The Dynamic Equine Embryo from Postfixation (Day 17) to the End of the Embryo Stage (Day 40)

Author(s):  
O.J. Ginther
Keyword(s):  
2021 ◽  
Vol 22 (11) ◽  
pp. 5499
Author(s):  
Veronica Corsetti ◽  
Carla Perrone-Capano ◽  
Michael Sebastian Salazar Intriago ◽  
Elisabetta Botticelli ◽  
Giancarlo Poiana ◽  
...  

Dorsal root ganglia (DRG) neurons synthesize acetylcholine (ACh), in addition to their peptidergic nature. They also release ACh and are cholinoceptive, as they express cholinergic receptors. During gangliogenesis, ACh plays an important role in neuronal differentiation, modulating neuritic outgrowth and neurospecific gene expression. Starting from these data, we studied the expression of choline acetyltransferase (ChAT) and vesicular ACh transporter (VAChT) expression in rat DRG neurons. ChAT and VAChT genes are arranged in a “cholinergic locus”, and several splice variants have been described. Using selective primers, we characterized splice variants of these cholinergic markers, demonstrating that rat DRGs express R1, R2, M, and N variants for ChAT and V1, V2, R1, and R2 splice variants for VAChT. Moreover, by RT-PCR analysis, we observed a progressive decrease in ChAT and VAChT transcripts from the late embryonic developmental stage (E18) to postnatal P2 and P15 and in the adult DRG. Interestingly, Western blot analyses and activity assays demonstrated that ChAT levels significantly increased during DRG ontogenesis. The modulated expression of different ChAT and VAChT splice variants during development suggests a possible differential regulation of cholinergic marker expression in sensory neurons and confirms multiple roles for ACh in DRG neurons, both in the embryo stage and postnatally.


Author(s):  
Yuanyuan Li ◽  
Ning-Hua Mei ◽  
Gui-Ping Cheng ◽  
Jing Yang ◽  
Li-Quan Zhou

Mitochondrion plays an indispensable role during preimplantation embryo development. Dynamic-related protein 1 (DRP1) is critical for mitochondrial fission and controls oocyte maturation. However, its role in preimplantation embryo development is still lacking. In this study, we demonstrate that inhibition of DRP1 activity by mitochondrial division inhibitor-1, a small molecule reported to specifically inhibit DRP1 activity, can cause severe developmental arrest of preimplantation embryos in a dose-dependent manner in mice. Meanwhile, DRP1 inhibition resulted in mitochondrial dysfunction including decreased mitochondrial activity, loss of mitochondrial membrane potential, reduced mitochondrial copy number and inadequate ATP by disrupting both expression and activity of DRP1 and mitochondrial complex assembly, leading to excessive ROS production, severe DNA damage and cell cycle arrest at 2-cell embryo stage. Furthermore, reduced transcriptional and translational activity and altered histone modifications in DRP1-inhibited embryos contributed to impeded zygotic genome activation, which prevented early embryos from efficient development beyond 2-cell embryo stage. These results show that DRP1 inhibition has potential cytotoxic effects on mammalian reproduction, and DRP1 inhibitor should be used with caution when it is applied to treat diseases. Additionally, this study improves our understanding of the crosstalk between mitochondrial metabolism and zygotic genome activation.


2002 ◽  
Vol 50 (6) ◽  
pp. 851-862 ◽  
Author(s):  
Yoshihiro Akimoto ◽  
Naomi Yamakawa ◽  
Kiyoshi Furukawa ◽  
Koji Kimata ◽  
Hayato Kawakami ◽  
...  

The expression and distribution of the long form of Type XII collagen were investigated histochemically during chicken corneal development using a monoclonal antibody (P3D11) raised against the N-terminal domain of chicken Type XII collagen. Specificity of the antibody was confirmed by immunoprecipitation before and after bacterial collagenase digestion. Immunofluorescent microscopic studies showed that during chicken cornea formation, the long form of Type XII collagen is initially detected on Day 3 embryo (stage 19) in the sub-epithelial matrix of the corneal periphery and in the matrix around the optic cup. On Day 5 embryo (stage 27) the long form was expressed in the primary stroma. Thereafter, as the secondary stroma was formed, the long form localized in the sub-epithelial and sub-endothelial matrices and in the anterior region of the limbus (corneoscleral junction) before the formation of Descemet's and Bowman's membranes. After hatching, the immunoreactivity decreased predominantly in the sub-epithelial and sub-endothelial matrices but remained at the anterior region of the limbus. Immunoelectron microscopic examination demonstrated that the long form localizes in the Descemet's and Bowman's membranes and along the collagen fibrils in the stroma with a periodic repeat. Based on the distribution of the long form of Type XII collagen in the sub-epithelial and sub-endothelial matrices and limbus, it was suggested that the long form of Type XII collagen is involved in formation of the Descemet's and Bowman's membranes and in stabilization of the limbus.


2019 ◽  
Vol 109 (6) ◽  
pp. 821-830 ◽  
Author(s):  
C.-X. Sun ◽  
Z.-X. Li

AbstractThe major component of aphid alarm pheromone is (E)-β-farnesene (EβF), but the molecular mechanisms of EβF synthesis are poorly understood. Here we established a biological model to study the modulation of EβF synthesis in the bird cherry-oat aphid Rhopalosiphum padi by using quantitative polymerase chain reaction, gas chromatography/mass spectrometry and RNA interference. Our results showed that the rearing conditions significantly affected the weight of adult and modulated EβF synthesis in a transgenerational manner. Specifically, the quantity of EβF per milligram of aphid was significantly reduced in the individually reared adult or 1st-instar nymphs derived from 1-day-old adult reared individually, but EβF in the nymph derived from 2-day-old adult that experienced collective conditions returned to normal. Further study revealed that the production of EβF started in embryo and was extended to early nymphal stage, which was modulated by farnesyl diphosphate synthase genes (RpFPPS1 and RpFPPS2) and rearing conditions. Knockdown of RpFPPS1 and RpFPPS2 confirmed the role played by FPPS in the biosynthesis of aphid alarm pheromone. Our results suggested that the production of EβF starts at the embryo stage and is modulated by FPPS and rearing conditions in R. padi, which sheds lights on the modulatory mechanisms of EβF in the aphid.


2019 ◽  
Vol 70 (19) ◽  
pp. 5101-5114 ◽  
Author(s):  
Xiangxiang Kong ◽  
Landi Luo ◽  
Jinjie Zhao ◽  
Qian Chen ◽  
Guanxiao Chang ◽  
...  

Root FRIGIDA activated the novel targets MAF4/5 to delay flowering; temporal expresson of FRIGIDA as early as at the pro-embryo stage is efficient in delaying flowering.


2018 ◽  
Vol 23 (2) ◽  
pp. 155-157 ◽  
Author(s):  
Alvaro Plaza Reyes ◽  
Fredrik Lanner

2006 ◽  
Vol 18 (2) ◽  
pp. 203
Author(s):  
C. Ponsart ◽  
H. Quinton ◽  
A. Rohou ◽  
J. Kelhembo ◽  
G. Bourgoin ◽  
...  

Previous studies have shown that the time between flushing and freezing of bovine embryos can influence pregnancy rates (PRs) following embryo transfer (ET). The aim of this study was to determine which time components can influence ET results. Time components between flushing of a superovulated donor and freezing of the collected embryos were investigated under field conditions. Embryos were frozen in 1.5 M ethylene glycol (EG) for direct transfer. During January 2003, ET technicians (EmbryoTop, Rennes cedex, France) recorded systematically times corresponding to each step comprising the time spent in vitro (TIV) from 153 recovery sessions (RS) with freezing: end of flushing, beginning and end of search of embryos, start of equilibration in EG, beginning and end of straw loading, introduction to −7°C in the freezer, and seeding. Numbers of donor cows and ET technicians doing the freezing (n = 5) were noted for each RS. Embryo (stage, quality) and recipient (breed, parity) characteristics were also noted. A total of 548 frozen embryos were transferred and PRs were assessed. Variability of time components was investigated (Bourgoin et al. 2004 Reprod. Fertil. Dev. 16, 207). The influence of time components and other variation factors was tested on PRs (t-tests and chi-square analysis). The TIV averaged 210 ± 80 min and did not influence PR (≤4 h = 51.9% (n = 393) vs. >4 h = 55.5% (n = 155); P > 0.05), as well as duration of flushing (32 ± 8 min), interval between end of flushing and search (31 ± 27 min), duration of search (45 ± 25 min) and interval between end of search and beginning of freezing (101 ± 63 min). Only significant factors were kept for further analysis. The effects of recipient parity, number of donor cows per RS, and interval between introduction of straw to −7°C, and seeding were tested in a multivariate logistic model. PR varied strongly with parity of recipient (+25% in heifers vs. cows; P = 0.001). PRs were higher when the interval between straw introduction in the freezer and seeding lasted at least 5 min (2–4 min = 48.0% (n = 254) vs. 5–8 min = 57.1% (n = 294); P = 0.009). Time and operator effects were confounded. Overall PR results for the two technicians who used mostly 2–4 min intervals averaged 47% (operator values = 35.6, 48.9, and 54.5) whereas PRs were 54.9 and 60.5% for those waiting 5 min or more before inducing seeding (n = 2). PRs were higher when at least two donor cows were collected per RS (1 donor cow = 49% (n = 259) vs. ≥2 donor cows = 56.4% (n = 289); P = 0.003). This was not in agreement with previous observations in fresh embryos (Bourgoin et al. 2004). However, the number of donor cows strongly influenced the number of viable embryos per RS (1 donor cow = 11 ± 5 vs. ≥2 donor cows = 18 ± 8.5; P < 0.05) and could permit the choice of more embryos to be frozen. These results show that good PR may be achieved with a delay of several hours between flushing and freezing, when heifers are used as recipients. Moreover, confirmed from higher numbers of operators, these data show that it is better to wait at least 5 min to achieve equilibration of the embryo before seeding.


2016 ◽  
Vol 28 (2) ◽  
pp. 183 ◽  
Author(s):  
L. C. Carrenho-Sala ◽  
R. V. Sala ◽  
M. Fosado ◽  
D. C. Pereira ◽  
S. Garcia ◽  
...  

A retrospective study was performed to evaluate factors that influence pregnancy per embryo transfer (P/ET) in an IVF-embryo transfer program. A total of 5026 fresh in vitro-produced embryos were transferred during 2014 and evaluated for effects of embryo quality, embryo stage, size of corpus luteum (CL; 18–19.9 mm or ≥20 mm), interval from GnRH to embryo transfer, number of previous embryo transfer (0, 1, 2, 3, ≥4); and interaction of embryo stage and interval from GnRH to embryo transfer. One group (n = 850) had detection of oestrus after prostaglandin F2α application but most heifers (n = 4176) received fixed timed embryo transfer after a 5-day CIDR-Synch protocol: Day –8 CIDR inserted; Day –3 CIDR removed and prostaglandin F2α; Day –2 prostaglandin F2α; Day 0 GnRH. Ultrasound was performed on Day 6 after GnRH or oestrus to measure CL size and on Day 32 and 60 to determine pregnancy. Data for P/ET were analysed by logistic regression (LOGISTIC procedure, SAS 9.4). Embryo quality influenced P/ET at Day 32 [Grade 1 48.4% (1273/2631) v. Grade 2 37.6% (900/2395); P < 0.01] and at Day 60 [Grade 1 38.9% (1023/2631) v. Grade 2 29.0% (694/2395); P < 0.01], and altered pregnancy loss [Grade 1 19.6% (250/1273) v. Grade 2 22.9% (206/900); P = 0.03]. Stage of the embryo also had an effect on P/ET at Day 32 [Stage 6 35.5%a (582/1641), Stage 7 46.3%b (1431/3092), and Stage 8 54.6%c (160/293); P < 0.01] and at Day 60 [Stage 6 28.2%a (462/1641), Stage 7 36.6%b (1131/3092), and Stage 8 41.6%b (122/293); P < 0.01], but did not affect pregnancy loss (P = 0.22). Interestingly, interval from GnRH (or oestrus) until embryo transfer did not affect P/ET at Day 32 (P = 0.10), 60 (P = 0.23), or pregnancy loss (P = 0.3), nor was there an interaction between interval and embryo stage at Day 32 (P = 0.77), 60 (P = 0.96) or pregnancy loss (P = 0.55). As shown in Table 1, embryo stage 6 was always the lowest and stage 8 always the greatest P/ET regardless of interval from GnRH to embryo transfer. Size of CL also did not affect P/ET at Day 32 (P = 0.09), 60 (P = 0.21), or pregnancy loss (P = 0.90). Number of previous embryo transfer also did not alter P/ET at Day 32 [0 = 43.3% (886/2046), 1 = 44.1% (639/1450), 2 = 43.4% (444/1024), 3 = 42.6% (146/343), and ≥4 = 35.6% (58/163); P = 0.33] or 60 (P = 0.51) or pregnancy loss (P = 0.12). In conclusion, embryo stage and quality are the major factors that impacted P/ET in this study, with surprisingly little effect of interval from GnRH to embryo transfer, size of the CL, and number of previous embryo transfer. Thus, recipient programs for IVF-embryo transfer can be designed with substantial flexibility. Table 1.Effect of embryo stage and recipient synchrony on pregnancies per embryo transfer on Day 32 in recipient dairy heifers


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