Physiology and modulation factors of ovulation in rabbit reproduction management

Rabbit is an induced ovulatory species, so ovulation takes place after mating. Traditionally, exogenous and synthetic hormonal factors (administered by intramuscular and intravaginal route) such as GnRH and analogues, or different physical procedures (i.e. stimulation by intravaginal cannula) have been used to induce ovulation in females when artificial insemination is applied in rabbit farms. Restriction and public rejection of the use of hormones is leading to the study of the seminal plasma components with potential action on ovulation induction. The aim of the present review is to collect and summarise the strategies used in recent years to trigger ovulation and improve rabbit fertility management with respect to more animal-friendly manipulation methods. Furthermore, special attention has been paid to the use of a semen component (as endogen molecule) such as beta nerve growth factor (β-NGF) in male and female rabbit reproductive physiology. This neurotrophin and its receptors (TrKA and p75NTR) are abundantly distributed in both male and female rabbit reproductive tracts, and it seems to have an important physiological role in sperm maturation and behaviour (velocity, apoptosis and capacitation), as well as a modulatory factor of ovulation. Endogen β-NGF is diluted in the seminal doses with the extenders; hence it could be considered an innovative and alternative strategy to avoid the current exogenous (by intramuscular route) and stressful hormonal treatments used in ovulation induction. Their addition in seminal dose could be more physiological and improve animal welfare in rabbit farms.

Effects of Variations in Hormonal Treatments Upon In vitro Regeneration Potential in Embryo Explants of Arenga pinnata

One of the challenges related to propagation of Arenga pinnata is its lengthy period of seed dormancy. In this study, in vitro regeneration was carried out to determine the effect of hormonal treatment on the embryo explant of Arenga pinnata. Embryos were surface sterilized and cultured into different media supplemented with various hormones concentrations and combinations. Each treatment contained of Kinetin (KN) hormone (1.0, 2.0, and 3.0 mg/l) and in combination with indole-3-acetic acid (IAA) of 0.1, 0.2, 0.3 mg/l. The height of plumule and length of radical was observed and recorded. Treatment 8 (3 mg/ml KN + 0.1 mg/ml IAA) showed 59.09% in plumule height increment while treatment 4 (1 mg/ml KN + 0.3 mg/ml IAA) showed the highest radical increments with 93.62%. The knowledge gained in this study consequently helps us to better understand the role of KN and IAA in the in vitro regeneration protocol. Since in vitro method able to produce higher number of in vitro seedlings at one time, it is important to establish the in vitro regeneration protocol for this plant.

New Therapeutics in Endometriosis: A Review of Hormonal, Non-Hormonal, and Non-Coding RNA Treatments

Endometriosis is defined as endometrial-like tissue outside the uterine cavity. It is a chronic inflammatory estrogen-dependent disease causing pain and infertility in about 10% of women of reproductive age. Treatment nowadays consists of medical and surgical therapies. Medical treatments are based on painkillers and hormonal treatments. To date, none of the medical treatments have been able to cure the disease and symptoms recur as soon as the medication is stopped. The development of new biomedical targets, aiming at the cellular and molecular mechanisms responsible for endometriosis, is needed. This article summarizes the most recent medications under investigation in endometriosis treatment with an emphasis on non-coding RNAs that are emerging as key players in several human diseases, including cancer and endometriosis.

Regulation of metamorphosis in neopteran insects is conserved in the paleopteran Cloeon dipterum (Ephemeroptera)

In the Paleozoic era, more than 400 Ma, a number of insect groups continued molting after forming functional wings. Today, however, flying insects stop molting after metamorphosis when they become fully winged. The only exception is the mayflies (Paleoptera, Ephemeroptera), which molt in the subimago, a flying stage between the nymph and the adult. However, the identity and homology of the subimago still is underexplored. Debate remains regarding whether this stage represents a modified nymph, an adult, or a pupa like that of butterflies. Another relevant question is why mayflies have the subimago stage despite the risk of molting fragile membranous wings. These questions have intrigued numerous authors, but nonetheless, clear answers have not yet been found. By combining morphological studies, hormonal treatments, and molecular analysis in the mayfly Cloeon dipterum, we found answers to these old questions. We observed that treatment with a juvenile hormone analog in the last nymphal instar stimulated the expression of the Kr-h1 gene and reduced that of E93, which suppress and trigger metamorphosis, respectively. The regulation of metamorphosis thus follows the MEKRE93 pathway, as in neopteran insects. Moreover, the treatment prevented the formation of the subimago. These findings suggest that the subimago must be considered an instar of the adult mayfly. We also observed that the forelegs dramatically grow between the last nymphal instar, the subimago, and the adult. This necessary growth spread over the last two stages could explain, at least in part, the adaptive sense of the subimago.

Hormonal treatments for endometriosis: The endocrine background

Endometriosis is a benign uterine disorder characterized by menstrual pain and infertility, deeply affecting women’s health. It is a chronic disease and requires a long term management. Hormonal drugs are currently the most used for the medical treatment and are based on the endocrine pathogenetic aspects. Estrogen-dependency and progesterone-resistance are the key events which cause the ectopic implantation of endometrial cells, decreasing apoptosis and increasing oxidative stress, inflammation and neuroangiogenesis. Endometriotic cells express AMH, TGF-related growth factors (inhibin, activin, follistatin) CRH and stress related peptides. Endocrine and inflammatory changes explain pain and infertility, and the systemic comorbidities described in these patients, such as autoimmune (thyroiditis, arthritis, allergies), inflammatory (gastrointestinal/urinary diseases) and mental health disorders.The hormonal treatment of endometriosis aims to block of menstruation through an inhibition of hypothalamus-pituitary-ovary axis or by causing a pseudodecidualization with consequent amenorrhea, impairing the progression of endometriotic implants. GnRH agonists and antagonists are effective on endometriosis by acting on pituitary-ovarian function. Progestins are mostly used for long term treatments (dienogest, NETA, MPA) and act on multiple sites of action. Combined oral contraceptives are also used for reducing endometriosis symptoms by inhibiting ovarian function. Clinical trials are currently going on selective progesterone receptor modulators, selective estrogen receptor modulators and aromatase inhibitors. Nowadays, all these hormonal drugs are considered the first-line treatment for women with endometriosis to improve their symptoms, to postpone surgery or to prevent post-surgical disease recurrence. This review aims to provide a comprehensive state-of-the-art on the current and future hormonal treatments for endometriosis, exploring the endocrine background of the disease.

P–195 The influence of hormones and initial cell number on the size of self-assembled embryo-like structures

Study question Do hormonal treatments and initial cell number influence the formation of embryo-like structures (ELS) during their development in regard to size? Summary answer The chosen initial cell number for ELS-assembly seems to influence the ELS size only until day 4, while hormones affect embryo size throughout their development. What is known already The initial cell number is an important parameter for the development of ELS, which might help to better understand how embryos regulate their size. Previous studies on differently sized natural murine embryos revealed that an initial difference in size at the early stage is compensated until E6.75. Normal-size embryos experience an increased mitotic activity before E6.75, whereas larger sized embryos show an increased apoptotic activity, indicating an important control point of cell turnover by adapting mitotic activity and cell survival. Embryo development is strongly dependent on appropriate β-estradiol and progesterone levels. Study design, size, duration The first set of experiments interrogated the influence of initial cell number (two conditions) on the size of formed ELS during the first 3 days (D1–3). The second set included two different hormonal treatments and the two conditions of initial cell number (the same as in the first experiments) generating four different groups. For each day one Aggrewell (generating 1200 ELS/well) per condition was harvested. Experiments were repeated at least three times. Participants/materials, setting, methods ELS are generated by self-assembly in microwell-chamber plates combining embryonic stem cells, trophoblast stem cells and extraembryonic endoderm stem cells. Cells were cultured with and without addition of β-estradiol and progesterone, starting with different initial cell numbers (106 vs. 42 cells/ELS). ELS were harvested, stained, and at least 40 randomly picked ELS per condition were measured and statistically analyzed with Two-way ANOVA and Tukey’s multiple comparison test. Results show the average area ± SD. Main results and the role of chance The results show a continuous increase in the size of ELS during the first three days of cultivation, with significant lower values (on D1-D3) when ELS were assembled from 42 initial cells (D1: 224.1±87.7 μm²; D3: 674.0± 84.4 μm²) compared to ELS formed with 106 initial cells (D1: 467.1±224.1 μm²; D3: 1275.0±348.0 μm²). Onward on the course of self-assembly, ELS with 42 initial cells were still smaller on D4 (1465.7±657.6 μm²) compared to ELS formed with 106 initial cells (2028.6±522.4 μm²). However, these differences could not be measured on D5 (106 initial cells: 1892.2±603.7 μm²; 42 initial cells: 1855±448.5 μm²), D6 (106 initial cells: 2143.3±622.1 μm²; 42 initial cells: 1788.4±585.5 μm²) and D7 (106 initial cells: 2146.7±628.1 μm²; 42 initial cells: 2319.5±778.8 μm²). Differences between the conditions with and without hormonal treatments (HT) could also be detected especially when ELS were generated with 42 cells: on D4 ELS with HT (1730.4±852.4 μm²) were significantly larger than without hormones (1201.2±462.9 μm²). In contrast, on D7 HT influenced the size of ELS distinctly depending on the initial cell number (42 cells: 1989.2±558.3 μm² with HT vs. 2649.7±999.4 μm² without HT; 106 cells: 2334.9±770.2μm² with HT vs. 1958.6±486.1 μm² without HT). Limitations, reasons for caution An even cell distribution is crucial for reproducible ELS-formation. Unfortunately, the used techniques for cell seeding led to an uneven distribution within the microwells. Moreover, different orientation of ELS during the size assessment might be an additional reason for the high variance of ELS size within one condition. Wider implications of the findings: Even if the results seem to be in accordance with the observations made with natural embryos regarding compensation of size until E6.75, additional experiments need to be conducted. Further investigations should be carried out by testing different culture formats to obtain a more even cell distribution during the cultivation. Trial registration number Not applicable