Nakayama S, Kaiser K, Aigaki T
Department of Biology, Tokyo Metropolitan University, Japan.
Mol Gen Genet. 1997 Apr 28;254(4):449-55. doi: 10.1007/s004380050438.
Sex-peptide (SP), which is secreted by the accessory gland of Drosophila males, is transferred to the female during copulation, thereby reducing her sexual appetite (receptivity to males) and stimulating ovulation/oviposition. SP is known to be taken up into the hemolymph of mated females, but it is not clear whether there are two separate target tissues, for behavioral changes and ovulation or only one target for both responses. We have employed the GAL4-UAS system to express SP transgene constructs, both in different tissues and in different cellular components of virgin females. A cytoplasmic form of SP lacking a signal sequence did not evoke any responses, even when expressed ubiquitously. In contrast, a membrane-bound form of SP induced typical post-mating behavior, indicating that SP must be outside the cell in order to exert its biological effects. A total of 204 randomly selected P[GAL4] enhancer-trap lines were screened for their ability to induce SP responses in combination with the membrane-bound SP expressed under GAL4 control. Thirty-three lines were associated with both behavioral change and stimulated ovulation. No line was associated with only one of the two responses, implying that the SP target(s) mediating the two responses are either identical, very closely located, or present in two distinct tissues with a common set of genetic determinants. Western blot analysis of head, thorax, and abdominal extracts revealed that the biological activity was correlated with expression in the head fraction.
性肽(SP)由果蝇雄性附腺分泌,在交配过程中传递给雌性,从而降低其性欲望(对雄性的接受度)并刺激排卵/产卵。已知SP会被交配过的雌性吸收到血淋巴中,但尚不清楚是否存在两个独立的靶组织,分别用于行为变化和排卵,还是这两种反应只有一个靶组织。我们利用GAL4-UAS系统在未交配雌性的不同组织和不同细胞成分中表达SP转基因构建体。一种缺乏信号序列的细胞质形式的SP即使在全身表达也不会引起任何反应。相比之下,一种膜结合形式的SP诱导了典型的交配后行为,这表明SP必须位于细胞外才能发挥其生物学作用。总共筛选了204个随机选择的P[GAL4]增强子陷阱系,以检测它们与在GAL4控制下表达的膜结合SP结合时诱导SP反应的能力。33个系与行为变化和刺激排卵都有关联。没有一个系只与两种反应中的一种有关联,这意味着介导这两种反应的SP靶组织要么相同,要么位置非常接近,要么存在于具有一组共同遗传决定因素的两个不同组织中。对头部、胸部和腹部提取物的蛋白质免疫印迹分析表明,生物活性与头部组分中的表达相关。