Functional and genetic characterization of neuropeptide Y-like receptors in Aedes aegypti.
Functional and genetic characterization of neuropeptide Y-like receptors in Aedes aegypti.
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DOI：
10.1371/journal.pntd.0002486
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发表时间：
2013
影响因子：
3.8
通讯作者：
Vosshall LB
中科院分区：
文献类型：
--
作者：
Liesch J;Bellani LL;Vosshall LB
雌性埃及伊蚊是登革热的主要传播媒介，每年造成 50-1 亿例感染，通过吸血在人与蚊子之间传播。艾。已知埃及伊蚊的宿主寻找行为在吸血后会被血淋巴携带的体液因子抑制三天。头肽-I 是介导这种抑制的候选肽，但该肽改变蚊子行为的机制以及它通过其发出信号的受体尚不清楚。头肽-I 显示出与短神经肽-F 肽 (sNPF) 的序列相似性，后者与进食行为有关，并且已知通过神经肽 Y (NPY) 样受体 (NPYLR) 发出信号。我们在 Ae 中发现了 8 个 NPYLR。埃及伊蚊基因组，并在基于细胞的钙成像测定中筛选每个肽，以检测其对一组肽的敏感性。四个Ae。埃及伊蚊 NPYLR 对一个或多个肽配体有反应，但只有 NYPLR1 对头肽-I 以及 sNPF 有反应。在莱姆病载体肩胛硬蜱的基因组中鉴定出的两个 NPYLR1 同源物也对头肽-I 敏感。注射合成头肽-I 和 sNPF-3 抑制非血液喂养雌性蚊子的宿主寻找行为，而注射缓冲液或无活性头肽-I [Cys10] 的对照注射则没有效果。为了探究 NPYLR1 是否是吸血诱导的宿主寻找抑制所必需的，我们使用锌指核酸酶生成了 5 个独立的 npylr1 无效突变株，并测试了它们的行为异常。 npylr1 突变体在运动、产卵、糖喂养、血液喂养、宿主寻找以及吸血后抑制宿主寻找方面表现出正常行为。在这项工作中，我们消除了四个 Ae 的孤儿。埃及伊蚊 NPYLR 并鉴定 NPYLR1 作为候选 sNPF 受体，该受体也对头肽-I 敏感。然而，npylr1 本身并不是宿主寻找抑制所必需的，我们得出的结论是其他受体、其他肽或两者都调节这一重要行为。雌性蚊子传播许多致命的传染病，包括疟疾、登革热和黄热病。这些蚊子需要吸血来产卵，并优先以人类为食，从而在进食时传播疾病。登革热媒介蚊子的雌性埃及伊蚊在吸血后会发生自然的行为变化，在三天多的时间内它们对人类失去吸引力。我们有兴趣了解这种自然的行为抑制，因为它可能提供控制蚊子吸血行为的机会。先前的研究表明，一种名为“头肽-I”的小蛋白质在注射到通常对人类表现出非常高的吸引力的非吸血雌性体内时可以模仿这种行为抑制。在这项工作中，我们着手寻找头肽-I 受体，并探究它是否会导致这种行为抑制。通过测试八种不同的候选受体，我们发现一种名为 NPYLR1 的受体对头肽 I 做出反应，但对另一种名为 sNPF-3 的肽更敏感。我们制作了缺乏 npylr1 基因的突变蚊子，发现突变体表现出正常的食糖和血液行为。我们得出的结论是，必须有额外的受体和/或肽共同导致雌性蚊子对人类吸引力的长期抑制。
Female Aedes aegypti mosquitoes are the principal vector for dengue fever, causing 50–100 million infections per year, transmitted between human and mosquito by blood feeding. Ae. aegypti host-seeking behavior is known to be inhibited for three days following a blood meal by a hemolymph-borne humoral factor. Head Peptide-I is a candidate peptide mediating this suppression, but the mechanism by which this peptide alters mosquito behavior and the receptor through which it signals are unknown. Head Peptide-I shows sequence similarity to short Neuropeptide-F peptides (sNPFs) that have been implicated in feeding behaviors and are known to signal through Neuropeptide Y (NPY)-Like Receptors (NPYLRs). We identified eight NPYLRs in the Ae. aegypti genome and screened each in a cell-based calcium imaging assay for sensitivity against a panel of peptides. Four of the Ae. aegypti NPYLRs responded to one or more peptide ligands, but only NYPLR1 responded to Head Peptide-I as well as sNPFs. Two NPYLR1 homologues identified in the genome of the Lyme disease vector, Ixodes scapularis, were also sensitive to Head Peptide-I. Injection of synthetic Head Peptide-I and sNPF-3 inhibited host-seeking behavior in non-blood-fed female mosquitoes, whereas control injections of buffer or inactive Head Peptide-I [Cys10] had no effect. To ask if NPYLR1 is necessary for blood-feeding-induced host-seeking inhibition, we used zinc-finger nucleases to generate five independent npylr1 null mutant strains and tested them for behavioral abnormalities. npylr1 mutants displayed normal behavior in locomotion, egg laying, sugar feeding, blood feeding, host seeking, and inhibition of host seeking after a blood meal. In this work we deorphanized four Ae. aegypti NPYLRs and identified NPYLR1 as a candidate sNPF receptor that is also sensitive to Head Peptide-I. Yet npylr1 alone is not required for host-seeking inhibition and we conclude that other receptors, additional peptides, or both, regulate this important behavior. Female mosquitoes are responsible for spreading many deadly infectious diseases including malaria, dengue fever, and yellow fever. These mosquitoes require a blood meal to produce eggs and preferentially feed on humans, thereby spreading disease as they feed. Females of the dengue vector mosquito Aedes aegypti undergo a natural change in behavior after a blood meal in which they lose attraction to humans for over three days. We are interested in understanding this natural behavioral inhibition because it may provide an opportunity to control mosquito blood-feeding behavior. Previous work showed that a small protein called Head Peptide-I could mimic this behavioral inhibition when injected into non-blood-fed females, which normally show very high attraction to humans. In this work, we set out to find the Head Peptide-I receptor and ask if it causes this behavioral inhibition. By testing eight different candidate receptors, we found one called NPYLR1 that responds to Head-Peptide I but is much more sensitive to another peptide called sNPF-3. We made mutant mosquitoes that lack the npylr1 gene and found that the mutants showed normal sugar- and blood-feeding behavior. We conclude that there must be additional receptors and/or peptides that together cause this long-lasting inhibition of female mosquito attraction to humans.
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DOI：
10.1073/pnas.0702726104
发表时间：
2007-05-15
影响因子：
11.1
作者：
Ja, William W.;Carvalho, Gil B.;Benzer, Seymour
通讯作者：
Benzer, Seymour
影响因子：
4.4
作者：
Predel, Reinhard;Neupert, Susanne;Garczynski, Stephen F.;Crim, Joe W.;Brown, Mark R.;Russell, William K.;Kahnt, Joerg;Russell, David H.;Nachman, Ronald J.
通讯作者：
Nachman, Ronald J.
DOI：
10.1016/s0006-291x(02)02351-3
发表时间：
2002-10-11
影响因子：
3.1
作者：
Mertens, I;Meeusen, T;Schoofs, L
通讯作者：
Schoofs, L
DOI：
10.1016/0020-1790(89)90073-5
发表时间：
1989-01-01
期刊：
INSECT BIOCHEMISTRY
影响因子：
--
作者：
MATSUMOTO, S;BROWN, MR;LEA, AO
通讯作者：
LEA, AO
影响因子：
4.8
作者：
OFFERMANNS, S;SIMON, MI
通讯作者：
SIMON, MI