Source: UNIVERSITY OF MISSOURI submitted to NRP
UTERINE GLAND FUNCTION IN RUMINANTS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1009276
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Mar 1, 2016
Project End Date
Feb 28, 2021
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF MISSOURI
(N/A)
COLUMBIA,MO 65211
Performing Department
Animal Sciences
Non Technical Summary
Infertility, pregnancy loss and intrauterine growth retardation are common problems that affect reproductive efficiency, health and development in livestock as well as humans. A large percentage of these problems may be attributed to uterine dysfunction during the critical windows of pregnancy establishment and implantation. Collectively, results from the proposed studies should identify uterine factors affecting embryonic survival, conceptus development and fetal/placental growth. Knowledge gained from the proposed studies will be useful to design management, biotechnology and genetic applications aimed at enhancing production efficiency of animal agriculture.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
30139101050100%
Knowledge Area
301 - Reproductive Performance of Animals;

Subject Of Investigation
3910 - Cross-commodity research--multiple animal species;

Field Of Science
1050 - Developmental biology;
Goals / Objectives
Long-term research goals are to: (1) determine the hormonal, cellular and molecular mechanisms regulating uterine development; (2) determine the role(s) of uterine glands in mammalian pregnancy; (3) identify and determine the cellular and molecular mechanisms regulating expression of genes in the uterine glands; and (4) identify and determine the physiological function(s) of genes expressed in the uterine glands during pregnancy. Accomplishment of these research goals should significantly advance our understanding of mammalian uterine function and provide new knowledge essential to solve reproductive problems that involve endometrial dysfunction, such as infertility, pregnancy loss and intrauterine growth retardation. Research in this proposal will utilize a novel uterine gland knockout (UGKO) ewe model to identify genes expressed in the endometrial glands and proteins present in the uterine lumen of the sheep uterus that affect the capacity of uterine endometrium to support conceptus survival and development. Many of the growth factors, cytokines and proteins produced by the endometrium of sheep are also found in the endometrium of other ruminants as well as humans. The central hypothesis is that substances secreted by the uterine glands are required for conceptus (embryo/fetus and associated placental membranes) survival and growth. In order to test this hypothesis, the differentiation of uterine glands has been epigenetically ablated in neonatal ewes, thereby creating a unique Uterine Gland Knock Out (UGKO) phenotype in adult ewes that does not exist in any other available mammalian model system. Available results from current research findings strongly support the central hypothesis that uterine glands are required for peri-implantation conceptus survival and growth in the ewe. Specific objectives of the research plan are to test the hypotheses that defects in conceptus survival and elongation in UGKO ewes are due to absence of: (1) specific genes encoding secretory proteins and molecular transporters expressed by the uterine glands; and (2) specific proteins and other biological active substances (amino acids, lipids, microvesicles, sugars, etc.) in the uterine lumen that are synthesized and secreted or transported by the endometrial glands.
Project Methods
OBJECTIVE ONE: Functional genomic analysis of endometrial GE gene expression utilizing the UGKO ewe model.Experiments in this objective will test the hypothesis that defects in conceptus survival and development in UGKO ewes are due to absence of specific genes encoding secretory proteins expressed by the uterine glands. The approach will be to use next generation sequencing analyses to profile and compare gene expression in endometrium from UGKO ewes on Day 14 post-mating with that in endometrium from Day 14 pregnant (Px) ewes and Day 14 non-Px control ewes. In addition, GE from endometrium of Day 14 Px ewes will be isolated by laser capture microdissection (LCM) and profiled by RNA sequencing analysis to ascertain which differentially expressed genes are specifically expressed by GE. (Please see the attached proposal for further details)OBJECTIVE TWO: Functional proteomic analysis of the uterine milieuExperiments in this objective will test the hypothesis that defects in conceptus survival and elongation in UGKO ewes are due to absence of specific proteins and other bioactive molecules produced or transported by the uterine glands into the uterine lumen. The approach is to analyze uterine flush proteins and other metabolites from Day 14 Px, non-Px and bred UGKO ewes using high throughput technologies. (Please see the attached proposal for further details)

Progress 10/01/20 to 02/28/21

Outputs
Target Audience:Target audience include scientists, educators, and producers. Efforts included presentation of seminars to deliver science based knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via Eleanore O'Neil who completed a Ph.D. in Animal Sciences and Kimberly Davenport who is a new postdoctoral research fellow working on various aspects of the project. How have the results been disseminated to communities of interest?The results have been published in peer-reviewed scientific journals and also presented at regional, national and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?Continue projects with cattle to identify the function of gland-derived factors on conceptus growth and development for pregnancy success in ruminants.

Impacts
What was accomplished under these goals? Many of these goals were accomplished and published in peer-reviewed scientific journals.

Publications


    Progress 03/01/16 to 02/28/21

    Outputs
    Target Audience:Target audience include scientists, educators, and producers. Efforts included presentation of seminars to deliver science based knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via several Ph.D. students and two postdoctoral research fellows. How have the results been disseminated to communities of interest?The results were published in peer-reviewed scientific journals and also presented at regional, national and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? The project identifiedspecific genes encoding secretory proteins and molecular transporters expressed by the uterine glands; and specific proteins and other biological active substances (amino acids, lipids, microvesicles, sugars, etc.) in the uterine lumen that are synthesized and secreted or transported by the endometrial glands in both sheep and cattle.

    Publications


      Progress 10/01/19 to 09/30/20

      Outputs
      Target Audience:Target audience include scientists, educators, and producers. Efforts included presentation of seminars to deliver science based knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via Eleanore O'Neil who completed a Ph.D. in Animal Sciences and Kimberly Davenport who is a new postdoctoral research fellow working on various aspects of the project. How have the results been disseminated to communities of interest?The results have been published in peer-reviewed scientific journals and also presented at regional, national and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?Continue projects with cattle to identify the function of gland-derived factors on conceptus growht and development for pregnancy success in ruminants.

      Impacts
      What was accomplished under these goals? Many of these goals were accomplished and published in peer-reviewed scientific journals.

      Publications

      • Type: Journal Articles Status: Published Year Published: 2020 Citation: O'Neil EV, Burns GW, Ferreira CR, Spencer TE. Characterization and regulation of extracellular vesicles in the lumen of the ovine uterus . Biol Reprod. 2020 Apr 24;102(5):1020-1032. doi: 10.1093/biolre/ioaa019. PMID: 32055841.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ali A, Stenglein MD, Spencer TE, Bouma GJ, Anthony RV, Winger QA. Trophectoderm-Specific Knockdown of LIN28 Decreases Expression of Genes Necessary for Cell Proliferation and Reduces Elongation of Sheep Conceptus. Int J Mol Sci. 2020 Apr 6;21(7):2549. doi: 10.3390/ijms21072549. PMID: 32268593; PMCID: PMC7177537.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Moraes JGN, Behura SK, Bishop JV, Hansen TR, Geary TW, Spencer TE. Analysis of the uterine lumen in fertility-classified heifers: II. Proteins and metabolites . Biol Reprod. 2020 Mar 13;102(3):571-587. doi: 10.1093/biolre/ioz197. PMID: 31616912; PMCID: PMC7331878.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Pfeiffer CA, Meyer AE, Brooks KE, Chen PR, Milano-Foster J, Spate LD, Benne JA, Cecil RF, Samuel MS, Ciernia LA, Spinka CM, Smith MF, Wells KD, Spencer TE, Prather RS, Geisert RD. Ablation of conceptus PTGS2 expression does not alter early conceptus development and establishment of pregnancy in the pig . Biol Reprod. 2020 Feb 14;102(2):475-488. doi: 10.1093/biolre/ioz192. PMID: 31616930; PMCID: PMC7523696.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Moraes JGN, Behura SK, Geary TW, Spencer TE. Analysis of the uterine lumen in fertility-classified heifers: I. Glucose, prostaglandins, and lipids . Biol Reprod. 2020 Feb 14;102(2):456-474. doi: 10.1093/biolre/ioz191. PMID: 31616913; PMCID: PMC7331873.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ortega MS, Kelleher AM, O'Neil E, Benne J, Cecil R, Spencer TE. NANOG is required to form the epiblast and maintain pluripotency in the bovine embryo. Mol Reprod Dev. 2020 Jan;87(1):152-160. doi: 10.1002/mrd.23304. Epub 2019 Dec 5. PMID: 31803983; PMCID: PMC6983337.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: O'Neil EV, Brooks K, Burns GW, Ortega MS, Denicol AC, Aguiar LH, Pedroza GH, Benne J, Spencer TE. Prostaglandin-endoperoxide synthase 2 is not required for preimplantation ovine conceptus development in sheep. Mol Reprod Dev. 2020 Jan;87(1):142-151. doi: 10.1002/mrd.23300. Epub 2019 Nov 20. PMID: 31746519.
      • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kelleher AM, DeMayo FJ, Spencer TE. Uterine Glands: Developmental Biology and Functional Roles in Pregnancy. Endocr Rev. 2019 Oct 1;40(5):1424-1445. doi: 10.1210/er.2018-00281. PMID: 31074826; PMCID: PMC6749889.
      • Type: Journal Articles Status: Published Year Published: 2019 Citation: S�nchez JM, Mathew DJ, Behura SK, Passaro C, Charpigny G, Butler ST, Spencer TE, Lonergan P. Bovine endometrium responds differentially to age-matched short and long conceptuses . Biol Reprod. 2019 Jul 1;101(1):26-39. doi: 10.1093/biolre/ioz060. PMID: 30977805; PMCID: PMC6614577.
      • Type: Journal Articles Status: Published Year Published: 2021 Citation: O'Neil EV, Spencer TE. Insights into the lipidome and primary metabolome of the uterus from day 14 cyclic and pregnant sheep . Biol Reprod. 2021 Jul 2;105(1):87-99. doi: 10.1093/biolre/ioab053. PMID: 33768235.
      • Type: Journal Articles Status: Published Year Published: 2021 Citation: Geisert RD, Meyer AE, Pfeiffer CA, Johns DN, Lee K, Wells KD, Spencer TE, Prather RS. Gene editing to investigate the role of conceptus factors in the establishment of pregnancy in the pig. Reproduction. 2021 Apr;161(4):R79-R88. doi: 10.1530/REP-20-0604. PMID: 33560267.
      • Type: Journal Articles Status: Published Year Published: 2021 Citation: Chaney HL, Grose LF, Charpigny G, Behura SK, Sheldon IM, Cronin JG, Lonergan P, Spencer TE, Mathew DJ. Conceptus-induced, interferon tau-dependent gene expression in bovine endometrial epithelial and stromal cells . Biol Reprod. 2021 Mar 11;104(3):669-683. doi: 10.1093/biolre/ioaa226. PMID: 33330929.
      • Type: Journal Articles Status: Published Year Published: 2022 Citation: McLean MK, Geary TW, Zezeski AL, Smith MF, Spencer TE, Pohler KG, Reese ST, Perry GA. Impact of preovulatory estradiol concentrations on subsequent luteal function in beef cattle. Syst Biol Reprod Med. 2022 Aug;68(4):286-297. doi: 10.1080/19396368.2022.2038717. Epub 2022 Apr 8. PMID: 35394393.
      • Type: Journal Articles Status: Published Year Published: 2022 Citation: Geisert RD, Johns DN, Pfeiffer CA, Sullivan RM, Lucas CG, Simintiras CA, Redel BK, Wells KD, Spencer TE, Prather RS. Gene editing provides a tool to investigate genes involved in reproduction of pigs. Mol Reprod Dev. 2022 Jun 23. doi: 10.1002/mrd.23620. Epub ahead of print. PMID: 35736243.
      • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Spencer TE, Wells KD, Lee K, Telugu BP, Hansen PJ, Bartol FF, Blomberg L, Schook LB, Dawson H, Lunney JK, Driver JP, Davis TA, Donovan SM, Dilger RN, Saif LJ, Moeser A, McGill JL, Smith G, Ireland JJ. Future of biomedical, agricultural, and biological systems research using domesticated animals. Biol Reprod. 2022 Apr 26;106(4):629-638. doi: 10.1093/biolre/ioac019. PMID: 35094055; PMCID: PMC9189970.
      • Type: Journal Articles Status: Published Year Published: 2022 Citation: Ali A, Iqbal MA, Abbas MW, Bouma GJ, Anthony RV, Spencer TE, Winger QA. Trophectoderm Transcriptome Analysis in LIN28 Knockdown Ovine Conceptuses Suggests Diverse Roles of the LIN28-let-7 Axis in Placental and Fetal Development. Cells. 2022 Apr 5;11(7):1234. doi: 10.3390/cells11071234. PMID: 35406798; PMCID: PMC8997724.
      • Type: Book Chapters Status: Published Year Published: 2021 Citation: Green JA, Geisert RD, Johnson GA, Spencer TE. Implantation and Placentation in Ruminants. Adv Anat Embryol Cell Biol. 2021;234:129-154. doi: 10.1007/978-3-030-77360-1_7. PMID: 34694480.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Recuero S, S�nchez JM, Mateo-Otero Y, Bag�s-Arnal S, McDonald M, Behura SK, Spencer TE, Kenny DA, Yeste M, Lonergan P, Fernandez-Fuertes B. Mating to Intact, but Not Vasectomized, Males Elicits Changes in the Endometrial Transcriptome: Insights From the Bovine Model. Front Cell Dev Biol. 2020 Jul 10;8:547. doi: 10.3389/fcell.2020.00547. PMID: 32766237; PMCID: PMC7381276.
      • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: O'Neil EV, Burns GW, Spencer TE. Extracellular vesicles: Novel regulators of conceptus-uterine interactions? Theriogenology. 2020 Jul 1;150:106-112. doi: 10.1016/j.theriogenology.2020.01.083. Epub 2020 Feb 19. PMID: 32164992; PMCID: PMC8559595.
      • Type: Journal Articles Status: Published Year Published: 2020 Citation: Bag�s-Arnal S, S�nchez JM, Fernandez-Fuertes B, McDonald M, Behura SK, Spencer TE, Fair T, Lonergan P. Location relative to the corpus luteum affects bovine endometrial response to a conceptus. Reproduction. 2020 May;159(5):643-657. doi: 10.1530/REP-19-0464. PMID: 32168470.


      Progress 10/01/18 to 09/30/19

      Outputs
      Target Audience:Target audience includes scientists, educators, and producers. Efforts included presentation of seminars to deliver sciencebased knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via Eleanore O'Neil and Greg Burns who are pursuing a Ph.D. in Animal Sciences and conducting experiments related to the project. How have the results been disseminated to communities of interest?The results have been published in peer-reviewed scientific journals and also presented at regional, national and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?Analysis of lipids and proteins as well as RNA sequencing data will be completed.

      Impacts
      What was accomplished under these goals? Tissues from normal and uterine gland knockout (UGKO) ewes were collected for analysis of lipids and proteins as well as single cell RNA sequencing. The samples are now being analyzed.

      Publications

      • Type: Journal Articles Status: Published Year Published: 2019 Citation: Spencer TE, Kelleher AM, Bartol FF. Development and Function of Uterine Glands in Domestic Animals. Annual Review of Animal Biosciences 2019; 7:125-147; PMID: 30183326, doi: 10.1146/annurev-animal-020518-115321
      • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kelleher AK, Demayo FJ, Spencer TE. Uterine Glands: Developmental Biology and Functional Roles in Pregnancy. Endocrine Reviews 2019; 40:1424-1445. doi.org/10.1210/er.2018-00281


      Progress 10/01/17 to 09/30/18

      Outputs
      Target Audience:Target audience includes scientists, educators, and producers. Efforts included presentation of seminars to deliver science-based knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via Eleanore O'Neil and Greg Burns who arepursuing a Ph.D. in Animal Sciences and conducting experiments related to the project. How have the results been disseminated to communities of interest?The results have been published in peer-reviewed scientific journals and also presented at regional, national and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?Analysis of lipids and proteins as well as RNA sequencing data will be completed.

      Impacts
      What was accomplished under these goals? Tissues from normal and uterine gland knockout (UGKO) ewes were collected for analysis of lipids and proteins as well as single cell RNA sequencing. The samples are now being analyzed.

      Publications

      • Type: Journal Articles Status: Published Year Published: 2018 Citation: Spencer TE, Kelleher AM, Bartol FF. Development and Function of Uterine Glands in Domestic Animals. Annu Rev Anim Biosci. 2018 Sep 5. doi: 10.1146/annurev-animal-020518-115321. [Epub ahead of print]
      • Type: Journal Articles Status: Published Year Published: 2018 Citation: Kelleher AM, Milano-Foster J, Behura SK, Spencer TE. Uterine glands coordinate on-time embryo implantation and impact endometrial decidualization for pregnancy success. Nat Commun. 2018 Jun 22;9(1):2435. doi: 10.1038/s41467-018-04848-8.


      Progress 10/01/16 to 09/30/17

      Outputs
      Target Audience:Target audience include scientists, educators, and producers. Efforts included presentation of seminars to deliver sciencebased knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via Eleanore O'Neil who is pursuing a Ph.D. in Animal Sciences and conducting experiments related to the project. How have the results been disseminated to communities of interest?The results have been published in peer-reviewed scientific journals and also presented at regional, national and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?Analysis of lipids and proteins will be completed and data extended to bovine uteroplacental tissues.

      Impacts
      What was accomplished under these goals? Tissues from normal and UGKO ewes were collected for analysis of lipids and proteins.

      Publications


        Progress 03/01/16 to 09/30/16

        Outputs
        Target Audience:Target audience includes scientists, educators, and producers. Efforts included presentation of seminars to deliver science-based knowledge to stakeholders and experiential learning opportunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training was primarily via Eleanore O'Neil who ispursuing a Ph.D. in Animal Sciences and conducted experiments related to the project. 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?During the past year, approximately 40 UGKO ewes were made using neonatal ewe lambs and another 20 UGKO ewes are being made a present. Those ewe lambs will not reach puberty until later in 2017 and then used for experiments.

        Impacts
        What was accomplished under these goals? During the past year, approximately 40 UGKO ewes were made using neonatal ewe lambs and another 20 UGKO ewes are being made a present. Those ewe lambs will not reach puberty until later in 2017 and then used for experiments.

        Publications