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Expressions of sugar transporters/trehalases in relation to PTTH-stimulated ecdysteroidogenesis in the silkworm, Bombyx mori 糖转运体/三卤酶的表达与 PTTH 刺激的家蚕蜕皮激素生成的关系
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-07 DOI: 10.1016/j.jinsphys.2024.104672

The prothoracic gland (PG) is the source of ecdysteoids in larval insects. Although numerous studies have been conducted on signaling networks involved in prothoracicotropic hormone (PTTH)-stimulated ecdysteroidogenesis in PGs, less is known about regulation of metabolism in PGs. In the present study, we investigated correlations between expressions of sugar transporter (St)/trehalase (Treh) genes and PTTH-stimulated ecdysteroidogenesis in Bombyx mori PGs. Our results showed that in vitro PTTH treatment stimulated expression of the St1 gene, but not other transporter genes. Expression of the Treh1 gene was also stimulated by PTTH treatment. An immunoblotting analysis showed that St1 protein levels in Bombyx PGs increased during the later stage of the last larval instar and were not affect by PTTH treatment. PTTH treatment enhanced Treh enzyme activity in a time-dependent manner. Blocking either extracellular signal-regulated kinase (ERK) signaling with U0126 or phosphatidylinositol 3-kinase (PI3K) signaling with LY294002 decreased PTTH-stimulated Treh enzyme activity, indicating a link from the ERK and PI3K signaling pathways to Treh activity. Treatment with the Treh inhibitor, validamycin A, blocked PTTH-stimulated Treh enzyme activity and partially inhibited PTTH-stimulated ecdysteroidogenesis. Treatment with either a sugar transport inhibitor (cytochalasin B) or a specific glycolysis inhibitor (2-deoxy-D-glucose, 2-DG) partially inhibited PTTH-stimulated ecdysteroidogenesis. Taken together, these results indicate that increased expressions of St1/Treh1 and Treh activity, which lie downstream of PTTH signaling, are involved in PTTH stimulation in B. mori PGs.

前胸腺(PG)是幼虫体内蜕皮激素的来源。尽管已有大量研究涉及前胸腺激素(PTTH)刺激前胸腺中类蜕皮激素生成的信号网络,但对前胸腺中新陈代谢的调控却知之甚少。在本研究中,我们研究了糖转运体(St)/曲卤酶(Treh)基因的表达与 PTTH 刺激的蜕皮激素生成之间的相关性。结果表明,体外 PTTH 处理刺激了 St1 基因的表达,但没有刺激其他转运体基因的表达。PTTH处理也刺激了Treh1基因的表达。免疫印迹分析表明,St1 蛋白水平在末龄幼虫后期在雌雄鹅膏蕈蚊 PGs 中增加,且不受 PTTH 处理的影响。PTTH 处理以时间依赖性的方式增强了 Treh 酶的活性。用U0126阻断细胞外信号调节激酶(ERK)信号传导或用LY294002阻断磷脂酰肌醇3-激酶(PI3K)信号传导都会降低PTTH刺激的Treh酶活性,这表明ERK和PI3K信号传导途径与Treh酶活性有关。用 Treh 抑制剂有效霉素 A 处理可阻断 PTTH 刺激的 Treh 酶活性,并部分抑制 PTTH 刺激的类固醇生成。用糖转运抑制剂(细胞松弛素 B)或特异性糖酵解抑制剂(2-脱氧-D-葡萄糖,2-DG)处理可部分抑制 PTTH 刺激的蜕皮激素生成。综上所述,这些结果表明,St1/Treh1表达量的增加和Treh活性(位于PTTH信号转导的下游)参与了B. mori PGs的PTTH刺激。
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引用次数: 0
Evolution and development of Drosophila melanogaster under different thermal conditions affected cell sizes and sensitivity to paralyzing hypoxia 黑腹果蝇在不同温度条件下的进化和发育影响了细胞大小和对麻痹性缺氧的敏感性。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-06 DOI: 10.1016/j.jinsphys.2024.104671

Environmental gradients cause evolutionary and developmental changes in the cellular composition of organisms, but the physiological consequences of these effects are not well understood. Here, we studied experimental populations of Drosophila melanogaster that had evolved in one of three selective regimes: constant 16 °C, constant 25 °C, or intergenerational shifts between 16 °C and 25 °C. Genotypes from each population were reared at three developmental temperatures (16 °C, 20.5 °C, and 25 °C). As adults, we measured thorax length and cell sizes in the Malpighian tubules and wing epithelia of flies from each combination of evolutionary and developmental temperatures. We also exposed flies from these treatments to a short period of nearly complete oxygen deprivation to measure hypoxia tolerance. For genotypes from any selective regime, development at a higher temperature resulted in smaller flies with smaller cells, regardless of the tissue. At every developmental temperature, genotypes from the warm selective regime had smaller bodies and smaller wing cells but had larger tubule cells than did genotypes from the cold selective regime. Genotypes from the fluctuating selective regime were similar in size to those from the cold selective regime, but their cells of either tissue were the smallest among the three regimes. Evolutionary and developmental treatments interactively affected a fly’s sensitivity to short-term paralyzing hypoxia. Genotypes from the cold selective regime were less sensitive to hypoxia after developing at a higher temperature. Genotypes from the other selective regimes were more sensitive to hypoxia after developing at a higher temperature. Our results show that thermal conditions can trigger evolutionary and developmental shifts in cell size, coupled with changes in body size and hypoxia tolerance. These patterns suggest links between the cellular composition of the body, levels of hypoxia within cells, and the energetic cost of tissue maintenance. However, the patterns can be only partially explained by existing theories about the role of cell size in tissue oxygenation and metabolic performance.

环境梯度会引起生物细胞组成的进化和发育变化,但这些影响的生理后果还不甚明了。在这里,我们研究了黑腹果蝇的实验种群,这些种群在三种选择性制度中的一种中进化:恒定16 °C、恒定25 °C或16 °C和25 °C之间的代际转换。每个种群的基因型在三种发育温度(16 °C、20.5 °C和25 °C)下饲养。成年后,我们测量了每种进化和发育温度组合中苍蝇的胸部长度以及马尔皮格小管和翅膀上皮细胞的大小。我们还让这些处理中的苍蝇在短时间内几乎完全缺氧,以测量其对缺氧的耐受性。对于来自任何选择系统的基因型,在较高温度下发育都会导致苍蝇体型变小,细胞变小,而与组织无关。在每个发育温度下,与冷选择系统的基因型相比,暖选择系统的基因型体型较小,翅细胞较小,但小管细胞较大。波动选择机制的基因型与冷选择机制的基因型大小相似,但其任一组织的细胞都是三种机制中最小的。进化和发育处理相互作用,影响了苍蝇对短期麻痹性缺氧的敏感性。来自低温选择系统的基因型在较高温度下发育后,对缺氧的敏感性降低。其他选择系统的基因型在较高温度下发育后对缺氧更敏感。我们的研究结果表明,热条件会引发细胞大小的进化和发育转变,同时还会导致体型和耐缺氧能力的变化。这些模式表明,身体的细胞组成、细胞内的缺氧水平和组织维持的能量成本之间存在联系。然而,细胞大小在组织氧合和新陈代谢中的作用的现有理论只能部分解释这些模式。
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引用次数: 0
Effects of age on oxidative stress and locomotion in the pollinator, Megachile rotundata 年龄对传粉昆虫 Megachile rotundata 的氧化应激和运动的影响
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-03 DOI: 10.1016/j.jinsphys.2024.104666

Despite numerous aging studies, the relationship between oxidative stress, aging, and decline in functions such as locomotion is still debated. Insects offer a promising model for analyzing the relationship between oxidative stress and aging, because they exhibit vast differences in lifespan that may be affected by the environment, social factors, levels of activity, and aging interventions. In this study, we explore the effects of aging on oxidative stress and locomotion using the pollinator, Megachile rotundata, a species that is very mobile and active in the adult stage. Across the adult lifespan of M. rotundata, we assessed changes in walking, flight, oxidative damage, and antioxidant defenses. Our results suggest that M. rotundata experience age-related declines in flight, but not walking. Additionally, we found that oxidative damage and antioxidant capacity initially increase with age and physical activity, but then levels are maintained. Overall, these data show that M. rotundata, like some other organisms, may not perfectly follow the free radical theory of aging.

尽管进行了大量衰老研究,但氧化应激、衰老和运动等功能下降之间的关系仍存在争议。昆虫为分析氧化应激和衰老之间的关系提供了一个很有前景的模型,因为昆虫的寿命表现出巨大的差异,这些差异可能会受到环境、社会因素、活动水平和衰老干预措施的影响。在本研究中,我们利用传粉昆虫 Megachile rotundata(一种在成虫阶段非常活跃的物种)来探讨衰老对氧化应激和运动的影响。我们评估了Megachile rotundata在整个成年期的行走、飞行、氧化损伤和抗氧化防御能力的变化。我们的研究结果表明,轮虫的飞行能力会随着年龄的增长而下降,但步行能力不会。此外,我们还发现,氧化损伤和抗氧化能力最初会随着年龄和体力活动而增加,但随后会保持在一定水平。总之,这些数据表明,轮虫和其他一些生物一样,可能并不完全遵循自由基衰老理论。
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引用次数: 0
Plasticity in moth mating behavior and ejaculate proteomic diversity induced by male competition 飞蛾交配行为的可塑性和雄性竞争诱导的射精蛋白质组多样性
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-01 DOI: 10.1016/j.jinsphys.2024.104673
Zheng Yahong , Shi Lingping , Zhang Jing , Li Huixia , Wu Lingli , Zhao Qian

In male competition, large and costly ejaculates are advantageous. Prior research on male accessory gland secretions in Plutella xylostella left open questions about how males modulate their mating behaviors and ejaculate composition allocation in response to varying levels of competition. The current study aimed to delve deeper into these unexplored facets. A totally of 928 ejaculate proteins were identified across males exposed to different competition conditions. Notably, males courting under non–, low-, and high-competition scenarios exhibited 867, 635, and 858 ejaculate proteins, respectively. Approximately 10% of these ejaculate proteins displayed variations that aligned with changes in competition intensity. Subsequent analyses focused on the proteins transferred to females, revealing that 44% of ejaculate proteins were transferred, with 37 proteins exhibiting differential expression. Functional analyses uncovered their crucial roles in sperm maturation, motility, and capacitation. Our findings reveal adaptive adjustments in ejaculate protein abundance and transmission in P. xylostella as a response to varying competition levels. Moreover, fluorescent sperm labeling indicated higher sperm transfer during low competition correlated with shorter sperm length. Furthermore, evidence suggests that males shorten their courtship duration and extend their mating duration when faced with competition. These results illustrate how competition drives ejaculate investment and behavioral plasticity, offering valuable insights for advancements in assisted reproductive technologies and pest management strategies.

在雄性竞争中,大而昂贵的射精是有利的。先前对木虱雄性附属腺分泌物的研究留下了一些悬而未决的问题,即雄性如何调节其交配行为和射精成分分配以应对不同程度的竞争。本研究旨在深入探讨这些尚未探索的方面。本研究鉴定了暴露于不同竞争条件下的雄性的928种射精蛋白质。值得注意的是,在非竞争、低竞争和高竞争条件下求偶的雄性分别表现出867、635和858种射精蛋白。在这些射精蛋白质中,约有 10%的蛋白质随着竞争强度的变化而变化。随后的分析集中于转移到雌性体内的蛋白质,结果发现44%的射精蛋白质被转移,其中37种蛋白质表现出差异表达。功能分析揭示了它们在精子成熟、运动和获能过程中的关键作用。我们的研究结果表明,作为对不同竞争水平的反应,木虱的射精蛋白丰度和传递发生了适应性调整。此外,荧光精子标记表明,在低竞争水平下,精子转移率较高,而精子长度较短。此外,有证据表明,雄性在面临竞争时会缩短求偶时间,延长交配时间。这些结果说明了竞争是如何驱动射精投资和行为可塑性的,为辅助生殖技术和害虫管理策略的进步提供了宝贵的启示。
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引用次数: 0
Interaction between females and males grapevine moth Lobesia botrana modifies further mating preference 雌性和雄性葡萄蠹蛾 Lobesia botrana 之间的相互作用改变了进一步的交配偏好。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-01 DOI: 10.1016/j.jinsphys.2024.104668
Léa Lorrain-Soligon , Karen Muller , Camille Delaby , Denis Thiéry , Jérôme Moreau

During reproduction, females may boost their fitness by being selective based on direct material benefits provided by the males, such as nuptial gifts. In Lepidoptera, male provides a spermatophore containing nutrients. However, virgin males produce a bigger spermatophore, containing spermatozoa and nutrients, allowing higher female fertility. Lepidoptera females that could detect the sexual status of males may thus prefer a male without previous mating experience (i.e. a virgin male). This mate selection could be achieved by the use of chemical indices, such as sexual pheromones and cuticular compounds, known to be possibly exchanged during reproduction, and which can be indicators of a previous mating experience and known to be possibly sources of information exchanged. In this study, we experimentally presented Lobesia botrana virgin males with females in order for them to be exposed to females’ natural sexual pheromones or cuticular compounds. 12 or 48 h after the exposure of males to either females’ sexual pheromones or cuticular compounds, these males were confronted to naïve females, which have a choice between them or a virgin non-exposed males. We highlighted that, despite producing a spermatophore of similar volume, all exposed virgin males were less likely to mate with females 12 h after exposure, while after 48 h of exposure this is only the case for virgin males exposed to sexual pheromones. L. botrana females may thus discriminate male sexual experience based on chemical cues (either from cues transferred directly from females to males, or from changes in the cuticular or pheromone males’ profile) indicating past mating experiences. Mating duration was longer for males exposed to sexual pheromones after 12 h only, and for males exposed to cuticular compounds after 48 h only. Pheromones signal might be more persistent over time and seems to more easily gather information for males. The physiological reasoning behind this result still needs to be investigated.

在繁殖过程中,雌性可能会根据雄性提供的直接物质利益(如新婚礼物)进行选择,从而提高自身的适应能力。在鳞翅目昆虫中,雄性提供含有营养物质的精囊。然而,处女雄性会产生更大的精囊,其中包含精子和营养物质,从而提高雌性的生育能力。鳞翅目雌虫可以检测雄虫的性状况,因此可能更喜欢没有交配经验的雄虫(即处男)。这种配偶选择可以通过使用化学指标来实现,如性信息素和角质化合物,已知它们可能在繁殖过程中进行交换,可以作为先前交配经历的指标,已知它们可能是信息交换的来源。在本研究中,我们通过实验将草履虫处女雄性与雌性放在一起,让它们接触雌性的天然性信息素或角质化合物。在雄性暴露于雌性的性信息素或角质化合物12或48小时后,这些雄性将面对天真的雌性,雌性可以在它们或未暴露的处女雄性之间做出选择。我们发现,尽管产生的精子体积相似,但所有暴露的处男在暴露12小时后与雌性交配的可能性都较低,而暴露48小时后,只有暴露于性信息素的处男才会出现这种情况。因此,L. botrana雌性可能会根据化学线索(或者是雌性直接传递给雄性的线索,或者是雄性角质层或信息素特征的变化)来辨别雄性的性经验,这些线索表明雄性有过去的交配经验。接触性信息素 12 小时后的雄性交配持续时间更长,接触角质化合物 48 小时后的雄性交配持续时间更长。信息素信号可能更持久,似乎更容易为雄性收集信息。这一结果背后的生理原因仍有待研究。
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引用次数: 0
The heat shock response in Polistes spp. brood from differing climates following heat stress 来自不同气候条件的 Polistes spp.
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-01 DOI: 10.1016/j.jinsphys.2024.104667
A.B. Amstrup , H. Kovac , H. Käfer , A. Stabentheiner , J.G. Sørensen

Temperature is a crucial factor in many physiological processes, especially in small ectotherms whose body temperature is highly influenced by ambient temperature. Polistes (paper wasps) is a genus of primitively eusocial wasps found in widely varying thermal environments throughout the world. Paper wasps construct open-faced combs in which the brood is exposed to varying ambient temperatures. The Heat Shock Response is a physiological mechanism that has been shown to help cope with thermal stress. We investigated the expression of heat shock proteins in different life stages of three species of Polistes from different climates with the aim of deducing adaptive patterns. This was done by assaying heat shock protein (hsp70, hsp83, hsc70) expression during control conditions (25 °C) or a heat insult (35 or 45 °C) in individuals collected from natural populations in Alpine, Temperate, or Mediterranean climates. Basal expression of hsc70 and hsp83 was found to be high, while hsp70 and hsp83 expression was found to be highly responsive to severe heat stress. As expression levels varied based on species, geographical origin, and life stage as well as between heat shock proteins, the Heat Shock Response of Polistes was found to be complex. The results suggest that adaptive utilization of the heat shock response contributes to the ability of Polistes spp. to inhabit widely different thermal environments.

温度是许多生理过程的关键因素,尤其是体温受环境温度影响很大的小型外温动物。纸黄蜂(Polistes)是原始社会性黄蜂的一个属,在世界各地广泛存在于不同的热环境中。纸黄蜂建造敞开式蜂巢,使雏蜂暴露在不同的环境温度下。热休克反应是一种有助于应对热应力的生理机制。我们研究了来自不同气候条件下的三种纸袋蜂不同生命阶段的热休克蛋白表达情况,旨在推断其适应模式。我们从阿尔卑斯、温带或地中海气候的自然种群中采集了个体,在对照条件(25 °C)或热胁迫(35 或 45 °C)下检测热休克蛋白(hsp70、hsp83、hsc70)的表达。研究发现,hsc70和hsp83的基础表达量较高,而hsp70和hsp83的表达量对严重的热胁迫反应灵敏。由于物种、地理起源、生命阶段以及不同热休克蛋白之间的表达水平不同,研究发现多刺鱼的热休克反应是复杂的。研究结果表明,对热休克反应的适应性利用有助于提高 Polistes 属栖息于不同热环境的能力。
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引用次数: 0
No evidence for the melanin desiccation hypothesis in a larval Lepidopteran 一种鳞翅目幼虫的黑色素干燥假说没有证据。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-01 DOI: 10.1016/j.jinsphys.2024.104669
Sarah Britton , Goggy Davidowitz

Water regulation is an important physiological challenge for insects due to their small body sizes and large surface area to volume ratios. Adaptations for decreasing cuticular water loss, the largest avenue of loss, are especially important. The melanin desiccation hypothesis states that melanin molecules in the cuticle may help prevent water loss, thus offering protection from desiccation. This hypothesis has much empirical support in Drosophila species, but remains mostly untested in other taxa, including Lepidoptera. Because melanin has many other important functions in insects, its potential role in desiccation prevention is not always clear. In this study we investigated the role of melanin in desiccation prevention in the white-lined Sphinx moth, Hyles lineata (Lepidoptera, Sphingidae), which shows high plasticity in the degree of melanin pigmentation during the late larval instars. We took advantage of this plasticity and used density treatments to induce a wide range of cuticular melanization; solitary conditions induced low melanin pigmentation while crowded conditions induced high melanin pigmentation. We tested whether more melanic larvae from the crowded treatment were better protected from desiccation in three relevant responses: i) total water loss over a desiccation period, ii) change in hemolymph osmolality over a desiccation period, and iii) evaporation rate of water through the cuticle. We did not find support for the melanin desiccation hypothesis in this species. Although treatment influenced total water loss, this effect did not occur via degree of melanization. Interestingly, this implies that crowding, which was used to induce high melanin phenotypes, may have other physiological effects that influence water regulation. There were no differences between treatments in cuticular evaporative water loss or change in hemolymph osmolality. However, we conclude that osmolality may not sufficiently reflect water loss in this case. This study emphasizes the context dependency of melanin’s role in desiccation prevention and the importance of considering how it may vary across taxa. In lepidopteran larvae that are constantly feeding phytophagous insects with soft cuticles, melanin may not be necessary for preventing cuticular water loss.

由于昆虫体型小、表面积与体积比大,水分调节是昆虫面临的一项重要生理挑战。减少最大的失水途径--角质层失水的适应性尤其重要。黑色素干燥假说认为,角质层中的黑色素分子可能有助于防止水分流失,从而提供干燥保护。这一假说在果蝇物种中得到了很多经验支持,但在包括鳞翅目昆虫在内的其他类群中大多仍未得到验证。由于黑色素在昆虫中还有许多其他重要功能,因此它在防止干燥方面的潜在作用并不总是很明确。在本研究中,我们研究了黑色素在白线斯芬克斯蛾(鳞翅目,Sphingidae)的干燥预防中的作用。我们利用这种可塑性,使用密度处理来诱导广泛的角质层黑色素化;独居条件下诱导低黑色素色素沉着,而拥挤条件下诱导高黑色素色素沉着。我们测试了以下三个相关反应:i) 干燥期的总失水率;ii) 干燥期的血淋巴渗透压变化;iii) 水分通过角质层的蒸发率。我们没有发现黑色素干燥假说在该物种中得到支持。虽然处理会影响总失水率,但这种影响并不是通过黑色素的程度产生的。有趣的是,这意味着用于诱导高黑色素表型的拥挤可能会产生其他影响水分调节的生理效应。不同处理之间在角质层蒸发失水或血淋巴渗透压变化方面没有差异。不过,我们得出结论,在这种情况下,渗透压可能不能充分反映失水情况。这项研究强调了黑色素在防止干燥方面所起作用的环境依赖性,以及考虑黑色素在不同类群中如何变化的重要性。在鳞翅目幼虫不断取食具有软角质层的植食性昆虫时,黑色素可能不是防止角质层失水所必需的。
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引用次数: 0
An improved method to assess the encapsulation response in arthropods 评估节肢动物包囊反应的改进方法。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-01 DOI: 10.1016/j.jinsphys.2024.104670
Lucas J. Kirschman, Hannah M. Eastman, Frank J. Irovic, Nathaniel A. Nix, Ly Tuan Kiet Bui, Sydney L. Blackmon, Jaylen S. Greenlee, Rubina Lamichhane, Jack D. Mabuce, Hannah K. McAllister, Lilly F. Nevill, Peyton Redinger, Nia I. Rivers, Jackson L. Sprague

Ecoimmunology explores how ecological factors and evolutionary processes influence immune responses across various taxa and how immune responses trade-off with other traits. Studying immune responses requires biologically meaningful immunoassays applicable to a broad range of taxa and are sensitive enough to detect changes in the immune response. Useful immunoassays should also correlate with immunocompetence and fitness. The encapsulation response, a complex immune mechanism in arthropods, serves as a robust method for ecoimmunological investigations. However, traditional methods to test the encapsulation response can require long training. This study introduces an innovative, cost-effective method for assessing the encapsulation immune response in arthropods, which simplifies the procedure by reducing the training time and skill required. Our modified device utilizes a pen and syringe assembly for inserting monofilaments into arthropod larvae. We compared our device against traditional methods. Despite the new method being 22% faster, it did not compromise the accuracy or effectiveness of the encapsulation response when compared with traditional techniques, demonstrating similar degrees of melanization and encapsulation. Our method allowed for more accessible participation by less experienced researchers, such as undergraduates, facilitating their involvement in ecoimmunological research.

生态免疫学探索生态因素和进化过程如何影响不同类群的免疫反应,以及免疫反应如何与其他性状进行权衡。研究免疫反应需要有生物意义的免疫测定方法,这些方法应适用于广泛的类群,并具有足够的灵敏度来检测免疫反应的变化。有用的免疫测定还应与免疫能力和适应性相关。包囊反应是节肢动物的一种复杂免疫机制,是生态免疫学研究的有力方法。然而,测试包囊反应的传统方法需要长时间的训练。本研究介绍了一种创新的、具有成本效益的方法,用于评估节肢动物的包囊免疫反应,该方法通过减少所需的培训时间和技能简化了操作过程。我们改进的装置利用笔和注射器组件将单丝插入节肢动物幼虫体内。我们将我们的设备与传统方法进行了比较。尽管新方法快了 22%,但与传统技术相比,它并没有降低封装反应的准确性或有效性,显示出相似的黑色化和封装程度。我们的方法使经验不足的研究人员(如本科生)更容易参与,促进了他们参与生态免疫学研究。
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引用次数: 0
Caste-specific development of the dopaminergic system in bumble bees (Bombus ignitus) 大黄蜂(Bombus ignitus)多巴胺能系统的种姓特异性发育。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-06-19 DOI: 10.1016/j.jinsphys.2024.104665
Takafumi Onuma, Ken Sasaki

The dopaminergic system is involved in caste-specific behaviors in eusocial bumble bees. However, little is known about how the caste differences in dopaminergic system are formed during pupal stages in the brains of bumble bees. Thus, we investigated the levels of dopamine-related substances and expression of genes encoding enzymes involved in dopamine synthesis and metabolism, dopamine receptors, and a dopamine transporter in the brain of female Bombus ignitus. The levels of dopamine and dopamine-related substances in the brain were significantly higher in gynes than in workers from the late pupal stage to emergence, but the dynamics were similar between the castes. The relative expression levels of genes encoding enzymes involved in dopamine synthesis (BigTh and BigDdc) and dopamine metabolism (BigNat) increased significantly from pupal stage to emergence, but there were no differences in the relative expression levels of these genes between castes. A similar pattern was seen in the relative expression levels of four dopamine receptor genes (BigDop1, BigDop2, BigDop3, and BigDopEcR) and a dopamine transporter gene (BigDat). Compared with the honey bee Apis mellifera, the caste-specific dopaminergic system in the bumble bee is less differentiated, which might reflect the degree of behavioral specialization in these two species.

多巴胺能系统参与了社会性熊蜂的种性特异行为。然而,人们对多巴胺能系统的种性差异是如何在熊蜂蛹期大脑中形成的知之甚少。因此,我们研究了雌性点火熊蜂脑内多巴胺相关物质的水平以及编码多巴胺合成和代谢酶、多巴胺受体和多巴胺转运体的基因的表达。从化蛹后期到出壳,雌鸟脑内多巴胺和多巴胺相关物质的含量明显高于工蜂,但不同种群之间的动态变化相似。编码多巴胺合成酶(BigTh和BigDdc)和多巴胺代谢酶(BigNat)的基因的相对表达水平从蛹期到出巢期显著增加,但这些基因的相对表达水平在不同种群之间没有差异。四个多巴胺受体基因(BigDop1、BigDop2、BigDop3和BigDopEcR)和一个多巴胺转运体基因(BigDat)的相对表达水平也出现了类似的模式。与蜜蜂Apis mellifera相比,大黄蜂种姓特异性多巴胺能系统的分化程度较低,这可能反映了这两个物种的行为特化程度。
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引用次数: 0
A sperm-activating trypsin-like protease from the male reproductive tract of Spodoptera litura: Proteomic identification, sequence characterization, gene expression profile, RNAi and the effects of ionizing radiation 一种来自斑翅虫雄性生殖道的精子活化胰蛋白酶样蛋白酶:蛋白质组鉴定、序列表征、基因表达谱、RNAi 和电离辐射的影响。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-06-17 DOI: 10.1016/j.jinsphys.2024.104664
Priya Yadav , Rakesh K. Seth , Stuart E. Reynolds

Like other lepidopteran insects, males of the tobacco cutworm moth, Spodoptera litura produce two kinds of spermatozoa, eupyrene (nucleate) and apyrene (anucleate) sperm. Formed in the testis, both kinds of sperm are released into the male reproductive tract in an immature form and are stored in the duplex region of the tract. Neither type of sperm is motile at this stage. When stored apyrene sperm from the duplex are treated in vitro with an extract of the prostatic region of the male tract, or with mammalian trypsin, they become motile; activation is greater and achieved more rapidly with increasing concentration of extract or enzyme. The activating effect of prostatic extract is blocked by soybean trypsin inhibitor (SBTI), also in a dose-dependent way. These results suggest that the normal sperm-activating process is due to an endogenous trypsin-like protease produced in the prostatic region.

Proteomic analysis of S. litura prostatic extracts revealed a Trypsin-Like Serine Protease, TLSP, molecular weight 27 kDa, whose 199-residue amino acid sequence is identical to that of a predicted protein from the S. litura genome and is highly similar to predicted proteins encoded by genes in the genomes of several other noctuid moth species. Surprisingly, TLSP is only distantly related to Serine Protease 2 (initiatorin) of the silkmoth, Bombyx mori, the only identified lepidopteran protein so far shown to activate sperm. TLSP has features typical of secreted proteins, probably being synthesized as an inactive precursor zymogen, which is later activated by proteolytic cleavage.

cDNA was synthesized from total RNA extracted from the prostatic region and was used to examine TLSP expression using qPCR. tlsp mRNA was expressed in both the prostatic region and the accessory glands of the male tract. Injection of TLSP-specific dsRNA into adult males caused a significant reduction after 24 h in tlsp mRNA levels in both locations. The number of eggs laid by females mated to adult males that were given TLSP dsRNA in 10 % honey solution, and the fertility (% hatched) of the eggs were reduced. Injecting pupae with TLSP dsRNA caused the later activation of apyrene sperm motility by adult male prostatic extracts to be significantly reduced compared to controls.

Exposure of S. litura pupae to ionizing radiation significantly reduced expression of tlsp mRNA in the prostatic part and accessory gland of irradiated males in both the irradiated generation and also in their (unirradiated) F1 progeny. The implications of these findings for the use of the inherited sterility technique for the control of S. litura and other pest Lepidoptera are discussed.

与其他鳞翅目昆虫一样,烟草切割蛾(Spodoptera litura)的雄性也会产生两种精子,一种是有核(eupyrene)精子,另一种是无核(apyrene)精子。这两种精子都在睾丸中形成,以未成熟的形式释放到雄性生殖道中,并储存在生殖道的二叠区。这两种精子在这一阶段都没有运动能力。在体外用男性生殖道前列腺区提取物或哺乳动物胰蛋白酶处理储存在双重区的芘精子时,它们会变得有活力;提取物或酶的浓度越高,激活效果越大,速度越快。前列腺提取物的激活作用会被大豆胰蛋白酶抑制剂(SBTI)阻断,这种阻断也是剂量依赖性的。这些结果表明,正常的精子激活过程是由前列腺区域产生的内源性胰蛋白酶样蛋白酶引起的。对S. litura前列腺提取物的蛋白质组分析发现了一种分子量为27 kDa的类胰蛋白酶丝氨酸蛋白酶(TLSP),其199个氨基酸序列与S. litura基因组中的一种预测蛋白相同,并且与其他几种夜蛾物种基因组中基因编码的预测蛋白高度相似。令人惊讶的是,TLSP 与丝蛾的丝氨酸蛋白酶 2(initiatorin)只有很远的亲缘关系,而丝蛾的丝氨酸蛋白酶 2 是迄今为止发现的唯一能激活精子的鳞翅目昆虫蛋白。从前列腺区域提取的总 RNA 合成了 cDNA,并用 qPCR 检测 TLSP 的表达。向成年雄性动物注射 TLSP 特异性 dsRNA 24 小时后,这两个部位的 tlsp mRNA 水平均显著下降。在 10% 的蜂蜜溶液中注射 TLSP dsRNA 的成年雄虫交配的雌虫产卵数量和卵的受精率(孵化率)均有所下降。与对照组相比,给蛹注射 TLSP dsRNA 会显著降低成年雄性前列腺提取物对芘精子活力的激活作用。将 S. litura 蛹暴露于电离辐射中会显著降低辐照雄性前列腺部分和附属腺体中 tlsp mRNA 的表达,无论是在辐照一代还是在其 F1 后代(未受辐照)中都是如此。本文讨论了这些发现对利用遗传不育技术控制 S. litura 和其他有害鳞翅目昆虫的影响。
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引用次数: 0
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Journal of insect physiology
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