ZnSO4 Protects against premature ovarian failure through PI3K/AKT/GSK3ß signaling pathway.

Zinc (Zn) is an essential trace element with anti-inflammatory and antioxidant effects and plays a crucial role in the female reproductive system. We aimed to investigate the protective effect of ZnSO4 on premature ovarian failure (POF) in SD rats and granulosa cells (GCs) treated with cisplatin. We also explored the underlying mechanisms. In vivo experiments showed that ZnSO4 increased the serum levels of Zn2+, increased estrogen (E2) secretion, and decreased follicle-stimulating hormone (FSH) secretion in rats. ZnSO4 increased ovarian index, protected ovarian tissues and blood vessels, reduced excessive follicular atresia, and maintained follicular development. At the same time, ZnSO4 inhibited apoptosis in the ovaries. In vitro experiments showed that ZnSO4 combination treatment restored the intracellular levels of Zn2+ and inhibited the apoptosis of GCs. ZnSO4 inhibited cisplatin-induced reactive oxygen species (ROS) production and preserved mitochondrial membrane potential (MMP). We also found that ZnSO4 protected against POF by activating the PI3K/AKT/GSK3ß signaling pathway and reducing apoptosis of GCs. These data suggest that ZnSO4 may be a potential therapeutic agent for protecting the ovaries and preserving fertility during chemotherapy.

Plasma metabolic differences in cows affected by inactive ovaries or normal ovarian function post partum.

Anestrus is essential to an unsuccessful pregnancy in dairy cows. One of the many factors that influences anestrus is the inactive ovary. To characterize in detail the plasma metabolic pro- file, anestrus cows suffering from inactive ovaries were compared with those with natural estrus. The Holstein cows 60 to 90 day postpartum in an intensive dairy farm were assigned into inactive ovaries groups (IO, n=20) and natural estrus group (CON, n=22) according to estrus signs and rectal palpation of ovaries. Plasma samples from two groups of cows were collected from the tail vein to screen differential metabolites using gas chromatography/mass spectrometry (GC/MS) techniques and multivariate statistical analysis and pathways. The results showed that 106 compounds were screened by GC/MS and 14 compounds in the IO group were decreased by analyzing important variables in the projection values and p values of MSA.Through pathway analysis, 14 compounds, mainly associated with carbohydrate metabolism and amino acid meta- bolism, were identified to results in IO, which may seriously affect follicular growth. Metabolo- mics profiling, together with MSA and pathway analysis, showed that follicular growth and development in dairy cows is related to carbohydrate and amino acid metabolism by a single or multiple pathway(s).

TMCO1 is essential for ovarian follicle development by regulating ER Ca2+ store of granulosa cells.

TMCO1 (transmembrane and coiled-coil domains 1) is an endoplasmic reticulum (ER) transmembrane protein that actively prevents Ca2+ stores from overfilling. To characterize its physiological function(s), we generated Tmco1-/- knockout (KO) mice. In addition to the main clinical features of human cerebrofaciothoracic (CFT) dysplasia spectrum, Tmco1-/- females manifest gradual loss of ovarian follicles, impaired ovarian follicle development, and subfertility with a phenotype analogous to the premature ovarian failure (POF) in women. In line with the role of TMCO1 as a Ca2+ load-activated Ca2+ channel, we have detected a supernormal Ca2+ signaling in Tmco1-/- granulosa cells (GCs). Interestingly, although spontaneous Ca2+ oscillation pattern was altered, ER Ca2+ stores of germinal vesicle (GV) stage oocytes and metaphase II (MII) arrested eggs were normal upon Tmco1 ablation. Combined with RNA-sequencing analysis, we also detected increased ER stress-mediated apoptosis and enhanced reactive oxygen species (ROS) level in Tmco1-/- GCs, indicating the dysfunctions of GCs upon TMCO1 deficiency. Taken together, these results reveal that TMCO1 is essential for ovarian follicle development and female fertility by maintaining ER Ca2+ homeostasis of GCs, disruption of which causes ER stress-mediated apoptosis and increased cellular ROS level in GCs and thus leads to impaired ovarian follicle development.

Reductions in the number of mid-sized antral follicles are associated with markers of premature ovarian senescence in dairy cows.

High-producing dairy cows are subfertile; however, the mechanisms responsible for the decreased fertility are unknown. The aim of the present study was to test the hypothesis that culled dairy cows (4-8 years old) characterised by 'Lo' ovaries (i.e. those with <10 mid-antral follicles) are affected by premature ovarian senescence. Cows in which both ovaries were 'Lo' ovaries represented 5% of the total population analysed, and exhibited reduced ovarian size (P<0.001) and increased perifollicular stroma (P<0.05) compared with age-matched controls (i.e. cows in which both ovaries had >10 mid-antral follicles; 'Hi' ovaries). The total number of follicles, including healthy and atretic primordial, primary, secondary and small antral follicles, was lower in Lo ovaries (P<0.01). Interestingly, the primordial follicle population in Lo ovaries was lower (P<0.05) than in the control. Finally, the follicular fluid of mid-antral follicles from Lo ovaries had reduced oestradiol and anti-Müllerian hormone levels (P<0.05), but increased progesterone concentrations (P<0.05). Together, these data account for the reduced fertility of cows with Lo ovaries and are in agreement with previous observations that oocytes isolated from Lo ovaries have reduced embryonic developmental competence. Cows with a specific Lo ovary condition may represent a suitable model to address the causes of low fertility in high-yielding dairy cows, as well as the condition of premature ovarian aging in single-ovulating species.

The progression of small-follicle reserves in the ovaries of wild African elephants (Loxodonta africana) from puberty to reproductive senescence.

This study aimed to determine whether the follicle reserve in the ovary of the African elephant declines progressively after puberty and whether its depletion constrains the fertility of older females. Elephant ovaries were fixed in 4% neutral buffered formalin and small-follicle counts made using stereological protocols. Excepting a slight rise in small-follicle numbers between 16 and 25 years of age, there was a trend for follicle numbers to fall from puberty to 70 years. Reproductive status did not impact significantly on small-follicle numbers (P=0.31). The number of early primary follicles, initially higher in number than true primary follicles, fell from post-puberty to nil at 45 years of age. Six of the seven oldest animals in the study showed signs of recent ovarian activity in the form of antral follicles, corpora lutea or large corpora nigra. The four oldest elephants (mean age 69 years) had a median small-follicle count of 11,113. In summary, it appears that the elephant ovary is capable of supplying oocytes for ovulation right up to the time of death at the age of maximum life expectancy, although the follicle reserve becomes depleted in some older elephants.

Relationships between the first ovulation postpartum and polymorphism in genes relating to function of immunity, metabolism and reproduction in high-producing dairy cows.

The decrease in fertility and conception rates of high-producing dairy cows is one of the major negative impacts for today's producers. The recovery of ovarian activity postpartum is affected by the status of immunity, metabolism and reproduction and plays a critical role in subsequent fertility after parturition in the cow. In the present study we investigated the relationships between polymorphisms in genes relating to the above functions and the first postpartum ovulation as a marker of the recovery of ovarian function in the cow. In immune function related-factors, the occurrence of first postpartum ovulation within 3 weeks in the C/C genotypes of tumor necrosis factor α (TNFα) exon (55.4%) and the A/G genotypes of TNFα promoter (55.4%) was significantly higher than that in T/T genotypes of TNFα exon (14.3%) and A/A genotypes of TNFα promoter (14.3%). Moreover, anovulatory cows with the T/T genotype of TNFα exon and the A/A genotype of TNFα promoter tended to have a prolonged days open compared with those of the other genotypes of TNFα polymorphisms. In metabolic function-related factors, ovulatory and anovulatory cows had a different distribution for alleles of the growth hormone receptor, but there were no significant differences in genotype and allele frequency of insulin-like growth factor-I polymorphism. No significant relationships were found between ovarian function after parturition and polymorphisms for reproduction-related genes. In conclusion, polymorphisms of TNFα gene both in exon and promoter regions have a strong association with the early first ovulation within 3 weeks after parturition in the high-producing dairy cow. Taken together, polymorphisms of TNFα gene could be strongly related to early first ovulation after parturition, thus being an effective tool of selection for improving reproductive performance in the high-producing dairy cow.