scholarly journals Long-Term Effects Following Fresh/Vitrified Embryo Transfer Are Transmitted by Paternal Germline in a Large Size Rabbit Cohort

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1272 ◽  
Author(s):  
Ximo Garcia-Dominguez ◽  
José Salvador Vicente ◽  
María P. Viudes-de-Castro ◽  
Francisco Marco-Jiménez

The concept of developmental programming suggests that the early life environment influences offspring phenotype in later life, whose effects may also be manifested in further generations. Valuable pieces of evidence come from the fields applying assisted reproductive technologies (ARTs), which deprive embryos of their optimal maternal environment and were thus associated with subsequent developmental deviations. Recently, we demonstrated that the in vitro manipulations during a vitrified embryo transfer procedure incurs a cumulative and transgenerational decline in the growth performance of the resulting offspring. Here, we provide a longitudinal study to investigate whether previous developmental deviations could be indistinctly paternally or maternally transmitted using crossbred mattings. Our findings revealed that early embryo manipulations through fresh and vitrified embryo transfer incurred paternally transmissible effects over the growth pattern and adult body weight, which seemed not inheritable via the female germline. Similar inheritable effects were observed after fresh and vitrified embryo transfer, suggesting that disturbing optimal embryo development through in vitro manipulations was the principal trigger of transmissible effects, rather than embryo cryopreservation per se.

Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1043 ◽  
Author(s):  
Ximo Garcia-Dominguez ◽  
Francisco Marco-Jiménez ◽  
David S. Peñaranda ◽  
José Salvador Vicente

Nowadays, assisted reproductive technologies (ARTs) are considered valuable contributors to our past, but a future without their use is inconceivable. However, in recent years, several studies have evidenced a potential impact of ART on long-term development in mammal species. To date, the long-term follow-up data are still limited. So far, studies have mainly focused on in vitro fertilization or in vitro culture, with information from gametes/embryos cryopreservation field being practically missing. Herein, we report an approach to determine whether a vitrified embryo transfer procedure would have long-term consequences on the offspring. Using the rabbit as a model, we compared animals derived from vitrified-transferred embryos versus those naturally conceived, studying the growth performance, plus the weight throughout life, and the internal organs/tissues phenotype. The healthy status was assessed over the hematological and biochemical parameters in peripheral blood. Additionally, a comparative proteomic analysis was conducted in the liver tissue to investigate molecular cues related to vitrified embryo transfer in an adult tissue. After vitrified embryo transfer, birth weight was increased, and the growth performance was diminished in a sex-specific manner. In addition, vitrified-transferred animals showed significantly lower body, liver and heart weights in adulthood. Molecular analyses revealed that vitrified embryo transfer triggers reprogramming of the liver proteome. Functional analysis of the differentially expressed proteins showed changes in relation to oxidative phosphorylation and dysregulations in the zinc and lipid metabolism, which has been reported as possible causes of a disturbed growth pattern. Therefore, we conclude that vitrified embryo transfer is not a neutral procedure, and it incurs long-term effects in the offspring both at phenotypic and molecular levels. These results described a striking example of the developmental plasticity exhibited by the mammalian embryo.


2014 ◽  
Vol 63 (4) ◽  
pp. 39-46 ◽  
Author(s):  
Yana Nikolayevna Kravchuk ◽  
Alla Stanislavovna Kalugina ◽  
Olga Vladimirovna Bystrova ◽  
Svetlana Aleksandrovna Shlykova

Background. Embryo cryopreservation is an essential part of ART programs today. In recent years vitrification method is used increasingly widely. Purposes and tasks. To compare the effectiveness of ART programs using vitrified and fresh embryos, as well as different endometrial preparation regimes for frozen\thawed embryo transfer (modified natural cycle (MNC) and the preparatory hormone therapy(PHT)). To analyze the course of pregnancy and perinatal outcomes after vitrified embryo transfer. Materials and methods. We prospectively assessed the ART programs effectiveness and perinatal outcomes in 153 patients (I group), who underwent vitrified embryo transfer in 2011-2013 year. To prepare the endometrium for thawed embryo transfer in 83 patients PHT (Ia subgroup) and MNC in 70 patients (Ib subgroup) were used. Control group consisted of 70 patients, who underwent fresh embryo transfer. Results. The clinical pregnancy rate, birth rate and “take home baby” rate were not significantly different between the I (47,5 %; 30,9 %; 30,9 %) and II (53,0 %; 34,9 %; 32,5 %) groups, and between Ia (48,3 %; 28,4 %; 28,4 %) and IIb (46,6 %; 34,1 %; 34,1 %) subgroups. Complications during pregnancy and delivery, birthweight, length, Apgar score, congenital malformation rate did not differ significantly after vitrified and fresh embryo transfer. Conclusion. Vitrification is an effective method to achieve clinical results, comparable to native cycles. Application of PHT and MNC results in similar clinical outcomes. Transfer Vitrified embryo transfer does not have a negative impact on obstetric and perinatal outcomes when compared with native cycles.


2019 ◽  
Author(s):  
Wei Chen ◽  
Yong Peng ◽  
Xinyi Ma ◽  
Siming Kong ◽  
Shuangyan Tang ◽  
...  

AbstractThe births of more than 8 million infants have been enabled globally through assisted reproductive technologies (ARTs), including conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with either fresh embryo transfer (ET) or frozen embryo transfer (FET). However, the potential for elevated risks of ART-related disorders persists in adult life, and the underlying epigenetic mechanisms are largely uncharacterized. Here, we recruited 100 nuclear families and profiled the DNA methylomes, genome-wide histone modifications and transcriptomes to clarify the inherent extra risks attributable to specific ART procedures. We discovered that IVF-ET seemed to introduce less disturbance into the infant epigenome than IVF-FET or ICSI-ET did. Furthermore, we noted approximately half of the DNA methylomic changes in ART-conceived offspring could be explained by parental background biases. Through removal of the parental effect, we confirmed that ART per se would introduce minor DNA methylation changes locally. More importantly, we found that ART-induced epigenomic alterations were highly enriched in the processes which might contribute to increased incidence of preeclampsia during pregnancy and metabolic syndrome in offspring. Overall, our study provides an epigenetic basis for the potential long-term health risks in ART-conceived offspring that reinforces the need to review all methods of human ART.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yukiko Abe ◽  
Harumi Nakao ◽  
Motoki Goto ◽  
Moe Tamano ◽  
Michinori Koebis ◽  
...  

AbstractGenetic engineering of non-human primates, which are most closely related to humans, has been expected to generate ideal animal models for human genetic diseases. The common marmoset (Callithrix jacchus) is a non-human primate species adequate for the production of genetically modified animals because of their small body size and high reproductive capacity. Autologous embryo transfer (AET) is routinely utilized in assisted reproductive technologies for humans but not for experimental animals. This study has developed a novel method for efficiently producing mutant marmosets using AET and CRISPR/Cas9 systems. The embryos were recovered from oviducts of naturally mated females, injected with Cas9/guide RNA, and transferred into the oviducts of the donors. This AET method can reduce the time for in vitro culture of embryos to less than 30 min. This method uses an embryo donor as the recipient, thus reducing the number of animals and allowing for “Reduction” in the 3R principles of humane experimental technique. Furthermore, this method can utilize nulliparous females as well as parous females. We applied our novel method and generated the 6 marmosets carrying mutations in the fragile X mental retardation 1 (FMR1) gene using only 18 females including 14 nulliparous females.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mutyala Satish ◽  
Sandhya Kumari ◽  
Waghela Deeksha ◽  
Suman Abhishek ◽  
Kulhar Nitin ◽  
...  

AbstractPhosphodiesterase (PDE) inhibitors, such as pentoxifylline (PTX), are used as pharmacological agents to enhance sperm motility in assisted reproductive technology (ART), mainly to aid the selection of viable sperm in asthenozoospermic ejaculates and testicular spermatozoa, prior to intracytoplasmic sperm injection (ICSI). However, PTX is reported to induce premature acrosome reaction (AR) and, exert toxic effects on oocyte function and early embryo development. Additionally, in vitro binding studies as well as computational binding free energy (ΔGbind) suggest that PTX exhibits weak binding to sperm PDEs, indicating room for improvement. Aiming to reduce the adverse effects and to enhance the sperm motility, we designed and studied PTX analogues. Using structure-guided in silico approach and by considering the physico-chemical properties of the binding pocket of the PDEs, designed analogues of PTX. In silico assessments indicated that PTX analogues bind more tightly to PDEs and form stable complexes. Particularly, ex vivo evaluation of sperm treated with one of the PTX analogues (PTXm-1), showed comparable beneficial effect at much lower concentration—slower AR, higher DNA integrity and extended longevity of  spermatozoa and  superior embryo quality. PTXm-1 is proposed to be a better pharmacological agent for ART than PTX for sperm function enhancement.


2014 ◽  
Vol 11 (2) ◽  
pp. 36-42
Author(s):  
P Erdenetogtokh ◽  
S Ganbat ◽  
Hiroshi Suzuki

Babesia infections occur mainly in animals, and are transmitted by ticks. The severity of the diseases varies considerably depending on the species of Babesia involved as well as the immune response of the infected animal. In Mongolia infection produced by Babesia parasites is widely spread, provoking severe damage to the agricultural and economic sectors. Currently, strategies to control and prevent the infection are inefficient. Indeed, the necessity to look for suitable and accessible strategies to obtain animals free from the infection is needed. Currently, assisted reproductive technologies (ART) are used for the improvement of productivity in livestock. Moreover, embryo transfer seams to be useful approach to obtain clean embryos obtained from infected animals. Therefore, by using a mice model (ICR) infected with Babesia microti, an alternative method to obtain animals free from infection was examined. ICR mice at 8 weeks old were challenged with 0.2 ml of 1x107 IRBC/ml by i.p injection. After infection, superovulation was induced and then embryos were obtained and washed. Then, their development stage along with their morphological characteristics were monitored. In vitro embryos obtained from uninfected mice were used as a control group. The results indicate that the infection does not have any influence on pre-implantation embryonic development and morphological characteristics. Thus, we suggest that embryos obtained from infected animals might be useful for embryo transfer in order to improve productivity of livestock and reduce the risk of congenital infection. In summary, ART such as embryo transfer might be an useful technique in countries where Babesiosis is an endemic disease. DOI: http://dx.doi.org/10.5564/mjas.v11i2.214 Mongolian Journal of Agricultural Sciences Vol.11(2) 2013 pp.36-42


2021 ◽  
Vol 22 (16) ◽  
pp. 8574
Author(s):  
Huixia Yang ◽  
Zhi Ma ◽  
Lin Peng ◽  
Christina Kuhn ◽  
Martina Rahmeh ◽  
...  

Epigenetics play a vital role in early embryo development. Offspring conceived via assisted reproductive technologies (ARTs) have a three times higher risk of epigenetic diseases than naturally conceived children. However, investigations into ART-associated placental histone modifications or sex-stratified analyses of ART-associated histone modifications remain limited. In the current study, we carried out immunohistochemistry, chip-sequence analysis, and a series of in vitro experiments. Our results demonstrated that placentas from intra-cytoplasmic sperm injection (ICSI), but not in vitro fertilization (IVF), showed global tri-methylated-histone-H3-lysine-4 (H3K4me3) alteration compared to those from natural conception. However, for acetylated-histone-H3-lysine-9 (H3K9ac) and acetylated-histone-H3-lysine-27 (H3K27ac), no significant differences between groups could be found. Further, sex -stratified analysis found that, compared with the same-gender newborn cord blood mononuclear cell (CBMC) from natural conceptions, CBMC from ICSI-boys presented more genes with differentially enriched H3K4me3 (n = 198) than those from ICSI-girls (n = 79), IVF-girls (n = 5), and IVF-boys (n = 2). We also found that varying oxygen conditions, RNA polymerase II subunit A (Polr2A), and lysine demethylase 5A (KDM5A) regulated H3K4me3. These findings revealed a difference between IVF and ICSI and a difference between boys and girls in H3K4me3 modification, providing greater insight into ART-associated epigenetic alteration.


2015 ◽  
Vol 27 (8) ◽  
pp. 1125 ◽  
Author(s):  
Michael J. Bertoldo ◽  
Yann Locatelli ◽  
Christopher O'Neill ◽  
Pascal Mermillod

The processes of assisted reproductive technologies (ART) involve a variety of interventions that impact on the oocyte and embryo. Critically, these interventions cause considerable stress and coincide with important imprinting events throughout gametogenesis, fertilisation and early embryonic development. It is now accepted that the IVM and in vitro development of gametes and embryos can perturb the natural course of development to varying degrees of severity. Altered gene expression and, more recently, imprinting disorders relating to ART have become a focused area of research. Although various hypotheses have been put forward, most research has been observational, with little attempt to discover the mechanisms and periods of sensitivity during embryo development that are influenced by the culture conditions following fertilisation. The embryo possesses innate survival factor signalling pathways, yet when an embryo is placed in culture, this signalling in response to in vitro stress becomes critically important in mitigating the effects of stresses caused by the in vitro environment. It is apparent that not all embryos possess this ability to adequately adapt to the stresses experienced in vitro, most probably due to an inadequate oocyte. It is speculated that it is important that embryos use their survival signalling mechanisms to maintain normal epigenetic programming. The seeming redundancy in the function of various survival signalling pathways would support this notion. Any invasion into the natural, highly orchestrated and dynamic process of sexual reproduction could perturb the normal progression of epigenetic programming. Therefore the source of gametes and the subsequent culture conditions of gametes and embryos are critically important and require careful attention. It is the aim of this review to highlight avenues of research to elucidate the effects of stress and the relationship with epigenetic programming. The short- and long-term health and viability of human and animal embryos derived in vitro will also be discussed.


2019 ◽  
Vol 31 (1) ◽  
pp. 169
Author(s):  
O. Sebastián ◽  
F. Guerrero ◽  
R. Romero ◽  
F. Muñoz ◽  
A. Parlange ◽  
...  

Assisted reproductive technologies (ART) continue to develop rapidly, allowing for the development of techniques to increase reproductive efficiency and contribute to the genetic improvement of cattle. Some of these techniques include in vitro production (IVP) of embryos and embryo transfer. These modern ART can help produce offspring with highly desirable characteristics. However, there is a lack of information on the percentage of pregnancies obtained following transfer (P/ET) of IVP embryos derived using semen of cloned Bos indicus bulls. The objective of this study was to compare embryo transfer results of IVP embryos created using frozen-thawed semen from 5 Brahman bulls (Bos indicus) with characteristics and genetics of high commercial value. The embryos were produced on two different dates, 45 days apart, using pooled oocytes harvested by ovum pickup from 15 Brahman cows at random stages of the oestrous cycle. Procedures for IVP were performed in a commercial laboratory (Genemex Internacional) in the state of Chiapas, Mexico. For IVF, conventional semen was used from 1 bull (B1) and his clone (B12), the grandson of B1 (B2), and from 2 nonrelated bulls (B3 and B4). A total of 100 embryos were transferred nonsurgically by a private practitioner on a ranch in the state of Campeche, Mexico. The recipients were commercial crossbred cows synchronized using a FTET program. On Day 0, recipients received an intravaginal device containing 1.9g of progesterone (CIDR) and 2mg of oestradiol benzoate IM. On Day 8, the CIDR was removed and cows received 25mg of dinoprost tromethamine, 200IU of eCG, and 0.5mg of oestradiol cypionate IM. Embryos were transferred on Day 17. The overall P/ET was 42% (42/100). The P/ET for IVP embryos produced with semen from bulls B1, B12, B2, B3, and B4 was 3/15 (20%), 3/8 (37%), 23/42 (55%), 8/20 (40%), and 5/15 (33%), respectively. The P/ET was numerically greater for embryos produced using semen from the cloned bull (37%; B12) compared to embryos produced using semen from the original noncloned bull (20%; B1), although this difference was not statistically significant (P=0.62, Fisher’s exact test). There was a significant difference (P<0.05) for the P/ET obtained with embryos produced using semen from bulls B1 and B2, but results for the other bulls were not significantly different. As far as we know, this is the first scientific report in Mexico concerning the use and comparison of semen from cloned and noncloned bulls for the production and transfer of bovine IVP embryos. In general, a wide numerical range of P/ET using the different bulls was observed (i.e. 20-55%). In this preliminary study, there was no impact of using frozen-thawed semen from a cloned bull for IVP on P/ET. The results from this research can contribute to the study and development of ART to improve P/ET obtained using Zebu IVP embryos. However, further research with a larger numbers of animals is required to confirm whether using semen from cloned and noncloned Bos indicus bulls for IVP impacts pregnancy success following embryo transfer.


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