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PubMed1991

Influence of short-term maternal zinc deficiency on the in vitro development of preimplantation mouse embryos.

In this study, we evaluated the use of mouse preimplantation embryos as a model to study zinc deficiency-induced abnormal development. In Experiment 1, the effect of culture medium Zn concentrations on blastocyst development was studied. Preimplantation embryos (2 and 4 cells) obtained from superovulated females developed normally in media containing 0.7-30 microM Zn for up to 72 hr; higher levels of medium Zn resulted in abnormal development. In Experiment 2A, females were fed diets containing 50 (+Zn) or 0.4 (-Zn) micrograms Zn/g (760 vs 6 nmol/g, respectively) from 1 day before to 1 day after mating (3 days total). Preimplantation embryos were removed from the dams and cultured for 72 hr in 0.7 microM Zn medium. Embryos from the -Zn dams were morphologically normal at time zero; however, over the 72-hr period, these embryos tended to develop at a slower rate than controls, although compaction and cavitation frequency were similar. By the end of the 72-hr culture period, embryos from -Zn dams had significantly fewer cells than did embryos from control dams. In Experiment 2B, an extended period of maternal Zn deprivation (6 days) was used to investigate the potential for further impairment of in vitro preimplantation embryo development observed in Experiment 2A. Results from this experiment were consistent with those from Experiment 2A, in addition to providing evidence that the developmental progress of embryos obtained from mice fed Zn-deficient diets for 6 days was significantly impaired. In Experiment 3, the potential for supplemental Zn in culture medium to overcome the impairment in development due to maternal Zn deficiency was investigated. Embryos from female mice subjected to the same dietary regimen described in Experiment 2A were cultured to the blastocyst stage in medium containing Zn at a concentration of either 0.7 or 7.7 microM. Medium Zn supplementation did not improve development of embryos from dams fed Zn-deficient diets. In summary, embryos from mice fed -Zn diets for a 3- or 6-day period encompassing oocyte maturation and fertilization exhibited impaired development in vitro. This impairment was not overcome by medium Zn supplementation.

Animals↗
PubMed1977

Growth and mineral content of cultured chick embryos.

In order to assess mineral uptake by chick embryos cultured outside of the shell, growth and mineral (Ca, Mg, K, Na) composition of control and cultured embryos and total egg contents (embryo plus residual egg contents) were measured from 8 through 21 days of incubation. At day nine and thereafter, cultured embryos contained significantly less Ca and Mg than did control embryos. At day 11 and thereafter, ash weight of cultured embryos was significantly less than that of controls. At 13 days and thereafter, wet weight, dry weight, toe length, K and Na content of cultured embryos were significantly less than in controls. To assess relative mineral uptake of cultured embryos, mineral (Ca, Mg, K, Na) content per gram embryo dry weight was plotted against incubation age. Values for the ratio of Mg, K and Na per gram dry weight were similar for control and cultured embryos. Control values for Ca per dry weight were consistently higher than those for cultured embryos during the period from 9 to 21 days. The data suggest that Ca may not be present in sufficient quantity, even on a relative growth basis, to perform its normal physiological functions in cultured embryos.

Animals↗
PubMed2003

Embryo donation at an Australian university in-vitro fertilisation clinic: issues and outcomes.

OBJECTIVES: To review the choices of couples relinquishing frozen embryos and the outcomes of embryo donation at a major in-vitro fertilisation (IVF) clinic. DESIGN AND SETTING: Retrospective audit of 11.5 years of data (1991-2002) from the Monash University IVF clinic, Melbourne. PARTICIPANTS: Couples who make decisions regarding the fate of their frozen embryos, and recipient couples taking part in embryo adoption. MAIN OUTCOME MEASURES: Couples' choices with regard to the fate of their frozen embryos, and the outcome of donated embryo treatment cycles. RESULTS: Of 1246 couples relinquishing frozen embryos, 1116 (89.5%) opted to discard rather than donate their embryos. Sixty-six per cent of donated embryos survived thawing. From donated-embryo transfer to 50 women in 92 cycles, a 17.4% pregnancy rate per transfer cycle was achieved, and 10 women delivered 11 healthy babies at term. At the time of our audit there were 98 couples on the waiting list to adopt embryos. CONCLUSIONS: It is worth considering how couples can be encouraged to donate rather than discard their surplus frozen embryos. An educational program on relevant legal, social and clinical issues may facilitate this.

Adult↗
PubMed1988

Lethality and bone alterations in chicken embryos and newly hatched chickens given bone-active agents.

Studies were undertaken to assess the chicken embryo and newly hatched chicken as models for studying the effects of bone-active agents. Initially, 1,25-dihydroxycholecaliferol (1,25[OH]2D3), sodium fluoride (NaF), parathyroid extract, epidermal growth factor, and prostaglandin E2, were tested for lethality over a broad dose range. One or 3 injections of 1,25(OH)2D3 into the yolk sac of chicken embryos resulted in death of embryos given greater than or equal to 0.1 ng/injection, whereas 0.01 ng was tolerated by the embryos. Administering 1,25(OH)2D3 intraperitoneally to newly hatched chickens as a single injection or weekly for 3 weeks resulted in no deaths at doses up to 50 ng. One or 3 IV injections of 800 micrograms of NaF were lethal to embryos, whereas injections of less than or equal to 400 micrograms were tolerated by the embryo. Giving chickens feed and water containing 2.4 g of NaF/kg was lethal, but no deaths occurred when chickens were given feed containing less than or equal to 1.2 g of NaF/kg. Mortality associated with the administration of epidermal growth factor to embryos was inconsistent, in that death occurred in embryos given a single injection of greater than or equal to 250 ng, but no deaths occurred in embryos given 3 injections at similar doses. Parathyroid extract and prostaglandin E2 were not lethal when administered to embryos and chickens in a single-injection or multiple-injection regimen. Overall, lethality in chicken embryos given a particular agent reflected the dose of bone-active agent injected, rather than the number of injections. Three of the bone-active agents were selected to characterize their microscopic bone effects in chicken embryos and chickens.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Cellular and nuclear volume of primitive red blood cells in digynic and diandric triploid and control diploid mouse embryos.

It has long been known that a relationship exists between ploidy and cellular and nuclear volume. This has been observed in amphibia and plants, and more recently in postimplantation tetraploid mouse embryos. We wished to establish whether a similar relationship exists in digynic and diandric triploid mouse embryos, as well as establishing whether the cellular or nuclear volume of primitive nucleated red blood cells of these two classes of triploids were the same. Spontaneously occurring digynic triploid embryos isolated from LT/Sv strain female mice result from the fertilisation of primary oocytes. These embryos develop to the forelimb-bud stage but invariably possess neural tube and cardiac abnormalities. Diandric triploid embryos were also analysed, and were produced experimentally by standard micromanipulatory techniques. They have a relatively normal morphology, and can survive up to the forelimb--bud stage. Primitive red blood cellular and nuclear volume was analysed in serially sectioned digynic and diandric triploid and in developmentally matched diploid control embryos isolated both as littermates of triploid embryos from LT/Sv strain mice, and from other genetically dissimilar diploid controls. The triploid and diploid embryos were analysed between 8-8.5 and 10-10.5 days of gestation, respectively. The cellular and nuclear volumes of the primitive red blood cells in the digynic and diandric triploid embryos were not significantly different, though they were significantly greater than comparable measurements for diploid embryos. We have therefore confirmed that a predictable relationship exists between cellular and nuclear volume and ploidy in the material analysed. The red blood cellular volume in triploid and diploid embryos increased in a predictable way over time, while their nuclear volume decreased in a predictable way over the same time period. A similar relationship has previously been observed when the nucleated red blood cells of diploid and tetraploid embryos were analysed morphometrically.

Animals↗
PubMed1995

Prolonged development of normal and parthenogenetic postimplantation mouse embryos in vitro.

Parthenogenetic mammalian embryos show reduced placental development and do not develop beyond the 25-somite stage. But non-parthenogenetic embryos in culture, without a functional placenta, can develop to 40 somites or more. We have therefore examined the possibility that parthenogenetic embryos might also show prolonged development in culture. After parthenogenetic activation and diploidization, 23% of CBA and 56-58% of hybrid (CBAxC57BL/6) F1 mouse eggs developed in culture to blastocysts. When transferred to pseudopregnant recipients: 60% of the CBA blastocysts implanted and 26% of these developed to somite stage embryos; 71-72% of the hybrid blastocysts implanted and 11-17% of these developed to somite stage embryos. Improved development of postimplantation embryos explanted into culture at about the 15-20 somite stage was obtained by opening the visceral yolk sac (without exteriorizing the embryo). All the normal (non-parthenogenetic) embryos cultured in this way developed to more than 35 somites and many reached 45-55 somites. Under the same conditions, 11/17 diploid parthenogenetic CBA embryos developed in culture to more than 35 somites and 5 of these reached 45 somites; and 9/28 diploid parthenogenetic (CBAxC57BL/6) F1 embryos developed to 35 somites or more and 5 of these reached 45 somites. The size and protein content of the parthenogenetic embryos after culture was less than that of the normal embryos of equivalent stages. These results raise new possibilities for the analysis of parthenogenesis and genomic imprinting, including studies of the effects of adding to the culture medium specific growth factors and demethylating agents.

Animals↗
PubMed1990

Factors affecting embryo implantation after human in vitro fertilization: a hypothesis.

In the clinical practice of human in vitro fertilization, pregnancy is dependent on embryo implantation. Pregnancy is a function of the number of embryos transferred, with multiple embryos resulting in a higher likelihood of pregnancy. We formulated a mathematic model of embryo implantation. This model describes embryo implantation as dependent on three factors--transfer efficiency, embryo quality, and endometrial receptivity. Application of existing embryo implantation data to this model allows the calculation of the approximate value of each of these factors. On the basis of historic data, data obtained from our in vitro fertilization program, and these theoretic considerations, it is our hypothesis that (1) there is an inherent inefficiency associated with the mechanical transfer of embryos into the uterine cavity, which limits the maximal embryo implantation rate; (2) the quality of embryos produced by controlled ovarian hyperstimulation, follicle aspiration, and in vitro fertilization is very high and approaches that of embryos produced in natural cycles in vivo; and (3) endometrial receptivity is markedly diminished in stimulated cycles and is the current rate-limiting step of pregnancy success of in vitro fertilization.

Embryo Implantation↗
PubMed2005

In vitro and in vivo viability of vitrified and non-vitrified embryos derived from eCG and FSH treatment in rabbit does.

This study aimed to evaluate the in vitro and in vivo viability of vitrified and non-vitrified embryos derived from eCG and FSH treatments in rabbit does. Ninety-six nulliparous does were randomly subjected to consecutive superovulation treatments with eCG (20 IU/kg body weight intramuscularly (i.m.), eCG group), FSH (3 x 0.6 mg/doe at 24 h intervals i.m., FSH group), or without superovulation treatment (control group). Does were artificially inseminated 3 days later and ovulation was induced immediately by hCG (75 IU/doe intravenous). Seven experimental groups were differentiated: first FSH and eCG treatment, second FSH and eCG treatment, eCG-interchanged group (does with previous FSH treatment), FSH-interchanged group (does with previous eCG treatments) and control group. Embryos were collected in vivo by laparoscopy 76-80 h post-insemination in the first and second recovery cycles and post mortem in the third recovery cycles. The ovulation rate was significantly higher in does treated with the first-FSH than in those treated with eCG or in control does (25.2+/-2.0 versus 19.2+/-1.4 to 11.0+/-1.5, and 12.2+/-1.2, first-FSH, first-eCG to second-eCG and control groups, respectively, P < 0.05). Significant differences were observed in the total recovery influenced by ovulation rate in each group (20.3+/-2.2 to 9.4+/-1.2, first-FSH to control groups). Embryo donor rate (donor with at least one normal embryo) was similar among groups with an overall of 75.1%. The number of normal embryos recovered per doe with at least one normal embryo increased significantly in relation to ovulation rate (17.7+/-2.2 to 8.41+/-3, first-FSH and control groups). The vitrification of embryos negatively affected their in vitro development to hatched blastocyst in all groups (88.1% versus 48%, P > 0.05). However, after embryo transfer, this negative effect was only observed in superovulated vitrified embryos (16.8 and 12.8% versus 39.4% total born rate from eCG, FSH and control vitrified groups, P < 0.05). Results indicated that the primary treatments with eCG or FSH increased the number of normal embryos recovered per donor doe, but these embryos are more sensitive to vitrification protocols.

Animals↗
PubMed1993

Relationship between embryo morphology and implantation rate after in vitro fertilization treatment in conception cycles.

OBJECTIVE: To investigate the relationship between the embryo number and morphology in conception cycles and the incidence of multiple pregnancies. DESIGN: The study is based on information received from a computerized data base. SETTING: In Vitro Fertilization Unit, Sapir Medical Center, Kfar Saba, Israel. PATIENTS: A total of 117 consecutive pregnancies resulted from replacement of fresh embryos in our IVF-ET program. MAIN OUTCOME MEASURES: The impact of embryo quality, as assessed by morphological parameters, on the multiple pregnancy rate (PR). RESULTS: Implantation rates positively correlated with the number and the quality of transferred embryos. However, no multiple pregnancies occurred when only two embryos were replaced. There were no multiple pregnancies when only embryos of low quality (grades 1 and 2) were transferred. Furthermore, there was no correlation between the number of replaced embryos of poor quality and the rate of implantation. The multiple PR increased from 10% when a mixture of high and low quality embryos were transferred to 30.76% when only embryos of highest quality were transferred. CONCLUSION: The implantation rate of transferred embryos is directly correlated with the morphological scoring. The results of the study suggest that the number of embryos transferred should be balanced against their morphological quality to reduce the rate of multiple pregnancies.

Adult↗
PubMed2000

[Morphological aspect of embryos obtained after fertilization in vitro for male factor or ICSI].

OBJECTIVE: In order to study the eventual impact on fertilization and embryo characteristics of the microinjection procedure we compared the quality of the embryos obtained by ICSI with those of in vitro fertilization with male factors (MF IVF). MATERIAL AND METHODS: One hundred thirty-four cycles of IVF treatment (group 1) were selected with oligoasthenozoospermia according to WHO criteria with a total number of motile spermatozoa between 500,000 and 1 million. One thousand eighty-eight mature oocytes and 486 embryos were obtained. One hundred forty-three cycles of intracytoplasmic sperm injection (group 2) were performed in couples whose in vitro fertilization was imparticable because of extreme sperm impairment. One thousand one hundred forty-seven mature oocytes were injected and 626 embryos were obtained. RESULTS: In group 1, the pregnancy rate per embryo transfer and the implantation rate were respectively 22.7% and 12.3%. In group 2, the pregnancy rate per embryo transfer was 37.1% and the implantation rate was 17%. The statistical analysis of the embryos obtained in the two different groups did not demonstrate any difference in the distribution of the more regular and less fragmented embryos (group A) and those of the more irregular and fragmented embryos (group B). No statistical difference was demonstrated in the chronology of the division of these embryos (groups 1 and 2). CONCLUSION: The pregnancy rate by cycle and by transfer reported by ICSI (p < 0.003 and p < 0.015 respectively) could be related to a significantly higher mean number of transferred embryos (2.65 vs 2.02) in probable relation with a higher cleavage rate (p < 0.00001).

Adult↗
PubMed2006

Secretion of soluble HLA-G by day 3 human embryos associated with higher pregnancy and implantation rates: assay of culture media using a new ELISA kit.

Identification of chemical markers in human embryo culture media that relate to embryo quality and implantation potential could be an invaluable tool in embryo selection for transfer. Embryonic secretion of soluble human leukocyte antigen G (sHLA-G) has been postulated to be a marker for 'embryonic competence'. This study examines sHLA-G concentrations in day 3 culture media droplets of embryos that were selected for transfer, as well as those being cryopreserved. A total of 712 embryo culture supernatants from 83 patients were assayed. Soluble HLA-G was detected in 306 of the 712 samples tested. In the 58 transfers in which at least one embryo selected for embryo transfer was positive for sHLA-G, the pregnancy rate was 64% (37/58) and the implantation rate per embryo transferred was 38%. In contrast, patients receiving only embryos that were negative for sHLA-G had both a lower pregnancy rate of 36% (9/25; P < 0.05) and a decreased implantation rate (19%; P < 0.05). Expression of sHLA-G was also related to increasing cell stage. Concentration of sHLA-G in embryo culture media was variable and in the low range (3-10 ng/ml). These data suggest an association between implantation potential and embryonic secretion of sHLA-G. The commercial assay kit utilized allowed for same day assessment of sHLA-G secretion. Addition of sHLA-G status to traditional morphological criteria may be useful as a clinical tool for embryo selection.

Adult↗
PubMed2000

Embryos with high implantation potential after intracytoplasmic sperm injection can be recognized by a simple, non-invasive examination of pronuclear morphology.

Embryos are conventionally selected for transfer based on the evaluation of the cleavage speed and extent of blastomere fragmentation. Here we examined whether the predictive value of these criteria, as indicators of the chance of embryo implantation, can be further potentiated by adding previously described criteria reflecting the regularity of pronuclear development. In a group of embryos selected for transfer in 380 fresh embryo transfer cycles according to the conventional criteria, the transfer of only those embryos that developed from zygotes judged normal at the pronuclear stage (pattern 0) gave significantly higher pregnancy (44.8%) and implantation (30.2%) rates compared with the pregnancy (22.1%; P < 0. 05) and implantation rates (11.2%; P < 0.001) for the transfers of only those embryos that developed from zygotes judged abnormal (non-pattern 0). The transfer of only one pattern 0 embryo was sufficient for the optimal chance of pregnancy (no differences in pregnancy rates after transfer of one, two or three pattern 0 embryos), whereas the transfer of two pattern 0 embryos mostly resulted in a twin pregnancy. The inclusion of the criteria based on pronuclear morphology can thus lead to the application of a single embryo transfer policy and optimize the selection of embryos for transfer and cryopreservation.

Cell Nucleus↗
PubMed2001

Elective single embryo transfer: the value of cryopreservation.

In-vitro fertilization is associated with a high rate of multiple pregnancies, a consequence of the number of embryos transferred. There is a challenge in avoiding even twin pregnancies in assisted reproduction, and this can be accomplished with elective single embryo transfer and a good cryopreservation programme. In our follow-up study, we analysed all our elective single embryo transfers during 1998-1999. In all these cycles at least one embryo was frozen. A total of 127 elective single embryo transfers were performed with a clinical pregnancy rate of 38.6%. The highest implantation rate was obtained with four-cell embryos with <10% fragmentation (39.8%). Thirty-four patients have delivered (26.8%), one of these being a monozygotic pregnancy. In total 129 frozen-thawed cycles have been achieved in 83 patients. One frozen-thawed embryo has been transferred in 46 cycles with a clinical pregnancy rate of 17.4%, and two embryos have been transferred in 83 cycles, with a clinical pregnancy rate of 37.3%. Up until now, 66 of 125 patients in our single embryo transfer programme have delivered or have on-going pregnancies, and 77 still have embryos frozen. The cumulative delivery rate per oocyte retrieval is 52.8% and the twin rate 7.6%. We conclude that elective single embryo transfer with a good cryopreservation programme results in very acceptable pregnancy rates with a low risk of twins. This is a cost-effective practice that substantially reduces all risks associated with multiple pregnancies and lowers the cost per delivery.

Cost-Benefit Analysis↗
PubMed2003

Study of low-temperature (4 degrees C) transport of mouse two-cell embryos enclosed in oviducts.

PURPOSE: We examined usefulness of a mouse embryo transportation system for low-temperature transport of oviducts containing mouse two-cell embryos. METHODS: Oviducts containing two-cell mouse embryos were stored at 4 degrees C for 36 h. After that, embryos were collected and cultured for 96 h in Potassium Simplex Optimized Medium (KSOM) medium and evaluated for their rate of development to hatched blastocysts. Embryos were transferred to recipients, and the rate of survival to live young was investigated. The oviducts were then transported from Yamagata to Kumamoto (distance of approx. 1,000 km). At the destination, embryos were implanted in recipient dams and were studied to evaluate their survival to live young. RESULTS: After preservation for 36 h at 4 degrees C, 68.3% of two-cell embryos developed to hatched blastocysts. As a result of transplanting 546 embryos into 25 recipients, 109 normal live young mice were obtained; the rate of development was 20.0%. Results of oviduct transport from Yamagata to Kumamoto indicated that 30.2% of transplanted embryos developed to live young. CONCLUSION: Low-temperature transport of two-cell embryos in oviducts is useful as a method of shipping mouse embryos between institutes.

Animals↗
PubMed2003

[Comparison of vitrification cryopreservation among different stages of mouse embryos].

The cryopreservation of different embryo stages collected from ICR, C57BL/6 and F1 of DBA*C57BL/6 was carried out by using vitrification method. The morphology, in vitro development and birth rates of these embryos were compared after frozen-thawed. The results showed that more than 75% of the morphology from 2-cell embryos to morula stages from different strains was normal, the normal morphology rates of 8-cell embryos being the highest, while those of blastulas being the lowest. The in vitro development rates became higher as the embryos developed. The morphology of in vivo and in vitro fertilized frozen 2-cell embryos showed no difference, but the development rate of in vivo fertilized frozen 2-cell embryos was significantly higher than that of in vitro ones. Embryos that underwent 3 times frozen-thawing remained normal morphology. The pregnant rate and birth rate of frozen 2-cell embryos after embryo transfer were 64% and 40% respectively, but lower than those of fresh 2-cell embryo transfer.

Animals↗
PubMed1998

Probing the morphological developmental path of plant embryos by image tracking

An image analysis and pattern recognition system was applied to track the morphological changes of individual plant somatic embryos during the course of their development into mature embryos. A Fourier descriptor was used to transform the morphological information into quantitative features (Fourier features), which are amenable to mathematical and statistical analysis. At a given time point, the status of each developing embryo is represented by a point in the multidimension feature space spanned by these Fourier features. Connecting each point representing the individual embryos over time gives a trajectory which depicts the embryos' developmental "path" or history. Large variations in embryo development were observed, which is consistent with the population heterogeneity seen in batch embryo cultures. The rate at which each embryo progresses in the feature space was measured by a developmental vector. For embryos in a given class or developmental stage, the magnitude of the developmental vector exhibited a wide distribution. The results revealed that embryos with a higher developmental rate during the early stage of development had a higher chance of reaching the mature stage in a relatively short time. This single embryo tracking method is potentially a valuable tool in developing a correlation between the embryo's morphological features during the early stage of development and its eventual developmental fate. Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗
PubMed1998

Micronuclei and developmental abnormalities in 4-day mouse embryos after paternal treatment with acrylamide.

The developmental consequences of paternal exposure to acrylamide (50 mg/kg i.p. for 5 days) were assessed in preimplantation embryos. There was a significant increase in the proportion of morphologically abnormal embryos after postmeiotic treatment during spermatogenesis (88.7% vs. 14.8% in control). Abnormal embryos had an average of 1.8 +/- 3.5 cells and > 80% had at least one fragmented nucleus. In addition, morphologically normal embryos were significantly delayed (34.3 +/- 12.8 cells per embryo vs. 57.6 +/- 15.7 in control, P < 0.001). Acrylamide caused 10- and 20-fold increases in frequencies of cells with micronuclei (MN) in morphologically normal and abnormal embryos, respectively (41 and 93 MN per 1,000 cells). Both centromere-negative (MN-) and centromere-positive (MN+) were induced. Nuclei of abnormal embryos were significantly larger (900 microm2 vs. 250 microm2) than controls. In addition, MN of abnormal embryos were larger than those of normal embryos (21.2 microm2 vs. 6.5 microm2, P < 0.01). Among control embryos, MN+ were significantly larger than MN- (P < 0.05). These findings suggest that the preimplantation embryo is a sensitive indicator of paternally transmitted effects on early development. Multiple mechanisms appear to be involved, including cytogenetic damage, proliferation arrest/delay, and fertilization failure. Future studies are needed to establish how induced cytological defects in preimplantation embryos contribute to birth defects and other postimplantation abnormalities.

Abnormalities, Drug-Induced↗
PubMed1996

Effect of heat stress on development in vitro and in vivo and on synthesis of heat shock proteins in porcine embryos.

The present study was conducted (1) to examine the effect of an acute increase in ambient temperature on the development of porcine day 6 embryos in culture and after transfer to recipient gilts, and (2) to analyze intracellular production of heat shock proteins (hsps). The viability of porcine day 6 embryos following a temporary acute elevation in ambient temperature (at 42 degrees-45.5 degrees C and for 10-180 min) was examined. Synthesis of 70 kDa hsp (hsp70) and 90 kDa hsp (hsp90) was determined by SDS-PAGE and Western blot analysis in porcine day 6 embryos subjected to heat stresses. Nonheat-stressed embryos were considered as control. Significantly higher numbers of viable nuclei were observed in treatment groups of 42 degrees C-10 min (236.6 +/- 71.4; P < 0.05) and 43 degrees C-30 min (276.8 +/- 89.4; P < 0.005) compared to control (173.9 +/- 53.9). The 42 degrees C-180 min group (158.0 +/- 27.1 microns) had a greater increase in diameter after 24 hr in culture following heat stress compared to control (82.5 +/- 47.3 microns), while heat stress with 43 degrees C for > or = 60 min, 44 degrees-44.5 degrees C for > or = 30 min, or 45 degrees-45.5 degrees C for > or = 10 min impaired their survival, as assessed by differences in number of viable nuclei. The embryos subjected to heat stresses under the conditions of 42 degrees C-180 min, 43 degrees C-10 min, 43 degrees C-30 min, 44 degrees C-10 min, or 45 degrees C-10 min developed to normal piglets after transfer to recipient gilts. Overall pregnancy rate was 75% (6/8), and farrowing rate 62.5% (5/8). Of heat-stressed embryos transferred, 59% (36/61) developed to normal piglets. Heat-stress conditions of 42 degrees C for 180 min, 43 degrees C for 30 min, 44 degrees C for 10 min, and 45 degrees C for 10 min were determined as critical with respect to the in vitro and in vivo survival of porcine embryos. Porcine day 6 embryos constitutively synthesized hsp70 even without heat stress, while hsp90 was detected only at trace level. Neither hsp70 nor hsp90 levels increased in the embryos subjected to heat stresses. In conclusion, porcine day 6 embryos could continue to develop in vivo or during in vitro culture after exposure to acute and temporary rise in temperature. However, no increase of hsp70 and hsp90 was observed in the heat-stressed porcine embryos, while hsp70 was detected in the nonheat-stressed porcine embryos. The precise mechanism of the thermotolerance was unclear.

Animals↗