Division of Epidemiology of Microbial Diseases, School of Public Health, Yale University, New Haven, CT 06511, United States.
J Insect Physiol. 2011 Nov;57(11):1553-61. doi: 10.1016/j.jinsphys.2011.08.009. Epub 2011 Aug 22.
During pregnancy in the viviparous tsetse fly, lipid mobilization is essential for the production of milk to feed the developing intrauterine larva. Lipophorin (Lp) functions as the major lipid transport protein in insects and closely-related arthropods. In this study, we assessed the role of Lp and the lipophorin receptor (LpR) in the lipid mobilization process during tsetse reproduction. We identified single gene sequences for GmmLp and GmmLpR from the genome of Glossinamorsitansmorsitans, and measured spatial and temporal expression of gmmlp and gmmlpr during the female reproductive cycle. Our results show that expression of gmmlp is specific to the adult fat body and larvae. In the adult female, gmmlp expression is constitutive. However transcript levels increase in the larva as it matures within the mother's uterus, reaching peak expression just prior to parturition. GmmLp was detected in the hemolymph of pregnant females and larvae, but not in the uterine fluid or larval gut contents ruling out the possibility of direct transfer of GmmLp from mother to offspring. Transcripts for gmmlpr were detected in the head, ovaries, midgut, milk gland/fat body, ovaries and developing larva. Levels of gmmlpr remain stable throughout the first and second gonotrophic cycles with a slight dip observed during the first gonotrophic cycle. GmmLpR was detected in multiple tissues, including the midgut, fat body, milk gland, spermatheca and head. Knockdown of gmmlp by RNA interference resulted in reduced hemolymph lipid levels, delayed oocyte development and extended larval gestation. Similar suppresion of gmmlpr did not significantly reduce hemolymph lipid levels or oogenesis duration, but did extend the duration of larval development. Thus, GmmLp function as the primary shuttle for lipids originating from the midgut and fat body to the ovaries and milk gland to supply resources for developing oocytes and larval nourishment, respectively. Once in the milk gland however, lipids are apparently transferred into the developing larva not by lipophorin but by another carrier lipoprotein.
在胎生采采蝇的怀孕过程中,脂类动员对于产生乳汁来喂养子宫内发育的幼虫至关重要。脂磷蛋白 (Lp) 是昆虫和密切相关的节肢动物中主要的脂质运输蛋白。在这项研究中,我们评估了 Lp 和脂磷蛋白受体 (LpR) 在采采蝇繁殖过程中的脂类动员中的作用。我们从 Glossina morsitans morsitans 的基因组中鉴定了 GmmLp 和 GmmLpR 的单个基因序列,并测量了 gmmlp 和 gmmlpr 在雌性生殖周期中的时空表达。我们的结果表明,gmmlp 的表达特异性存在于成年脂肪体和幼虫中。在成年雌性中,gmmlp 的表达是组成型的。然而,随着幼虫在母体内成熟,转录水平在幼虫中增加,在分娩前达到峰值表达。GmmLp 在怀孕雌性和幼虫的血淋巴中被检测到,但不在子宫液或幼虫肠道内容物中被检测到,这排除了 GmmLp 直接从母亲传递给后代的可能性。gmmlpr 的转录本在头部、卵巢、中肠、乳腺/脂肪体、卵巢和发育中的幼虫中被检测到。gmmlpr 的水平在第一和第二生殖周期中保持稳定,在第一生殖周期中略有下降。GmmLpR 在多个组织中被检测到,包括中肠、脂肪体、乳腺、精囊和头部。通过 RNA 干扰敲低 gmmlp 导致血淋巴脂质水平降低、卵母细胞发育延迟和幼虫妊娠延长。类似地抑制 gmmlpr 并没有显著降低血淋巴脂质水平或卵母细胞发生持续时间,但确实延长了幼虫发育的持续时间。因此,GmmLp 作为从中肠和脂肪体起源的脂质的主要穿梭蛋白,分别转运到卵巢和乳腺,为发育中的卵母细胞和幼虫的营养提供资源。然而,一旦进入乳腺,脂质显然不是通过脂磷蛋白而是通过另一种载脂蛋白进入正在发育的幼虫。