯尼亚非洲食品卫生昆虫科学研究院、坦桑尼亚伊法卡拉卫生研究院和冈比亚医学研究委员会实验室的科学家们也参与了这一项目。

在此之前，科学家们已经利用青蛙卵研究了飞蛾、蜜蜂和果蝇的嗅觉感受器。将为昆虫感受器编码的DNA注入青蛙卵并给以足够的时间以生产蛋白并使其定位。结果青蛙卵的表面被布满了蚊子的嗅觉感受器。一个经过基因改造的青蛙卵被放入电压钳系统，然后一种有气味的物质被溶入有青蛙卵的缓冲液中。如果蚊子的嗅觉感受器对这种物质有反应，那么青蛙卵的电学特性将会改变并可被检测出。“青蛙卵系统反应相对比较迅速并且敏感性高，并且还可以让我们很准确的测量气味反应，” Zwiebel实验室的高级研究员Guirong Wang说。他是这项PNAS研究的主要作者并且作了上千条的卵/气味记录。“但是，我们只把它称为有中等生产能力的系统，因为尽管它建立起来较快，但我们不得不手工制备气味溶液，而这就相对较慢。”通过比较发现，耶鲁大学的果蝇系统是个比较低效的系统，因为它需要大约三个月的时间才能将蚊子的气味感受器基因转入果蝇的触角内。最初由卡尔森实验室开发的这一系统使用缺失了一个嗅觉感受器的变异果蝇。耶鲁实验室的研究生Allison Carey一次只能将蚊子基因系统性的植入一只果蝇体内，以让蚊子气味感受器能在缺失的感受器的位置进行表达。尽管这一方法与青蛙卵方法相比有些慢，但它仍有一些优点：最突出的一点是它可以与挥发性气味剂发生反应，因而它可以与不易溶入水的气味化合物一起进行研究。它在探测抑制感受器而非激动感受器的化合物方面也非常有效。

“两个团队均使用同样的按蚊的72种嗅觉感受器和同样的110种气味剂来进行实验，”Wang说。范德尔比特大学研究团队在检测了6300个气味-受体结合后在青蛙卵的37个嗅觉感受器上发现有反应。“两个系统的研究结果非常相似，仅有很小的差异。”两组研究均发现大多数蚊子嗅觉感受器可以对许多不同的气味产生反应而少数的感受器仅对一种或数种气味起反应。研究者们发现在某些情况下一种气味可能触发数种感受器，而在另外的情况下，感受器只对特定的气味起反应。尤为特别的是，他们发现27种按蚊感受器会对人体汗液中的化合物产生强烈反应。“我们目前正在筛选那些可以与这些感受器互相作用的化合物。我们把它们称为行为中断嗅觉化合物（BDOCs）。” Zwiebel说。“可以激动某些感受器的化合物可以帮助诱捕或驱赶蚊子，而那些阻断感受器活性的化合物可以帮助保护人们免受蚊子干扰。最终我们希望能得到一种含多种化合物并具有驱蚊效果的混合体。”这一研究项目已经研发了一种可以吸引蚊子的BDOCs混合物，并且已经申请了专利，他们也确认了几种具有驱蚊效果的BDOCs。目前正在与几家私营公司就产品开发进行讨论。

2010-02-24 16:44

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