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沙漠蝗(Schistocerca gregaria)不同组织中与阶段相关的6 kDa肽(PRP)的定位——免疫细胞化学和质谱分析方法

Localization of the phase-related 6-kDa peptide (PRP) in different tissues of the desert locust Schistocerca gregaria--immunocytochemical and mass spectrometric approach.

作者信息

Rahman M M, Breuer M, Begum M, Baggerman G, Huybrechts J, De Loof A

机构信息

Laboratory for Developmental Physiology, Genomics and Proteomics, Zoological Institute, Katholieke Universiteit Leuven, Naamsestraat 59, 3000 Leuven, Belgium.

出版信息

J Insect Physiol. 2008 Mar;54(3):543-54. doi: 10.1016/j.jinsphys.2007.12.003. Epub 2007 Dec 8.

Abstract

A 6-kDa phase-related peptide (PRP) was recently identified from the hemolymph of the desert locust Schistocerca gregaria. Its presence in much higher concentrations in the crowd-reared (gregarious) phase than in the isolated-reared (solitarious) one suggests a role in phase polyphenism. However, when tested in a variety of classical bioassays, no activity could be found. We hoped that uncovering its site(s) of synthesis might yield hints as to possible functions. An antiserum was raised against the C-terminal 16 aa part of PRP for use in immunocytochemistry. No immunoreactivity was recorded in the fat body, midgut, or Malpighian tubules. The strongest positive immunostaining was observed in the follicle cells of the ovary and in the seminal vesicle tubes of the male accessory gland complex. Also, positive were a pair of large neurosecretory cells in the subesophageal ganglion, the storage part of the corpora cardiaca and some nerve fibers in the brain- and abdominal regions. An additional mass spectrometric analysis was successfully done in combination with a BLAST search to detect possible false positive staining. This confirmed the presence of genuine PRP in most of the immunopositive tissues. Additional experiments are needed to unravel the role of PRP.

摘要

最近,从沙漠蝗虫(Schistocerca gregaria)的血淋巴中鉴定出一种6 kDa的阶段相关肽(PRP)。它在群体饲养(群居)阶段的浓度比单独饲养(独居)阶段高得多,这表明它在阶段多型性中发挥作用。然而,在各种经典生物测定中进行测试时,未发现其活性。我们希望揭示其合成位点可能会为其可能的功能提供线索。制备了针对PRP C末端16个氨基酸部分的抗血清,用于免疫细胞化学。在脂肪体、中肠或马氏管中未记录到免疫反应性。在卵巢的卵泡细胞和雄性附腺复合体的精囊管中观察到最强的阳性免疫染色。此外,咽下神经节中的一对大型神经分泌细胞、心侧体的储存部分以及脑区和腹部区域的一些神经纤维也呈阳性。结合BLAST搜索成功进行了额外的质谱分析,以检测可能的假阳性染色。这证实了大多数免疫阳性组织中存在真正的PRP。需要进行更多实验来阐明PRP的作用。

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