Manfrin Chiara, Tom Moshe, De Moro Gianluca, Gerdol Marco, Guarnaccia Corrado, Mosco Alessandro, Pallavicini Alberto, Giulianini Piero Giulio
Dipartimento di Scienze della Vita, Università di Trieste, Trieste, Italy.
PLoS One. 2013 Jun 19;8(6):e65176. doi: 10.1371/journal.pone.0065176. Print 2013.
The crustacean Hyperglycemic Hormone (cHH) is a neuropeptide present in many decapods. Two different chiral isomers are simultaneously present in Astacid crayfish and their specific biological functions are still poorly understood. The present study is aimed at better understanding the potentially different effect of each of the isomers on the hepatopancreatic gene expression profile in the crayfish Pontastacus leptodactylus, in the context of short term hyperglycemia. Hence, two different chemically synthesized cHH enantiomers, containing either L- or D-Phe(3), were injected to the circulation of intermolt females following removal of their X organ-Sinus gland complex. The effects triggered by the injection of the two alternate isomers were detected after one hour through measurement of circulating glucose levels. Triggered changes of the transcriptome expression profile in the hepatopancreas were analyzed by RNA-seq. A whole transcriptome shotgun sequence assembly provided the assumedly complete transcriptome of P. leptodactylus hepatopancreas, followed by RNA-seq analysis of changes in the expression level of many genes caused by the application of each of the hormone isomers. Circulating glucose levels were much higher in response to the D-isoform than to the L-isoform injection, one hour from injection. Similarly, the RNA-seq analysis confirmed a stronger effect on gene expression following the administration of D-cHH, while just limited alterations were caused by the L-isomer. These findings demonstrated a more prominent short term effect of the D-cHH on the transcription profile and shed light on the effect of the D-isomer on specific functional gene groups. Another contribution of the study is the construction of a de novo assembly of the hepatopancreas transcriptome, consisting of 39,935 contigs, that dramatically increases the molecular information available for this species and for crustaceans in general, providing an efficient tool for studying gene expression patterns in this organ.
甲壳类高血糖激素（cHH）是一种存在于许多十足目动物中的神经肽。螯虾体内同时存在两种不同的手性异构体，其具体生物学功能仍知之甚少。本研究旨在更好地理解在短期高血糖情况下，每种异构体对小龙虾Pontastacus leptodactylus肝胰腺基因表达谱的潜在不同影响。因此，在去除间 molt 期雌性小龙虾的 X 器官 - 窦腺复合体后，将两种不同的化学合成 cHH 对映体（分别含有 L - 或 D - Phe(3)）注射到其循环系统中。注射一小时后，通过测量循环葡萄糖水平来检测注射两种交替异构体所引发的效应。通过 RNA - seq 分析肝胰腺中转录组表达谱的触发变化。全转录组鸟枪法序列组装提供了假定完整的 P. leptodactylus 肝胰腺转录组，随后对每种激素异构体应用后许多基因表达水平的变化进行 RNA - seq 分析。注射后一小时，与注射 L - 异构体相比，注射 D - 异构体后循环葡萄糖水平要高得多。同样，RNA - seq 分析证实，施用 D - cHH 后对基因表达的影响更强，而 L - 异构体仅引起有限的变化。这些发现表明 D - cHH 在转录谱上具有更显著的短期效应，并揭示了 D - 异构体对特定功能基因组的影响。该研究的另一个贡献是构建了肝胰腺转录组的从头组装，由 39,935 个重叠群组成，这极大地增加了该物种以及一般甲壳类动物可用的分子信息，为研究该器官中的基因表达模式提供了一个有效的工具。
