Journal of Reproduction and Development最新文献

The study of the size of cells and organelles has a long history, dating back to the 1600s when cells were defined. In particular, various methods have elucidated the size of the nucleus and the mitotic spindle in several species. However, little research has been conducted on oocyte size and organelles in mammals, and many questions remain to be answered. The appropriate size is essential to cell function properly. Oocytes have a very large cytoplasm, which is more than 100 times larger than that of general somatic cells in mammals. In this review, we discuss how oocytes acquire an enormous cytoplasmic size and the adverse effects of a large cytoplasmic size on cellular functions.

To improve the accuracy of ultrasonographic assessment of luteal function, we investigated the relationship between ovarian ultrasonographic findings on Day 7 (Day 1 = ovulation) and plasma progesterone (P₄) concentration, nutritional metabolic factors, and pregnancy outcome. A total of 47 spontaneous estrus events were investigated in 38 lactating Holstein cows (artificial insemination, n = 31; embryo transfer, n = 16). Transrectal ultrasonography was performed on Days 0 and 7 to measure the pre-ovulatory follicle area on Day 0 and the luteal tissue area (LTA), luteal blood flow area (LBF), relative LBF (rLBF) (= LBF/LTA), and dominant follicle area (DFA) on Day 7. Blood samples were collected on Day 7 to measure plasma P₄, insulin-like growth factor-I (IGF-I), insulin, and metabolites. Plasma P₄ concentration was positively correlated with LTA but was not associated with LBF or rLBF. Plasma P₄ concentration was positively correlated with blood glucose and IGF-I and negatively correlated with blood urea nitrogen and free fatty acid, and no significant relationship was found between the ultrasonographic findings of the corpus luteum (CL) and these blood metabolites. Pregnant cows had smaller DFA than non-pregnant cows. In conclusion, LTA measurement can help predict plasma P₄ concentration, but it was difficult to detect variations in plasma P₄ concentration in relation to changes in energy status by evaluating the CL ultrasonographically. A combined assessment of CL and first-wave dominant follicle may be important in evaluating fertility.

We examined various methods to enhance the accessibility of intracytoplasmic sperm injection (ICSI) technology to more users by making the technique easier, more efficient, and practical. First, the methods for artificially removing the mouse sperm tail were evaluated. Trypsin treatment was found to efficiently remove the sperm tails. The resultant sperm cells had a lower oocyte activation capacity; however, the use of activated oocytes resulted in the same fecundity as that of fresh, untreated sperm. Pre-activated oocytes were more resistant to physical damage, showed higher survival rates, and required less time per injection. Testing this method in rats yielded similar results, although the oocyte activation method was different. Remarkably, this method resulted in higher birth rates of rat progeny than with conventional methods of rat ICSI. Our method thereby streamlines mouse and rat ICSI, making it more accessible to laboratories across many disciplines.

The vitrification of zygotes is important for their use as donors for generating genome-edited mice. We previously reported the successful vitrification of mouse zygotes using carboxylated ε-poly-L-lysine (COOH-PLL). However, this vitrification solution contains fetal calf serum (FCS), which contains unknown factors and presents risks of pathogenic viral and microbial contamination. In this study, we examined whether polyvinyl alcohol (PVA) can be used as an alternative to FCS in vitrification solutions for mouse zygotes. When COOH-PLL was added to the vitrification solutions, zygotes vitrified with solutions containing 0.01% PVA (PV0.01) and those vitrified in a control solution containing FCS (75.6%) developed into blastocysts (78.4%). In addition, there were no significant differences in the ability to develop to term between the control solution (46.6%) and PV0.01 (44.1%) groups. In conclusion, we clearly demonstrated that PVA can replace FCS in our vitrification solution supplemented with COOH-PLL for mouse zygotes.

It has been suggested that domestication has turned cattle from seasonal breeders to annual breeders. This study examined the seasonal differences in early postpartum ovulation and subsequent reproductive performance in 542 Holstein cows. Cows displaying corpora lutea in the ovary at 26 days postpartum were defined as early ovulators. Factors affecting the occurrence of early ovulation were analyzed, and subsequent reproductive traits were compared between cows with and without early ovulation. During the summer season, 70.6% of calving cows showed early ovulation, whereas 48.7, 39.2, and 47.2% presented this condition in autumn, winter, and spring, respectively (P < 0.01). Third parity cows showed early ovulation more often than their first parity counterparts (P < 0.05). Cows with a 2.50 to 3.00 or > 3.00 body condition score (BCS) more frequently became early ovulators than those with BCSs < 2.50 (P < 0.01). Calving year was a risk factor, and uterine abnormalities were also often risk factors for early ovulation. The survival analysis showed that seasonal differences in the occurrence of early ovulation did not completely affect the time to first service and pregnancy. Proportional hazard regression analysis revealed that calving year, parity, and early ovulation were risk factors for the time to first service and that calving year was a risk factor for the time to pregnancy. In conclusion, domesticated dairy cows maintain seasonality in postpartum ovarian activity but not in subsequent fertility.

Oocyte quality is the limiting factor in female fertility. It is well known that maternal nutrition plays a role in reproductive function, and manipulating nutrition to improve fertility in livestock has been common practice in the past, particularly with respect to negative energy balance in cattle. A deficiency in nicotinamide adenine dinucleotide (NAD⁺) production has been associated with increased incidences of miscarriage and congenital defects in humans and mice, while elevating NAD⁺ through dietary supplements in aged subjects improved oocyte quality and embryo development. NAD⁺ is consumed by Sirtuins and poly-ADP-ribose polymerases (PARPs) within the cell and thus need constant replenishment in order to maintain various cellular functions. Sirtuins and PARPs play important roles in oocyte maturation and embryo development, and their activation may prove beneficial to in vitro embryo production and livestock breeding programs. This review examines the roles of NAD⁺, Sirtuins and PARPs in aspects of fertility, providing insights into the potential use of NAD⁺-elevating treatments in livestock breeding and embryo production programs.

Oogenesis depends on close interactions between oocytes and granulosa cells. Abnormal signaling between these cell types can result in infertility. However, attempts to manipulate oocyte-granulosa cell interactions have had limited success, likely due to the blood-follicle barrier (BFB), which prevents the penetration of exogenous materials into ovarian follicles. Here, we used adenoviruses (AVs) to manipulate the oocyte-granulosa cell interactions. AVs penetrated the BFB and transduced granulosa cells through ovarian microinjection. Although AVs caused transient inflammation, they did not impair fertility in wild-type mice. Introduction of Kitl-expressing AVs into congenitally infertile Kitl^Sl-t/Kitl^Sl-t mutant mouse ovaries, which contained only primordial follicles because of a lack of Kitl expression, restored fertility through natural mating. The offspring showed no evidence of AV integration and exhibited normal genomic imprinting patterns for imprinted genes. These results demonstrate the usefulness of AVs for manipulating oogenesis and suggest the possibility of gene therapies for human female infertility.

The effect of confirmation of ovulation by rectal palpation and ultrasonography after artificial insemination (AI) on the development of the corpus luteum (CL) and conception rate was investigated in cows. A total of 90 clinically healthy Holstein-Friesian dairy cows were examined in this study. After AI, the cows were divided into three groups (30 cows per group). In Group I, ovulation was confirmed by rectal palpation at 24 h after AI. In Group II, ovulation was confirmed using transrectal ultrasonography 24 h after AI. In Group III, ovulation was not confirmed after AI. Day 0 was defined as the day when ovulation was confirmed in Groups I and II, and as the day after AI was performed in group III. Transrectal ultrasonography was performed on days 3, 5, 7, and 14 to measure the CL diameter, tissue area, and CL blood flow area, and the ratio of CL blood flow area to CL tissue area was calculated. On the day of CL measurement, blood samples were collected to determine the plasma concentrations of progesterone (P₄) and estradiol-17β (E₂). Pregnancy was diagnosed at 28 and 60 days after AI. A high conception rate of approximately 80% was achieved in Groups I and II, in which confirmation of ovulation was conducted. There were no differences in the diameter, tissue area, or blood flow area of the CL between the three groups. These results indicate that the confirmation of ovulation by rectal palpation and transrectal ultrasonography did not affect the formation and function of the CL or conception rate.

In mammals, the corpus luteum (CL) is a transient organ that secretes progesterone (P4). In the absence of pregnancy, the CL undergoes regression (luteolysis), which is a crucial preparation step for the next estrous cycle. Luteolysis, initiated by uterine prostaglandin F_2α (PGF) in cattle, is usually divided into two phases, namely functional luteolysis characterized by a decline in P4 concentration and structural luteolysis characterized by the elimination of luteal tissues from the ovary. Programmed cell death (PCD) of luteal cells, including luteal steroidogenic cells (LSCs) and luteal endothelial cells (LECs), plays a crucial role in structural luteolysis. The main types of PCD are caspase-dependent apoptosis (type 1), autophagic cell death (ACD) via the autophagy-related gene (ATG) family (type 2), and receptor-interacting protein kinase (RIPK)-dependent programmed necrosis (necroptosis, type 3). However, these PCD signaling pathways are not completely independent and interact with each other. Over the past several decades, most studies on luteolysis have focused on apoptosis as the principal mode of bovine luteal cell death. Recently, ATG family members were reported to be expressed in bovine CL, and their levels increased during luteolysis. Furthermore, the expression of RIPKs, which are crucial mediators of necroptosis, is reported to increase in bovine CL during luteolysis and is upregulated by pro-inflammatory cytokines in bovine LSCs and LECs. Therefore, apoptosis, ACD, and necroptosis may contribute to bovine CL regression. In this article, we present the recent findings regarding the mechanisms of the three main types of PCD and the contribution of these mechanisms to luteolysis.