scholarly journals C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1424
Author(s):  
Sofia Lamas ◽  
Filipa Franquinho ◽  
Marlene Morgado ◽  
João R. Mesquita ◽  
Fátima Gärtner ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Genetically Engineered ◽  
Invasive Technique ◽  
Success Rates ◽  
Embryo Survival ◽  
Common Procedure ◽  
Genetically Engineered Mice ◽  
Lower Success Rate ◽  
Uterine Environment ◽  
Embryo Number

Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were transferred after overnight culture of freshly collected zygotes using different conditions: unilateral transfers using a total of 6, 8, 12, 15, 20 and 25 embryos were performed initially; then, this strain was also used for bilateral transfers using a total of 6, 12 and 20 embryos equally divided by the two oviducts. Groups of 25 embryos were not tested for the bilateral technique, since this condition produced the lower success rate when using the unilateral technique and 20 embryos would still represent a large number of embryos. A group of 2-cell B6129F1 embryos was also transferred using unilateral and bilateral ET with 6, 12 and 20 embryos. Crl:CD1(ICR) were used as recipient females for non-reciprocal transfers and C57BL/6J were used to test reciprocal transfers (only tested for six C57BL/6J unilateral transfers). Unilateral transfers using C57BL/6J mice produced higher success rates using six embryos, compared to the other groups transferred unilaterally (p-values between 0.0001 and 0.0267), but the mean number of pups per litter was not different among groups. Bilateral transfer produced higher number of pups when 20 embryos were divided by the two oviducts compared to six (p = 0.0012) or 12 (p = 0.0148) embryos, but with no differences in success rates. No statistical differences were found between the groups of B6129F1, but better results were obtained on bilateral transfers using a total of six embryos. For the strain tested (C57BL/6J), the uterine environment (Crl:CD1(ICR) or C57BL/6J recipient) does not impact the outcome of the technique. These results complement previous work published using genetically engineered mice strains and show that unilateral transfers using low number of embryos (6), produce better outcomes when compared to bilateral or unilateral transfers using more embryos. It also highlights differences between the outcome of bilateral transfers in the two strains tested. A set of historical data of genetically engineered mice at a C57BL/6J background was also included, confirming that lower embryo numbers are related to higher success rates. Together, the outcome of these experiments can be important to reduce the number of recipient and donor females, optimize embryo transfers and improve animal welfare discouraging the use of a more invasive technique.

100 EMBRYO SURVIVAL AND CONCEPTUS ELONGATION FOLLOWING ASYNCHRONOUS EMBRYO TRANSFER IN CATTLE

2015 ◽  
Vol 27 (1) ◽  
pp. 143
Author(s):  
F. Randi ◽  
B. Fernandez ◽  
M. McDonald ◽  
C. Johnson ◽  
N. Forde ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Prostaglandin F2α ◽  
Early Embryo ◽  
Embryo Survival ◽  
Treatment Groups ◽  
Length Data ◽  
Uterine Environment ◽  
To Receive ◽  
Intravaginal Device

Maternal progesterone (P4) regulates early conceptus growth and development in ruminants. Early embryo transfer studies in sheep and cattle demonstrated a need for close synchrony between the embryo and the uterine environment of the recipient. However, manipulating P4 may be one way of strategically regulating the temporal changes that normally occur in the uterine environment in order to allow flexibility in the timing of embryo transfer. For example, previous studies have demonstrated that P4 administration during the first few days of the oestrous cycle facilitates pregnancy establishment with older embryos. The aim of this study was to examine the effect of embryo-uterine synchrony on conceptus elongation in cattle. Oestrous cycles of crossbred beef heifers were synchronised using an 8-day P4-Releasing Intravaginal Device (PRID Delta®, CEVA, Mountain View, CA, USA) with administration of a prostaglandin F2α analogue (Enzaprost®, CEVA; 5 mL equivalent to 25 mg of dinoprost) given on the day before PRID removal. Heifers were checked for signs of oestrus 4 times per day commencing 30 h after PRID withdrawal. Only those seen in standing oestrus (n = 50) were randomly assigned to 1 of 5 treatment groups to receive Day 7 in vitro-produced blastocysts (n = 10 per recipient) (1) on Day 5 post-oestrus; (2) on Day 5, with P4 supplementation via PRID from Day 3 to 5 + 750 IU of eCG at PRID insertion; (3) on Day 5, PRID Delta from Day 3 to 5 plus 3000 IU of hCG at PRID insertion; (4) on Day 7, or (5) on Day 9. At embryo age Day 14, all heifers were slaughtered and the uterus was flushed to recover and measure conceptuses. Data are summarised in Table 1. Fewer recipients yielded conceptuses (P < 0.05) and fewer conceptuses overall were recovered (P < 0.05) following transfer on Day 5 compared with Day 7 or Day 9. Supplementation with P4 resulted in short cycles (evidenced by corpus luteum regression and/or a recent ovulation at slaughter) in 33.3 to 54.5% of recipients receiving embryos on Day 5. Mean conceptus length was greater (P < 0.05) following transfer to an advanced uterus. In conclusion, transfer of embryos to a retarded (Day 5) uterine environment results in poor embryo survival. Supplementation with P4 shortened the interoestrous period in a significant number of heifers. Transfer to an advanced uterine environment promotes conceptus elongation, presumably driven by P4. Table 1.Embryo survival and conceptus length data

Melatonin Treatment of Donor and Recipient Ewes in an Embryo Transfer Programme: Effects on Ovulation Rate and Embryo Survival During Normal Seasonal Anoestrus

1994 ◽  
Vol 1994 ◽  
pp. 78-78
Author(s):  
J.J. Robinson ◽  
T.G. McEvoy ◽  
R.P. Aitken ◽  
I. Robertson
Keyword(s):  
Embryo Transfer ◽  
Intrauterine Insemination ◽  
Survival Rates ◽  
Ovulation Rate ◽  
Fertilization Failure ◽  
Melatonin Treatment ◽  
Embryo Survival ◽  
Seasonal Differences ◽  
Uterine Environment ◽  
Expected Values

Conception rates and litter sizes in ewes are often higher during the peak of the breeding season than at the beginning or end. How much of this is due to seasonal differences in ovulation rate (Montgomery et al 1988) and ram fertility (Colas, 1983) is uncertain. Using laparoscopic intrauterine insemination which virtually eliminates fertilization failure, Aitken et al (1990) obtained estimates of embryo survival for Greyface ewes induced to ovulate in mid-June (mid anoestrus) that were 25, 19 and 13% below the expected values for 2, 3 and 4 ovulations respectively. While this implies that embryos transferred to recipient ewes at an induced oestrus during seasonal anoestrus may have sub-optimal survival rates, it does not identify whether the cause lies solely with the embryo or with the uterine environment or with both. The present study employs reciprocal embryo transfer in conjunction with melatonin treatment to identify the influence of seasonality on the inherent viability of the embryos and the ability of the uterine environment to sustain pregnancy.

Effect of long-term selection for early postnatal growth rate on survival and prenatal development of transferred mouse embryos

Reproduction ◽  
2000 ◽  
pp. 205-210 ◽  
Author(s):  
CA Ernst ◽  
BK Rhees ◽  
CH Miao ◽  
WR Atchley
Keyword(s):  
Body Weight ◽  
Embryo Transfer ◽  
Maternal Effects ◽  
Body Weight Gain ◽  
Tail Length ◽  
Prenatal Development ◽  
Control Line ◽  
Embryo Survival ◽  
Uterine Environment ◽  
Transfer Procedures

Reciprocal embryo transfer procedures were performed among mouse selection lines to examine prenatal maternal effects on survival and development of transferred embryos. Mice were from generations 28 and 29 of an experiment to select for (i) increased body weight again from 0 to 10 days (E+); (ii) decreased body weight gain from 0 to 10 days (E-); or (iii) a randomly bred control line (C). A total of 118 embryo transfer procedures performed 12 h after conception resulted in 983 progeny born to 89 litters. There was a 39% overall embryo survival rate and 75% overall pregnancy success rate. Response to superovulation and oestrous synchronization was significantly lower (P < 0.01) in the E+ line. E+ individuals that did superovulate produced an average of 37 oocytes per flush, which was significantly higher than in the control line mice (29 oocytes per flush; P < 0.01). The ability to complete pregnancy successfully was not influenced by uterine environment or embryo-uterine interaction. In contrast, embryo survival in successful pregnancies was significantly affected by uterine environment. There were large maternal effects for body weight and tail length at birth; E+ recipients produced pups that were significantly larger than E- recipient pups (P < 0.01), which in turn were significantly larger than pups gestated by control recipients (P < 0.01).

scholarly journals Toxicity modelling of Plk1-targeted therapies in genetically engineered mice and cultured primary mammalian cells

2011 ◽  
Vol 2 (1) ◽  
Author(s):  
Monika Raab ◽  
Sven Kappel ◽  
Andrea Krämer ◽  
Mourad Sanhaji ◽  
Yves Matthess ◽  
...  
Keyword(s):  
Targeted Therapies ◽  
Mammalian Cells ◽  
Genetically Engineered ◽  
Genetically Engineered Mice

scholarly journals Tenascin-C in Heart Diseases—The Role of Inflammation

2021 ◽  
Vol 22 (11) ◽  
pp. 5828
Author(s):  
Kyoko Imanaka-Yoshida
Keyword(s):  
Ventricular Remodeling ◽  
Heart Diseases ◽  
Genetically Engineered ◽  
Matricellular Protein ◽  
Myocardial Repair ◽  
Inflammatory Reactions ◽  
Tenascin C ◽  
Genetically Engineered Mice

Tenascin-C (TNC) is a large extracellular matrix (ECM) glycoprotein and an original member of the matricellular protein family. TNC is transiently expressed in the heart during embryonic development, but is rarely detected in normal adults; however, its expression is strongly up-regulated with inflammation. Although neither TNC-knockout nor -overexpressing mice show a distinct phenotype, disease models using genetically engineered mice combined with in vitro experiments have revealed multiple significant roles for TNC in responses to injury and myocardial repair, particularly in the regulation of inflammation. In most cases, TNC appears to deteriorate adverse ventricular remodeling by aggravating inflammation/fibrosis. Furthermore, accumulating clinical evidence has shown that high TNC levels predict adverse ventricular remodeling and a poor prognosis in patients with various heart diseases. Since the importance of inflammation has attracted attention in the pathophysiology of heart diseases, this review will focus on the roles of TNC in various types of inflammatory reactions, such as myocardial infarction, hypertensive fibrosis, myocarditis caused by viral infection or autoimmunity, and dilated cardiomyopathy. The utility of TNC as a biomarker for the stratification of myocardial disease conditions and the selection of appropriate therapies will also be discussed from a clinical viewpoint.

scholarly journals Characterization and visualization of murine coagulation factor VIII-producing cells in vivo

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Morisada Hayakawa ◽  
Asuka Sakata ◽  
Hiroko Hayakawa ◽  
Hikari Matsumoto ◽  
Takafumi Hiramoto ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Factor Viii ◽  
Fluorescent Protein ◽  
Coagulation Factor ◽  
Coagulation Factors ◽  
Genetically Engineered ◽  
Coagulation Factor Viii ◽  
Liver Sinusoidal Endothelial Cells ◽  
Genetically Engineered Mice

AbstractCoagulation factors are produced from hepatocytes, whereas production of coagulation factor VIII (FVIII) from primary tissues and cell species is still controversial. Here, we tried to characterize primary FVIII-producing organ and cell species using genetically engineered mice, in which enhanced green fluorescent protein (EGFP) was expressed instead of the F8 gene. EGFP-positive FVIII-producing cells existed only in thin sinusoidal layer of the liver and characterized as CD31high, CD146high, and lymphatic vascular endothelial hyaluronan receptor 1 (Lyve1)+. EGFP-positive cells can be clearly distinguished from lymphatic endothelial cells in the expression profile of the podoplanin− and C-type lectin-like receptor-2 (CLEC-2)+. In embryogenesis, EGFP-positive cells began to emerge at E14.5 and subsequently increased according to liver maturation. Furthermore, plasma FVIII could be abolished by crossing F8 conditional deficient mice with Lyve1-Cre mice. In conclusion, in mice, FVIII is only produced from endothelial cells exhibiting CD31high, CD146high, Lyve1+, CLEC-2+, and podoplanin− in liver sinusoidal endothelial cells.

scholarly journals Immune Competency of a Hairless Mouse Strain for Improved Preclinical Studies in Genetically Engineered Mice

2010 ◽  
Vol 9 (8) ◽  
pp. 2354-2364 ◽  
Author(s):  
Beverly S. Schaffer ◽  
Marcia H. Grayson ◽  
Joy M. Wortham ◽  
Courtney B. Kubicek ◽  
Amanda T. McCleish ◽  
...  
Keyword(s):  
Mouse Strain ◽  
Hairless Mouse ◽  
Genetically Engineered ◽  
Preclinical Studies ◽  
Genetically Engineered Mice ◽  
Immune Competency

scholarly journals miR-146a is a significant brake on autoimmunity, myeloproliferation, and cancer in mice

2011 ◽  
Vol 208 (6) ◽  
pp. 1189-1201 ◽  
Author(s):  
Mark P. Boldin ◽  
Konstantin D. Taganov ◽  
Dinesh S. Rao ◽  
Lili Yang ◽  
Jimmy L. Zhao ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Immune Cell ◽  
Myeloid Cell ◽  
Genetically Engineered ◽  
Biological Processes ◽  
Targeted Deletion ◽  
Genetically Engineered Mice ◽  
Immune Defects ◽  
Molecular Brake

Excessive or inappropriate activation of the immune system can be deleterious to the organism, warranting multiple molecular mechanisms to control and properly terminate immune responses. MicroRNAs (miRNAs), ∼22-nt-long noncoding RNAs, have recently emerged as key posttranscriptional regulators, controlling diverse biological processes, including responses to non-self. In this study, we examine the biological role of miR-146a using genetically engineered mice and show that targeted deletion of this gene, whose expression is strongly up-regulated after immune cell maturation and/or activation, results in several immune defects. Collectively, our findings suggest that miR-146a plays a key role as a molecular brake on inflammation, myeloid cell proliferation, and oncogenic transformation.

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