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Developmental programming of reproduction in the female animal 雌性动物的生殖发育程序
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-16 DOI: 10.1016/j.anireprosci.2024.107456
Vahid Akbarinejad , Robert A. Cushman

Successful reproduction is a cornerstone in food animal industry in order to sustain food production for human. Therefore, various methods focusing on genetics and postnatal environment have been identified and applied to improve fertility in livestock. Yet there is evidence indicating that environmental factors during prenatal and/or neonatal life can also impact the function of reproductive system and fertility in the animals during adulthood, which is called the developmental programming of reproduction. The current review summarizes data associated with the developmental origins of reproduction in the female animals. In this regard, this review focuses on the effect of plane of nutrition, maternal body condition, hypoxia, litter size, maternal age, parity, level of milk production and milk components, lactocrine signaling, stress, thermal stress, exposure to androgens, endocrine disrupting chemicals, mycotoxins and pollutants, affliction with infection and inflammation, and maternal gut microbiota during prenatal and neonatal periods on the neuroendocrine system, puberty, health of reproductive organs and fertility in the female offspring. It is noteworthy that these prenatal and neonatal factors do not always exert their effects on the reproductive performance of the female by compromising the development of organs directly related to reproductive function such as hypothalamus, pituitary, ovary, oviduct and uterus. Since they can impair the development of non-reproductive organs and systems modulating reproductive function as well (e.g., metabolic system and level of milk yield in dairy animals). Furthermore, when these factors affect the epigenetics of the offspring, their adverse effects will not be limited to one generation and can transfer transgenerationally. Hence, pinpointing the factors influencing developmental programming of reproduction and considering them in management of livestock operations could be a potential strategy to help improve fertility in food animals.

成功繁殖是食用畜牧业维持人类粮食生产的基石。因此,人们发现并应用了各种以遗传学和产后环境为重点的方法来提高家畜的繁殖力。然而,有证据表明,产前和/或新生儿期的环境因素也会影响动物生殖系统的功能和成年后的生育能力,这就是所谓的生殖发育程序。本综述总结了与雌性动物生殖发育起源有关的数据。在这方面,本综述侧重于营养平面、母体身体状况、缺氧、窝产仔数、母体年龄、奇偶数、产乳水平和乳汁成分、泌乳激素信号传导、应激、热应激、暴露于雄激素、内分泌干扰化学物、雌激素和雄激素等因素的影响、内分泌干扰化学物质、霉菌毒素和污染物、感染和炎症以及产前和新生儿期母体肠道微生物群对雌性后代的神经内分泌系统、青春期、生殖器官健康和生育能力的影响。值得注意的是,这些产前和新生儿期因素并不总是通过损害与生殖功能直接相关的器官(如下丘脑、垂体、卵巢、输卵管和子宫)的发育来影响女性的生殖能力。因为它们也会损害非生殖器官和调节生殖功能的系统的发育(如奶牛的新陈代谢系统和产奶量水平)。此外,当这些因素影响到后代的表观遗传学时,其不利影响将不仅限于一代人,还可能跨代转移。因此,找出影响繁殖发育程序的因素,并在家畜饲养管理中加以考虑,可能是帮助提高食用动物繁殖力的潜在策略。
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引用次数: 0
Factors affecting the success of laparoscopic artificial insemination in sheep 影响绵羊腹腔镜人工授精成功的因素
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-14 DOI: 10.1016/j.anireprosci.2024.107453
E.A. Spanner, S.P. de Graaf, J.P. Rickard

Successful artificial breeding underpins rapid genetic and production gains in animal agriculture. In sheep, artificial insemination with frozen semen is performed via intrauterine laparoscopy as frozen-thawed spermatozoa do not traverse the cervix in sufficient numbers for high fertility and transcervical insemination is anatomically impossible in most ewes. Historically, laparoscopic artificial insemination has always been considered reasonably successful, but recent anecdotal reports of poor fertility place it at risk of warning adoption. Understanding the male, female and environmental factors that influence the fertility of sheep is warranted if the success of artificial insemination is to be improved and genetic progress maximised for the sheep industry. This review details the current practice of laparoscopic AI in sheep. It explores the effects of semen quantity and quality, the ewe, her preparation, and environmental conditions, on the fertility obtained following laparoscopic artificial insemination.

成功的人工繁殖是畜牧业快速提高遗传和产量的基础。在绵羊中,冷冻精液的人工授精是通过宫腔腹腔镜进行的,因为冷冻解冻的精子没有足够的数量穿过子宫颈以获得高受精率,而且从解剖学角度来看,大多数母羊不可能进行经子宫颈人工授精。从历史上看,腹腔镜人工授精一直被认为是相当成功的,但最近关于受精率低的轶事报道使其面临被警告采用的风险。要想提高人工授精的成功率,最大限度地促进绵羊产业的遗传进步,就必须了解影响绵羊繁殖力的公羊、母羊和环境因素。本综述详细介绍了当前绵羊腹腔镜人工授精的实践。它探讨了精液的数量和质量、母羊、母羊的准备工作以及环境条件对腹腔镜人工授精后的受胎率的影响。
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引用次数: 0
Cold-induced damage in boar sperm cooled to 5°C in Lactose Egg Yolk (LEY) medium with different Glycerol concentrations 在不同甘油浓度的乳糖蛋黄(LEY)培养基中冷却至 5°C 的公猪精子的冷诱导损伤
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-13 DOI: 10.1016/j.anireprosci.2024.107455
Pablo Torres, Daniela Marina Malcervelli, Jonathan Hernán Cartelle, María Laura Fischman, Humberto Osvaldo Cisale

Boar sperm is highly susceptible to cold damage. When temperature drops to 5°C, the plasmatic membrane is destabilized. The freezing process causes a reduction of the fertility window because frozen/thawed boar sperm has less survivability. The aim of this work was to analyze the effect on sperm characteristics and response to capacitation stimuli of cooling to 5°C using a controlled protocol. Also, we evaluated if the addition of Glycerol 2% or 3% at 5°C was able to modify these parameters. For this purpose, we assessed motility, plasmatic membrane integrity and acrosomal membrane status. Capacitation was induced using Tyrode´s capacitating medium (TCM) and assessed by chlortetracycline stain and induction of acrosomal reaction with Progesterone. Motility patterns were analyzed using a CASA system. These tests were performed at three different points of the freezing curve: 37°C; 17°C and 5°C. Response to TCM vs TBM was only significant at 37°C. While at 37°C and 17°C capacitated sperm was below 20%, at 5°C reached 50% both in the TBM and TCM. CASA analysis showed that spermatozoa exposed to TCM had higher LIN and WOB than those in TBM. All parameters were similar in the Glycerol concentrations studied. These results suggest that the chilling process may be causing an effect similar to cryocapacitation along the cooling curve, starting subtle at 17°C and reaching 50% of the sperm population at 5°C, being independent of Glycerol concentration.

公猪精子极易受到低温损伤。当温度降至 5°C 时,质膜会失去稳定性。由于冷冻/解冻后的公猪精子存活率较低,因此冷冻过程会导致受精窗口期缩短。这项工作的目的是通过一个可控方案,分析降温至 5°C 对精子特性和获能刺激反应的影响。此外,我们还评估了在 5°C 时添加 2% 或 3% 的甘油是否能改变这些参数。为此,我们评估了运动能力、质膜完整性和顶体膜状态。使用泰罗德电容培养基(TCM)诱导电容,并通过金霉素染色和黄体酮诱导顶体反应进行评估。用 CASA 系统分析运动模式。这些测试在冷冻曲线的三个不同点进行:37°C、17°C 和 5°C。中药与 TBM 的反应仅在 37°C 时显著。在 37°C 和 17°C 时,获能精子低于 20%,而在 5°C 时,TBM 和 TCM 的获能精子均达到 50%。CASA 分析表明,与 TBM 相比,接触中药的精子具有更高的 LIN 和 WOB。在研究的甘油浓度下,所有参数都相似。这些结果表明,沿着冷却曲线,冷冻过程可能会产生与低温螯合类似的效果,从17°C开始微弱,到5°C时精子数量达到50%,且与甘油浓度无关。
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引用次数: 0
PPP2R2A promotes Hu sheep pituitary cell proliferation and gonadotropin secretion associated with prolificacy PPP2R2A 促进绵羊垂体细胞增殖和促性腺激素分泌
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-13 DOI: 10.1016/j.anireprosci.2024.107457
Hui Xu , Yu Cai , Hua Yang , Shanglai Li , Peiyong Chen , Zongyou Wei , Feng Wang , Ziyu Wang , Yanli Zhang

The anterior pituitary plays a critical role in the endocrine system, contains gonadotrophs, which regulate reproductive efficiency by secreting follicle-stimulating hormone (FSH) and luteinizing hormone (LH). PPP2R2A is a serine-threonine phosphatase that regulates reproductive functions in both females and males, its function in pituitary cells remain unclear. Hu sheep is a highly prolific breed, which makes it suitable for studying reproductive mechanisms. In this study, the relative abundances of PPP2R2A mRNA expression were higher in the pituitary of high-prolificacy (HF) Hu sheep compared to those of low-prolificacy (LF) Hu sheep. Additionally, we demonstrated that PPP2R2A promotes pituitary cell proliferation and gonadotropin secretion using the EdU assay and ELISA, respectively. Moreover, it inhibits pituitary cell apoptosis using flow cytometry. Furthermore, PPP2R2A may affect pituitary cell function by regulating the AKT/mTOR signaling pathway. In summary, our findings suggest that PPP2R2A may play a role in regulating pituitary function and influencing the secretion of gonadotropins.

垂体前叶在内分泌系统中发挥着关键作用,它含有促性腺激素,通过分泌卵泡刺激素(FSH)和黄体生成素(LH)来调节生殖效率。PPP2R2A 是一种丝氨酸-苏氨酸磷酸酶,可调节雌性和雄性的生殖功能,但它在垂体细胞中的功能仍不清楚。胡羊是一个高产品种,因此适合研究其生殖机制。在这项研究中,与低繁殖力(LF)胡羊相比,高繁殖力(HF)胡羊垂体中 PPP2R2A mRNA 表达的相对丰度更高。此外,我们还利用 EdU 试验和酶联免疫吸附试验分别证明了 PPP2R2A 能促进垂体细胞增殖和促性腺激素分泌。此外,它还通过流式细胞术抑制了垂体细胞的凋亡。此外,PPP2R2A 可能通过调节 AKT/mTOR 信号通路来影响垂体细胞的功能。总之,我们的研究结果表明,PPP2R2A 可能在调节垂体功能和影响促性腺激素分泌方面发挥作用。
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引用次数: 0
Maternal recognition of pregnancy in the pig: A servomechanism involving sex steroids, cytokines and prostaglandins 猪母体对怀孕的识别:涉及性类固醇、细胞因子和前列腺素的服务机制
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-12 DOI: 10.1016/j.anireprosci.2024.107452
Rodney D. Geisert , Fuller W. Bazer , Caroline G. Lucas , Caroline A. Pfeiffer , Ashley E. Meyer , Riley Sullivan , Destiny N. Johns , Mariana Sponchiado , Randall S. Prather

Maternal recognition of pregnancy (MRP) is a term utilized in mammals to describe pathways in which the conceptus alters the endometrial environment to prevent regression of corpora lutea to ensure continued production of progesterone (P4) required for establishment and maintenance of pregnancy. For nearly 40 years after publication of the endocrine/exocrine theory, conceptus estrogen (E2) was considered the primary maternal recognition signal in the pig. Conceptus production of prostaglandin E2 (PGE2) was also considered to be a major factor in preventing luteolysis. An addition to E2 and PGE2, pig conceptuses produce interleukin 1B2 (IL1B2) and interferons (IFN) delta (IFND) and gamma (IFNG). The present review provides brief history of the discovery of E2, PGs and IFNS which led to research investigating the role of these conceptus secreted factors in establishing and maintaining pregnancy in the pig. The recent utilization of gene editing technology allowed a more direct approach to investigate the in vivo roles of IL1B2, E2, PGE2, AND IFNG for establishment of pregnancy. These studies revealed unknown functions for IFNG and ILB2 in addition to PGE2 and E2. Thus, pregnancy recognition signal is via a servomechanism in requiring sequential effects of P4, E2, IL1B2, PGE2 and IFNG. Results indicate that the original established dogma for the role of conceptus E2 and PGs in MRP is a far too simplified model that involves the interplay of numerous mechanisms for inhibiting luteolysis, inducing critical elongation of the conceptuses and resolution of inflammation in pigs.

母体识别妊娠(MRP)是哺乳动物的一个术语,用于描述受体改变子宫内膜环境以防止黄体退化,从而确保持续产生建立和维持妊娠所需的孕酮(P4)的途径。在内分泌/外分泌理论发表后的近 40 年中,受体雌激素(E2)一直被认为是猪的主要母体识别信号。受体产生的前列腺素 E2(PGE2)也被认为是防止黄体溶解的主要因素。除 E2 和 PGE2 外,猪的受体还能产生白细胞介素 1B2 (IL1B2) 和干扰素 (IFN) δ (IFND) 及 γ (IFNG)。本综述简要介绍了发现 E2、PGs 和 IFNS 的历史,这些发现促使人们开始研究这些受体分泌因子在建立和维持猪妊娠中的作用。最近,基因编辑技术的使用使得研究 IL1B2、E2、PGE2 和 IFNG 在建立妊娠中的作用有了更直接的方法。这些研究揭示了除 PGE2 和 E2 外,IFNG 和 ILB2 的未知功能。因此,妊娠识别信号是通过一种伺服机制,需要 P4、E2、IL1B2、PGE2 和 IFNG 的相继作用。研究结果表明,最初关于受体 E2 和 PGs 在 MRP 中的作用的教条是一个过于简化的模型,它涉及到抑制黄体溶解、诱导受体临界伸长和解决猪的炎症等多种机制的相互作用。
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引用次数: 0
Ovarian lavage / artificial insemination method as an alternative approach for controlled reproduction of the northern pike (Esox lucius) 卵巢灌洗/人工授精法作为北方梭子鱼(Esox lucius)控制繁殖的替代方法
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-12 DOI: 10.1016/j.anireprosci.2024.107454
Erfan Akbari Nargesi , Danial Gorouhi

This study aimed to compare the effectiveness of the ovarian lavage / artificial insemination method with the traditional hormonal administration and fertilization methods over the artificial reproduction of the northern pike (Esox lucius). For this purpose, groups of five females were treated as follows: intraperitoneal injection of saline (C1); ovarian lavage with saline (C2); intraperitoneal injection of carp pituitary extract (CPE, T1); ovarian lavage with CPE (T2); intraperitoneal injection of CPE and ovarian lavage with semen after 72 h (T3); ovarian lavage with CPE and ovarian lavage with semen after 72 h (T4). According to the results, no fish ovulated in the control groups (C1 and C2). There were no significant differences (n.s.) among experimental treatments (P > 0.05, n.s.) in the reproductive parameters, such as latency time, ovulation rate, stripped egg amount, and pseudo-gonadosomatic index. The lowest fertilization rate (54.8%) was observed in the T4 treatment and significantly differed from the T1 and T2 treatments (P < 0.05). Moreover, the highest survival at swim-up stage was measured in the T4 treatment with a significant difference compared to the T1 group (P < 0.05). The survival at the eyed-egg stage, hatching rate, and malformations were similar (P > 0.05, n.s.) in all applied strategies. The results demonstrated that the ovarian lavage / artificial insemination method could be applied to control northern pike reproduction, like the traditional fertilization method. Consequently, this novel technique can be suggested as an alternative strategy to facilitate the hatchery operations in the controlled reproduction of this species.

本研究旨在比较卵巢灌洗/人工授精法与传统的激素管理和受精法对北方梭鱼人工繁殖的效果。为此,对每组 5 尾雌鱼进行了以下处理:腹腔注射生理盐水(C1);用生理盐水进行卵巢灌洗(C2);腹腔注射鲤鱼垂体提取物(CPE,T1);用 CPE 进行卵巢灌洗(T2);腹腔注射 CPE 并在 72 小时后用精液进行卵巢灌洗(T3);用 CPE 进行卵巢灌洗并在 72 小时后用精液进行卵巢灌洗(T4)。结果显示,对照组(C1 和 C2)没有鱼排卵。各试验处理之间的生殖参数,如潜伏时间、排卵率、剥离卵量和假性腺指数,均无明显差异(> 0.05, n.s.)。T4 处理的受精率最低(54.8%),与 T1 和 T2 处理相比差异显著(< 0.05)。此外,T4 处理的幼鱼在上游期的存活率最高,与 T1 组相比差异显著(< 0.05)。在所有应用策略中,卵裂期存活率、孵化率和畸形率相似( > 0.05, n.s.)。结果表明,卵巢灌洗/人工授精法与传统的受精法一样,可用于控制北方梭鱼的繁殖。因此,这项新技术可作为一种替代策略,用于促进孵化场控制该物种的繁殖。
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引用次数: 0
The opioid peptide leucine enkephalin modulates hypothalamic-hypophysial axis in the cichlid fish Oreochromis mossambicus 阿片肽亮氨酸脑啡肽调节慈鲷的下丘脑-生理下轴
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-08 DOI: 10.1016/j.anireprosci.2024.107451
Deepak Shinde, Shilpa K. Bhat, C.B. Ganesh

In vertebrates, opioid peptides are thought to be involved in the regulation of reproduction; however, the significance of enkephalins in testicular function remains unclear. We examined the influence of δ-opioid receptor agonist leucine enkephalin (L-ENK) on the hypophysial-testicular axis of the cichlid fish Oreochromis mossambicus. Treatment with a low dose of L-ENK (60 µg) caused a significant increase in the numbers of primary and secondary spermatocytes and early and late spermatids, concomitant with intense immunolabelling of testicular androgen receptors, but did not significantly alter serum luteinizing hormone (LH) and 11-ketotestosterone (11-KT) levels compared to those of controls. Nevertheless, treatment with a high dose of L-ENK (200 µg) caused a significant reduction in the numbers of secondary spermatocytes as well as late spermatids associated with marginal immunolabelling of androgen receptors and significantly lower concentrations of serum 11-KT and LH compared to controls. In addition, the serum cortisol level was not affected in low-dose L-ENK-treated fish, but its level was significantly increased in the high-dose L-ENK-treated group. Together, these findings indicate that a low dose of L-ENK stimulates the germ cells at the meiosis stage and promotes further stages of spermatogenesis, whereas a high concentration of L-ENK inhibits spermatogenesis at the advanced stages. This effect appears to be mediated through the suppression of testicular steroidogenesis and the reduction of LH release in the pituitary gland of tilapia. The findings also suggest that elevated L-ENK levels in teleosts may exert their inhibitory influence on the hypophysial-testicular axis via glucocorticoids.

在脊椎动物中,阿片肽被认为参与了生殖调节;然而,脑啡肽在睾丸功能中的意义仍不清楚。我们研究了δ-阿片受体激动剂亮氨酸脑啡肽(L-ENK)对慈鲷鱼(Oreochromis mossambicus)生理下轴-睾丸轴的影响。与对照组相比,低剂量(60 µg)L-ENK 能显著增加初级和次级精母细胞以及早期和晚期精子的数量,同时对睾丸雄激素受体进行强烈的免疫标记,但不会显著改变血清促黄体生成素(LH)和 11-酮睾酮(11-KT)的水平。然而,与对照组相比,高剂量 L-ENK 处理(200 µg)导致次级精母细胞和晚期精子数量显著减少,雄激素受体免疫标记边缘化,血清 11-KT 和 LH 浓度显著降低。此外,低剂量 L-ENK 处理的鱼血清皮质醇水平未受影响,但高剂量 L-ENK 处理组的皮质醇水平明显升高。这些发现共同表明,低剂量的 L-ENK 可刺激减数分裂阶段的生殖细胞,并促进精子发生的后续阶段,而高浓度的 L-ENK 则会抑制精子发生的后期阶段。这种作用似乎是通过抑制睾丸类固醇生成和减少罗非鱼垂体释放 LH 来实现的。研究结果还表明,远东鱼类体内 L-ENK 水平的升高可能是通过糖皮质激素对睾丸生精轴线产生抑制作用。
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引用次数: 0
In vitro characteristics of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and TLR7/8 ligand R848 用牛血清白蛋白(BSA)柱和 TLR7/8 配体 R848 分选的 X 型和 Y 型公羊精子的体外特征
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-03-02 DOI: 10.1016/j.anireprosci.2024.107450
Stanimir Yotov , Desislava Abadjieva , Anatoli Atanasov , Boyana Ivanova , Paulina Taushanova , Ivan Fasulkov , Elena Kistanova

The quality of the separated fractions in sex-sorted semen is very important for the success of the artificial insemination. This study aimed to evaluate some in vitro characteristics (DNA quantity, kinematic parameters and enzymes activity) of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and toll-like receptors (TLR)7/8 ligand R848. The ejaculates from six rams were collected by artificial vagina and subjected to a computer-assisted semen analysis (CASA). Total motility and percentage of the sperms with rapid and medium progressivity or non-progressivity in whole ejaculates and in X and Y fractions were analyzed. Activity of the enzymes ALP, GGT, CK, LDH and accumulation of lactate in the seminal plasma of ejaculates and in the environmental fluid of sexed spermatozoa were measured by biochemical analyzer. DNA was isolated from precipitated spermatozoa, and its quantity was measured. For both protocols the DNA mass from X-bearing fractions was higher, than from Y-bearing fractions. The high total motility of X- and Y-bearing spermatozoa as well as greater percent sperms with progressive motility were observed after use of BSA protocol. The application of TLR7/8 ligand R848 protocol led to reducing of Y-sperm motility and enhancement of non-progressivity in both fractions, which corresponded to the determined high amount of the extracellular lactate. For both methods, the significantly reduced activity of enzymes in the X and Y spermatozoa environmental fluids was established. Both protocols produce X- and Y-sperm fractions with satisfactory quality (over 80% total motility and over 50% rapid and medium progressive spermatozoa in each fraction).

性别分选精液中分离部分的质量对人工授精的成功非常重要。本研究旨在评估用牛血清白蛋白(BSA)柱和收费样受体(TLR)7/8配体 R848 分选的 X 型和 Y 型公羊精子的一些体外特征(DNA 数量、运动参数和酶活性)。通过人工阴道采集了六只公羊的射精,并对其进行了计算机辅助精液分析(CASA)。分析了整个精液以及 X 和 Y 部分精子的总运动能力、快速和中等进展性或非进展性精子的百分比。生化分析仪测量了射精的精浆和有性别精子环境液中 ALP、GGT、CK、LDH 等酶的活性和乳酸的积累。从沉淀的精子中分离出 DNA 并测量其数量。在两种方案中,X 型精子的 DNA 质量都高于 Y 型精子。使用 BSA 方案后,X 型和 Y 型精子的总运动能力较高,具有渐进运动能力的精子百分比也较高。使用 TLR7/8 配体 R848 方案会降低 Y 型精子的运动能力,并提高两个馏分中的非运动能力,这与确定的高细胞外乳酸盐含量相符。对于这两种方法,X 精子和 Y 精子环境液中酶的活性都明显降低。两种方法都能产生质量令人满意的X和Y精子馏分(每个馏分中总运动率超过80%,快速和中等进展精子超过50%)。
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引用次数: 0
Simple method for isolation and culture of primary buffalo (Bubalus bubalis) endometrial epithelial cells (pBuEECs) and its characterization using high throughput proteomics approach 分离和培养原代水牛(Bubalus bubalis)子宫内膜上皮细胞(pBuEECs)的简便方法,以及利用高通量蛋白质组学方法对其进行表征
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-29 DOI: 10.1016/j.anireprosci.2024.107449
Shradha Jamwal , Nikunj Tyagi , Jaideep Kumar , Jai Kumar Kaushik , Sudarshan Kumar , Ashok Kumar Mohanty

Early embryonic mortality resulting from insufficient interaction between the embryo and the uterus leads to the failure of pregnancy in livestock animals. Thus, it is imperative to comprehend the multifaceted process of implantation at molecular levels, which requires synchronized feto-maternal interaction. The in-vitro models serve as valuable tools to investigate the specific stages of implantation. The present study was undertaken to develop a simple method to isolate and culture the primary buffalo endometrial epithelial cells (pBuEECs), followed by proteome profiling of the proliferating cells. Collagenase I was used to separate uterine epithelial cells (UECs) from the ipsilateral uterine horn, and then the cells were separated using a cell strainer. After being seeded on culture plates, UECs developed colonies with characteristic epithelial shape and expressed important markers such as cytokeratin 18 (KRT18), progesterone receptor (PGR), β-estrogen receptor (ESR1), and leukemia inhibitory factor (LIF), which were confirmed by PCR. The purity of epithelial cells was assessed using cytokeratin 18 immunostaining, which indicated approximately 99% purity in cultured cells. The proteome profiling of pBuEECs via high-throughput tandem mass spectrometry (MS), identified a total of 3383 proteins. Bioinformatics analysis revealed enrichment in various biological processes, including cellular processes, metabolic processes, biological regulation, localization, signaling, and developmental processes. Moreover, the KEGG pathway analysis highlighted associations with the ribosome, proteosome, oxidative phosphorylation, spliceosome, and cytoskeleton regulation pathways. In conclusion, these well characterized cells offer valuable in-vitro model to enhance the understanding of implantation and uterine pathophysiology in livestock animals, particularly buffaloes.

由于胚胎与子宫之间的相互作用不足,胚胎早期死亡导致家畜妊娠失败。因此,当务之急是在分子水平上理解植入的多方面过程,这需要胎儿与母体的同步互动。模型是研究着床特定阶段的宝贵工具。本研究旨在开发一种简单的方法来分离和培养原代水牛子宫内膜上皮细胞(pBuEECs),然后对增殖细胞进行蛋白质组分析。用胶原酶 I 从同侧子宫角分离子宫上皮细胞(UECs),然后用细胞过滤器分离细胞。在培养板上播种后,UECs 形成了具有特征性上皮形状的菌落,并表达了细胞角蛋白 18(KRT18)、孕酮受体(PGR)、β-雌激素受体(ESR1)和白血病抑制因子(LIF)等重要标记物,这些标记物均通过 PCR 得到证实。使用细胞角蛋白 18 免疫染色法评估了上皮细胞的纯度,结果显示培养细胞的纯度约为 99%。通过高通量串联质谱(MS)对 pBuEECs 进行蛋白质组分析,共鉴定出 3,383 种蛋白质。生物信息学分析表明,这些蛋白质富集于各种生物过程,包括细胞过程、代谢过程、生物调控、定位、信号转导和发育过程。此外,KEGG 通路分析还强调了与核糖体、蛋白质组、氧化磷酸化、剪接体和细胞骨架调控通路的关联。总之,这些特征良好的细胞为加深对家畜(尤其是水牛)植入和子宫病理生理学的了解提供了宝贵的模型。
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引用次数: 0
Integrated omics analysis reveals a correlation between gut microbiota and egg production in captive African penguins (Spheniscus demersus) 综合全局分析揭示了圈养非洲企鹅肠道微生物群与产卵量之间的相关性
IF 2.2 2区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-02-27 DOI: 10.1016/j.anireprosci.2024.107448
Jingle Jiang, Di Hu, Enle Pei

The egg production of captive African penguins differs considerably between individuals. An understanding of the physiological differences in African penguins with relatively greater and lesser egg production is meaningful for the captive breeding program of this endangered species. The objective of this study was to investigate differential microbial composition and metabolites in captive African penguins with different egg production. Fecal samples were collected from captive female African penguins during the breeding season. The results of 16 S rRNA gene sequencing showed that African penguins with different egg production had similar microbial diversities, whereas a significant difference was observed between their microbial community structure. African penguins with relatively greater egg production exhibited a higher relative abundance of Alphaproteobacteria, Rhizobiales, Bradyrhizobiaceae, Bradyrhizobium and Bosea. Meanwhile, penguins with relatively lesser egg production had an increased proportion of Klebsiella and Plesiomonas. We further identified a total of 1858 metabolites in female African penguins by liquid chromatography-mass spectrometry analysis. Among these metabolites, 13 kinds of metabolites were found to be significantly differential between African penguins with different egg production. In addition, the correlation analysis revealed that the egg production had significant correlations with most of the differential microbial bacteria and metabolites. Our findings might aid in understanding the potential mechanism underlying the phenomenon of abnormal egg production in captive African penguins, and provide novel insights into the relationship between gut microbiota and reproduction in penguins.

人工饲养的非洲企鹅的产卵量在个体之间存在很大差异。了解产蛋量相对较高和较低的非洲企鹅的生理差异,对这一濒危物种的人工繁殖计划很有意义。本研究的目的是调查不同产蛋量的圈养非洲企鹅体内不同的微生物组成和代谢物。研究人员在繁殖季节收集了圈养雌性非洲企鹅的粪便样本。16 S rRNA基因测序结果表明,产卵量不同的非洲企鹅具有相似的微生物多样性,但其微生物群落结构存在显著差异。产蛋量相对较高的非洲企鹅表现出较高的Alphaproteobacteria、Rhizobiales、Bradyrhizobiaceae、Bradyrhizobium和Bosea的相对丰度。与此同时,产卵量相对较少的企鹅体内克雷伯氏菌和普莱西单胞菌的比例有所增加。通过液相色谱-质谱分析,我们进一步鉴定了雌性非洲企鹅体内的1858种代谢物。在这些代谢物中,有13种代谢物在不同产蛋量的非洲企鹅之间存在显著差异。此外,相关分析表明,产蛋量与大多数不同的微生物细菌和代谢物有明显的相关性。我们的研究结果可能有助于理解圈养非洲企鹅异常产蛋现象的潜在机制,并为企鹅肠道微生物群与繁殖之间的关系提供了新的见解。
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Animal Reproduction Science
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