東方果實蠅NMDA受器cDNA選殖及轉錄表現

cDNA Cloning and Transcriptional Expression of the N-methyl-D-aspartate Receptor in the Oriental Fruit Fly, Bactrocera dorsalis

簡靖倚(Ching-Yi Chien) ； 陳美娥(Mei-Er Chen)

《台灣昆蟲》 32卷4期 (2012/12) Pp. 303-316

https://doi.org/10.6661/TESFE.2012019

摘要

本研究利用RT-PCR及RACE選殖東方果實蠅（Bactrocera dorsalis）N-methyl-D-aspartate（NMDA）受器，得到全長3,643 bp之cDNA，含有一個2,958 bp之開放讀架，轉譯一個985個胺基酸之蛋白。利用basic local alignment search tool（BLAST）運算法分析序列，結果與Drosophila melanogaster NMDA receptor 1類源關係相近（85%相同度），將之命名為Bactrocera dorsalis NMDA receptor subunit 1（BdNR1）。以RT-PCR偵測BdNR1於不同發育時期之表現，結果顯示BdNR1於幼蟲期大量表現，蛹早期不表現，蛹晚期有少量表現，於0～10日齡雌成蟲則皆有表現。進一步測試BdNR1表現之組織專一性，結果顯示在成蟲各組織中以在飛行肌肉的表現量最高。以real-time PCR偵測0～10日齡雌成蟲飛行肌肉BdNR1的表現量，發現每個日齡皆有表現且表現量無明顯差異。東方果實蠅NMDA受器次單位1（BdNR1）在活動力相對較活躍的幼蟲期及成蟲期有表現，且於雌成蟲主要表現於飛行肌肉，而在不活動的蛹前期則不表現。因此我們推測BdNR1可能影響肌肉運動。另外，於其他昆蟲研究結果顯示阻斷NMDA受器會抑制卵巢發育。本研究注射NMDA受器通道阻斷劑MK-801於東方果實蠅雌成蟲，以測試其對卵巢發育之影響。結果顯示注射MK-801之實驗組卵巢大小與對照組比較並無差異。

關鍵字

東方果實蠅 ； NMDA受器 ； 飛行肌肉 ； 肌肉運動

並列摘要

In this study the Bactrocera dorsalis N-methyl-D-aspartate receptor (NMDA receptor) subunit 1 (BdNR1) cDNA was cloned using RT-PCR and RACE. The complete BdNR1 cDNA had a length of 3,643 bp including an open reading frame which is 2,958 bp to encode a 985-amino acid protein. The BLAST analysis revealed a high identity (85%) with the Drosophila melanogaster NMDA receptor 1. RT-PCR was applied to detect the transcriptional expression of BdNR1 in different developmental stages. The results showed that BdNR1 was highly expressed in the larvae. In pupae, there was no expression in the early period and only a little in the late period. In adults, BdNR1 was expressed in from 0- to 10-day-old female adults. Further, the highest transcriptional expression was found in the flight muscle in adults. Real-time PCR analysis revealed that BdNR1 was expressed in the flight muscle of 0- to 10-day-old female adults, and the expression level was not different. In conclusion, BdNR1 was expressed in vivacious larvae and adults, and not in immovable pupae. In addition, BdNR1 was highly expressed in the flight muscle. This suggests that BdNR1 may affect muscle contraction. However, blocking the NMDA receptor inhibits the ovarian development in other insects. In this study, the NMDA receptor antagonist, MK-801, was injected into female adults to detect the effect of the NMDA receptor on the ovarian development in B. dorsalis. The results showed that the size of the ovaries has no significant difference between the control group and the insects injected with MK-801.

並列關鍵字

Bactrocera dorsalis ； NMDA receptor ； flight muscle ； muscle contraction