Progress 05/15/20 to 04/11/24
Outputs
Target Audience:reearch scientists, dairy and beef nutrition consultants, veterinarians, and producers Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Three graduate students participated in the project. Lane Haimon will receive her PhD in May 2024 and publish 2-3 papers from the grant. Quinn Hoorn will receivean MS degree in May 2024. Masroor Sagheer is currently working on the PhD (expected graduation is December 2025) and will publish one paper from the grant. How have the results been disseminated to communities of interest?the PI has made presentations about the research to the following conferences focused on nutritional consultants and dairy producers: Methyl donors and epigenetic regulation of the early embryo. Southwestern Nutrition, Conference, Chandler, AZ, February 9, 2022. Methyl donors and epigenetic regulation of the early embryo. Tristate Dairy Nutrition Conference, Fort Wayne, IN, April 11, 2022. Methyl donors and epigenetic regulation of the early embryo. Florida Ruminant Nutrition Conference, Gainesville, FL, May 9, 2022. Dialing in on the role of methionine in enhancing reproductive performance, fetal programming, and the consequences for lifetime performance: can we use methyl donors to affect postnatal phenotype. Adisseo VIP Seminar, My 22-24, 2023, Fort Worth Texas. In addition, the presentation at Fort Wayne was broadcast through the Real Science Podcast sponsored by Balchem. The PI has also presented results of the research to scientists by giving seminars as follows: University of California-Davis: Adventures in developmental programming during the preimplantation period in the cow. University of California Davis, College of Biological Sciences Storer Lectureship, November 18, 2021. Impact of the life experiences of a bovine embryo from the zygote to blastocyst stage of development on its subsequent fate. University of Missouri Division of Animal Sciences, Inaugural R.M. Roberts Lecture, August 19, 2022. Some challenges and unrealized opportunities towards widespread use of the in vitro produced embryo in cattle production. Eric Lamming Memorial Lecture, International Ruminant Reproduction Symposium, Galway, Ireland, May 28 to June 1, 2023.65. From zygote to blastocyst: setting the stage for subsequent development. Gilbert S. Greenwald Symposium on Reproductive and Developmental Sciences, Kansas University Medical Center, Kansas City, KS, November 1-4 2023. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
In Experiment 1, we aimed to confirm, and expand on previous findings, how choline impacts embryos from the first 7 days of development to affect postnatal phenotype. Bos indicus embryos were cultured in a choline-free medium or medium supplemented with 1.8 mM choline. Blastocyst-stage embryos were transferred into crossbred recipients. Once born, calves were evaluated at birth, 94 d, 178 d and at weaning (average age=239 d). Following weaning, all calves were enrolled into a feed efficiency trial before being separated by sex, with males being slaughtered at approximately 580 d of age and females followed until their first pregnancy check. Results confirm that exposure of 1.8 mM choline chloride during the first 7 d of development alters postnatal characteristics of the resultant calves. Calves of both sexes from choline-treated embryos were consistently heavier through weaning. There was sex-dependent alterations in DNA methylation in whole blood due to choline treatment. After weaning, feed efficiency was affected by an interaction with sex, with choline calves being more efficient for males and less efficient for females. Calves from choline-treated embryos were heavier, or tended to be heavier, than calves from control embryos. Carcass weight was heavier for choline calves and the cross-sectional area of the Longissumus dorsi muscle was increased by choline. Few females became pregnant during the experiment although numerically more choline females were pregnant than control females. Results confirm that exposure of the preimplantation embryo to 1.8 mM choline altered phenotypes and epigenetic landscape of the animal after birth including up to two years after exposure. This study highlights the importance and influence of the first few days of development on phenotype throughout the entire life cycle of an animal. A concentration of 1.8 mM choline is much higher than that found in blood. In the second experiment, it was hypothesized here that choline exerts similar effects on the developmental program of the embryo when added at concentrations similar to those in peripheral blood. Oocytes were collected via ovum pick up and embryos were produced in vitro Embryos were cultured for 7 days in medium with 4 mMcholine chloride, or, as a vehicle control, with an additional 4 mMsodium chloride. Grade 1 blastocysts were transferred into recipients and pregnancy was diagnosed at approximately 28 days of gestation. Subsequent calves were weighed at birth and at weaning. Addition of choline to culture medium did not affect the proportion of embryos that became blastocysts or the proportion of transferred blastocysts that produced a diagnosed pregnancy. Birth weight was unaffected by treatment but calves derived from choline-treated embryos were heavier at time of weaning and gained more per day from birth until weaning than calves derived from embryos treated with vehicle. Results demonstrate that choline can act on the preimplantation embryo at a physiologically-relevant concentration to alter postnatal phenotype. Observations are further evidence for the importance of the first days of embryonic development for the phenotype of the resulting calf. The goal of the third experiment was to evaluate whether feeding rumen-protected choline (RPC) during the peri-conceptional period also programs fetal development. A total of 211 postpartum first service, suckled beef cows were blocked by breed and body weight and assigned randomly to RPC or vehicle group. Cows were synchronized with a 7-day Cosynch-CIDR protocol for timed artificial insemination (TAI) and individually fed 60 g RPC in 454 g ground corn gluten or 454 g ground corn gluten (vehicle) through the Super SmartFeed® machine (C-Lock, Rapid City, SD, USA) from one day before until 7 days after TAI. Pregnancy diagnosis was performed with ultrasonography at days 28 and 42 post-TAI, and calves were followed until weaning. There was no effect of RPC supplementation on pregnancy rate and pregnancy loss. The mean gestation length did not differ between vehicle and RPC . Similarly, RPC did not affect calf birth weight. At an average age of 118 days (range 75-150 days; age included in the statistical model), vehicle calves were heavier than RPC . There was no difference in withers height, so the weight/height ratio was greater for vehicle than RPC. Paired testes weight measured at castration at an average age of 118 days (range 75-150 days) was similar between vehicle and RPC . Differences in body weight persisted at weaning (248 days, range 205-280 days) with vehicle being heavier than RPC. Vehicle calves also tendedto be taller at weaningbut weight/height ratio was greaterfor vehicle. In conclusion, supplementation of RPC during the periconceptional period programmed development to alter calf phenotype in the postnatal period. The net result, reduced body weight, was the opposite of the phenotype caused by addition of choline to culture medium. Thus, choline administered to cows as RPC exerts different actions on the embryo than choline provided to embryo culture medium. The reasons for these differences need to be evaluated.
Publications
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Progress 05/15/22 to 05/14/23
Outputs
Target Audience:Scientists, nutrition consultants, veterinarians and farmers Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Three graduate students participated in the project. Lane Haimon will receive her PhD in May 2024 and publish 2-3 papers from the grant. Quinn Hoorn will receivean MS degree in May 2024. Masroor Sagheer is currently working on the PhD (expected graduation is December 2025) and will publish one paper from the grant. How have the results been disseminated to communities of interest?the PI has made presentations about the research to the following conferences focused on nutritional consultants and dairy producers: Methyl donors and epigenetic regulation of the early embryo. Florida Ruminant Nutrition Conference, Gainesville, FL, May 9, 2022. Dialing in on the role of methionine in enhancing reproductive performance, fetal programming, and the consequences for lifetime performance: can we use methyl donors to affect postnatal phenotype. Adisseo VIP Seminar, My 22-24, 2023, Fort Worth Texas. In addition, the presentation at Fort Wayne was broadcast through the Real Science Podcast sponsored by Balchem. The PI has also presented results of the research to scientists by giving seminars as follows: Impact of the life experiences of a bovine embryo from the zygote to blastocyst stage of development on its subsequent fate. University of Missouri Division of Animal Sciences, Inaugural R.M. Roberts Lecture, August 19, 2022. Some challenges and unrealized opportunities towards widespread use of the in vitro produced embryo in cattle production. Eric Lamming Memorial Lecture, International Ruminant Reproduction Symposium, Galway, Ireland, May 28 to June 1, 2023.65. From zygote to blastocyst: setting the stage for subsequent development. Gilbert S. Greenwald Symposium on Reproductive and Developmental Sciences, Kansas University Medical Center, Kansas City, KS, November 1-4 2023. What do you plan to do during the next reporting period to accomplish the goals?submit all papers for the grant
Impacts
What was accomplished under these goals?
In Experiment 1, we aimed to confirm, and expand on previous findings, how choline impacts embryos from the first 7 days of development to affect postnatal phenotype.Bos indicusembryos were cultured in a choline-free medium or medium supplemented with 1.8 mM choline. Blastocyst-stage embryos were transferred into crossbred recipients. Once born, calves were evaluated at birth, 94 d, 178 d and at weaning (average age=239 d). Following weaning, all calves were enrolled into a feed efficiency trial before being separated by sex, with males being slaughtered at approximately 580 d of age and females followed until their first pregnancy check. Results confirm that exposure of 1.8 mM choline chloride during the first 7 d of development alters postnatal characteristics of the resultant calves. Calves of both sexes from choline-treated embryos were consistently heavier through weaning. There was sex-dependent alterations in DNA methylation in whole blood due to choline treatment. After weaning, feed efficiency was affected by an interaction with sex, with choline calves being more efficient for males and less efficient for females. Calves from choline-treated embryos were heavier, or tended to be heavier, than calves from control embryos. Carcass weight was heavier for choline calves and the cross-sectional area of theLongissumus dorsimuscle was increased by choline. Few females became pregnant during the experiment although numerically more choline females were pregnant than control females. Results confirm that exposure of the preimplantation embryo to 1.8 mM choline altered phenotypes and epigenetic landscape of the animal after birth including up to two years after exposure. This study highlights the importance and influence of the first few days of development on phenotype throughout the entire life cycle of an animal. A concentration of 1.8 mM choline is much higher than that found in blood. In the second experiment, it was hypothesized here that choline exerts similar effects on the developmental program of the embryo when added at concentrations similar to those in peripheral blood. Oocytes were collected via ovum pick up and embryos were producedin vitroEmbryos were cultured for 7 days in medium with 4 mMcholine chloride, or, as a vehicle control, with an additional 4 mMsodium chloride. Grade 1 blastocysts were transferred into recipients and pregnancy was diagnosed at approximately 28 days of gestation. Subsequent calves were weighed at birth and at weaning. Addition of choline to culture medium did not affect the proportion of embryos that became blastocysts or the proportion of transferred blastocysts that produced a diagnosed pregnancy. Birth weight was unaffected by treatment but calves derived from choline-treated embryos were heavier at time of weaning and gained more per day from birth until weaning than calves derived from embryos treated with vehicle. Results demonstrate that choline can act on the preimplantation embryo at a physiologically-relevant concentration to alter postnatal phenotype. Observations are further evidence for the importance of the first days of embryonic development for the phenotype of the resulting calf. The goal of the third experiment was to evaluate whether feeding rumen-protected choline (RPC) during the peri-conceptional period also programs fetal development. A total of 211 postpartum first service, suckled beef cows were blocked by breed and body weight and assigned randomly to RPC or vehicle group. Cows were synchronized with a 7-day Cosynch-CIDR protocol for timed artificial insemination (TAI) and individually fed 60 g RPC in 454 g ground corn gluten or 454 g ground corn gluten (vehicle) through the Super SmartFeed® machine (C-Lock, Rapid City, SD, USA) from one day before until 7 days after TAI. Pregnancy diagnosis was performed with ultrasonography at days 28 and 42 post-TAI, and calves were followed until weaning. There was no effect of RPC supplementation on pregnancy rate and pregnancy loss. The mean gestation length did not differ between vehicle and RPC . Similarly, RPC did not affect calf birth weight. At an average age of 118 days (range 75-150 days; age included in the statistical model), vehicle calves were heavier than RPC . There was no difference in withers height, so the weight/height ratio was greater for vehicle than RPC. Paired testes weight measured at castration at an average age of 118 days (range 75-150 days) was similar between vehicle and RPC . Differences in body weight persisted at weaning (248 days, range 205-280 days) with vehicle being heavier than RPC. Vehicle calves also tendedto be taller at weaningbut weight/height ratio was greaterfor vehicle. In conclusion, supplementation of RPC during the periconceptional period programmed development to alter calf phenotype in the postnatal period. The net result, reduced body weight, was the opposite of the phenotype caused by addition of choline to culture medium. Thus, choline administered to cows as RPC exerts different actions on the embryo than choline provided to embryo culture medium. The reasons for these differences need to be evaluated.
Publications
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Progress 05/15/21 to 05/14/22
Outputs
Target Audience:Choline is a micronutrient that plays important roles in DNA methylation (through oxidation to betaine), lipid metabolism (as a precursor for phosphatidylcholine) and intercellular signaling (through acetylation to form acetylcholine). Typically, choline is absent in defined embryo culture media. It was hypothesized that addition of choline chloride to culture medium for the first 7 days of development would alter embryonic function in a manner that programs subsequent fetal development. Bovine embryos were produced by in vitro fertilization using oocytes from a total of 20 Brahman donors and one of three Brahman sires. Donors were randomly assigned to produce embryos cultured with either 1.8 mM choline chloride (equivalent to the total amount of choline metabolites in blood in postpartum cows) or vehicle. Choline was present in culture medium through day 7 of development. Blastocysts were transferred to suckled beef cattle recipients. There was no effect of choline on pregnancy rate at day 28 or 56 or on calving rate. The fraction and percent of embryos resulting in a live calf were 27/65 (41.5%) for choline and 26/66 (39.4%) for vehicle. There was no difference in gestation length between choline (292.4 + 1.0 days) and vehicle (291.3 + 1.2 days). Birth weight, however, was greater (P=0.0443) for calves derived from choline-treated embryos (37.4 +/- 0.8 kg) than for calves from control embryos (34.8 + 0.9 kg). Results indicate that provision of choline chloride to embryo culture medium can alter the developmental program of the embryo to result in calves of larger weight at birth. Further investigation will be performed to evaluate if effects seen at birth are sustained later in the postnatal period Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Two graduate students, Lane Haimon and Quinn Hoorn, have participated in the project and a third student, Masroor Sagheer, has started work on the project for 2022. How have the results been disseminated to communities of interest?One paper has been published. In addition, the PI has made presentations about the research to the following conferences focused on nutritional consultants and dairy producers: Methyl donors and epigenetic regulation of the early embryo. Southwester Nutrition, Conference, Chandler, AZ, February 9, 2022. Methyl donors and epigenetic regulation of the early embryo. Tristate Dairy Nutrition Conference, Fort Wayne, IN, April 11, 2022 In addition, the presentation at Fort Wayne will be broadcast through the Real Science Podcast sponsored by Balchem. The PI hasalso presented results of the research to scientists at the University of California-Davis: Adventures in developmental programming during the preimplantation period in the cow. University of California Davis, College of Biological Sciences Storer Lectureship, November 18, 2021. What do you plan to do during the next reporting period to accomplish the goals?We will continue monitoring the calves produced for both objectives as described in the grant.
Impacts
What was accomplished under these goals?
Both experiments have been initated. Calves for Objective 1 have been born and all matings for Objective 2 have been completed.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Estrada-Cort�s E, Ortiz W, Rabaglino MB, Block J, Rae O, Jannaman EA, Xiao Y, Hansen PJ. Choline acts during preimplantation development of the bovine embryo to program postnatal growth and alter muscle DNA methylation. FASEB J. 2021 Oct;35(10):e21926. doi: 10.1096/fj.202100991R.
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Progress 05/15/20 to 05/14/21
Outputs
Target Audience:The primary audience is the community of scientists involved in animal reproduction. Other audiences are the pharmaceutical industry, veterinary practitioners, livestock advisors, and dairy farmers. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One graduate student, Lane Haimon, did most of the work on the project. She was assisted by two other graduate students, Thiago Amaral and Dani Heredia. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?We will complete collection of blood samples for the embryo transfer projects and start collecting data on the resultant calves. We will also initiate the choline feeding trial (Objective 2) and continue studies on the actions of choline on the characteristics of the blastocyst.
Impacts
What was accomplished under these goals?
To date, we have completed the transfer of embryos for Objective 1 as outlined in the grant. In addition, we initiated a second embryo transfer trial where embryos were treated with a lower concentration of choline (0.004 mM as compared to 1.8 mM in the first transfer experiment). This experiment is being conducted because 1) preliminary data indicate the lower dose of choline has effects on development not seen with the higher dose) and 2) we had an unforeseen opportunity to transfer embryos into an additional herd of cows. We have also completed the cow work needed for the pilot experiment to determine the optimal amounf of choline to feed to beef cows. We will analyze choline concentrations in blood later in the year. Another series of experiments are starting to determine how choline affects characteristics of the embryo. Experiments for year 1 focused on possible actions of acetlycholine, a derivative of choline. There were no effects of choline on competence of embryos to develop to the blastocyst stage.
Publications
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