Robotic Precisely Oocyte Blind Enucleation Method

Oocyte enucleation is a critical procedure for somatic cell nuclear transfer. Yet, the main steps of oocyte enucleation are still manually operated, which presents several drawbacks such as low precision, high repetition error, and long training time for operators. For improving the operation efficiency and success rate, a robotic precise oocyte blind enucleation method is presented in this paper. The proposed method involves the following key techniques: oocyte translation control, oocyte immobilization and penetration control, and enucleation volume control based on the adaptive slide mode. Compared with the manual blind enucleation method, the proposed robotic blind enucleation method reduced the operation time by 44.5% (manual method: 62 s vs. proposed method: 34.4 s), increased the accuracy of enucleation by 83.1% (manual method: 30.7 vs. proposed method: 5.2), increased the success rate from 80% to 93.3%, and increased the cleavage rate from 41.7% to 63.3%.

The effects of in-vitro pH decrease on the gametogenesis of the red tree coral, Primnoa pacifica

Primnoa pacifica is the most ecologically important coral species in the North Pacific Ocean where it provides important habitat for commercially important fish and invertebrates. Ocean acidification (OA) is more rapidly increasing in high-latitude seas because anthropogenic CO2 uptake is greater in these regions. This is due to the solubility of CO2 in cold water and the reduced buffering capacity due to low alkalinity of colder waters. Primnoa pacifica colonies were cultured for six to nine months in either pH 7.55 (predicted 2100 pH levels) or pH 7.75 (control). Oocyte development and fecundity in females, and spermatocyst stages in males were measured to assess the effects of pH on gametogenesis. Oocyte diameters were 13.6% smaller and fecundities were 30.9% lower in the Year 2100 samples, indicating that OA may limit oocyte formation, potentially through lipid limitation. A higher proportion of vitellogenic oocytes (65%) were also reabsorbed (oosorption) in the Year 2100 treatment. Lowered pH appeared to advance the process of spermatogenesis with a higher percentage of later stage sperm compared to control controls. There was a laboratory effect observed in all measurement types, however these only significantly affected the analyses of spermatogenesis. These results indicate that reproduction may not be possible in an acidified ocean, or that if spawning could occur, spawned oocytes would not be sufficiently equipped to support the normal development of larvae.

Effects of puberty and gonadotropins on the molecular events controlling meiotic resumption of mouse oocytes

Although studies suggest that the low competence of oocytes from prepubertal animals is due to their insufficient cytoplasmic maturation and that FSH improves oocyte maturation possibly by retarding meiotic progression and allowing more time for cytoplasmic maturation, the mechanisms by which puberty and gonadotropins regulate meiotic progression require additional detailed studies. For the first time, we observed that while meiotic progression was significantly slower, the maturation-promoting factor (MPF) activity of oocytes was significantly higher in prepubertal than in adult mice. To resolve this contradiction, we specified the molecules regulating the MPF activity and their localization during oocyte maturation in prepubertal and adult mice primed with or without gonadotropins. Our tests using corresponding enzyme regulators suggested that while activities of protein kinase A were unaffected, the activity of adenylate cyclase (ADCY) and phosphodiesterase increased while cell division cycle 2 homolog A (CDC2A) decreased significantly after puberty. While most of the adult oocytes had CDC2A protein concentrated in the germinal vesicle (GV) region, the majority of prepubertal oocytes showed no nuclear concentration of CDC2A. Maximally priming mice with equine chorionic gonadotropin brought the above parameters of prepubertal oocytes close to those in adult oocytes. Together, the results suggest that puberty and gonadotropin control oocyte meiotic progression mainly by regulating the ADCY activity and the concentration of the activated MPF toward the GV region.

Awakening the oocyte: controlling primordial follicle development

Oocytes are sequestered in primordial follicles before birth and remain quiescent in the ovary, often for decades, until recruited into the growing pool throughout the reproductive years. Therefore, activation of follicle growth is a major biological checkpoint that controls female reproductive potential. However, we are only just beginning to elucidate the cellular mechanisms required for either maintenance of the quiescent primordial follicle pool or initiation of follicle growth. Understanding the intracellular signalling systems that control oocyte maintenance and activation has significant implications for improving female reproductive productivity and longevity in mammals, and has application in domestic animal husbandry, feral animal population control and infertility in women.

Emerging Technologies to Control Oocyte Apoptosis Are Finally Treading on Fertile Ground

As the medical community strives to improve on the efficacy of anticancer treatments, a critical issue not to be overlooked since the overall quantity of life has been substantially increased in many cancer survivors is the quality of that life post-therapy. Indeed, one of the most worrisome side effects of conventional cancer treatments is damage to the gonads. This problem is compounded in females since the ovaries, unlike the testes, are incapable of germ cell renewal in postnatal life. As a consequence, the inappropriate destruction of female germ cells (oocytes) following exposure to chemotherapeutic drugs and radiation is irreparable, often leading to premature menopause and infertility [1]. Considering recent estimates that 1 in 52 human females between birth and age 39 (i.e., the pre-reproductive and reproductive years) will be diagnosed with, and presumably treated for, cancer [2], new strategies to minimize or prevent gonadal damage during such treatments would have a profound positive impact on millions of lives.

In Vitro Oocyte Maturation and Ovulation in Rainbow Trout (Salmo gairdneri), Northern Pike (Esox lucius), and Goldfish (Carassius auratus)

The endocrine processes which control oocyte maturation (resumption of meiosis) and ovulation have been studied in vitro in the trout Salmo gairdneri. Follicular maturation is ultimately under the control of a pituitary gonadotropin which induces the follicle to synthesize specific steroids; these steroids act in turn directly on the oocyte to promote maturation. The systematic study of the in vitro efficiency of various steroids have shown that 17α-hydroxy-20β-dihydroprogesterone plays a preferential role in initiating maturation; this steroid has a high affinity for a plasma protein system. The efficiency of this steroid, similarly to the efficiency of the gonadotropin, can be modulated by other circulating steroids. The precise chronology of some events of follicle maturation have been defined using inhibitors of protein and RNA synthesis.The ovulatory process (sensu stricto: expulsion of matured oocyte from the follicular envelopes) has been experimentally dissociated from oocyte maturation, and some mediators likely to act on ovulation have been identified.These data permit the consideration of novel means of intervention at the ovarian level to synchronize maturation and ovulation in fish, in order to give new tools for progress in aquaculture.