scholarly journals MicroRNA-210 Regulates Endoplasmic Reticulum Stress and Apoptosis in Porcine Embryos

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 221
Author(s):  
Muhammad Rosyid Ridlo ◽  
Eui Hyun Kim ◽  
Geon A. Kim
Keyword(s):  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Embryo Development ◽  
Caspase 3 ◽  
Formation Rate ◽  
Embryo Production ◽  
Embryo Survival ◽  
Expression Levels ◽  
In Vitro Embryo Production

Endoplasmic reticulum (ER) stress can be triggered during in vitro embryo production and is a major obstacle to embryo survival. MicroRNA (miR)-210 is associated with cellular adaptation to cellular stress and inflammation. An experiment was conducted to understand the effects of miR-210 on in vitro embryo development, ER stress, and apoptosis; to achieve this, miR-210 was microinjected into parthenogenetically activated embryos. Our results revealed that miR-210 inhibition significantly enhanced the cleavage rate, blastocyst formation rate, and total cell number (TCN) of blastocysts, and reduced expression levels of XBP1 (p < 0.05). miR-210 inhibition greatly reduced the expression of ER stress-related genes (uXBP1, sXBP1, ATF4, and PTPN1) and Caspase 3 and increased the levels of NANOG and SOX2 (p < 0.05). A miR-210-mimic significantly decreased the cleavage, blastocyst rate, TCN, and expression levels of XBP1 compared with other groups (p < 0.05). The miR-210-mimic impaired the expression levels of uXBP1, sXBP1, ATF4, PTPN1, and Caspase 3 and decreased the expression of NANOG and SOX2 (p < 0.05). In conclusion, miR-210 plays an essential role in porcine in vitro embryo development. Therefore, we suggest that miR-210 inhibition could alleviate ER stress and reduce apoptosis to support the enhancement of in vitro embryo production.

106 COMPARISON OF IN VITRO EMBRYO PRODUCTION IN PANAMA USING HOLSTEIN OOCYTES WITH BOS INDICUS SEXED SEMEN FROM DONORS IN DIFFERENT LOCATIONS: FIELD DATA

2016 ◽  
Vol 28 (2) ◽  
pp. 183
Author(s):  
S. J. R. Rodriguez ◽  
Y. E. Ramirez ◽  
E. Gomes ◽  
L. F. Nasser ◽  
J. H. F. Pontes ◽  
...  
Keyword(s):  
Embryo Development ◽  
Bos Indicus ◽  
Embryo Production ◽  
University Of Illinois ◽  
Oocyte Collection ◽  
Sexed Semen ◽  
The Usa ◽  
In Vitro Embryo Production ◽  
The University

The objective of this work was to compare in vitro embryo production of Bos taurus × Bos indicus cross embryos using oocytes from Holstein donors under different production and environment systems. This study also examined the possibility for in vitro production using oocytes imported and transported fresh between the USA and Panama. All animals were mature Holstein cows going through a normal lactation. The first group of donors was from the University of Illinois dairy herd and went through 3 ovum pickup sessions. The second group of donors were Holstein cows already adapted to Panama and went through 10 ovum pickup sessions. The Panamanian herd of Holstein donors were born and raised in Panama in an area of mountains, on average 1300 m above sea level. This environment does not have the typical hot and humid tropical weather seen in other regions of Panama. Both groups of donors were aspirated without stimulation during the years 2013 and 2014. Oocytes recovered from donors in Illinois were imported fresh under a special sanitary research protocol between Panama and the University of Illinois. The transport of fresh oocytes from the USA to Panama was done using a portable incubator set at 39°C (Minutube of America). Oocytes were matured during transport in 5-mL tubes (~30–35 oocytes per tube) containing 400 µL of maturation media (TCM-199) that had been equilibrated with 5% CO2. Oocytes recovered from donors in Panama were matured using the same media. For both groups, oocytes were inseminated 24 h after ovum pickup using sexed semen from the same bull. All embryo production procedures followed the protocols of the In vitro Brasil™ commercial system. At 72 h postinsemination, cleavage was evaluated. On Day 7 after insemination, embryo development to the blastocyst stage (early to expanded) was recorded. Data were analysed using Chi-squared. As shown in Table 1, there was no effect of oocyte collection location on embryo development. These results indicate that it is possible to produce a viable in vitro-produced embryo using fresh oocytes collected and transported from different countries. This work opens the possibility to access superior genetics and improve herds in countries seeking to increase their production systems and potentially improve their quality of life. Table 1.Effect of oocyte collection location on embryo development This project was supported by Programa de Competitividad ProCom Senacyt, Panama.

206 EFFECT OF FOLLICULAR WAVE SYNCHRONIZATION AND SUPERSTIMULATION ON IN VITRO EMBRYO PRODUCTION

2008 ◽  
Vol 20 (1) ◽  
pp. 182 ◽  
Author(s):  
K. Imai ◽  
Y. Inaba ◽  
H. Yoshioka ◽  
Y. Aikawa ◽  
M. Ohtake ◽  
...  
Keyword(s):  
Formation Rate ◽  
Cumulus Cell ◽  
Oocyte Quality ◽  
Annual Conference ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Follicular Wave ◽  
The Mean ◽  
In Vitro Embryo Production

We previously reported that follicular wave synchronization, by removal of the dominant follicle on Day 5 after ovum pickup (OPU), was effective in increasing oocyte quality in the developing follicles (Imai et al. 2006 32th Annual Conference of the IETS, poster presentation no. 277). The current study was designed to examine the effect of superstimulatory treatment to induce subsequent follicular wave synchronization on embryo production by OPU and IVM-IVF-IVC in Holstein dry cows. Cows were reared under the same feeding and environmental conditions, and 2 OPU sessions were conducted in each cow. In the first session, OPU was performed in 8 cows on arbitrary days of the estrous cycle by using a 7.5-MHz linear transducer with needle (Cova needle, Misawa Medical, Tokyo, Japan) connected to an ultrasound scanner (SSD-1200, Aloka, Tokyo, Japan). Follicles larger than 8 mm in diameter were then aspirated and a CIDR was inserted on Day 5 (the day of first OPU session = Day 0). Cows then received 30 mg of FSH (Antrin-R10; Kawasaki Mitaka Pharmaceutical Co., Tokyo, Japan) twice a day from Days 7 to 10 in decreasing doses (6, 6, 4, 4, 3, 3, 2, 2 mg) by i.m. injection. Cloprostenol (PGF; Clopromate C; Sumitomo Pharmaceuticals Co., Tokyo, Japan; 0.75 mg) was administered in the morning of Day 9 (third day of superstimulation). The second OPU session was performed 48 h after PGF administration (Day 11), and only follicles larger than 5 mm in diameter were aspirated. The CIDR was removed from the cows just before OPU. Collected oocytes were evaluated by their cumulus cell morphology, cytoplasmic color, and density. Grades 1 and 2 COC were matured, fertilized, and cultured as described by Imai et al. [2006 J. Reprod. Dev. 52(Suppl.), S19–S29]. Embryo development was assessed by the cleavage rate on Day 2 and by the blastocyst formation rate on Days 7 to 8 (the day of insemination = Day 0). Data were analyzed by Student's t-test. There were no differences in the mean (� SD) number of aspirated follicles or collected oocytes between the first (32.5 � 6.8 and 26.0 � 12.7, respectively) and second (29.3 � 10.4 and 19.0 � 9.4, respectively) OPU sessions (P > 0.1). The percentage of Grade 1 and 2 oocytes for the second OPU session (90.5 � 13.8%) was significantly higher (P < 0.01) than for the first OPU session (63.1 � 6.3%), and significant differences were found for cleavage (79.4 � 14.1, 61.8 � 25.1, P < 0.01) and blastocyst rates (68.1 � 16.7, 24.2 � 22.3, P < 0.001) between sessions. The mean numbers of blastocysts obtained per session were 4.3 � 2.9 and 12.8 � 8.7 in the first and second sessions, respectively (P < 0.01). These results indicate that superstimulatory treatment and subsequent follicular wave synchronization were effective on in vitro embryo production by increasing the oocyte quality.

238 KNOCKING DOWN OF THYROID HORMONE RECEPTORS INHIBITS DEVELOPMENT OF EARLY BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 267
Author(s):  
N. Y. Rho ◽  
F. A. Ashkar ◽  
T. Revay ◽  
P. Madan ◽  
W. A. King
Keyword(s):  
Embryo Development ◽  
Reference Genes ◽  
Messenger Rna ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Blastocyst Formation ◽  
Embryo Production ◽  
Mrna Transcripts ◽  
In Vitro Embryo Production

Thyroid hormones (TH) play an important role in the physiology of vertebrates, ranging from the regulation of metabolic processes to cell proliferation, differentiation, and embryo development. We have previously shown a beneficial effect of supplementing TH in in vitro embryo production media. Recently, detection of TH receptors (TR) in oocytes and early stages of pre-implantation embryos indicated a possible regulatory role for TH in these stages (unpublished data). The objective of this study was to investigate the importance of TR expression in the pre-attachment bovine embryo in vitro. Bovine embryos, produced by standard in vitro embryo production procedures, were microinjected at the zygote stage with small interfering RNA (siRNA) specifically designed for knocking down either TR-α or TR-β. In addition, groups of zygotes were microinjected with scrambled siRNA (SI) or were not injected (NI), and these groups served as controls. Embryo developmental rates were assessed using light microscopy for blastocyst formation rates and expression of TR messenger RNA (mRNA) transcripts at the blastocyst stage was assessed by quantitative PCR across all groups. Expression of TR mRNA was normalized against glyceraldehyde 3-phosphate dehydrogenase, H2a, and 18S as reference genes. There was a significant decrease in blastocyst formation rates in both embryo groups injected with either TR-α (P < 0.002) and TR-β (P < 0.001) siRNA compared with the NI and SI groups. Moreover, the TR-β knockdown group exhibited a lower developmental rate than the TR-α knockdown group, which indicates a stronger inhibitory role for TR-β. Quantification of the level of TR mRNA expression in four groups normalized with three different reference genes shows a consistent significant reduction in the levels of TR-α (P < 0.05) and TR-β (P < 0.02) mRNA transcripts compared with the NI and SI groups. However, TR-β expression was inhibited more than was TR-α expression. In conclusion, the results indicate that knocking down either TR-α or TR-β restrains embryo development. This suggests that TH play a vital role in the regulation of embryo development through their receptors during bovine early embryogenesis. The specific role of each of these receptors and their mechanism of action in mediating development needs to be further elucidated. Funding was provided by CRC, NSERC, and the EmbryoGENE network.

335 EFFECT OF MEM VITAMINS AND FORSKOLIN ON IN VITRO EMBRYO PRODUCTION AND SOPS-VITRIFICATION ABILITY OF IN VITRO DERIVED PORCINE BLASTOCYSTS

2010 ◽  
Vol 22 (1) ◽  
pp. 324 ◽  
Author(s):  
C. Cuello ◽  
C. Almiñana ◽  
J. Gomis ◽  
M. A. Gil ◽  
C. Maside ◽  
...  
Keyword(s):  
Least Squares ◽  
Factorial Design ◽  
Formation Rate ◽  
Dilution Method ◽  
Minimum Essential Medium ◽  
Cleavage Rate ◽  
Embryo Production ◽  
One Step ◽  
In Vitro Embryo Production

The aims of this study were, first to investigate the effect of minimum essential medium (MEM) vitamins (VITs) during IVM of porcine oocytes on in vitro embryo production of porcine embryos and second, to determine if the addition of VITs during IVM and the chemical delipidation with forskolin improve the vitrification ability of in vitro-derived porcine blastocysts. COCs were divided in two groups and matured in NCSU-23 for 44 h with 0.05% VITs (V group) or without VITs (NV group). Matured oocytes were inseminated with frozen-thawed spermatozoa. Presumptive zygotes were cultured for 16 h to assess IVF parameters (N = 767) or for 6 days (N = 2858) to evaluate the in vitro embryo development (Day 0 = day of IVF). On Day 5, some embryos from NV and V groups were cultured for 24 h with 10 μM forskolin (NVF and VF groups). The remaining embryos were cultured without forskolin (NVNF and VNF groups). On Day 6, embryos from the four experimental groups were assessed for blastocysts formation and some blastocysts (N = 414) were vitrified using superfine open pulled straws (SOPS). Vitrified blastocysts were warmed (one-step dilution method) and cultured for 24 h to assess their viability. Blastocysts with totally or partially reformed blastocoel cavity and normal or excellent morphology were considered viable. Five replicates of a 2 × 2 factorial design were conducted. Data was analyzed with the MIXED procedure. The threshold for significance was set at P < 0.05. Results are expressed as least squares means ± SEM. No differences were observed in the maturation of COCs treated with VITs (92.7 ± 1.3%) and non-treated COCs (94.3 ± 1.3%). The NV and V maturation groups showed similar penetration (83.0 ± 3% and 82.6 ± 3%, respectively) and monospermy (48.9 ± 6%, and 48.3 ± 6%, respectively) rates. The rate of monosermic oocytes related to the total of analyzed oocytes was similar for NV (38.1 ± 3.7%) and V (38.4 ± 3.7%) groups. The values of cleavage rate on Day 2 were similar for NV (66.7 ± 1.5%) and V (69.6 ± 1.6%) embryos. The addition of VITs to IVM medium improved (P < 0.01) up to 10 points the blastocysts formation rate, but the addition of forskolin at Day 5 did not affect this parameter. The V group showed a higher (P < 0.01) blastocysts rate (45.1 ± 2.5%) than the NV group (38.4 ± 2.3%).The addition of VITs did not affect the survival of in vitro-derived blastocysts after SOPS-vitrification on Day 6. However, the blastocysts cultured for 24 h with forskolin showed higher (P < 0.05) viability (44.0 ± 7.9%) after vitrification and warming than those embryos cultured without forskolin (34.1 ± 7.8%). In conclusion, the addition of MEM vitamins to IVM medium improves the blastocysts formation rate and the chemical delipidation with forskolin improve the cryosurvival of SOPS-vitrified porcine in vitro-derived blastocysts. This study was supported by the Seneca foundation of Murcia (GERM 04543/07), MICINN (AGL2009-12091 and RC-2007), and CARM (2I05SU0012).

186 SUPPLEMENTATION WITH LOW DOSES OF DIMETHYL SULFOXIDE DURING IN VITRO MATURATION RESULTS IN IMPROVED IN VITRO EMBRYO PRODUCTION IN CATTLE

2017 ◽  
Vol 29 (1) ◽  
pp. 201
Author(s):  
A. E. Ynsaurralde ◽  
M. Suvá ◽  
R. Bevacqua ◽  
S. Munilla ◽  
C. Luchetti ◽  
...  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Predictive Equation ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
First Polar Body ◽  
In Vitro Embryo Production

Oocyte in vitro maturation (IVM) is crucial for subsequent in vitro embryo production. It involves acquisition of competence for fertilization and embryo development. Therefore, its optimization could have a direct impact on in vitro embryo development. Dimethyl sulfoxide (DMSO) is commonly used as solvent or vehicle, but also increases the membrane permeability and behaves as a scavenger of cytotoxic free radicals. The aim of this study was to evaluate the effect of DMSO supplementation during bovine oocyte maturation on subsequent in vitro embryo development and to determine the optimal usage dose with no toxic effect. To this aim, cumulus-oocyte complexes were collected from slaughterhouse ovaries and IVM in TCM 199 containing 10% fetal bovine serum, 10 µg mL−1 of FSH, 0.3 mM sodium pyruvate, 100 mM cysteamine, and 2% antibiotic-antimycotic. The oocytes were incubated for 24 h at 6.5% CO2 in humidified air at 38.5°C. For Experiment 1, IVM medium was supplemented with DMSO at concentrations of 0, 0.1, 0.5, 1, or 10% (vol/vol) DMSO (n = 241, 195, 42, 192, 172 oocytes) and IVM rate was determined by presence of the first polar body. For Experiment 2, 0, 0.1, 0.25, 0.5, 0.75, 1, or 10% (vol/vol) DMSO (n = 446, 322, 65, 194, 77, 250, 39 oocytes) was supplemented to IVM medium and cleavage and blastocyst rates were determined to establish the optimal usage dose. In vitro fertilization was performed according to Brackett and Oliphant (1975), with 16 × 106 spermatozoa/mL for 5 h. Afterwards, presumptive zygotes were cultured in SOF for 7 days at 38.5°C and 5% O2. Cleavage and blastocyst rates were determined on Days 2 and 7, respectively. Results were statistically analysed using Fisher’s exact test by GraphPad Prism software (GraphPad Software Inc., La Jolla, CA, USA). Also, the percentage of blastocyst was adjusted to DMSO concentration using the R software quadratic regression model. The optimum usage dose was determined by calculating the maximum of the estimated predictive equation. In vitro maturation in 10% DMSO resulted in significantly lower first polar body extrusion rates (0% = 74%a, 0.1% = 73%a, 0.5% = 83%a, 1% = 66%a, and 10% = 8%b; different letters indicate statistical differences) and lower cleavage rates (0% = 75%a, 0.1% = 77%a, 0.25% = 80%a, 0.5% = 79%a, 0.75% = 78%a, 1% = 77%a, and 10% = 3%b) than the other treatments. Furthermore, blastocyst production was higher for the 0.25 and 0.5% (vol/vol) supplemented DMSO groups (0% = 26%b, 0.1% = 37%ab, 0.25% = 40%a, 0.5% = 41%a, 0.75% = 34%ab, 1% = 23%b, and 10% = 0%c). The predictive equation results indicate that the maximum percentage of blastocysts is obtained with a concentration of 0.458% (vol/vol) of DMSO. In conclusion, DMSO supplementation during IVM of bovine oocytes had a positive effect on in vitro development. Further studies will be carried out to elucidate its mechanism of action.

145 Inhibitory effects of gossypol on bovine in vitro embryo production

2019 ◽  
Vol 31 (1) ◽  
pp. 197
Author(s):  
L. K. Hatamoto-Zervoudakis ◽  
M. F. Duarte Jr ◽  
T. F. Motheo ◽  
P. P. Tsuneda ◽  
J. T. Zervoudakis
Keyword(s):  
Embryo Development ◽  
Free Gossypol ◽  
Wilcoxon Test ◽  
Free Form ◽  
Bovine Embryo ◽  
Sodium Pyruvate ◽  
Embryo Production ◽  
Parametric Data ◽  
In Vitro Embryo Production

Cottonseed and its derivatives are frequently used in cattle feed as an effective dietary fibre supply and high protein and energy food source. However, the cotton plant contains gossypol, which in its free form induces male and female infertility. Therefore, this study aimed to evaluate the effect of gossypol supplementation on bovine in vitro embryo production. Ovaries were retrieved from slaughterhouses, and cumulus-oocyte complexes (COC) were recovered by follicular puncture. Based on free gossypol concentration present on the in vitro maturation, sperm capacitation, IVF and in vitro culture media, grades I, II and III COC (n=646) were divided in 3 treatments: 0μg mL−1 (control), 5μg mL−1 (G5) and 10μg mL−1 (G10). The COC were matured under a humidified atmosphere of 5% CO2 in air at 38.5°C for 24h in 90-μL droplets containing TCM-199 supplemented with 10% FCS, 0.2mM sodium pyruvate, LH, FSH, 75μg mL−1 amikacin, 17β-oestradiol. Each droplet corresponded to one replicate (n=14) and contained 15 to 18 COC. Matured COC and sperm were co-incubated in droplets (8-13 COC per 90μL) of TALP-IVF media supplemented with 6mg mL−1 BSA, 0.2mM sodium pyruvate, 30μg mL−1 heparin, 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine, 75μg mL−1 amikacin under a 5% CO2 humidified atmosphere at 38.5°C, for 20h. For IVF, non-sexed frozen-thawed semen was selected with Percoll® gradient. The resulting pellet was subjectively evaluated for motility and concentration and then diluted to final concentration of sperm mL−1 with fertilization medium. Presumptive zygotes were then cultured in 90-μL droplets of SOFaaci medium supplemented with 2.7mM myo-inosytol, 0.2mM pyruvate, 2.5% FCS (v/v), 5mg mL−1 BSA, 75μg mL−1 amikacin, and maintained for 8 days at 38.5°C in a humidified atmosphere with 5% CO2 in air. Cleavage, blastocysts production and hatching rates were evaluated at Days 3, 7 and 8, respectively. Data were submitted to ANOVA for parametric data and Wilcoxon test for non-parametric variables using the SAS software (SAS Institute Inc., Cary, NC, USA). Significance level was set at 5%. Cleavage rates of the control (81.05%) and G5 (71.85%) were higher compared with G10 (19.64%; P &lt; 0.0001). Blastocyst production was lower in G5 (12.18%) compared with control (30.35%), and the addition of 10μg mL−1 of free gossypol (G10) completely inhibited embryo development (0%; P &lt; 0.0001). As for the percentage of hatched blastocysts, the control (66.75%) had greater values compared with G5 (34.52%; P &lt; 0.0001). Thus, the addition of 5 and 10μg mL−1 of free gossypol are extremely hazardous for in vitro bovine embryo development. Whether these deleterious effects take place in a similar fashion during in vivo embryo production remains to be investigated.

133 In Vitro Embryo Production from Oocytes Collected from Non-Supertimulated Wood Bison (Bison bison athabascae) Following Maturation In Vitro Using Portable Incubators

2018 ◽  
Vol 30 (1) ◽  
pp. 206
Author(s):  
M. P. Cervantes ◽  
G. P. Adams ◽  
M. Anzar ◽  
J. M. Palomino ◽  
G. F. Mastromonaco
Keyword(s):  
Embryo Development ◽  
Transvaginal Ultrasound ◽  
Calf Serum ◽  
Cumulus Cells ◽  
Bison Bison ◽  
Embryo Production ◽  
Commercial Medium ◽  
Wood Bison ◽  
In Vitro Embryo Production

This study was done to determine the feasibility of in vitro embryo production in wood bison during the anovulatory season, without ovarian superstimulation or follicle wave synchronization, to simulate collection conditions in a wild or field setting. The experiment provided the opportunity to compare embryo development using 2 different maturation media and incubator systems. The cumulus-oocyte complexes (COC) were collected by transvaginal ultrasound-guided follicle aspiration during May from non-superstimulated bison. Compact COC were allocated to 2 groups and matured in standard maturation medium using a portable gassed incubator, or in commercial medium using a portable non-gassed incubator. In the former (Standard), the COC were placed in a round-bottomed tube containing TCM-199 medium with 5% calf serum, 5 μg mL−1 LH, 0.5 μg mL−1 FSH, and 0.05 μg mL−1 gentamicin, and the tube was placed in a portable incubator with 5% CO2. In the latter (Commercial), COC were placed in a round-bottom tube containing the commercial medium (Boviteq, Saint-Hyacinthe, QC, Canada), and placed in a portable incubator without CO2. After 24 h of maturation, oocytes were fertilized in vitro (Day 0) in Brackett-Oliphant medium at 38.5°C in a conventional incubator with 5% CO2 humidified atmosphere. Presumptive zygotes were cultured in CR1aa plus 5% calf serum, at 38.5°C and in 5% CO2, 5% O2, and 90% N2 and high humidity. Cleavage was recorded on Day 3 and embryo development was recorded on Day 7. Cleavage and transferable embryo rates (calculated from the total number of oocytes submitted to IVF) were compared between groups by chi-squared test. No difference in cleavage rates was observed between Standard and Commercial treatment groups [68.1 (32/47) v. 79.2% (57/72), respectively; P = 0.25], nor in morula plus blastocyst rates on Day 7 (36.2 v. 45.8%, respectively; P = 0.39). However, the rate of transferable embryos (grade 1 and grade 2) on Day 7 was higher in the Commercial group (38.9 v. 12.8%; P < 0.01). Of the COC in the Commercial group, a higher number of morula plus blastocyst were observed to be compact good COC (>3 layers of cumulus cells) than compact regular COC (1-3 layers of cumulus cells) (66.7 v. 31.0% respectively; P < 0.05), along with a higher number of transferable embryos on Day 7 (60.0 v. 23.8%, respectively; P < 0.05). In conclusion, wood bison oocytes collected during the anovulatory season from non-superstimulated, non-synchronized bison and matured in vitro using portable incubators were competent to develop to the morula and blastocyst stages following IVF and culture. These results are important for future plans that require transporting oocytes from remote collection sites to the IVF laboratory, particularly with respect to the effectiveness of commercial maturation media which does not require CO2 supplementation. Research was supported by the Natural Sciences and Engineering Research Council of Canada.

scholarly journals Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production

2014 ◽  
Vol 94 (1) ◽  
pp. 35-42 ◽  
Author(s):  
L. L. Vianna ◽  
J. Pradieé ◽  
E. C. S. Santos ◽  
A. O. Gonçalves ◽  
L. F. M. Pfeifer ◽  
...  
Keyword(s):  
Sex Ratio ◽  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Sperm Selection ◽  
Selection Methods ◽  
Embryo Production ◽  
Embryo Survival ◽  
In Vitro Embryo Production

Vianna, L. L., Pradieé, J., Santos, E. C. S., Gonçalves, A. O., Pfeifer, L. F. M., Rheingantz, M. G. T., Dode, M. A. N., Vieira, A. D., Lima, V. F. H., Correa, M. N. and Pegoraro, L. M. C. 2014. Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production. Can. J. Anim. Sci. 94: 35–42. The objective of this study was to evaluate alternatives in small volumes to conventional gradient of Percoll® on semen quality, in vitro embryo production, sex ratio and embryo survival after vitrification. Thawed semen was randomly allocated to one of four density gradient selection methods: (1) conventional Percoll® (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), and (4) MiniOptiprep (MO). Sperm kinetics and quality were evaluated. Use of P, MP and MI gradients did not affect sperm motility (P>0.05). However, there was a decrease in total and progressive sperm motility in MO (70.8 and 51.3% vs. 87.3 and 69.5% for P; 87.3 and 73% for MP; 92.3 and 78.8% for MI; P<0.05). The MO had lower membrane integrity compared with P, MP and MI (39.7 vs. 70.5, 72.3, 63.8%, respectively, P<0.05). The percentage of blastocysts produced was higher in MI than in MP and MO (21.1 vs. 16.1 and 16.9%, P<0.05) and similar to P (18.4%; P>0.05). Sex ratio and embryo survival after vitrification were similar among groups (P>0.05). Semen selected by Isolate and Optiprep gradient, at the concentrations and small volumes used, demonstrated similar characteristics and in vitro embryo production to conventional Percoll® gradient.

The life span of short-lived plasma cells is partly determined by a block on activation of apoptotic caspases acting in combination with endoplasmic reticulum stress

Blood ◽  
2010 ◽  
Vol 116 (18) ◽  
pp. 3445-3455 ◽  
Author(s):  
Holger W. Auner ◽  
Christine Beham-Schmid ◽  
Niall Dillon ◽  
Pierangela Sabbattini
Keyword(s):  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Caspase 3 ◽  
Plasma Cells ◽  
Myeloma Cell ◽  
Immunoglobulin Secretion ◽  
Effector Caspase ◽  
Induced Apoptosis

Abstract Apoptosis of short-lived plasma cells after a few days of intense immunoglobulin secretion is critical for maintaining a controlled humoral immune response. The mechanisms that regulate this process are poorly understood. Here we report that the key apoptotic caspases, caspase-3 and caspase-9, become resistant to activation by apoptotic stimuli when B cells differentiate into short-lived plasma cells. As a consequence, apoptosis of most short-lived plasma cells in vitro and in vivo is effector caspase-independent. We also show that a triaspartic acid repeat that normally prevents activation of caspase-3 becomes stabilized in short-lived plasma cells and myeloma cell lines. The block on caspase activation occurs before the accumulation of intracellular immunoglobulins and a progressive rise in secretory stress in the endoplasmic reticulum (ER). Plasma cells show increased susceptibility to ER stress–induced apoptosis and activate the ER-associated caspase-12, which is required specifically for nuclear apoptotic events. In nonlymphoid cells that cannot activate effector caspases, programmed cell death is delayed in response to ER stress. These observations suggest that the block on activation of key apoptotic caspases has evolved in short-lived plasma cells to prolong survival under conditions of ER stress resulting from high-level immunoglobulin secretion.

scholarly journals Inhibition of Endoplasmic Reticulum Stress by 4-Phenyl Butyric Acid Presents Therapeutic Effects on Periodontitis: Experimental Studies In Vitro and in Rats

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Yang Feng ◽  
Rong Zhang ◽  
Yi-rong Wang ◽  
Fei Chen ◽  
Qiang Luo ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Butyric Acid ◽  
Alveolar Bone ◽  
Experimental Studies ◽  
Therapeutic Effects ◽  
Periodontal Ligament Stem Cells ◽  
Alizarin Red ◽  
Expression Levels

This study investigated the probable mechanisms of endoplasmic reticulum (ER) stress involved in periodontitis in vitro and in vivo. We isolated periodontal ligament stem cells from periodontitis patients and healthy controls (P-PDLSCs and H-PDLSCs). To further simulate the periodontal microenvironment in patients, lipopolysaccharide (LPS) was used to treat H-PDLSCs. The results showed that periodontitis-related inflammation gave rise to the upregulated expression levels of ER stress representative genes including GRP78, PERK, ATF4, and CHOP. In contrast, the treatment of 4-phenyl butyric acid (4-PBA) remarkably suppressed ER stress and supported cell viability. The increased secretion of proinflammatory factors like TNF-α, IL-1β, and IL-6 and the activation of NF-κB pathway were also attenuated by 4-PBA treatment. Moreover, 4-PBA treatment restored the impaired osteogenic differentiation ability of PDLSCs, as demonstrated by the upregulated expression levels of Runx2 and OCN as well as the enhanced Alizarin red staining. Local administration of 4-PBA could rescue alveolar bone resorption of LPS-induced periodontitis rats. Thus, our findings suggested ER stress might act as a promising therapeutic target against periodontitis.

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