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Mechanistic study of transcription factor Sox18 during heart development 心脏发育过程中转录因子 Sox18 的机制研究
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-17 DOI: 10.1016/j.ygcen.2024.114472
Jianxin Liang , Imtiaz Ul Hassan , Man Yee Cheung , Lei Feng , Yi-jyun Lin , Qi Long , Chengdong Wang , Yuyue Ding , Ziqing Wang , Yuan Zhang , Yulong Li , Donghao Guo , Xiaofang Guo , Thomas Chi Bun Wong , Muhammad Kaleem Samma , Zixin Rong , Xufeng Qi , Dongqing Cai , Sai-Ming Ngai , Hui Zhao

Heart development is a delicate and complex process regulated by coordination of various signaling pathways. In this study, we investigated the role of sox18 in heart development by modulating Wnt/β-Catenin signaling pathways. Our spatiotemporal expression analysis revealed that sox18 is mainly expressed in the heart, branchial arch, pharyngeal arch, spinal cord, and intersegmental vessels at the tailbud stage of Xenopus tropicalis embryo. Overexpression of sox18 in the X. tropicalis embryos causes heart edema, while loss-of-function of sox18 can change the signal of developmental heart marker gata4 at different stages, suggesting that sox18 plays an essential role in the development of the heart. Knockdown of SOX18 in human umbilical vein endothelial cells suggests a link between Sox18 and β-CATENIN, a key regulator of the Wnt signaling pathway. Sox18 negatively regulates islet1 and tbx3, the downstream factors of Wnt/β-Catenin signaling, during the linear heart tube formation and the heart looping stage. Taken together, our findings highlight the crucial role of Sox18 in the development of the heart via inhibiting Wnt/β-Catenin signaling.

心脏发育是一个微妙而复杂的过程,受各种信号通路的协调调控。本研究探讨了sox18通过调节Wnt/β-Catenin信号通路在心脏发育中的作用。我们的时空表达分析发现,在热带爪蟾胚胎的尾芽期,sox18主要在心脏、支弓、咽弓、脊髓和节间血管中表达。在X. tropicalis胚胎中过表达sox18会导致心脏水肿,而sox18功能缺失会改变不同阶段心脏发育标志物gata4的信号,这表明sox18在心脏发育中起着至关重要的作用。敲除人脐静脉内皮细胞中的SOX18表明,Sox18与Wnt信号通路的关键调节因子β-CATENIN之间存在联系。在线性心管形成和心脏环形阶段,Sox18 负向调节 Wnt/β-Catenin 信号转导的下游因子 islet1 和 tbx3。综上所述,我们的研究结果凸显了 Sox18 通过抑制 Wnt/β-Catenin 信号在心脏发育过程中的关键作用。
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引用次数: 0
RNA-Seq analysis offers insight into the TBBPA-DHEE-induced endocrine-disrupting effect and neurotoxicity in juvenile zebrafish (Danio rerio) RNA-Seq分析有助于深入了解TBBPA-DHEE诱导的幼年斑马鱼(Danio rerio)内分泌干扰效应和神经毒性。
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-14 DOI: 10.1016/j.ygcen.2024.114469
Emmanuel Sunday Okeke , Weiwei Feng , Mengna Luo , Guanghua Mao , Yao Chen , Ting Zhao , Xiangyang Wu , Liuqing Yang

Tetrabromobisphenol A bis(2-hydroxyethyl) ether (TBBPA-DHEE) is the major TBBPA derivative. It has been detected in different environmental samples. Previous studies show that TBBPA-DHEE caused neurotoxicity in rats. In this study, juvenile zebrafish were exposed to various concentrations of TBBPA-DHEE to ascertain the potential neurotoxicity of TBBPA-DHEE, the chemical, and its possible molecular mechanism of action. Behavioral analysis revealed that TBBPA-DHEE could significantly increase the swimming distance and speed in the 1.5 mg/L group compared to the control. In contrast, the swimming distance and speed were significantly reduced in the 0.05 and 0.3 mg/L groups, affecting learning, memory, and neurodevelopment. Similarly, TBBPA-DHEE exposure caused a concentration-dependent significant increase in the levels of excitatory neurotransmitters, namely, dopamine, norepinephrine, and epinephrine, which could be attributed to the change observed in zebrafish behavior. This demonstrates the neurotoxicity of TBBPA-DHEE on juvenile zebrafish. The concentration-dependent increase in the IBR value revealed by the IBR index reveals the noticeable neurotoxic effect of TBBPA-DHEE. Transcriptomic analysis shows that TBBPA-DHEE exposure activated the PPAR signaling pathways, resulting in a disturbance of fatty acid (FA) metabolism and changes in the transcript levels of genes involved in these pathways, which could lead to lipotoxicity and hepatotoxicity. Our findings demonstrate a distinct endocrine-disrupting response to TBBPA-DHEE exposure, possibly contributing to abnormal behavioral alterations. This study provides novel insights into underlying the mechanisms and effects of TBBPA-DHEE on aquatic organisms, which may be helpful for environmental/human health risk assessments of the emerging pollutant.

四溴双酚 A 双(2-羟乙基)醚(TBBPA-DHEE)是四溴双酚 A 的主要衍生物。它已在不同的环境样本中被检测到。先前的研究表明,TBBPA-DHEE 会对大鼠造成神经毒性。在这项研究中,幼年斑马鱼暴露于不同浓度的 TBBPA-DHEE 中,以确定 TBBPA-DHEE 这种化学品的潜在神经毒性及其可能的分子作用机制。行为分析表明,与对照组相比,1.5 mg/L 组的 TBBPA-DHEE 能显著增加游泳距离和速度。相比之下,0.05 毫克/升和 0.3 毫克/升组的游泳距离和速度则明显降低,从而影响学习、记忆和神经发育。同样,暴露于 TBBPA-DHEE 会导致兴奋性神经递质(即多巴胺、去甲肾上腺素和肾上腺素)水平的浓度依赖性显著增加,这可能是斑马鱼行为发生变化的原因。这证明了 TBBPA-DHEE 对幼年斑马鱼的神经毒性。IBR指数显示的IBR值随浓度的增加而增加,这表明TBBPA-DHEE具有明显的神经毒性作用。转录组分析表明,TBBPA-DHEE暴露激活了PPAR信号通路,导致脂肪酸(FA)代谢紊乱以及这些通路中相关基因转录水平的变化,从而可能导致脂肪毒性和肝毒性。我们的研究结果表明,暴露于 TBBPA-DHEE 会产生独特的内分泌干扰反应,这可能是导致异常行为改变的原因之一。这项研究为了解 TBBPA-DHEE 对水生生物的潜在机制和影响提供了新的见解,可能有助于对这种新出现的污染物进行环境/人类健康风险评估。
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引用次数: 0
The chondrichthyan glucagon-like peptide 3 regulates hepatic ketone metabolism in the Pacific spiny dogfish Squalus suckleyi 软骨鱼类胰高血糖素样肽 3 调节太平洋刺狗鱼 Squalus suckleyi 的肝酮代谢。
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-10 DOI: 10.1016/j.ygcen.2024.114470
Alyssa M Weinrauch , Ian A Bouyoucos , J Michael Conlon , W Gary Anderson

Chondrichthyans have a novel proglucagon-derived peptide, glucagon-like peptide (GLP)-3, in addition to GLP-1 and GLP-2 that occur in other vertebrates. Given that the GLPs are important regulators of metabolic homeostasis across vertebrates, we sought to investigate whether GLP-3 displays functional actions on metabolism within a representative chondrichthyan, the Pacific spiny dogfish Squalus suckleyi. There were no observed effects of GLP-3 perfusion (10 nM for 15 min) on the rate of glucose or oleic acid acquisition at the level of the spiral valve nor were there any measured effects on intermediary metabolism within this tissue. Despite no effects on apparent glucose transport or glycolysis in the liver, a significant alteration to ketone metabolism occurred. Firstly, ketone flux through the perfused liver switched from a net endogenous production to consumption following hormone application. Accompanying this change, significant increases in mRNA transcript abundance of putative ketone transporters and in the activity of β-hydroxybutyrate dehydrogenase (a key enzyme regulating ketone flux in the liver) were observed. Overall, while these results show effects on hepatic metabolism, the physiological actions of GLP are distinct between this chondrichthyan and those of GLP-1 on teleost fishes. Whether this is the result of the particular metabolic dependency on ketone bodies in chondrichthyans or a differential function of a novel GLP remains to be fully elucidated.

除了存在于其他脊椎动物体内的 GLP-1 和 GLP-2 外,软骨鱼类还有一种新的胰高血糖素衍生肽--胰高血糖素样肽 (GLP)-3。鉴于 GLPs 是脊椎动物新陈代谢平衡的重要调节剂,我们试图研究 GLP-3 是否对具有代表性的软骨鱼类--太平洋刺狗鱼(Squalus suckleyi)的新陈代谢产生功能性作用。GLP-3 灌注(10 nM,15 分钟)对螺旋瓣膜水平的葡萄糖或油酸获取速度没有观察到任何影响,对该组织内的中间代谢也没有测量到任何影响。尽管对肝脏中明显的葡萄糖转运或糖酵解没有影响,但酮体代谢发生了显著变化。首先,在应用激素后,通过灌注肝脏的酮通量从内源性净产生转为消耗。伴随这一变化,还观察到推定酮体转运体的 mRNA 转录本丰度和 β-羟丁酸脱氢酶(调节肝脏酮体通量的一种关键酶)的活性显著增加。总之,虽然这些结果显示了对肝脏代谢的影响,但 GLP 的生理作用在这种软骨鱼类和 GLP-1 对远洋鱼类的作用之间是不同的。这究竟是软骨鱼类对酮体的特殊代谢依赖,还是一种新型 GLP 的不同功能,仍有待全面阐明。
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引用次数: 0
Coping with extreme free cortisol levels: Seasonal stress axis changes in sympatric North American flying squirrels 应对极高的游离皮质醇水平:同域北美鼯鼠的季节性应激轴变化。
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-09 DOI: 10.1016/j.ygcen.2024.114467
Samantha M. Stead , Phoebe D. Edwards , Rebekah Persad , Rudy Boonstra , Julie A. Teichroeb , Rupert Palme , Jeff Bowman

Most environments exhibit predictable yearly changes, permitting animals to anticipate them. The hypothalamic–pituitary–adrenal (HPA) axis is a key physiological pathway that enables animals to cope with such changes. Monitoring glucocorticoid (the end products of the HPA axis) levels in wild animals throughout the year can improve our understanding of how this pathway responds to different conditions. For this study, we collected 18 months of data on two species of North American flying squirrels (Glaucomys sabrinus and G. volans) living in a southern Ontario forest where temperature and food availability fluctuate dramatically throughout the year. These squirrels are active year-round, nest communally, and rely on scatter hoarded foods in the winter months. Flying squirrels have extremely high levels of free plasma cortisol relative to other mammals, but it is unknown how these levels are affected by environmental and reproductive factors. For both species, our goals were to (1) validate an enzyme immunoassay (EIA) to measure their fecal glucocorticoid metabolite (FGM) concentrations and (2) assess yearly differences, seasonal changes, and the influence of sex, reproduction, and ambient temperature on FGM concentrations in each species. In the lab, we successfully validated the use of antibody 5α-pregnane-3β, 11β, 21-triol-20-one EIA for FGM analysis in both species. In the field, neither sex nor reproductive status (breeding condition or not) were linked to FGM concentrations in either species. FGM concentrations were higher in autumn compared to the spring and summer. There were no other seasonal differences. We discuss possible explanations for the autumn peak in FGM concentrations (increased energy expenditure and social nesting changes), as well as outline possible avenues for future research. Understanding how individuals and populations respond to environmental change is a critical goal in evolutionary ecology, particularly in the context of a rapidly changing Anthropocene.

大多数环境每年都会发生可预测的变化,使动物能够预测这些变化。下丘脑-垂体-肾上腺(HPA)轴是动物应对这种变化的关键生理途径。对野生动物体内糖皮质激素(HPA 轴的最终产物)水平进行全年监测,可以加深我们对这一途径如何应对不同条件的理解。在这项研究中,我们收集了生活在安大略省南部森林中的两种北美鼯鼠(Glaucomys sabrinus 和 G. volans)18 个月的数据。这些鼯鼠全年都在活动,集体筑巢,冬季则依靠分散囤积的食物过冬。与其他哺乳动物相比,鼯鼠的游离血浆皮质醇水平极高,但这些水平如何受到环境和繁殖因素的影响尚不清楚。对于这两个物种,我们的目标是:(1)验证一种酶免疫测定(EIA)方法,以测量其粪便中糖皮质激素代谢物(FGM)的浓度;(2)评估每个物种的年度差异、季节变化以及性别、繁殖和环境温度对 FGM 浓度的影响。在实验室中,我们成功地验证了使用抗体 5α-孕甾-3β,11β,21-三醇-20-酮 EIA 分析这两个物种的 FGM。在野外,性别和繁殖状态(是否处于繁殖状态)都与这两个物种的胎毛膜浓度无关。与春季和夏季相比,秋季的胎毛膜浓度更高。没有其他季节性差异。我们讨论了秋季 FGM 浓度达到峰值的可能原因(能量消耗增加和社会筑巢变化),并概述了未来研究的可能途径。了解个体和种群如何应对环境变化是进化生态学的一个重要目标,尤其是在人类世快速变化的背景下。
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引用次数: 0
Gonadotropin expression, pituitary and plasma levels in the reproductive cycle of wild and captive-reared greater amberjack (Seriola dumerili) 野生和人工饲养的大琥珀鱼(Seriola dumerili)生殖周期中促性腺激素的表达、垂体和血浆水平。
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-08 DOI: 10.1016/j.ygcen.2024.114465
Ioannis Fakriadis , Iris Meiri-Ashkenazi , Chen Bracha , Hanna Rosenfeld , Aldo Corriero , Rosa Zupa , Chrysovalentinos Pousis , Maria Papadaki , Constantinos C. Mylonas

We compared the endocrine status of the pituitary-gonad axis of wild and captive-reared greater amberjack (Seriola dumerili) during the reproductive cycle (April – July), reporting on the expression and release of the two gonadotropins for the first time in the Mediterranean Sea. Ovaries from wild females were characterized histologically as DEVELOPING in early May and SPAWNING capable in late May-July, the latter having a 3 to 4-fold higher gonadosomatic index (GSI). SPAWNING capable wild females exhibited an increase in pituitary follicle stimulating hormone (Fsh) content, plasma testosterone (T) and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P), while almost a 10-fold increase was observed in pituitary luteinizing hormone (Lh) content. An increasing trend of plasma 17β-estradiol (E2) was also recorded between the two reproductive stages in wild females. Captive-reared females sampled during the reproductive cycle exhibited two additional reproductive categories, with REGRESSED females having extensive follicular atresia and fish in the REGENERATING stage having only primary oocytes in their ovaries. Pituitary content of Fsh and Lh, fshb and lhb expression and plasma levels of Fsh and Lh remained unchanged among the four reproductive stages in captive females, in contrast with plasma E2 and T that decreased in the REGENERATING stage, and 17,20β-P which increased after the DEVELOPING stage. In general, no significant hormonal differences were recorded between captive-reared and wild DEVELOPING females, in contrast to SPAWNING capable females, where pituitary Lh content, plasma Fsh and T were found to be lower in females in captivity. Overall, the captive females lagged behind in reproductive development compared to the wild ones and this was perhaps related to the multiple handling of the sea cages where all the sampled fish were maintained. Between wild males in the DEVELOPING and SPAWNING capable stages, pituitary Lh content, plasma T and 17,20β-P, and GSI exhibited 3 to 4-fold increases, while an increasing trend of pituitary Fsh content, lhb expression levels and plasma 11-ketotestosterone (11-KT) was also observed, and an opposite trend was observed in plasma Lh. Captive males were allocated to one more category, with REGRESSED individuals having no spermatogenic capacity. During the SPAWNING capable phase, almost all measured parameters were lower in captive males compared to wild ones. More importantly, captive males showed significant differences from their wild counterparts throughout the reproductive season, starting already from the DEVELOPING stage. Therefore, it appears that captivity already exerted negative effects in males prior to the onset of the study and the multiple handling of the cage where sampled fish were reared. Overall, the present study demonstrated that female greater amberjack do undergo full vitellogenesis in captivity, albeit with some dysfunctions t

我们比较了野生大琥珀鱼(Seriola dumerili)和人工饲养大琥珀鱼(Seriola dumerili)在生殖周期(4月至7月)垂体-性腺轴的内分泌状况,首次报告了地中海两种促性腺激素的表达和释放情况。从组织学角度分析,野生雌鱼的卵巢在 5 月初处于发育期,在 5 月下旬至 7 月具有破卵能力,后者的促性腺激素指数(GSI)比前者高 3 至 4 倍。具有卵巢发育能力的野生雌性垂体促卵泡激素(Fsh)含量、血浆睾酮(T)和17,20β-二羟基-4-孕烯-3-酮(17,20β-P)含量均有所增加,而垂体黄体生成素(Lh)含量几乎增加了10倍。野生雌性动物血浆中的 17β-雌二醇(E2)含量在两个繁殖阶段之间也呈上升趋势。人工饲养的雌鱼在生殖周期中的采样显示出两种额外的生殖类型,其中REGRESSED雌鱼有广泛的卵泡闭锁,而处于REGENERATING阶段的鱼卵巢中只有初级卵母细胞。人工饲养的雌鱼在四个生殖阶段的垂体中Fsh和Lh的含量、fshb和lhb的表达以及血浆中Fsh和Lh的水平保持不变,而血浆中的E2和T在REGENERATING阶段下降,17,20β-P在DEVELOPING阶段后上升。总体而言,人工饲养雌性和野生发育期雌性在激素方面没有发现明显差异,而哺乳期雌性的垂体Lh含量、血浆Fsh和T均低于人工饲养雌性。总体而言,人工饲养雌鱼的生殖发育落后于野生雌鱼,这可能与饲养所有采样鱼的海笼的多次处理有关。处于发育期和哺乳期的野生雄鱼的垂体Lh含量、血浆T和17,20β-P以及GSI均增加了3至4倍,垂体Fsh含量、lhb表达水平和血浆11-酮睾酮(11-KT)也呈上升趋势,而血浆Lh则呈相反趋势。人工饲养的雄性个体又被划分为一个类别,REGRESSED个体没有生精能力。与野生雄性动物相比,人工饲养雄性动物在有精子生成能力阶段的几乎所有测量参数都较低。更重要的是,人工饲养的雄性个体在整个繁殖季节都表现出与野生雄性个体的显著差异,从发育阶段就开始了。由此看来,在研究开始之前,人工饲养就已经对雄鱼产生了负面影响,而且对饲养采样鱼的网箱进行了多次处理。总之,本研究表明,雌性大琥珀鱼在人工饲养条件下确实进行了完全的卵黄发生,尽管有些功能障碍可能与实验的饲养方式有关,而另一方面,雄性大琥珀鱼甚至在研究开始前就可能受到人工饲养的严重影响。
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引用次数: 0
Adiponectin and visfatin expression profile in extra-embryonic annexes and role during embryo development in layer and broiler chickens 脂肪连接蛋白和粘蛋白在蛋鸡和肉鸡胚胎外附属物中的表达概况以及在胚胎发育过程中的作用。
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-06 DOI: 10.1016/j.ygcen.2024.114466
Ophélie Bernardi , Christelle Ramé , Maxime Reverchon , Joëlle Dupont

Some evidence showed differences between layer and broiler embryo development. We recently showed that two adipokines, adiponectin and visfatin are expressed in the extra embryonic membranes and fluids. However, their role in the embryo development is unknown. Thus, our objectives were 1. to compare the expression of AdipoQ and its receptors AdipoR1 and AdipoR2 and visfatin in extra-embryonic annexes in broiler and layer breeders during the embryo development and 2. to investigate the role of two adipokines in embryo development in both broiler and layer breed after in ovo injection of blocking antibodies against chicken adiponectin or visfatin. We found that adiponectin, AdipoR1, AdipoR2 and visfatin were mainly more expressed in the allantoic that in amniotic membranes. In addition, these expressions increased according the stage of embryo development. We observed a higher expression in layer than in broiler of AdipoQ in allantoic membranes at ED14 and ED18, of AdipoR1 and AdipoR2 in both allantoic and amniotic membranes at ED7 and ED14 and of visfatin only in allantoic membrane from ED7 to ED18. AdipoQ and visfatin were absent in amniotic fluid at ED7 but present at ED14 or ED18 where higher concentrations were detected in layer than in broiler. Interestingly, we showed a strong positive correlation between Adipo and visfatin concentration in amniotic fluid and the body weight of embryo in both breeds. However, after in ovo injection of Adipo antibodies we did not observe any effect on the embryo mortality whereas injection of visfatin antibodies increased in a dose dependent manner the embryo mortality in both breeds. Taken together, Adipo and visfatin are higher expressed in layer than broiler in extra-embryonic membranes and amniotic fluid. Our data suggest also that visfatin could be a main regulator of embryo development.

一些证据表明,蛋鸡和肉鸡的胚胎发育存在差异。我们最近发现,胚胎外膜和体液中表达了两种脂肪因子--脂肪连素和粘蛋白。然而,它们在胚胎发育过程中的作用尚不清楚。因此,我们的目标是:1.比较肉种鸡和蛋种鸡胚胎发育过程中胚胎外附属物中 AdipoQ 及其受体 AdipoR1 和 AdipoR2 以及粘脂蛋白的表达情况;2.研究肉种鸡和蛋种鸡在卵内注射鸡脂肪连接蛋白或粘脂蛋白的阻断抗体后,两种脂肪因子在胚胎发育过程中的作用。我们发现,脂肪连通素、AdipoR1、AdipoR2 和 visfatin 主要在尿囊膜中的表达高于羊膜。此外,这些表达随胚胎发育阶段的不同而增加。我们观察到,在ED14和ED18阶段,蛋鸡尿囊膜中AdipoQ的表达量高于肉鸡;在ED7和ED14阶段,尿囊膜和羊膜中AdipoR1和AdipoR2的表达量均高于肉鸡;而在ED7至ED18阶段,只有尿囊膜中的粘蛋白表达量高于肉鸡。在 ED7 羊水中不存在 AdipoQ 和 visfatin,但在 ED14 或 ED18 羊水中存在,其中在蛋鸡中检测到的浓度高于肉鸡。有趣的是,我们发现羊水中 Adipo 和 visfatin 的浓度与这两个品种胚胎的体重有很强的正相关性。然而,在卵内注射 Adipo 抗体后,我们没有观察到对胚胎死亡率有任何影响,而注射 visfatin 抗体后,两个品种的胚胎死亡率都会以剂量依赖的方式增加。综上所述,在胚胎外膜和羊水中,脂肪素和粘蛋白在蛋鸡中的表达高于肉鸡。我们的数据还表明,粘蛋白可能是胚胎发育的主要调节因子。
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引用次数: 0
α1 and β3 adrenergic receptor-mediated excitatory effects of adrenaline on the caudal neurosecretory system (CNSS) in olive flounder, Paralichthys olivaceus 肾上腺素介导的α1和β3肾上腺素能受体对橄榄鲽尾神经分泌系统(CNSS)的兴奋作用
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-05 DOI: 10.1016/j.ygcen.2024.114468
Mengmeng Shi , Cheng Liu , Yeyang Qin , Lin Yv , Weiqun Lu

Adrenaline is one of the most important neurotransmitters in the central nervous system and is produced during stress. In this study, we investigated the modulatory role of adrenaline and adrenergic receptors on the neuroendocrine Dahlgren cells in the caudal neurosecretory system (CNSS) of olive flounder. Ex vivo electrophysiological recordings revealed that adrenaline significantly increased the firing frequency and altered the firing pattern of Dahlgren cells. Moreover, treatment with adrenaline led to a significant upregulation of ion channels and major hormone secretion genes in CNSS at the mRNA levels. Additionally, treatment with adrenaline resulted in a significantly elevation in the expression levels of α1- and β3-adrenergic receptors. Furthermore, the β3-adrenergic receptor antagonist exerts a significant inhibitory effect on adrenaline-induced enhancement firing activities of Dahlgren cells, whereas the α1-adrenergic receptor antagonist displays a comparatively weaker inhibitory effect. Additionally, the enhanced firing activity induced by adrenaline could be effectively suppressed by both α1- and β3-adrenergic receptor antagonists. Taken together, these findings provide strong evidence in favor of the excitatory effects of adrenaline through α1 and β3 adrenergic receptors in CNSS to stimulate the secretion of stress-related hormones, β3-adrenergic receptor plays a more dominant role in the modulation of firing activities of Dahlgren cells by adrenaline and thereby regulates the stress response in olive flounder.

肾上腺素是中枢神经系统中最重要的神经递质之一,在应激时产生。本研究探讨了肾上腺素和肾上腺素能受体对橄榄鲽尾部神经分泌系统(CNSS)中神经内分泌达尔格伦细胞的调节作用。体外电生理记录显示,肾上腺素能显著提高达尔格伦细胞的发射频率并改变其发射模式。此外,肾上腺素还能在 mRNA 水平上显著上调 CNSS 中的离子通道和主要激素分泌基因。此外,肾上腺素还导致α1-和β3-肾上腺素能受体的表达水平明显升高。此外,β3-肾上腺素能受体拮抗剂对肾上腺素诱导的大鼠细胞发射活动增强有明显的抑制作用,而α1-肾上腺素能受体拮抗剂的抑制作用相对较弱。此外,α1-肾上腺素能受体拮抗剂和β3-肾上腺素能受体拮抗剂都能有效抑制肾上腺素诱导的发射活动增强。综上所述,这些研究结果有力地证明了肾上腺素通过CNSS中的α1和β3肾上腺素能受体产生兴奋作用,从而刺激应激相关激素的分泌,而β3肾上腺素能受体在肾上腺素对大肠细胞发射活动的调节中起着更主要的作用,从而调节橄榄鲽的应激反应。
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引用次数: 0
Characterization of insulin-like growth factor 3 and its potential role in the spotted steed Hemibarbus maculatus ovary development 胰岛素样生长因子 3 的特征及其在斑点骏马 Hemibarbus maculatus 卵巢发育中的潜在作用
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-03 DOI: 10.1016/j.ygcen.2024.114464
Huifen Liu , Qi Guo , Xinyu Wang , Xiao Ma , Xuejun Li , Xue Tian

As a new member of the insulin-like growth factors (Igfs), Igf3 was reported to play a vital role in fish reproduction. However, in spotted steed, the function of Igf3 remains largely unknown. In the present study, we identified and characterized Igf3 gene in spotted steed. Structural analysis showed that Igf3 contained five domains (B, C, A, D, E) and six conserved cysteine residues. The expression of Igf3 mRNA and protein were increased during ovary development and peaked in the maturation stage. The subcellular localization of IGF3 was highly expressed in granulosa cells and theca cells. Furthermore, recombinant IGF3 protein was produced and in vitro treatment with ovarian follicles significantly promoted the germinal vesicle breakdown (GVBD) rates of spotted steed follicles. The mRNA expression of cdc2 and cyclinB genes were significantly increased after IGF3 treatment, which were main components of maturation promoting factor (MPF). In addition, transcription levels of 3β-hsd, 20β-hsd, Cyp17a and Cyp19a1a were also changed. Taken together, these findings suggest that Igf3 is essential for ovary steroidogenesis and maturation in spotted steed.

据报道,作为胰岛素样生长因子(Igfs)的新成员,Igf3在鱼类繁殖中发挥着重要作用。然而,在斑点叉尾鮰中,Igf3的功能在很大程度上仍然未知。在本研究中,我们发现并鉴定了斑点叉尾鮰的 Igf3 基因。结构分析表明,Igf3包含五个结构域(B、C、A、D、E)和六个保守的半胱氨酸残基。Igf3 mRNA和蛋白质的表达量在卵巢发育过程中增加,并在成熟期达到峰值。IGF3 在颗粒细胞和卵巢细胞中高表达。此外,重组IGF3蛋白的产生和体外处理卵泡可显著促进斑点骏马卵泡的生殖泡破裂率(GVBD)。IGF3处理后,促成熟因子(MPF)的主要成分cdc2和cyclinB基因的mRNA表达量明显增加。此外,3β-hsd、20β-hsd、Cyp17a和Cyp19a1a的转录水平也发生了变化。综上所述,这些研究结果表明,Igf3对斑点叉尾骡卵巢类固醇的生成和成熟至关重要。
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引用次数: 0
Zebrafish prss59.1 is involved in chorion development 斑马鱼 prss59.1 参与绒毛发育。
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-27 DOI: 10.1016/j.ygcen.2024.114453
Md. Rezanujjaman , Theeranukul Pachoensuk , Md. Forhad Hossain , Md. Maisum Sarwar Jyoti , Md. Rubel Rana , Eisei Tsutsumi , Takumi Mouri , Maria Bramastri Susilo , Klangnurak Wanlada , Chihiro Yamamoto , Md. Hasan Ali , Toshinobu Tokumoto

The prss59.1 gene was identified as one of 11 genes that were highly upregulated during the induction of ovulation in zebrafish by using an in vivo ovulation assay. Previously, we conducted biochemical characterization of Prss59.1 and revealed it to be a trypsin-like proteolytic enzyme. In this study, we established a prss59.1 gene knockout strain using the CRISPR/Cas9 system. Phenotypic analysis of prss59.1 knockout fish showed that prss59.1 is associated with chorion elevation, a prominent event in egg activation during fertilization. The chorions of heterozygous and homozygous prss59.1 mutant zebrafish were smaller than those of the wild type. The results suggested that Prss59.1 is necessary for chorion expansion. The homozygous prss59.1 mutant strain, with a small chorion, showed an extremely low survival rate. Fiber-supported knob-like structures (KS) on the chorion showed an abnormal structure in prss59.1 mutants. Prss59.1 was detected in the KS on the chorion. The pores on the chorion were smaller in the prss59.1 mutants than in the wild type. Transmission electron microscopy (TEM) observations of the cross sections of the chorions showed abnormalities in the chorion structure in prss59.1 mutants. These results demonstrated that Prss59.1 is involved in chorion elevation and in proper formation of the chorion, which is necessary for embryo development.

通过体内排卵试验,我们发现prss59.1基因是斑马鱼排卵诱导过程中高度上调的11个基因之一。此前,我们对 Prss59.1 进行了生化鉴定,发现它是一种类似胰蛋白酶的蛋白水解酶。在本研究中,我们利用 CRISPR/Cas9 系统建立了一个 prss59.1 基因敲除株系。对prss59.1基因敲除鱼的表型分析表明,prss59.1与绒毛膜隆起有关,而绒毛膜隆起是受精卵活化过程中的一个重要事件。杂合子和同合子prss59.1突变斑马鱼的绒毛比野生型的小。结果表明,Prss59.1 是绒毛膜扩张的必要条件。绒毛较小的同源prss59.1突变株存活率极低。在prss59.1突变体中,绒毛上的纤维支撑旋钮状结构(KS)显示出异常结构。在绒毛膜上的 KS 中检测到了 Prss59.1。与野生型相比,prss59.1突变体绒毛上的孔更小。对绒毛横截面的透射电子显微镜(TEM)观察显示,prss59.1突变体的绒毛结构异常。这些结果表明,Prss59.1参与了绒毛膜的升高和绒毛膜的正常形成,而绒毛膜的形成是胚胎发育所必需的。
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引用次数: 0
Investigations of the function of AMH in granulosa cells in hens 母鸡颗粒细胞中 AMH 功能的研究
IF 2.7 3区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-22 DOI: 10.1016/j.ygcen.2024.114454
Laurie Francoeur, Deena M. Scoville, Patricia A. Johnson

Anti-mullerian hormone (AMH) plays a crucial role in follicle regulation in mammals by preventing premature primordial follicle activation and restricting follicle development through reduction of FSH sensitivity and inhibition of FSH-induced increase of steroidogenic enzymes. AMH is produced by granulosa cells from growing follicles and expression declines at the time of selection in both mammalian and avian species. The role of AMH in chicken granulosa cells remains unclear, as research is complicated because mammalian AMH is not bioactive in chickens and there is a lack of commercially available chicken AMH. In the current experiments, we used RNA interference to study the role of AMH on markers of follicle development in the presence and absence of FSH. Cultured chicken granulosa cells from 3–5 mm follicles and 6–8 mm follicles, the growing pool from which follicle selection is thought to occur, were used. Transfection with an AMH-specific siRNA significantly reduced AMH mRNA expression in granulosa cells from 3–5 mm and 6–8 mm follicles. Genes of interest were only measured in granulosa cells of 3–5 mm follicles due to low expression of AMH mRNA at the 6–8 mm follicle stage. Knockdown of AMH mRNA did not affect markers of follicle development (follicle stimulating hormone receptor, FSHR; steroidogenic acute regulatory protein, STAR; cytochrome P450 family 11 subfamily A member 1, CYP11A1; bone morphogenetic protein receptor type 2, BMPR2) or FSH responsiveness in granulosa cells from 3–5 mm follicles, indicating that AMH does not regulate follicle development directly by affecting markers of steroidogenesis, FSHR or BMPR2 at this follicle stage in chickens.

抗苗勒氏管激素(AMH)通过降低 FSH 敏感性和抑制 FSH 诱导的类固醇生成酶的增加,防止原始卵泡过早活化并限制卵泡发育,从而在哺乳动物的卵泡调节中发挥关键作用。AMH由生长卵泡中的颗粒细胞产生,在哺乳动物和禽类物种中,AMH的表达量在选择时都会下降。AMH在鸡颗粒细胞中的作用仍不清楚,因为哺乳动物的AMH在鸡体内没有生物活性,而且缺乏市售的鸡AMH,所以研究很复杂。在目前的实验中,我们使用RNA干扰来研究AMH在FSH存在和不存在的情况下对卵泡发育标志物的作用。我们使用了来自3-5毫米卵泡和6-8毫米卵泡的培养鸡颗粒细胞,这两个卵泡被认为是进行卵泡选择的生长池。转染AMH特异性siRNA可明显降低3-5毫米和6-8毫米卵泡颗粒细胞中AMH mRNA的表达。由于6-8毫米卵泡阶段的AMH mRNA表达量较低,因此只在3-5毫米卵泡的颗粒细胞中测量了相关基因。敲除 AMH mRNA 并不影响卵泡发育的标志物(促卵泡激素受体 FSHR;类固醇生成急性调节蛋白 STAR;细胞色素 P450 家族 11 亚家族 A 成员 1 CYP11A1;骨形态发生蛋白受体类型 CYP11A1);这表明在鸡的这一卵泡阶段,AMH 不会通过影响类固醇生成标志物、FSHR 或 BMPR2 直接调节卵泡发育。
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引用次数: 0
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General and comparative endocrinology
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