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Serum concentrations of anti-Müllerian hormone modulate ovarian response to different doses of follicle-stimulating hormone in Japanese Black donors. 抗缪勒氏管激素的血清浓度可调节日本黑人捐献者卵巢对不同剂量卵泡刺激素的反应。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-12-13 Epub Date: 2024-11-10 DOI: 10.1262/jrd.2024-067
Hiroaki Okawa, Norihiro Yukiyama, Oky Setyo Widodo, Kanae Niimi, Yuta Chuman, Yasuo Fushimi, Mitsuhiro Takagi

Anti-Müllerian hormone (AMH) is a marker for predicting embryo production in cows subjected to superovulation; however, it remains to be established as a reliable predictor of reproductive performance. We hypothesized that the serum AMH concentration of donors affects the ovarian response to follicle-stimulating hormone (FSH) treatment during superovulation. Herein, we retrospectively investigated the association between different FSH doses and AMH concentrations in donor Japanese Black cows in a superovulation program and analyzed the number of total and transferable embryos recovered. The number of transferable embryos recovered from donors with high AMH levels was significantly higher than that recovered from donors with low AMH levels. Additionally, it increased further with a reduction in the FSH dose. These results illustrate that the AMH concentration is a useful marker for predicting embryo production after superovulation, and donors with high AMH levels produce more transferable embryos at low FSH doses than at high doses.

抗缪勒氏管激素(AMH)是预测超排卵奶牛胚胎生产的一个指标,但它作为繁殖性能的可靠预测指标仍有待确定。我们假设供体的血清 AMH 浓度会影响超级排卵期间卵巢对促卵泡激素(FSH)治疗的反应。在此,我们回顾性地研究了超级排卵计划中不同FSH剂量与供体日本黑牛AMH浓度之间的关系,并分析了回收的总胚胎数和可移植胚胎数。AMH水平高的供体回收的可移植胚胎数量明显高于AMH水平低的供体。此外,随着 FSH 剂量的减少,可移植胚胎数也进一步增加。这些结果说明,AMH 浓度是预测超排卵后胚胎产量的有效指标,AMH 水平高的供体在低 FSH 剂量下比高剂量下产生的可移植胚胎更多。
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引用次数: 0
Addition of granulocyte macrophage colony stimulating factor (GM-CSF) during in vitro oocyte maturation improves embryo development in a mouse model of advanced maternal age. 在体外卵母细胞成熟过程中加入粒细胞巨噬细胞集落刺激因子(GM-CSF)可改善高龄产妇小鼠模型的胚胎发育。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-12-13 Epub Date: 2024-11-09 DOI: 10.1262/jrd.2024-066
Anmol Saini, Nicole O McPherson, Mark B Nottle

Oocyte developmental competence declines in women aged 35 and older resulting in many women resorting to IVF. The present study determined whether adding Granulocyte-macrophage colony-stimulating factor (GM-CSF) during in vitro oocyte maturation (IVM) could improve oocyte developmental competence in a mouse model of advanced maternal age. Oocytes from 12-14 month C57BL6 J × CBA mice were treated with 10 ng/ml of GM-CSF during IVM, and embryo development, mitochondrial activity, spindle formation and chromosomal alignment were examined. The addition of GM-CSF tended to increase fertilisation rates (76.19 vs. 82.03%; P = 0.07) but did not affect cumulus expansion compared with control. The addition of GM-CSF also increased blastocysts rates (51.10 vs. 61.52%; P < 0.01) and the number of good quality blastocysts (33.31 vs. 44.13%; P < 0.05) present at 96 h of culture as well as inner cell mass (12.64 vs. 15.62 ; P < 0.01) and total cell number (42.98 vs. 48.78 ; P < 0.05). GM-CSF treatment also increased mitochondrial membrane potential two to three fold in the outer (2.86 vs. 0.97; P < 0.001), intermediate (3.25 vs. 0.89; P < 0.001) and peri nuclear areas (3.62 vs. 1.08; P < 0.001). GM-CSF treatment did not influence spindle formation or chromosomal alignment. Together our results indicate that the addition of GM-CSF during IVM may improve oocyte quality in women of advanced maternal age.

35 岁及以上妇女的卵母细胞发育能力下降,导致许多妇女不得不进行体外受精。本研究确定了在体外卵母细胞成熟(IVM)过程中添加粒细胞-巨噬细胞集落刺激因子(GM-CSF)是否能提高高龄产妇小鼠模型的卵母细胞发育能力。在体外卵母细胞成熟过程中用 10 纳克/毫升 GM-CSF 处理 12-14 个月 C57BL6 J × CBA 小鼠的卵母细胞,并检测胚胎发育、线粒体活性、纺锤体形成和染色体排列。与对照组相比,添加 GM-CSF 有提高受精率的趋势(76.19% vs. 82.03%;P = 0.07),但不影响积聚体的扩大。添加 GM-CSF 还能提高囊胚率(51.10 vs. 61.52%;P <0.01)和培养 96 小时后优质囊胚的数量(33.31 vs. 44.13%;P <0.05),以及内细胞质量(12.64 vs. 15.62;P <0.01)和总细胞数(42.98 vs. 48.78;P <0.05)。GM-CSF 处理还能使外层(2.86 对 0.97;P<0.001)、中间层(3.25 对 0.89;P<0.001)和核周区(3.62 对 1.08;P<0.001)的线粒体膜电位增加两到三倍。GM-CSF 处理不会影响纺锤体的形成或染色体的排列。我们的研究结果表明,在 IVM 期间添加 GM-CSF 可改善高龄产妇的卵母细胞质量。
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引用次数: 0
Assessment of anti-Müllerian hormone levels as a reproductive indicator in Japanese Black cattle. 评估作为日本黑牛繁殖指标的抗缪勒氏管激素水平。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-12-13 Epub Date: 2024-10-17 DOI: 10.1262/jrd.2024-047
Takeshi Koyama, Hiromi Suzuki, Miki Shimizu, Riuru Mizuno, Ayaha Ishigami, Nobuyuki Kamidate, Yoshihisa Otani, Michiko Okubo, Kousaku Souma, Hiroki Hirayama

The size of the ovarian reserve, an indicator of the number of primordial follicles, varies widely among individuals, and declines with age. However, the association between the ovarian reserve and fertility remains unclear. Therefore, in the present study, we analyzed the relationship between plasma concentrations of anti-Müllerian hormone (AMH), a marker of ovarian reserve, and reproductive outcomes in Japanese Black cattle. AMH level quartiles were positively associated with pregnancy following artificial insemination (AI), and the median number of days to pregnancy in Q4 (13 days, 95% confidence interval [CI] = 7-18 days) was significantly shorter than that in Q1 (21 days, 95% CI = 15-46 days). The odds ratio for the predicted pregnancy rate by logistic regression analysis in Q4 (4.06, 95% CI = 1.54-10.67) was also significantly higher than that in Q1. Plasma AMH concentrations were significantly higher in summer (June-August) than in winter (December-February). Furthermore, a strong correlation (r = 0.856, P < 0.001) was observed between plasma AMH concentrations at 2 and 14 months of age. Calves with plasma AMH concentrations of > 700 pg/ml at 2 months old showed a transient increase and maximum AMH concentration within 5 months of birth. Overall, the results of this study indicate that the plasma AMH concentration serves as a predictive marker for the probability of conception following AI in Japanese Black cattle. The current findings contribute to the reliable assessment of AMH production and the early prediction of reproductive performance in sexually mature heifers.

卵巢储备功能是原始卵泡数量的指标,其大小因人而异,并随着年龄的增长而下降。然而,卵巢储备功能与生育能力之间的关系仍不明确。因此,在本研究中,我们分析了卵巢储备标志物抗缪勒氏管激素(AMH)的血浆浓度与日本黑牛繁殖结果之间的关系。AMH水平四分位数与人工授精(AI)后的妊娠呈正相关,Q4的中位妊娠天数(13天,95%置信区间[CI] = 7-18天)明显短于Q1(21天,95%置信区间 = 15-46天)。通过逻辑回归分析,第四季度预测妊娠率的几率比(4.06,95% CI = 1.54-10.67)也明显高于第一季度。血浆 AMH 浓度在夏季(6 月至 8 月)明显高于冬季(12 月至 2 月)。此外,2 月龄和 14 月龄犊牛的血浆 AMH 浓度之间存在很强的相关性(r = 0.829,P < 0.001)。2月龄时血浆AMH浓度大于700 pg/ml的犊牛在出生后5个月内AMH浓度出现短暂上升并达到最高值。总之,本研究结果表明,血浆 AMH 浓度可作为日本黑牛人工授精后受孕概率的预测指标。目前的研究结果有助于可靠地评估AMH的产生和早期预测性成熟母牛的繁殖性能。
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引用次数: 0
AVPV Kiss1 neuron-specific knockdown of purinergic P2X2 receptor suppresses LH surge and ovulation in Kiss1-Cre rats. AVPV Kiss1神经元特异性敲除嘌呤能P2X2受体可抑制Kiss1-Cre大鼠的LH激增和排卵。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-12-13 Epub Date: 2024-10-20 DOI: 10.1262/jrd.2024-046
Safiullah Hazim, Shunsuke Seki, Ryoya Yabushita, Mayuko Nagae, Hitomi Tsuchida, Masumi Hirabayashi, Yoshihisa Uenoyama, Hiroko Tsukamura, Naoko Inoue

Ovulation disorders are a major cause of low pregnancy rates and infertility in humans and livestock. Kisspeptin neurons located in the anteroventral periventricular nucleus (AVPV) are responsible for the generation of the gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) surge and the consequent ovulation in female rodents. The present study aimed to examine whether purinergic neurons are direct upstream stimulators of AVPV kisspeptin neurons that trigger the GnRH/LH surge and consequent ovulation in Kiss1-Cre rats. We specifically knocked down the mRNA expression of the P2rx2 purinergic receptor in AVPV kisspeptin neurons by administering an adeno-associated virus (AAV) vector containing Cre-dependent P2rx2 short hairpin RNA (shRNA) into the AVPV region of ovariectomized (OVX) Kiss1-Cre rats treated with a proestrus level of estradiol-17β (OVX + high E2) or ovary-intact Kiss1-Cre rats. The E2-induced afternoon LH surge was significantly suppressed by AVPV kisspeptin neuron-specific knockdown of P2rx2 in OVX + high E2 Kiss1-Cre rats compared with scrambled shRNA-treated control OVX + high E2 Kiss1-Cre rats. Furthermore, the specific knockdown of P2rx2 in AVPV kisspeptin neurons largely disrupted the estrous cycle, spontaneous LH surge, and ovulation in ovary-intact Kiss1-Cre rats. These findings suggest that purinergic neurons directly stimulate AVPV kisspeptin neurons via P2X2 receptors (P2RX2) to induce the GnRH/LH surge and consequent ovulation in female rats.

排卵障碍是人类和家畜受孕率低和不育的主要原因。位于雌性啮齿类动物前腹腔周围核(AVPV)的基斯普汀神经元负责促性腺激素释放激素(GnRH)/黄体生成素(LH)激增的产生以及随之而来的排卵。本研究旨在探讨嘌呤能神经元是否是 AVPV 吻肽神经元的直接上游刺激因子,从而触发 Kiss1-Cre 大鼠的 GnRH/LH 激增和随之而来的排卵。我们通过向卵巢切除(OVX)的 Kiss1-Cre 大鼠或卵巢未切除的 Kiss1-Cre 大鼠的 AVPV 区域注射含有 Cre 依赖性 P2rx2 短发夹 RNA(shRNA)的腺相关病毒(AAV)载体,特异性地敲除了 AVPV 吻肽素神经元中 P2rx2 嘌呤能受体 mRNA 的表达。与乱码 shRNA 处理的对照 OVX + 高 E2 Kiss1-Cre 大鼠相比,AVPV 吻肽神经元特异性敲除 P2rx2 能显著抑制 E2 诱导的午后 LH 激增。此外,特异性敲除 AVPV Kisspeptin 神经元中的 P2rx2 在很大程度上破坏了与卵巢接触的 Kiss1-Cre 大鼠的发情周期、自发性 LH 激增和排卵。这些研究结果表明,嘌呤能神经元通过P2X2受体(P2RX2)直接刺激AVPV吻肽素神经元,诱导雌性大鼠的GnRH/LH激增,进而诱导排卵。
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引用次数: 0
Application of bovine progesterone intravaginal controlled-release formulation for estrus synchronization treatment in goats. 将牛孕酮阴道内控释制剂用于山羊的发情同步治疗。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-12-13 Epub Date: 2024-11-10 DOI: 10.1262/jrd.2024-083
Kotaro Saito, Tomomi Tanaka, Natsumi Endo

Progesterone (P4)-impregnated controlled internal drug-releasing device for goats and sheep (CIDR-G) is used in the estrus synchronization protocol; however, it is not commercially available in Japan. In this study, we investigated whether a modification of CIDR-B, designed for cattle, could be used as a P4 release device for synchronizing estrus in goats. An in vitro analysis showed that the dissolution rate was not significantly different between CIDR-G and modified CIDR-B (CIDR-M). The administration of CIDR-G or CIDR-M to dairy goats resulted in plasma P4 concentrations being maintained at the functional luteal phase levels (>2 ng/ml) in both groups for 12 days after administration. However, P4 concentrations at 2, 4, and 8 days after the administration of CIDR-G were significantly higher than those after the administration of CIDR-M. These results suggest that the CIDR-M examined in this study can maintain P4 levels for 12 days after its administration as a P4 release device.

用于山羊和绵羊的浸渍了孕酮(P4)的可控体内药物释放装置(CIDR-G)被用于发情同步方案;但是,这种装置在日本还没有商业化。在这项研究中,我们探讨了为牛设计的 CIDR-B 的改进型是否可用作山羊同步发情的 P4 释放装置。体外分析表明,CIDR-G 和改良型 CIDR-B(CIDR-M)的溶解率没有明显差异。给奶山羊服用 CIDR-G 或 CIDR-M 后,两组的血浆 P4 浓度在 12 天内都保持在功能性黄体期水平(>2 纳克/毫升)。然而,给药后 2、4 和 8 天的 P4 浓度明显高于给药后的 CIDR-M。这些结果表明,本研究中的 CIDR-M 作为一种 P4 释放装置,可以在给药后 12 天内维持 P4 水平。
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引用次数: 0
Fertilization and developmental competence of in vitro fertilized embryos from C57BL/6J mice of different ages and the impact of vitrification. 不同年龄 C57BL/6J 小鼠体外受精胚胎的受精和发育能力以及玻璃化的影响。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-12-13 Epub Date: 2024-10-25 DOI: 10.1262/jrd.2024-082
Seiji Kito

Prepubertal animals are often preferred as sources of oocytes for assisted reproductive technologies (ARTs) in laboratory mice, but the normality and developmental competence of these oocytes remain controversial. This study systematically examined in vitro fertilization competence, embryo development, and fetal development after embryo transfer (ET) using oocytes from C57BL/6J mice aged 3 to 10 weeks. Oocytes were collected from superovulated females, fertilized, and cultured in vitro for 96 h or transferred at 2-cell stage to recipient females. Additionally, fetal development was compared between unfrozen and frozen-thawed in vitro-fertilized 2-cell embryos across different age groups. The number of ovulated oocytes per animal decreased with age, while the percentage of morphologically normal oocytes was highest in 3-week-old mice (99%) compared to older ages (70-86%, P < 0.05). Although fertilization percentages were consistently high (≥ 97%), blastocyst development in vitro, the nuclear counts of blastocysts and fetal development after ET were lowest for embryos from 3-week-old mice. Development of frozen-thawed embryos to fetuses was significantly reduced compared to unfrozen embryos in all age groups, except for those from 10-week-old mice. These findings suggest that oocytes from prepubertal mice, particularly from 3-week-old mice, are less developmentally competent than those from older mice. Therefore, the age of animals for oocyte source should be carefully considered based on the specific requirements of the research or ART applications.

青春期前的动物通常是实验室小鼠辅助生殖技术(ART)的首选卵母细胞来源,但这些卵母细胞的正常性和发育能力仍存在争议。本研究使用 3 至 10 周龄的 C57BL/6J 小鼠的卵母细胞,对体外受精能力、胚胎发育和胚胎移植(ET)后的胎儿发育进行了系统检测。卵母细胞取自超排卵雌鼠,受精后体外培养96小时,或在2细胞期移植到受体雌鼠体内。此外,还比较了不同年龄组未冷冻和冷冻解冻的体外受精 2 细胞胚胎的胎儿发育情况。每只动物的排卵卵母细胞数量随年龄的增长而减少,而 3 周龄小鼠的卵母细胞形态正常的比例最高(99%),年龄较大的小鼠为 70-86%(P < 0.05)。虽然受精率一直很高(≥ 97%),但 3 周龄小鼠胚胎的体外囊胚发育、囊胚核计数和 ET 后胎儿发育都是最低的。除 10 周龄小鼠的胚胎外,所有年龄组的冷冻解冻胚胎发育成胎儿的几率都明显低于未冷冻胚胎。这些研究结果表明,青春期前小鼠的卵母细胞,尤其是 3 周龄小鼠的卵母细胞,发育能力低于较大年龄小鼠的卵母细胞。因此,应根据研究或 ART 应用的具体要求,仔细考虑卵母细胞来源动物的年龄。
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引用次数: 0
Autophagy in the corpus luteum correlates with tissue growth in pregnant rats. 黄体中的自噬与怀孕大鼠的组织生长有关。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-10-01 Epub Date: 2024-07-07 DOI: 10.1262/jrd.2024-019
Yasuaki Oishi, Koji Asakawa, Yuri Ishiwata, Shota Oka, Ryota Terashima, Makoto Sugiyama, Keiichiro Kizaki, Mitsumori Kawaminami, Shiro Kurusu

The developmental activation of the corpus luteum (CL) structurally and functionally is critical for the temporally regulated establishment, maintenance, and termination of pregnancy in rats. In this study, we have investigated the possible involvement of autophagy in the regulation of the CL during pregnancy in rats. The expression ratio of microtubule-associated protein light chain 3 (LC3)-II/-I, a widely used indicator of autophagic activity, in the CL remained relatively stable until day 15 of pregnancy. Subsequently, it progressively increased until day 21, and then declined until day 3 postpartum. This fluctuation was closely associated with the tissue weight of the CL rather than progesterone (P4) production activity. Light and electron microscopy revealed the presence of immunoreactive LC3 aggregates and irregularly shaped autolysosome-like microstructures in the cytoplasm of luteal cells during late pregnancy. Notably, a bolus intrabursal injection of the autophagy inhibitor bafilomycin A1 on day 15 of pregnancy resulted in a significant reduction in luteal cell size and disrupted the normal alteration of circulating P4 levels. Consequently, treatment with this inhibitor increased the likelihood of the varied timing (both advanced and delayed) of delivery and led to reduced body weight in neonates when compared with the vehicle-treated control group. Our findings suggest that autophagy in the rat CL contributes to luteal tissue growth, influences P4 production, and thereby fine-tunes the regulation of gestation length in rats.

黄体(CL)在结构和功能上的发育激活对于大鼠妊娠的建立、维持和终止的时间调控至关重要。在这项研究中,我们探讨了自噬可能参与大鼠妊娠期黄体的调控。微管相关蛋白轻链 3(LC3)-II/-I 是一种广泛应用的自噬活性指标,其在 CL 中的表达比在妊娠第 15 天之前保持相对稳定。随后,它逐渐增加,直到第 21 天,然后下降,直到产后第 3 天。这种波动与CL的组织重量而非孕酮(P4)的产生活性密切相关。光镜和电子显微镜显示,在妊娠晚期,黄体细胞的细胞质中存在免疫活性 LC3 聚集体和不规则形状的自溶体样微结构。值得注意的是,在妊娠的第 15 天,向腔内注射自噬抑制剂巴佛洛霉素 A1 会导致黄体细胞体积显著缩小,并破坏循环 P4 水平的正常变化。因此,与用药物治疗的对照组相比,用这种抑制剂治疗增加了不同时间(提前和延迟)分娩的可能性,并导致新生儿体重下降。我们的研究结果表明,大鼠CL中的自噬作用有助于黄体组织的生长,影响P4的产生,从而微调大鼠妊娠期的长短。
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引用次数: 0
Dairy cow parity affects relationships among nutritional parameters in the blood of dams, umbilical cords, and calves and placental development at calving. 奶牛奇数影响母牛、脐带和犊牛血液中的营养参数与产犊时胎盘发育之间的关系。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-10-01 Epub Date: 2024-07-07 DOI: 10.1262/jrd.2024-010
Riku Mashimo, Hanon Ohban, Yuka Kumazaki, Sayaka Ito, Tomono Katagiri, Nobuyuki Kusaba, Chiho Kawashima

Heifer growth and milk production in lactating cows may diminish the nutrient supply to the fetus. This study aimed to analyze the characteristics of the nutrient supply to the fetus in primiparous and multiparous cows. We investigated maternal, umbilical cord, and calf blood glucose and amino acid levels, as well as placental development in 28 primiparous (PP) and 30 multiparous (MP) Holstein cows. Although the total cotyledonary weight and surface area showed no significant differences, the MP group exhibited larger individual cotyledons (P < 0.01) and fewer medium-sized cotyledons (P < 0.05). Within the PP group, total cotyledonary weight and surface area positively correlated with blood glucose (r = 0.71-0.77; P < 0.01) and total essential amino acid (r = 0.55; P < 0.05) concentrations in the umbilical veins. However, no significant correlation was observed in the MP group. Blood glucose and amino acid concentrations in the umbilical vein, umbilical artery, and calf were significantly lower in the MP group (P < 0.05), although no difference was observed in the dams between the groups. In conclusion, the nutrient status of primiparous cows can alter fetal nutrient supply. Moreover, multiparous cows have larger individual cotyledons as an adaptive response to increased milk production during pregnancy. However, this adaptive response in multiparous cows did not completely restore nutrient supply to the fetus to the same extent as that in primiparous cows. Therefore, the nutritional management of multiparous cows during pregnancy must be reconsidered.

泌乳奶牛的小母牛生长和产奶量可能会减少胎儿的营养供应。本研究旨在分析初产母牛和多产母牛的胎儿营养供应特点。我们调查了28头初产(PP)和30头多产(MP)荷斯坦奶牛的母体、脐带和犊牛血糖和氨基酸水平以及胎盘发育情况。虽然子叶总重量和表面积没有显著差异,但 MP 组的子叶单个较大(P < 0.01),中等大小的子叶较少(P < 0.05)。在 PP 组中,子叶总重量和表面积与脐静脉中的血糖(r = 0.71-0.77; P < 0.01)和总必需氨基酸(r = 0.55; P < 0.05)浓度呈正相关。然而,在 MP 组中没有观察到明显的相关性。MP组脐带血管、脐动脉和小牛体内的血糖和氨基酸浓度明显低于MP组(P < 0.05),但各组间母牛的血糖和氨基酸浓度无差异。总之,初产母牛的营养状况会改变胎儿的营养供应。此外,多胎奶牛的子叶个体较大,这是对孕期产奶量增加的一种适应性反应。然而,多胎奶牛的这种适应性反应并不能完全恢复与初产奶牛相同的胎儿营养供应。因此,必须重新考虑多胎奶牛妊娠期的营养管理问题。
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引用次数: 0
Cpne1 deficiency preserves sperm motility under Ca2+ channel blockade. 在 Ca2+ 通道阻断的情况下,Cpne1 的缺乏可保持精子活力。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-10-01 Epub Date: 2024-07-14 DOI: 10.1262/jrd.2024-027
Qiang Xie, Hanbin Zhang, Yuge Zhuang, Jinsheng Liu, Zicong Huang, Xiaoyuan Zhang, Ke Ma, Wenyuan Liu, Minyu Xie, Chuyu Huang, Xiaojing Zhong, Feilong Chen, Feng Zou, Wansong Zhang, Chunming Qiu, Canbiao Sun, Xiangjin Kang, Zhenguo Chen, Guofei Zhang

Calcium ions (Ca2+) play crucial roles in sperm motility and fertilization. The copine (CPNE) family comprises several Ca2+-dependent phospholipid-binding proteins. Of these, CPNE1 is extensively expressed in mammalian tissues; however, its precise role in testicular development and spermatogenesis is yet to be fully characterized. In this study, we used proteomics to analyze testicular biopsies and found that levels of CPNE1 were significantly reduced in patients with non-obstructive azoospermia (defective spermatogenesis) compared to those in patients with obstructive azoospermia (physiological spermatogenesis). In mice, CPNE1 is expressed at various stages of germ cell development and is associated with the Golgi apparatus. Ultimately, CPNE1 is expressed in the flagella of mature sperms. To further examine the role of CPNE1, we developed a Cpne1 knockout mouse model. Analysis showed that the loss of Cpne1 did not impair testicular development, spermatogenesis, or sperm morphology and motility in physiological conditions. When treated with gadolinium (III) chloride or 2-aminoethoxydiphenyl borate, known inhibitors of store-operated Ca2+ entry, Ca2+ signals and sperm motility were significantly compromised in wild-type mice; however, both mechanisms were conserved in KO mice. These results suggested that CPNE1 is dispensable for testicular development, spermatogenesis or sperm motility in physiological conditions. In addition, CPNE1 may represent a target of Ca2+ channel inhibitors and may therefore be implicated in the regulation of Ca2+ signaling and sperm motility.

钙离子(Ca2+)在精子运动和受精过程中起着至关重要的作用。copine(CPNE)家族由几种依赖于钙离子的磷脂结合蛋白组成。其中,CPNE1 在哺乳动物组织中广泛表达;然而,它在睾丸发育和精子生成中的确切作用尚未完全确定。在这项研究中,我们利用蛋白质组学分析了睾丸活检组织,发现与阻塞性无精子症(生理性精子发生)患者相比,非阻塞性无精子症(精子发生缺陷)患者体内的 CPNE1 水平明显降低。在小鼠体内,CPNE1 在生殖细胞发育的不同阶段均有表达,并与高尔基体相关。最终,CPNE1 在成熟精子的鞭毛中表达。为了进一步研究 CPNE1 的作用,我们建立了一个 Cpne1 基因敲除小鼠模型。分析表明,在生理条件下,Cpne1 的缺失不会影响睾丸发育、精子生成、精子形态和活力。当使用氯化钆(III)或2-氨基甲氧基二苯基硼酸盐(已知的贮存操作 Ca2+ 进入抑制剂)处理时,野生型小鼠的 Ca2+ 信号和精子活力会受到显著影响;然而,这两种机制在 KO 小鼠中都得到了保留。这些结果表明,在生理条件下,CPNE1 对睾丸发育、精子形成或精子运动是不可或缺的。此外,CPNE1 可能是 Ca2+ 通道抑制剂的靶标,因此可能与 Ca2+ 信号传导和精子运动的调节有关。
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引用次数: 0
Perilipin2 depletion causes lipid droplet enlargement in the ovarian corpus luteum in mice. 消耗 Perilipin2 会导致小鼠卵巢黄体脂滴增大。
IF 1.9 4区 生物学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Pub Date : 2024-10-01 Epub Date: 2024-07-15 DOI: 10.1262/jrd.2024-023
Megumi Ibayashi, Takayuki Tatsumi, Satoshi Tsukamoto

Lipid droplets (LDs) are endoplasmic reticulum-derived organelles that store neutral lipids (mostly triglycerides and cholesterol esters) within a phospholipid monolayer and appear in most eukaryotic cells. Perilipins (PLINs, comprising PLIN1-5) are abundant LD-associated proteins with highly variable expression levels among tissues. Although PLINs are expressed in the mammalian ovaries, little is known about their subcellular localization and physiological functions. In this study, we investigated the localization of PLIN1-3 and their relationship with LD synthesis using mCherry-HPos reporter mice, thereby enabling the visualization of LD biogenesis in vivo. PLIN2 and PLIN3 were localized as puncta in granulosa cells with low levels of LD synthesis in developing follicles. This localization pattern was quite different from that of PLIN1, which was mainly localized in the theca and interstitial cells with high levels of LD synthesis. In the corpus luteum, where LD synthesis is highly induced, PLIN2 and PLIN3 are abundant in the particulate structures, whereas PLIN1 is poorly distributed. We also generated global Plin2-deficient mice using the CRSPR/Cas9 system and demonstrated that the lack of PLIN2 did not alter the distribution of PLIN1 and PLIN3 but unexpectedly induced LD enlargement in the corpus luteum. Collectively, our results suggest that the localization of PLIN1-3 is spatiotemporally regulated and that PLIN2 deficiency influences LD mobilization in the corpus luteum within the ovaries.

脂滴(LDs)是内质网衍生的细胞器,在磷脂单层中储存中性脂质(主要是甘油三酯和胆固醇酯),出现在大多数真核细胞中。长链胰蛋白(PLINs,包括 PLIN1-5)是丰富的低密度脂蛋白相关蛋白,在不同组织中的表达水平差异很大。尽管PLINs在哺乳动物卵巢中也有表达,但人们对其亚细胞定位和生理功能知之甚少。在本研究中,我们利用 mCherry-HPos 报告小鼠研究了 PLIN1-3 的定位及其与 LD 合成的关系,从而实现了体内 LD 生物发生的可视化。在发育中的卵泡中,PLIN2和PLIN3以点状定位在低水平LD合成的颗粒细胞中。这种定位模式与PLIN1的定位模式截然不同,PLIN1主要定位在LD合成水平较高的囊胚和间质细胞中。在LD合成被高度诱导的黄体中,PLIN2和PLIN3大量存在于颗粒结构中,而PLIN1的分布则很不均匀。我们还利用 CRSPR/Cas9 系统生成了全基因 Plin2 缺失小鼠,结果表明,缺乏 PLIN2 不会改变 PLIN1 和 PLIN3 的分布,但却意外地诱导了黄体中 LD 的增大。总之,我们的研究结果表明,PLIN1-3的定位受时空调控,PLIN2的缺乏会影响卵巢内黄体的LD动员。
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Journal of Reproduction and Development
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