Reciprocal Embryo Transfer Research Articles

Oocyte cryopreservation and reciprocal in vitro fertilization in a transgender man on long term testosterone gender-affirming hormone therapy: a case report

To report a successful case of oocyte cryopreservation and subsequent invitro fertilization (IVF) in a transgender male receiving continued testosterone gender-affirming hormone therapy, followed by reciprocal embryo transfer (ET). A case report of a rare case of fertility preservation in a transgender man with concomitant use of testosterone therapy for 4 years before and during ovarian stimulation. Private fertility clinic with university affiliation. A 26-year-old transgender man undergoing oocyte cryopreservation before gender-affirming surgery. Fertility preservation using oocyte cryopreservation and IVF with reciprocal fresh ET into a cisfemale partner. Successful oocyte cryopreservation, oocyte thawing, and reciprocal IVF cycle. Oocyte cryopreservation of 29 mature oocytes. Sixteen mature oocytes survived the thaw, and 12 were fertilized with intracytoplasmic sperm injection. A fresh ET of an advanced blastocyst resulted in a clinical pregnancy and live birth. Fertility preservation with oocyte cryopreservation or IVF with embryo cryopreservation is feasible for patients on continued long-term testosterone gender-affirming therapy. Future studies on egg quality and reproductive outcomes are required. Our case report demonstrates a promising outcome in this patient population.

Embryonic factors mediate the maternal age-induced programming of offspring postnatal behavior in mice†.

Advanced maternal age is associated with adverse pregnancy and offspring outcomes, including neurodevelopmental disorders. While age-related oocyte and embryonic abnormalities may underlie this association, the aged maternal uterine environment also plays an important role in offspring development and survival. The aim of this study was to evaluate the contribution of maternal age-related embryonic and uterine factors on pregnancy and offspring behavior, by using a model of reciprocal embryo transfer between old and young female mice. Pregnancies were obtained by transferring embryos collected from either old (9-14months) or young (3-4months) C57BL/6J female mice to either young or old recipients. The results showed that embryos from old and young donors have comparable developmental potential when transferred to young recipients, whereas no pregnancies were obtained by transferring embryos of young females to old recipients. Moreover, the offspring conceived by aged females displayed altered ultrasonic vocalization and learning skills compared to the progeny of young females, even though they were both prenatally and postnatally fostered by young recipients. These results indicate that maternal factors mostly determine the occurrence of age-related pregnancy complications, whereas the long-term effects of maternal aging on the offspring's behavior could be already established at pre-implantation stages and depend on embryonic factors.

First Kiso pony foal produced via transfer of long-distance shipped fresh embryo to Hokkaido native pony.

Japanese native horses, which consists of 8 breeds, are threatened with extinction. Embryo transfer (ET) is used to reproduce endangered animals in various mammalian species. We aimed to perform ET using native ponies from Kiso and Hokkaido as donors and recipients, respectively. ET operation included long-distance transport of non-cryopreserved embryos from Nagano Prefecture to Hokkaido. Embryos were transported 1500 km over 9 h in a container maintained at 22°C. After transferring two embryos to two recipients, one mare delivered a healthy live foal. These results demonstrated that reciprocal ET with long-distance transportation of fresh embryos between the isolated breeds may allow for the proliferation of Japanese native horses.

Effects of fetal and maternal genotype on placentome morphology in sheep

Both maternal and fetal genotypes contribute to conceptus development. The objective was to determine how placentome number, size, and type and fetal weight was influenced after reciprocal embryo transfer in Columbia and Romanov sheep. Reciprocal embryo transfer was conducted between Columbia and Romanov ewes where a single embryo was transferred into Romanov and Columbia recipients [Romanov embryo in a Romanov uterus (RinR, n = 9); Romanov embryo in a Columbia uterus (RinC, n = 7); Columbia embryo in a Columbia uterus (CinC, n = 8); Columbia embryo in a Romanov uterus (CinR, n = 4)]. On day 130 of gestation, fetuses were weighed and placentomes were morphologically typed, weighed, and measured.Regardless of maternal genotype, Romanov fetuses were smaller (P < 0.05) compared to Columbia fetuses. Moreover, CinC fetuses were larger (P < 0.05) than CinR fetuses. There was a tendency (P = 0.12) for a fetal by maternal genotype interaction on total placentome weight, but main effects were significant for fetal genotype (P = 0.04) and maternal genotype (P < 0.01). The number of Type A placentomes was greater than any other types. Type A placentomes had a greater (P < 0.05) contribution to total placentome weight within the Romanov uterus, or when associated with a Romanov fetus, than within the Columbia breed, in which placentome type was evenly distributed. The hypothesis that the Romanov uterus would limit the growth of a Columbia conceptus is accepted; however, the Romanov conceptus did not experience augmented growth when transferred into a Columbia uterus as predicted.

Pregnancy environment, and not preconception, leads to fetal growth restriction and congenital abnormalities associated with diabetes

Maternal diabetes can lead to pregnancy complications and impaired fetal development. The goal of this study was to use a mouse model of reciprocal embryo transfer to distinguish between the preconception and gestational effects of diabetes. To induce diabetes female mice were injected with a single high dose of streptozotocin and 3 weeks thereafter used as oocyte donors for in vitro fertilization (IVF) and as recipients for embryo transfer. Following IVF embryos were cultured to the blastocyst stage in vitro or transferred to diabetic and non-diabetic recipients. Diabetic and non-diabetic females did not differ in regard to the number of oocytes obtained after ovarian stimulation, oocytes ability to become fertilized, and embryo development in vitro. However, diabetic females displayed impaired responsiveness to superovulation. Reciprocal embryo transfer resulted in similar incidence of live fetuses and abortions, and no changes in placental size. However, fetuses carried by diabetic recipients were smaller compared to those carried by non-diabetic recipients, regardless hyperglycemia status of oocyte donors. Congenital abnormalities were observed only among the fetuses carried by diabetic recipients. The findings support that the diabetic status during pregnancy, and not the preconception effect of diabetes on oogenesis, leads to fetal growth restriction and congenital deformities.

378 Awardee Talk - Impact of fetal versus maternal contributions of Bos indicus and Bos taurus genetics on embryonic and fetal development

Abstract To evaluate how the inclusion of Bos indicus genotype influences early fetal development in cattle, a reciprocal embryo transfer approach was used in a completely randomized design with a 2×2×2 factorial arrangement of treatments in order to generate 55 pregnancies (n = 55). Recipient cows were randomly assigned to 1) a diet that met daily energy maintenance requirements (MAINT), or 2) a diet that restricted intake to 70% of the energy maintenance requirements (RESTR). Angus (AN) and Brangus (BN) embryo donors were superovulated and artificially inseminated with female sexed-sorted semen from the same breed. Embryos were then randomly transferred to either AN or BN recipients fed their respective diets for 28d. Recipients remained on the dietary scheme until d91 of gestation, and were then comingled and fed a common diet that met their requirements. Measurements included pregnancy establishment at d28 of gestation, pregnancy-associated glycoproteins (PAG; using 2 commercial [A1 and A2] and 1 in-house assay), and fetal size (CRL). Recipients in the RESTR diet had lower BW and BCS (diet×day; P &amp;lt; 0.01) than MAINT recipients. Energy-restricted AN recipients experienced greater (recipient breed×diet, P &amp;lt; 0.01) pregnancy failure by d28 than the other recipient breed × diet combinations. Restricted recipients that received AN embryos experienced greater pregnancy failure than RESTR recipients receiving BN embryos (embryo breed×diet; P = 0.03). Brangus embryos resulted in greater plasma concentrations of PAG in both A1 (embryo breed×day, P &amp;lt; 0.01) and A2 (embryo breed P &amp;lt; 0.01). Alternatively, recipients that received AN embryos had greater plasma concentrations of PAG for the in-house assay (embryo breed×day; P &amp;lt; 0.01). In addition, fetuses from AN recipients had greater CRL on d91 (breed×day, P &amp;lt; 0.01). In summary, Bos taurus cows experienced greater pregnancy failure when nutrient restricted. Furthermore, fetal size and profile of PAG production during early gestation differed between Bos indicus-influenced and Bos taurus cattle.

Impact of fetal vs. maternal contributions of Bos indicus and Bos taurus genetics on embryonic and fetal development1

To evaluate how the inclusion of Bos indicus genotype influences early fetal development in cattle, a reciprocal embryo transfer approach was used in a completely randomized design with a 2 × 2 × 2 factorial arrangement of treatments to generate 55 pregnancies over 2 consecutive years (n = 55). Recipient cows were randomly assigned to (i) a diet that met daily energy maintenance requirements (MAINT) or (ii) a diet that restricted intake to 70% of the energy maintenance requirements (RESTR). Angus (AN) and Brangus (BN) embryo donors were superovulated and artificially inseminated with female sexed-sorted semen from the same breed. Embryos were then randomly transferred to either AN or BN recipients fed their respective diets for 28 d. Recipients remained on the dietary scheme until day 91 of gestation and were then comingled and fed a common diet that met their energy requirements until calving. Measurements included pregnancy establishment at day 28 of gestation, interferon-stimulated genes (ISG) expression in peripheral blood leukocytes, pregnancy-associated glycoproteins (PAG; using two commercial [A1 and A2] and one in-house assay), and fetal crown-to-rump length (CRL). Recipients in the RESTR diet had lower BWs and BCS (diet × day; P < 0.01) than MAINT recipients. Energy-restricted AN recipients experienced greater (recipient breed × diet, P < 0.01) pregnancy failure by day 28 than the other recipient breed × diet combinations. Restricted recipients that received AN embryos experienced greater pregnancy failure than RESTR recipients receiving BN embryos (embryo breed × diet; P = 0.03). No relevant differences were observed in ISG expression (P > 0.10). Recipients that received BN embryos had greater plasma concentrations of PAG in both A1 (embryo breed × day, P < 0.01) and A2 (embryo breed; P < 0.01). Alternatively, recipients that received AN embryos had greater plasma concentrations of PAG for the in-house assay (embryo breed × day; P < 0.01). In addition, fetuses from AN recipients had greater CRL on day 91 (breed × day, P < 0.01). In summary, Bos taurus cows experienced greater pregnancy failure when nutrient restricted. Furthermore, fetal size and the profile of PAG production during early gestation differed between B. indicus-influenced and B. taurus cattle.

Reproductive failure in mice expressing transgenic follicle-stimulating hormone is not caused by loss of oocyte quality.

Human female reproductive aging features declining ovarian follicle reserve and oocyte quality, and rising levels of circulating follicle-stimulating hormone (FSH). We determined the effects of elevated FSH on oocyte-embryo development in mature mice exhibiting premature infertility caused by progressively rising transgenic human FSH (TgFSH) levels. Oocyte-embryo developmental competence and quality were examined using oocyte maturation and aneuploidy rates, biomarkers of oocyte quality, and reciprocal embryo transfers assessed for implantation and pregnancy. In vitro maturation suggested that TgFSH exposure only hindered oocyte developmental competence in old females, as significantly more oocytes from ≥12-month-old TgFSH females remained at germinal vesicle stage compared with age-matched control oocytes. Aneuploidy rates were equivalent in oocytes from aging TgFSH compared with wildtype females. Cumulus cell expression levels of candidate biomarker Inhba, Egfr, and Rgs2 transcripts were elevated in associated aneuploid vs euploid oocytes from both TgFSH and wildtype females. In vivo, embryos transferred from subfertile 6-month-old TgFSH females to wildtype recipients yielded normal implantation rates and more pups born compared with controls. Transfer of wildtype embryos rescued the fertility of 6-month-old TgFSH-recipient females, although pup birth weight was reduced in TgFSH vs wildtype recipients. Our current findings show that elevated FSH had minimal disruption of either embryo developmental capacity or uterine function when examined in isolation, and the subfertility of TgFSH female mice was not caused by altered oocyte aneuploidy or quality.

Maternal insulin sensitivity in midpregnancy does not determine birth weight after embryo transfer between large and small breed sheep

Embryo transfer of large sheep breed embryos (Suffolk) into small breed ewes (Cheviot) constrains birth size, but the maternal factors influencing fetal growth restriction are unknown. We hypothesized that reciprocal embryo transfer crosses between breeds of divergent size would affect pregnancy-related development of maternal insulin resistance in midgestation, thereby influencing fetal growth. Following superovulation, embryos were surgically collected 6 d postmating and transferred to recipients on the same day. Between- and within-breed transfers were performed. Between 60 and 70 d of pregnancy overnight-fasted ewes underwent hyperinsulinemic-euglycemic clamps for assessment of insulin sensitivity. Maternal insulin sensitivity did not vary with transferred lamb breed. Overall, Cheviot ewes tended to have higher fasting glucose (P = 0.068), fasting insulin (P = 0.052), and steady-state glucose (P = 0.065) concentrations than Suffolk ewes at the stage of pregnancy studied. As expected, transferred between-breed Suffolk lambs were born lighter (P = 0.014), and transferred between-breed Cheviot lambs tended to be heavier at birth (P = 0.056) than respective lambs transferred within breed. Midgestation insulin sensitivity does not appear to be a major factor constraining growth of large breed sheep fetus transferred into smaller breed or a factor in releasing constraint in growth of a small breed fetus within a larger breed ewe. However, as embryo size is already different between transferred groups by 19 d, factors other than maternal gestational insulin resistance may determine fetal growth in this embryo transfer paradigm.

Blastocyst implantation failure relates to impaired translational machinery gene expression

Oocyte quality is a well-established determinant of embryonic fate. However, the molecular participants and biological markers that affect and may predict adequate embryonic development are largely elusive. Our aim was to identify the components of the oocyte molecular machinery that part take in the production of a healthy embryo. For this purpose, we used an animal model, generated by us previously, the oocytes of which do not express Cx43 (Cx43(del/del)). In these mice, oogenesis appears normal, fertilisation does occur, early embryonic development is successful but implantation fails. We used magnetic resonance imaging analysis combined with histological examination to characterise the embryonic developmental incompetence. Reciprocal embryo transfer confirmed that the blastocyst evolved from the Cx43(del/del) oocyte is responsible for the implantation disorder. In order to unveil the genes, the impaired expression of which brings about the development of defective embryos, we carried out a genomic screening of both the oocytes and the resulting blastocysts. This microarray analysis revealed a low expression of Egr1, Rpl21 and Eif4a1 in Cx43(del/del) oocytes and downregulation of Rpl15 and Eif4g2 in the resulting blastocysts. We propose that global deficiencies in genes related to the expression of ribosomal proteins and translation initiation factors in apparently normal oocytes bring about accumulation of defects, which significantly compromise their developmental capacity. The blastocysts resulting from such oocytes, which grow within a confined space until implantation, may be unable to generate enough biological mass to allow their expansion. This information could be implicated to diagnosis and treatment of infertility, particularly to IVF.

The impact of maternal in utero environment on salt‐induced hypertension in the SS rat (1120.4)

There is considerable interest in mechanisms by which environmental factors impact disease phenotypes later in life. We previously demonstrated that SS (Dahl‐salt sensitive) male rats fed a grain‐based diet (G) show attenuated salt‐induced blood pressure increase and renal damage than animals fed a casein‐based protein diet, AIN‐76a (A). We hypothesized that differences in dietary‐induced susceptibility may be driven by the in utero environment of pregnant females fed these different diets. Using SS rats fed the casein‐based (SSA), or a grain‐based (SSG) diets for many generations, we performed reciprocal embryo transfers, weaned the offspring to a 0.4% NaCl AIN‐76A diet for 6 weeks, then switched the rats to a 4.0% NaCl AIN‐76A diet for mean arterial pressure (MAP) and urinary albumin excretion (UAE) measurements. Compared to control male F1 generation rats generated by the transfer of SSA embryos into SSA females (SSA&gt;A), F1 males generated by transfer of SSA embryos into SSG female recipients (SSA&gt;G) demonstrated significantly reduced pressure (MAP, 157±6 mmHg vs. 138±4; n=8‐14/group) and renal damage (UAE, 204±16 mg/day vs. 96±13 mg/day; n=12‐18/group) after 21 days of exposure to a 4.0% NaCl diet. Conversely, reciprocal embryo transfer of SSG embryos into SSA recipients (SSG&gt;A) promoted hypertension (MAP, 166±5 mmHg, n=21) and renal disease (UAE, 187±27 mg/day, n=25) in male offspring. These data support the hypothesis that the maternal in utero environment is a significant driving factor in the hypertensive phenotype of the SS rat. Interestingly, following breeding the of the protected SSA&gt;G F1 animals on the A diet, the disease attenuation was lost in the F2 generation (MAP=152±5 mmHg; Ualb=145±25 mg/day, n=12‐13/group), suggesting that; 1) that the protective effect of the G diet is not transgenerational and 2) that severe salt‐induced hypertension disease phentoypes can be restored to SS rats fed the grain‐based diet by switching the breeders to the pro‐hypertensive AIN‐76A diet.

Matrix metalloproteinase-9 deficiency phenocopies features of preeclampsia and intrauterine growth restriction

The pregnancy complication preeclampsia (PE), which occurs in approximately 3% to 8% of human pregnancies, is characterized by placental pathologies that can lead to significant fetal and maternal morbidity and mortality. Currently, the only known cure is delivery of the placenta. As the etiology of PE remains unknown, it is vital to find models to study this common syndrome. Here we show that matrix metalloproteinase-9 (MMP9) deficiency causes physiological and placental abnormalities in mice, which mimic features of PE. As with the severe cases of this syndrome, which commence early in gestation, MMP9-null mouse embryos exhibit deficiencies in trophoblast differentiation and invasion shortly after implantation, along with intrauterine growth restriction or embryonic death. Reciprocal embryo transfer experiments demonstrated that embryonic MMP9 is a major contributor to normal implantation, but maternal MMP9 also plays a role in embryonic trophoblast development. Pregnant MMP9-null mice bearing null embryos exhibited clinical features of PE as VEGF dysregulation and proteinuria accompanied by preexisting elevated blood pressure and kidney pathology. Thus, our data show that fetal and maternal MMP9 play a role in the development of PE and establish the MMP9-null mice as a much-needed model to study the clinical course of this syndrome.

Preovulatory estradiol and the establishment and maintenance of pregnancy in suckled beef cows1

In postpartum beef cows, GnRH-induced ovulation of small dominant follicles decreased pregnancy rates and increased late embryonic/fetal mortality. In Exp. 1, single ovulation reciprocal embryo transfer (ET) was used to examine the relationship between preovulatory serum concentrations of estradiol at GnRH-induced ovulation in donor and recipient cows and establishment and maintenance of pregnancy. Suckled beef cows (n = 1,164) were administered GnRH (GnRH1, 100 μg) on d -9 (GnRH1), PGF(2α) on d -2, and GnRH2 (GnRH2, 100 μg) on d 0 (CO-Synch protocol) either with (donors; n = 810) or without (recipients; n = 354) AI. Single embryos (n = 394) or oocytes (n = 45) were recovered from the donor cows (d 7; ET) and all live embryos were transferred into recipients. Serum concentration of estradiol at GnRH2 was positively correlated with follicle size at GnRH2 (r = 0.45, P < 0.01) and progesterone at ET (r =0.34, P < 0.01). Donor cows with greater estradiol at GnRH2 were more likely to yield an embryo than an unfertilized oocyte (P < 0.01). Donor and recipient cows were retrospectively divided into 4 groups [low estradiol (<8.4 pg/mL) or high estradiol (≥8.4 pg/mL)] based on serum concentration of estradiol at GnRH2. Pregnancy rate at d 27 for low-low (n = 78), low-high (n = 80), high-low (n = 91), and high-high (n = 101) groups (donor-recipient, respectively) was 45, 65, 43, and 61% respectively (P < 0.02). Because recipient cows with greater estradiol concentration at GnRH2 had greater pregnancy rates in Exp. 1, the objective of Exp. 2 was to evaluate the effect of estradiol supplementation on pregnancy rate. Ovulation was synchronized in suckled beef cows (n = 600) using the CO-Synch protocol with the insertion of a controlled internal drug release (CIDR; intravaginal progesterone supplement) from d -9 until d -2. Approximately one-half of the cows (n = 297) received an injection of estradiol cypionate (ECP; 0.5 mg intramuscularly) 24 h before AI. Compared with the no treatment (Control) cows, ECP treatment increased (P < 0.01) pregnancy rates of cows induced to ovulate smaller dominant follicles (<12.2 mm). In conclusion, GnRH-induced ovulation of small dominant follicles was associated with reduced serum estradiol, fertilization rate (donor cows), and pregnancy establishment (recipient cows). Furthermore, ECP supplementation during the preovulatory period increased pregnancy rates in cows induced to ovulate smaller dominant follicles.

Pregnancy establishment and maintenance in cattle1

A single ovulation, reciprocal embryo transfer study was used to investigate effects of oocyte competence and maternal environment on pregnancy establishment and maintenance in beef cows. Estrous cycles were synchronized in suckled beef cows and embryo donors were inseminated on d 0 (n = 810). Cows were classified on d 0 as having a small (<12.5 mm) or large (≥12.5 mm) ovulatory follicle and randomly chosen as donors or recipients to remove confounding effects of ovulatory follicle size on fertility. Embryos (n = 393) or oocytes (n = 44) were recovered on d 7, and all viable embryos were transferred into recipients (n = 354). All statistical analyses were conducted using the GLM procedure of SAS. Path analysis (with significance set at P < 0.10) was used to examine potential cause-effect relationships among the measured variables. Greater donor cow BW, circulating estradiol concentration at insemination, postpartum interval, and ovulatory follicle size directly increased (P < 0.10) fertilization success. Greater donor cow age was the only factor that directly decreased (P < 0.10) fertilization success. Viability of d-7 embryos was directly inhibited (P < 0.10) by rapid follicular growth rate from d -2 to 0 and heavier BW. Direct beneficial effects to embryo viability were increased serum progesterone concentration on d -2 and ovulatory follicle size. Pregnancy maintenance from d 7 to 27 was enhanced (P < 0.10) by increased serum estradiol concentration on d 0 and progesterone concentration on d 7 in the recipient cow. Increased follicular diameter in the recipient cow on d 0 was detrimental to pregnancy maintenance from d 7 to 27. This manuscript defines the complex interplay and relative contributions of endocrine and physical factors both prior and subsequent to fertilization that influence both oocyte competence and maternal environment and their roles in establishment and maintenance of pregnancy.

Pregnancy rates, prenatal and postnatal survival of offspring, and litter sizes after reciprocal embryo transfer in DBA/2JHd, C3H/HeNCrl and NMRI mice

Success of embryo transfer is often a limiting factor in transgenic procedures and rederivation efforts, and depends on the genetic background of the donor and recipient strains used. Here we show that embryo transfer to DBA/2J females is possible, and present data on pre- and postnatal success rates after reciprocal embryo transfer using the inbred DBA/2J and C3H/HeN, and outbred NMRI strains. The highest embryo yield was achieved in outbred NMRI females, but embryo yields were similar in DBA/2J and C3H/HeN mice following superovulation despite poor estrus cycle synchronization in DBA/2J females. In-strain transfer of DBA/2J blastocysts (transfer of embryos to recipients from the same strain) resulted in pregnancy rates (57.1%) similar to those obtained following in-strain transfer of C3H/HeN (60.0%) and NMRI mice (83.3%), although the prenatal survival rate of blastocysts was low. Moreover, from the pups born only half survived the postnatal period after transfer of DBA/2J and C3H/HeN blastocysts to DBA/2J recipients. These problems were not observed when transferring NMRI-blastocysts to C3H/HeN and DBA/2J mothers. The number of blastocysts transferred also had a positive effect on the success of embryo transfer. In conclusion, C3H/HeN and DBA/2J females can be used as recipients for embryo transfer procedures for certain donor strains like NMRI, as one major determinant seems to be the genetic background of the embryos transferred. We also recommend to increase the number of DBA/2J blastocysts transferred, and to foster the DBA/2J pups to other DBA/2J mothers postnatally for in-strain transfer of DBA/2J mice.

Contributions of the maternal uterine environment and piglet genotype on weaning survivability potential: I. Development of neonatal piglets after reciprocal embryo transfers between Meishan and White crossbred gilts1,2

In commercial pigs, the greatest susceptibility for pre-weaning mortality occurs in low birth-weight piglets. Despite their overall decreased birth weight, Meishan (MS) piglets have decreased pre-weaning mortality rates compared with contemporary Western breeds. The objective of the current study was to determine the contributions of the maternal uterine environment, piglet genotype, and their interaction on the development of neonatal piglets pertaining to pre-weaning survivability using reciprocal embryo transfer between MS and White crossbred (WC) pigs. Twenty-five successful pregnancies were produced from 2 farrowing seasons, generating litters of maternal uterine environment (MUE) by piglet genotype (PigG) combinations; MS × MS (n = 4 litters), MS × WC (n = 7 litters), WC × MS (n = 7 litters), and WC × WC (n = 7 litters). At approximately 24 h of age (Day 1), piglets (n = 173) were weighed and a blood sample was taken. Hematocrit, hemoglobin, glucose, plasma urea nitrogen, albumin, NEFA, lactate, and cortisol were measured in all blood samples. Representative piglets (n = 46) from each litter were harvested and body measurements (i.e., organ weights, tissue glycogen content, and body composition) were determined. Piglet data were analyzed by ANOVA using MIXED model procedures. Both MUE (P < 0.001) and PigG (P < 0.01) affected piglet BW, illustrating that piglets gestated in WC gilts were heavier than piglets gestated in MS gilts, and WC piglets were heavier than MS piglets. Serum albumin concentrations were increased (P < 0.05) in MS piglets compared with WC piglets, indicating greater liver maturity. Significant MUE × PigG interactions were observed for hematocrit and hemoglobin, in which the greatest concentrations were observed in MS piglets gestated in MS and WC gilts, and the lowest concentrations were observed in WC piglets gestated in WC gilts, demonstrating increased oxygen-carrying capability. The percentage of fat and nitrogen, as well as the GE of the body, were greater (P < 0.05) in MS piglets, indicating greater energy stores. Liver, bicep femoris, and LM glycogen concentrations were greater (P < 0.01) in WC piglets compared with MS piglets, demonstrating increased glycogen catabolism in MS piglets. This study demonstrated limited interactions between the maternal uterine environment and piglet genotype on weaning survivability potential, suggesting that the MS piglet is a viable model for pre-weaning survivability.

148 PIGLET GROWTH AND BLOOD COMPONENTS DURING LACTATION FOLLOWING RECIPROCAL EMBRYO TRANSFER BETWEEN MEISHAN AND WHITE CROSSBRED GILTS

Sow productivity has a significant economic impact on the swine industry and is influenced by several factors, including preweaning piglet mortality. In Western breeds, low birth weight piglets exhibit the greatest susceptibility to preweaning mortality. In contrast, Meishan (MS) piglets have decreased birth weights but lower preweaning mortality rates, suggesting that birth weight is not the sole component of preweaning survival. The objective of the current study was to determine the contributions of the maternal and piglet breed and their interactions on piglet growth and blood components pertaining to survivability during lactation following reciprocal embryo transfer between MS and White crossbred (WC) gilts. Twenty-five successful pregnancies were produced by embryo transfer in 2 farrowing seasons that represented all maternal and piglet breed combinations; MS × MS (n = 4 litters), MS × WC (n = 7 litters), WC × MS (n = 7 litters), and WC × WC (n = 7 litters). At Day 1, 10, and weaning (average weaning age = 18 days), piglets (n = 147, 97, and 94, respectively at Day 1, 10, and weaning) were weighed and blood samples were taken. Hematocrit, hemoglobin, glucose, nitrogen, nonesterified fatty acids, albumin, and cortisol were measured in all blood samples. All data were analysed for ANOVA using mixed model procedures. Piglet weights were greater (P &lt; 0.001) throughout lactation in piglets from WC dams regardless of piglet breed. As a result, average daily gains from Days 1 to 10 and weaning were greater (P &lt; 0.05) in piglets from WC dams. There were significant (P &lt; 0.001) maternal × piglet × day interactions for hematocrit and hemoglobin levels in which levels were greatest at Day 1 in MS piglets from WC dams and at Day 10 in MS piglets from MS dams but decreased in WC piglets from WC dams at Day 1. Glucose was greater (P &lt; 0.05) at Day 1 in piglets from WC dams regardless of piglet breed but was greater (P &lt; 0.05) at weaning in WC piglets regardless of maternal breed. Nitrogen was similar at Day 1 for all maternal and piglet breed combinations, but at Day 10 and weaning, nitrogen levels were greater (P &lt; 0.001) in MS piglets regardless of maternal breed. Nonesterified fatty acid was greater throughout lactation in piglets from MS dams irrespective of piglet breed. Albumin was greater (P &lt; 0.05) in MS piglets throughout lactation regardless of maternal breed. Cortisol was not different between the maternal and piglet breed combinations throughout lactation, but cortisol was greater (P &lt; 0.001) at Day 1 compared with Day 10 and weaning. This study demonstrated that piglet growth during lactation was influenced more by maternal breed in favor of WC dams, which supports previous crossbreeding studies. However, blood components pertaining to survivability displayed complex interactions between the piglet and maternal breed, which may signify possible mechanisms for improved preweaning survivability of MS pigs.

Maternal and Genetic Effects on the Acoustic Startle Reflex and its Sensitization in C3H/HeN, DBA/2JHd and NMRI Mice Following Blastocyst Transfer

In the present study, reciprocal embryo transfers were conducted to examine genetic and maternal effects on the baseline and fear-sensitized acoustic startle response (ASR) in the two inbred strains C3H/HeN and DBA/2JHd and the outbred strain NMRI. The largest differences in the ASR were found in untreated strains (effect size 0.6). The transfer procedure per se had a significant effect on the behavior of NMRI mice resulting in a reduction in the baseline, and an increase in the fear-sensitized ASR. In contrast, there were no significant effects of the transfer procedure in the two inbred strains. Autosomal genetic effects had a stronger impact on the amplitude of the ASR (effect sizes 0.5) than sex (effect sizes 0.06) as revealed by reciprocal embryo transfer. Nevertheless, the genetic effects on the fear-sensitized ASR were somewhat more variable and strain-dependent (effect sizes 0.1-0.2). Global maternal effects were detected after embryo transfer into NMRI mothers resulting in a larger reduction of the ASR in the offspring of DBA and NMRI donors than C3H donors (effect sizes 0.1-0.2). An additional fostering procedure was introduced to dissect uterine and postnatal maternal effects in NMRI offspring. Uterine factors changed the baseline ASR of the offspring in direction of the recipient mother strain. Surprisingly, postnatal maternal effects on the ASR were contrary to the behavior of the rearing mother. In conclusion, both genetic and prenatal/postnatal maternal factors persistently influenced the ASR of the offspring, whereas the fear-sensitized ASR was mainly influenced by genetic factors. Our study shows that uterine and postnatal maternal influences deserve more attention when determining the phenotype of genetically engineered mice at least in the first generation following embryo transfer.

Differential effects of embryo transfer and maternal factors on anxiety-related behavior and numbers of neuropeptide Y (NPY) and parvalbumin (PARV) containing neurons in the amygdala of inbred C3H/HeN and DBA/2J mice

The effects of reciprocal embryo transfers were studied on anxiety-related behavior of inbred C3H/HeN and DBA/2J mice on the elevated plus maze (EPM), and related to amygdaloid neuropeptide Y (NPY)- and parvalbumin (PARV)-immunoreactive neurons. Embryo transfer significantly reduced closed arm entries in in-stain-transferred C3H mice, and maternal factors influenced open arm entries only in interaction with genetic background and sex. In DBA/2J-mice, embryo transfer resulted in a reduced number of NPY-immunoreactive (NPY-ir) neurons, while PARV-immunoreactive (PARV-ir) cells were not affected. In C3H/HeN mice, however, in-strain embryo transfer only resulted in a reduction of the number of PARV-immunoreactive neurons. Maternal factors mainly induced changes in the number of NPY-ir neurons in the basolateral complex of the amygdala either directly or in interaction with genetic factors. In summary, in-strain embryo transfer had a minor effect on the behavior of C3H/HeN mice, and a differential influence on the numbers of amygdaloid NPY-ir and PARV-ir neurons of inbred C3H/HeN and DBA/2J mice. Maternal factors had a stronger impact on the numbers of NPY-ir neurons than PARV-ir neurons. The present results indicate that alterations in behavior and amygdala morphology induced by embryo transfer or maternal factors depend on the genetic background of the mouse strains used.

Maternal and Genetic Effects on Anxiety-Related Behavior of C3H/HeN, DBA/2J and NMRI Mice in a Motility-Box Following Blastocyst Transfer

Reciprocal embryo transfers were conducted to examine genetic and maternal effects on the behavior of inbred C3H/HeN and DBA/2J mice, and outbred NMRI mice using a motility-box. The behavioral variables measured were (i) horizontal locomotor activity assessed as the path and time spent during traveling; (ii) vertical activity assessed as the time spent with and numbers of rearings/leanings; (iii) and the time spent in the more anxiogenic central field. The transfer procedure per se resulted in a minor increase in vertical activity of inbred C3H/HeN mice, but had no effect in inbred DBA/2J mice. In contrast, outbred NMRI mice displayed a lower central field activity following embryo transfer indicating a higher anxiety level. Moreover, genetic differences between the mouse strains studied remained stable following embryo transfer for locomotor and vertical activity, but not central field activity depending on the recipient mother strain. Maternal effects were found for (i) vertical activity in the two inbred mouse strains, (ii) all behavioral variables studied in outbred NMRI mice, and (iii) an interaction with gender for the time spent in the anxiogenic central field. An additional fostering procedure revealed that the vertical activity of NMRI mice was modified towards the behavior of the recipient C3H/HeN strain by uterine factors, whereas the postnatal maternal effect of C3H/HeN mothers was the opposite. In summary, the effects of the embryo transfer procedure per se, stability of genetic characteristics following embryo transfer and maternal effects were related to the mouse strains used as donators and recipients, and the behavioral variables studied.