Progress 07/21/16 to 07/14/21
Outputs
Target Audience:The target audience for this research includes agricultural producers who are interested in increasing reproductive efficiency in livestock and others interested in the impact of nursing on development and the importance of milk as a means of continuing maternal/neonatal communication after birth. This research project serves as the basis for training graduate and undergraduate students in hypothesis testing, experimental design, research methodologies and data analysis. These studies also involve additional informal teaching about lactation, maternal programming of development and endocrine disruption. Students also gain the opportunity for experiential learning about pregnancy, parturition and lactation in swine. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research has provided research training opportunities for many students over the five-year period of the award. Students who have conducted hands on research on the farm and at the bench on this project include: Ashley George, Chelsee Holloway, Nina Paranjpe (Graduate students); Dorothy Knipple, Julie Wang, Dana Tsuchida, Yamirka Gonzalez , Katie Fafara, Jenna Lorippo, Erin Miners, Nicole Sadlier, Samantha Weitzman, Nicolina Zappley (Undergraduate students). In addition, Dr. Teh Ho, Research Associate in my laboratory has participated in the animal and laboratory work involved in these research studies. In addition, I teach a course entitled Animal Reproduction each Fall semester. I have provided research training to undergraduates by presenting this research to students majoring in Animal Sciences, Biotechnology and Biology (more than 500 students over 5 years). How have the results been disseminated to communities of interest?The results of this project have been disseminated in abstracts presented at local, national and international scientific meetings by students and staff. In addition, research data has been reported in invited talks given by the PI at a local high school, other universities in the US and at international meetings. Results have also been reported in publications in peer-reviewed journals. 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 goal of these studies is to improve reproductive efficiency in domestic animals by learning how nursing impacts programming of reproductive tract tissues that may ultimately have consequences for reproductive health and performance. A. To determine the effects of age and nursing on the neonatal porcine microRNA transcriptome and the miRNA x mRNA interactome from birth (PND 0) to PND 2 using next generation sequencing. This Objective 1 was completed in 2016 (RNAseq data) and 2017 (miRNAseq data) and published in two manuscripts in Biology of Reproduction. B. To investigate the long-term effects of low vs high colostrum consumption on the uterus at PND 14 and in the adult during the periattachment period in early pregnancy. For Objective 2, weare collaboratingwith Drs. Jeff Vallet, Clay Lents and Jeremy Miles at the USDA Meat Animal Research Center in Clay Center, Nebraska to evaluate the long-term effects of nursing on uterine development at pregnancy day (PxD) 13, the time of implantation in adult pigs.Dr. Vallet has identified low and high colostrum consuming gilts on their day of birth (postnatal day [PND]0) using an immunocrit assay. We have used this pig model system to study the effects of lactocrine deficiency in both the neonate at birth and PND 14 as well as in adult gilts around the time of attachment/implantation on PxD 13. In the current reporting period (FY 2020) data for the effects of lactocrine deficiency in adults with respect to changes in endometrial gene expression in early pregnancy were presented by George et al in 2019 in Biology of Reproduction. In addition, we published a 2019 review in the journal Animal describing the impact of low and high ingestion of colostrum on maternal programming of reproductive tract development in adult pigs. We are currently analyzing postnatal day (PND) 14 uterine and testicular tissues for effects of lactocrine deficiency. This collaboration is enabling us to address the question of short-term andlong-term effects of low and high colostrum ingestion on reproductive performance in pigs. This is a physiologically-relevantmodel system to better understand hownaturally occurring lactocrine deficiency through maternal (e.g., mastitis, agalactia) as well as neonatal factors (e.g., within litter competition for teat position, birth rank and low birth weight) can influence development. C. To identify whether epigenetic changes are occurring in response to age and nursing from birth by monitoring (a) global uterine DNA methylation patterns as well as (b) DNA methylation of promotor regions of specific genes involved in neonatal uterine development. In our initial work on Objective 3 reported last year, we validated several differentially expressed (DE) genes that have been identified as epigenetic-related from data generatedusing RNAseq. Our databases for uterine gene expression in the neonate (e.g.,PND 0, PND 2 nursed and PND 2 replacer-fed) and endometrial gene expression in the adult(PxD 13, high vs low immunocrit) have been screened with respect to either age or immunocrit status (high vs low immunocrit).Next steps in this research are to study epigenetic changes including histone modifications that may be involved in regulating uterine gene expression in response to lactocrine deficiency. However, these studies were delayed due to COVID 19 restrictions for students and staff conducting research in the laboratory. For the final year of this project through July 2021, the impact of lactocrine deficiency from birth on endometrial morphology, uterine gland development and forkhead homeobox A2 (FOXA2) immunostaining patterns on PND 14 were studied. FOXA2, a transcription factor essential for the differentiation and development of uterine glands, marks uterine glandular epithelial (GE) cells in humans, mice and sheep, but has not been reported in pigs. Immunoreactive FOXA2 protein was localized consistently and uniquely in porcine GE cells of both low and high iCrit gilts at PND 14. The increased number of uterine GE cells/mm2 in high iCrit gilts indicated an overall increase in FOXA2 expression. These data based on a group of 5-6 PND 14 gilts in each low and high iCrit group formed the basis of Nina Paranjpe's Masters thesis research. This year an undergraduate honors student in the laboratory, Jenna Lorippo, has followed up on these studies and completed the analysis of an additional six animals/ group. Together, these results indicate that lactocrine deficiency from birth alters endometrial development resulting in fewer GE cells and an associated decrease in overall expression of FOXA2. A manuscript based on these and other related findings is currently in preparation. In addition, we have started on the analysis of changes in testicular endpoints in response to lactocrine deficiency in boar piglets at PND 3-5. Testes have been collected and will be subjected to RNAseq analysis to identify global changes in gene expression in low and high iCrit boars. In addition, immunohistochemical localization studies to detect testicular proteins of interest that may be influenced by lactocrine deficiency will be studied.
Publications
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2021
Citation:
Maternal lactocrine programming of reproductive development. American Society for Reproductive Medicine, Baltimore, MD
- Type:
Theses/Dissertations
Status:
Submitted
Year Published:
2021
Citation:
Lactocrine deficiency alters postnatal porcine uterine development. Masters Thesis by Nina Paranjpe
|
Progress 10/01/19 to 09/30/20
Outputs
Target Audience:During this reporting period, these studies involved informal teaching about lactation, maternal programming of development and endocrine disruption. Students gained the opportunity for experiential learning about pregnancy, parturition and lactation in swine. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research project has provided research training opportunities from October 1, 2019 through Sept 30, 2020 for a graduate student in my laboratory, Nina Paranjpe. In addition, Dr. Teh Ho, Research Associate in my laboratory has participated in the animal and laboratory work involved in these research studies. Hard-working undergraduate students worked on this project includingJennifer Lorippo and Yamirka Gonzalez who gained valuable hands-on research experience in conducting these studies. In addition, since I teach a course entitled Animal Reproduction each Fall semester, in Fall 2019, during the COVID 19 pandemic I provided research training to undergraduates by presenting this research project to my class of ~ 120 undergraduate students majoring in Animal Sciences, Biotechnology and Biology. However, due to the COVID 19 pandemic, the opportunity for one-on-one research mentoring from mid-March 2020, forward was limited given the restrictions for on-campus work in laboratories. How have the results been disseminated to communities of interest?Results were disseminated in abstracts presented at national and international meetings and invited talks given by the PI at a local high school, another US university in the US and at an international meeting. Results have also been reported in publications in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we plan to work on Objective 2 and 3 of this project.Together with our collaborators at the USDA MARC, we are in the process of analyzing uterine and testicular tissuesfrom low and high immunocrit gilts at birth (PND 0), PND 14 and at pregnancy day 13 for histology, immunohistochemical and epigenetics analyses. With respect to Objective 3, we plan start by subjecting PxD 13 endometrial samples, from low and high immunocrit pigs, to chromatin immunoprecipitation (ChIP) analyses. From these studies we hope to learn more about chromatin remodeling and histone modifications in the adult endometrium in response to lactocrine deficiency during the neonatal period.
Impacts
What was accomplished under these goals?
The goal of these studies is to improve reproductive efficiency in domestic animals by learning how nursing impacts programming of reproductive tract tissues that may ultimately have consequences for reproductive health and performance. A. To determine the effects of age and nursing on the neonatal porcine microRNA transcriptome and the miRNA x mRNA interactome from birth (PND 0) to PND 2 using next generation sequencing. This Objective 1 was completed in 2016 (RNAseq data) and 2017 (miRNAseq data) and published in two manuscripts in Biology of Reproduction. B. To investigate the long-term effects of low vs high colostrum consumption on the uterus at PND 14 and in the adult during the periattachment period in early pregnancy. For Objective 2, weare collaboratingwith Drs. Jeff Vallet, Clay Lents and Jeremy Miles at the USDA Meat Animal Research Center in Clay Center, Nebraska to evaluate the long-term effects of nursing on uterine development at pregnancy day (PxD) 13, the time of implantation in adult pigs.Dr. Vallet has identified low and high colostrum consuming gilts on their day of birth (postnatal day [PND]0) using an immunocrit assay. We have used this pig model system to study the effects of lactocrine deficiency in both the neonate at birth and PND 14 as well as in adult gilts around the time of attachment/implantation on PxD 13. In the current reporting period (FY 2020) data for the effects of lactocrine deficiency in adults with respect to changes in endometrial gene expression in early pregnancy were presented by George et al in 2019 in Biology of Reproduction. In addition, we published a 2019 review in the journal Animal describing the impact of low and high ingestion of colostrum on maternal programming of reproductive tract development in adult pigs. We are currently analyzing postnatal day (PND) 14 uterine and testicular tissues for effects of lactocrine deficiency. This collaboration is enabling us to address the question of short-term andlong-term effects of low and high colostrum ingestion on reproductive performance in pigs. This is a physiologically-relevantmodel system to better understand hownaturally occurring lactocrine deficiency through maternal (e.g., mastitis, agalactia) as well as neonatal factors (e.g., within litter competition for teat position, birth rank and low birth weight) can influence development. C. To identify whether epigenetic changes are occurring in response to age and nursing from birth by monitoring (a) global uterine DNA methylation patterns as well as (b) DNA methylation of promotor regions of specific genes involved in neonatal uterine development. In our initial work on Objective 3 reported last year, we validated several differentially expressed (DE) genes that have been identified as epigenetic-related from data generatedusing RNAseq. Our databases for uterine gene expression in the neonate (e.g.,PND 0, PND 2 nursed and PND 2 replacer-fed) and endometrial gene expression in the adult(PxD 13, high vs low immunocrit) have been screened with respect to either age or immunocrit status (high vs low immunocrit).Next steps in this research are to study epigenetic changes including histone modifications that may be involved in regulating uterine gene expression in response to lactocrine deficiency. However, these studies were delayed due to COVID 19 restrictions for students and staff conducting research in the laboratory. Instead focus was directed to completing the analysis of other studies initiated by graduate student Nina Paranjpe. Objectives were to determine effects lactocrine deficiency from birth on endometrial morphology, uterine gland development and forkhead homeobox A2 (FOXA2) immunostaining patterns on PND 14. FOXA2, a transcription factor essential for the differentiation and development of uterine glands, marks uterine glandular epithelial (GE) cells in humans, mice and sheep, but has not been reported in pigs. Immunoreactive FOXA2 protein was localized consistently and uniquely in GE cells of both low and high iCrit gilts at PND 14. The increased number of uterine GE cells/mm2 in high iCrit gilts indicated an overall increase in FOXA2 expression. These results indicate that lactocrine deficiency from birth alters endometrial development resulting in fewer GE cells and an associated decrease in overall expression of FOXA2. A manuscript based on these and other related findings is currently in preparation.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
George AF, Ho T-Y, Prasad N, Keel BN, Miles JR, Vallet JL, Bartol FF, Bagnell CA. 2019. Neonatal lactocrine deficiency affects the adult porcine endometrial transcriptome at pregnancy day 13. Biol Reprod 100(1): 71-85.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Bagnell CA, Bartol FF. 2019. Review: Maternal programming of development in the pig and the lactocrine hypothesis. Animal, Dec; 13(12):2978-2985.
|
Progress 10/01/18 to 09/30/19
Outputs
Target Audience:The target audience for this research includes agricultural producers who are interested in increasing reproductive efficiency in livestock and others interested in the impact of nursing on development and the importance of milk as a means of continuing maternal/neonatal communication after birth. This research project serves as the basis for training graduate and undergraduate students in hypothesis testing, experimental design, research methodologies and data analysis. These studies also involve additional informal teaching about lactation, maternal programming of development and endocrine disruption. Students also gain the opportunity for experiential learning about pregnancy, parturition and lactation in swine. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research project has provided research training opportunities from October 1, 2018 through Sept 30, 2019 for the following graduate students:Nina Paranjpe. In addition, Dr. Teh Ho, Research Associate in my laboratory has participated in the animal and laboratory work involved in these research studies. Hard-working undergraduate students worked on this project including: Dorothy Knipple, Dana Tsuchida, Julie Wang, Nardine Nasr, Katherine Farfara and Josee Westrichwho gained valuable hands-on research experience in conducting these studies. In addition, since I teach a course entitled Animal Reproduction each Fall semester, in Fall 2019 I provided research training to undergraduates by presenting this research project to my class of ~ 120 undergraduate students majoring in Animal Sciences, Biotechnology and Biology. How have the results been disseminated to communities of interest?Results were disseminated in abstracts presented at national and international meetings and invited talks given by the PI at other universities in the US and at international meetings. Results have also been reported in publications in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we plan to work on Objective 2 and 3 of this project.Together with our collaborators at the USDA MARC, we are in the process of analyzing uterine and testicular tissuesfrom low and high immunocrit gilts at birth (PND 0), PND 14 and at pregnancy day 13 for histology, immunohistochemical and epigenetics analyses. With respect to Objective 3, we plan start by subjecting PxD 13 endometrial samples, from low and high immunocrit pigs, to chromatin immunoprecipitation (ChIP) analyses. From these studies we hope to learn more about chromatin remodeling and histone modifications in the adult endometrium in response to lactocrine deficiency during the neonatal period.
Impacts
What was accomplished under these goals?
Impact Statement: Consumption of 'first milk' termed colostrum soon after birth delivers nutrients and milk-borne bioactive factors (MbFs) to nursing young that can affect postnatal tisue development.Thus, milk is more than food. Female reproductive tissues undergo development postnatally although little is known about MbFs and how they affect development. The term lactocrine was coined to describe the delivery of MbFs in colostrumto offspring as a consequence of nursing. Thelactocrine hypothesis for maternal programming of postnatal development suggests that disruption of lactocrine signaling will alter the programming and trajectory of development withshort-term organizational effectsand long-term consequences on the adult phenotype.Understanding mechanisms through which lactocrine signaling, enabled by colostrum consumption shortly after birth, affects development and function of reproductive system tissues will permit refinement of recommendations for care of human infants. This will also be important for improvements in husbandry guidelines designed to optimize reproductive performance in the pig, an economically important domestic animal. The goal of these studies is to improve reproductive efficiency in domestic animals by learning how nursing impacts programming of reproductive tract tissues that may ultimately have consequences for reproductive health and performance. A. To determine the effects of age and nursing on the neonatal porcine microRNA transcriptome and the miRNA x mRNA interactome from birth (PND 0) to PND 2 using next generation sequencing. This Objective 1 was completed in 2016 (RNAseq data) and 2017 (miRNAseq data) and published in two manuscripts in Biology of Reproduction. B. To investigate the long-term effects of low vs high colostrum consumption on the uterus at PND 14 and in the adult during the periattachment period in early pregnancy. For Objective 2, weare collaboratingwith Drs. Jeff Vallet, Clay Lents and Jeremy Miles at the USDA Meat Animal Research Center in Clay Center, Nebraska to evaluate the long-term effects of nursing on uterine development at pregnancy day (PxD) 13, the time of implantation in adult pigs.Dr. Vallet has identified low and high colostrum consuming gilts on their day of birth (postnatal day [PND]0) using an immunocrit assay. We have used this pig model system to study the effects of lactocrine deficiency in both the neonate at birth and PND 14 as well as in adult gilts around the time of attachment/implantation on PxD 13. Data for the effects of lactocrine deficiency in adults with respect to changes in endometrial gene expression in early pregnancy were presented by George et al in 2018 in Biology of Reproduction. In the current reporting period, we are analyzing PND 14 uterine and testicular tissues for effects of lactocrine deficiency. This collaboration is enabling us to address the question of short-term andlong-term effects of low and high colostrum ingestion on reproductive performance in pigs. This is a physiologically-relevantmodel system to better understand hownaturally occurring lactocrine deficiency through maternal (e.g., mastitis, agalactis) as well as neonatal factors (e.g., within litter competition for teat position, birth rank and low birth weight) can influence development. C. To identify whether epigenetic changes are occurring in response to age and nursing from birth by monitoring (a) global uterine DNA methylation patterns as well as (b) DNA methylation of promotor regions of specific genes involved in neonatal uterine development. In our initial work on Objective 3 this year, we have validated several differentially expressed (DE) genes that have been identified as epigenetic-related from data generatedusing RNAseq. Our databases for uterine gene expression in the neonate (e.g.,PND 0, PND 2 nursed and PND 2 replacer-fed) and endometrial gene expression in the adult(PxD 13, high vs low immunocrit) have been screened with respect to either age or immunocrit status (high vs low immunocrit).Next steps in this research are to study epigenetic changes including histone modifications that may be involved in regulating uterine gene expression in response to lactocrine deficiency. Histone modification are post-translational changes to histone proteins that include methylation, phosphorylation, acetylation or ubiquitylation. We will begin by studying DE uterine genes in the neonate and adult and determine whether histone marks are altered in these tissues over time.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Bagnell CA and Bartol FF. 2019. Relaxin and the Milky Way: The lactocrine hypothesis and maternal programming of development. Molec Cellular Endocrinology 487:18-23.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Maternal lactocrine programming of reproductive tract development Dept of Biology,Seton Hall University, South Orange, NJ, November 9, 2018.
|
Progress 10/01/17 to 09/30/18
Outputs
Target Audience:The target audience for this research includes agricultural producers who are interested in increasing reproductive efficiency in livestock and others interested in the impact of nursing on development and the importance of milk as a means of continuing maternal/neonatal communication after birth. This research project serves as the basis for training graduate and undergraduate students in hypothesis testing, experimental design, research methodologies and data analysis. These studies also involve additional informal teaching about lactation, maternal programming of development and endocrine disruption. Students also gain the opportunity for experiential learning about pregnancy, parturition and lactation in swine. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research project has provided research training opportunities from Oct 1, 2017 through Sept 30, 2018 for the following graduate students: Ashley George, Nina Paranjpe. In addition, Dr. Teh Ho, Research Associate in my laboratory has participated in the animal and laboratory work involved in these research studies. Hard-working undergraduate students worked on this project including: Dorothy Knipple, Dana Tsuchida, Julie Wang, Nardine Nasr and Katherine Farfara who gained valuable hands-on research experience in conducting these studies. In addition, since I teach a course entitled Animal Reproduction each Fall semester, in Fall 2018 I provided research training to undergraduates by presenting this research project to my class of ~ 120 undergraduate students majoring in Animal Sciences, Biotechnology and Biology. How have the results been disseminated to communities of interest?Results were disseminated in abstracts presented at national and international meetings and invited talks given by the PI at other universities in the US and at international meetings. Results have also been reported in publications in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we plan to complete Objective 2. We will analyze the proteomic data from the uterine flushings at PxD 13 with the assistance of the Rutgers University bioinformatics specialists. Together with our collaborators at the USDA MARC, we are in the process of collecting additional tissues from low and high immunocrit gilts at birth (PND 0), PND 2 and at PND 14 for histology, immunohistochemical and epigenetics analyses. With respect to Objective 3, we plan start by subjecting PxD 13 endometrial samples, from low and high immunocrit pigs, to chromatin immunoprecipitation (ChIP) analyses. From these studies we hope to learn more about chromatin remodeling and histone modifications in the adult endometrium in response to lactocrine deficiency during the neonatal period.
Impacts
What was accomplished under these goals?
For newborn mammals, both human and animal, consumption of 'first milk' (colostrum) soon after birth delivers nutrients and milk-borne factors (MbFs) to infants that can affect the way that reproductive tissues develop and function in adults. Little is known about MbFs and how they affect development. Colostrum and milk are produced during lactation and delivery of MbFs from mother to infant via nursing is called lactocrine transmission. This research, using a pig model system, tests the lactocrine hypothesis for maternal programming of reproductive system development. The lactocrine hypothesis predicts that neonates deprived of colostrum will have reduced uterine capacity to support conceptus development as adults due to disruption of the uterine developmental program shortly after birth. Understanding mechanisms through which lactocrine signaling, enabled by colostrum consumption shortly after birth, affects development and function of reproductive system tissues will permit refinement of recommendations for care of human infants. This will also be important for improvements in husbandry guidelines designed to optimize reproductive performance in the pig, an economically important domestic animal. The goal of these studies is to improve reproductive efficiency in domestic animals by learning how nursing impacts programming of reproductive tract tissues that may ultimately have consequences for reproductive health and performance. A. To determine the effects of age and nursing on the neonatal porcine microRNA transcriptome and the miRNA x mRNA interactome from birth (PND 0) to PND 2 using next generation sequencing. This Objective 1 was completed in 2016 (RNAseq data) and 2017 (miRNAseq data) and published in two manuscripts in Biology of Reproduction. B. To investigate the long-term effects of low vs high colostrum consumption on the uterus at PND 14 and in the adult during the periattachment period in early pregnancy. For Objective 2, we have collaborated with Drs. Jeff Vallet, Clay Lents and Jeremy Miles at the USDA Meat Animal Research Center in Clay Center, Nebraska to evaluate the long-term effects of nursing on uterine development at pregnancy day 13, the time of implantation in adult pigs. Dr. Vallet has identified low and high colostrum consuming gilts at postnatal day (PND) 1 using an immunocrit assay. Low and high immunocrit gilts were allowed to mature and bred in order to collect tissues at pregnancy (PxD) 13 (around the time of attachment/implantation). We isolated RNA from uterine tissues for both RNAseq and miRNAseq analysis. Results from these studies indicated that more than 1150 differentially expressed endometrial mRNAs were detected on PxD 13 in high as compared to low iCrit gilts. These affected genes had functions related to solute transport, endometrial receptivity and immune response. Six differentially expressed microRNAs targeting more than 60 differentially expressed mRNAs were also identified. Results indicate that lactocrine deficiency from birth has long-term consequences for endometrial function as reflected by changes in the endometrial transcriptome during early pregnancy. These data were presented at the 2018 Meeting of the Society for the Study of Reproduction and published in a manuscript by George et al, 2018 in Biology of Reproduction. We also subjected uterine flush samples from PxD 13 gilts to proteomic analyses. This will enable us to see the effect of high or low colostrum intake during the neonatal period on both gene expression and translation to secreted proteins in adults. In addition, endometrial tissue from low and high immunocrit gilts on PND 14 were evaluated for histological changes as well as estrogen receptor alpha (ESR1) expression. The results indicated that lactocrine deficiency from birth, reflected by low PND 0 iCrit, affects patterns of neonatal uterine development at PND 14. Effects of lactocrine deficiency were similar to those observed for uteri obtained on PND 14 after experimental imposition of a lactocrine-null state by milk replacer feeding of neonatal gilts for two days from birth. Results support the idea that disruption of lactocrine input sufficient to affect the neonatal uterine developmental program, and to have long-term negative consequences for uterine capacity in adults, can occur in nursing gilts maintained under normal husbandry conditions in the nursery. This collaboration is enabling us to address the question of long-term effects of low and high colostrum ingestion on reproductive performance in pigs. C. To identify whether epigenetic changes are occurring in response to age and nursing from birth by monitoring (a) global uterine DNA methylation patterns as well as (b) DNA methylation of promotor regions of specific genes involved in neonatal uterine development. Our initial work on Objective 3 this year has been to search our RNAseq databases in both the neonate (PND 0 and PND 2) and adult (PxD 13) to identify epigenetic-related genes that are differentially-expressed (DE) with respect to either age or immunocrit status (high vs low immunocrit). We are in the process of validating those DEGs.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
George AF, Rahman KM, Miller DM, Wiley AA, Camp ME, Bartol FF, Bagnell CA. 2018. Effects of colostrum, feeding method and oral IGF1 on porcine uterine development. Reproduction 155:259-271. doi: 10.1530/REP-17-0658.
- Type:
Book Chapters
Status:
Published
Year Published:
2018
Citation:
George AF and Bagnell CA. 2018. Relaxin. In: TE Spencer and JA Flaws (ed). Encyclopedia of Reproduction, 2nd Edition, Volume 2, Female Reproduction - Pregnancy, Chapter 90, pp 591-596, Elsevier Inc., Waltham, MA.
- Type:
Book Chapters
Status:
Published
Year Published:
2018
Citation:
Bartol FF and Bagnell CA. 2018. Lactocrine Programming. In: TE Spencer and JA Flaws (ed). Encyclopedia of Reproduction, 2nd Edition, Volume 2, Female Reproduction - Lactation, pp 814-820, Elsevier Inc., Waltham, MA.
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
George AF, Ho T-Y, Prasad N, Keel BN, Miles JR, Vallet JL, Bartol FF, Bagnell CA. 2018. Neonatal lactocrine deficiency affects the adult porcine endometrial transcriptome at pregnancy day 13. Biol Reprod 1-15. doi: 10.1093/biolre/ioy180
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Relaxin and the Milky Way: the Lactocrine Hypothesis Plenary Lecture. 8th International Conference on Relaxin and Related Peptides, Cabo San Lucas, Mexico, May 6-11, 2018
|
Progress 10/01/16 to 09/30/17
Outputs
Target Audience:The target audience for this research includes agricultural producers who are interested in increasing reproductive efficiency in livestock and others interested in the impact of nursing on development and the importance of milk as a means of continuing maternal/neonatal communication after birth. This research project serves as the basis for training graduate and undergraduate students in hypothesis testing, experimental design, research methodologies and data analysis. These studies also involve additional informal teaching about lactation, maternal programming of development and endocrine disruption. Students also gain the opportunity for experiential learning about pregnancy, parturition and lactation in swine. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research project has provided research training opportunities from October 1, 2016 through Sept 30, 2017 for the following graduate students: Ashley George, Chelsee Holloway. In addition, Dr. Teh Ho, Research Associate in my laboratory has participated in the animal and laboratory work involved in these research studies. In addition, hard-working undergraduate students worked on this project including: Dorothy Knipple, Dana Tsuchida and Julie Wang who gained valuable hands-on research experience in conducting these studies. In addition, since I teach a course entitled Animal Reproduction each Fall semester, in Fall 2017 I provided research training to undergraduates by presenting this research project to my class of ~ 120 undergraduate students majoring in Animal Sciences, Biotechnology and Biology. How have the results been disseminated to communities of interest?Results were disseminated in abstracts presented at national and international meetings and invited talks given by the PI at other universities in the US and at international meetings. Results have also been reported in publications in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we plan to complete Objective 2, evaluating the long-term effects of nursing on uterine development at PND 14 and at pregnancy day 13, the time of implantation in adult pigs. The RNAseq and miRNAseq analyses of uterine tissues will be completed and prepared for publication. We are also subjecting uterine flush samples to proteomic analyses so that we can see the relationship between gene expression and translation to proteins secreted by these uterine samples. Together with our collaborators at the USDA MARC, we are in the process of collecting additional tissues from low and high immunocrit gilts at Pxd 13 for histology and immunohistochemical analysis. We also plan to begin tissue collection for the epigenetics studies described in Objective 3.
Impacts
What was accomplished under these goals?
Non-technical summary - Impact For newborn mammals, both human and animal, consumption of 'first milk' (colostrum) soon after birth delivers nutrients and milk-borne factors (MbFs) to infants that can affect the way that reproductive tissues develop and function in adults. Little is known about MbFs and how they affect development. Colostrum and milk are produced during lactation and delivery of MbFs from mother to infant via nursing is called lactocrine transmission. Proposed research, using a pig model system, will test the lactocrine hypothesis for maternal programming of reproductive system development. The lactocrine hypothesis predicts that neonates deprived of colostrum will have reduced uterine capacity to support conceptus development as adults due to disruption of the uterine developmental program shortly after birth. Understanding mechanisms through which lactocrine signaling, enabled by colostrum consumption shortly after birth, affects development and function of reproductive system tissues will permit refinement of recommendations for care of human infants. This will also be important for improvements in husbandry guidelines designed to optimize reproductive performance in the pig, an economically important domestic animal. The goal of these studies is to improve reproductive efficiency in domestic animals by learning how nursing impacts programming of reproductive tract tissues that may ultimately have consequences for reproductive health and performance. With respect to Objective 1, experiments focused on defining the age- and lactocrine-sensitive neonatal porcine uterine transcriptome using RNA sequencing (RNAseq) and microRNA sequencing (miRNAseq) were completed. Since miRNAs play an important role in regulating gene expression, we were interested in miRNA-mRNA interactions in the neonatal uterus. The analyses focused on miRNAs expressed in uteri (n = 4/group) obtained at birth [postnatal day (PND) 0] and from gilts that were either nursed ad libitum for two days from birth (PND 2N) or fed a commercial milk replacer formula (PND 2R) in lieu of nursing over this same period of time. We coupled this miRNA information with RNAseq data for uterine gene expression in the same animals. In this manner we defined miRNA-mRNA interactions in the neonatal porcine uterus with respect to age and nursing. Biological processes predicted to be affected by age and nursing included cell-to-cell signaling, cell morphology and tissue morphology. These data were published in Biology of Reproduction in Dec 2016 (RNAseq data) and Feb, 2017 (miRNAseq data). In addition, other studies were conducted to evaluate (1) lactocrine effects on uterine development as early as 12 h postnatally; (2) the impact of a single dose of colostrum at birth on postnatal development at 12 h postnatal; and (3) the effects of oral insulin-like growth factor 1 (IGF1) on uterine growth in this 12 h period after birth. Results showed that both endometrial thickness and stromal cell nuclear density were similar in nursed and replacer-fed gilts at 12 h postnatal. However, stromal and epithelial PCNA labeling indices were greater (P < 0.05) in nursed gilts compared to those fed milk replacer from birth. While uterine WNT7A expression was not affected by treatment, WNT5A, HOXA10, RXFP1 and VEGFA expression was greater (P < 0.05) in nursed gilts. Similarly, both proMMP9 and MMP9 protein levels were greater (P < 0.001) at 12 h postnatal in nursed gilts. Consistently, TIMP1 protein was detected in uterine tissues at 12 h postnatal in nursed but not in replacer-fed gilts. Relative levels of uterine proMMP2 and MMP2 proteins, detected in all tissues, did not differ between treatment groups. Present results show that nursing for 12 h from birth supports rapid establishment of a uterine developmental program. This is illustrated by patterns of endometrial cell proliferation, expression of genes associated with uterine wall development, and establishment of a MMP9/TIMP1 system that evolves by 12 h postnatally. These data were presented at the Society for the Study of Reproduction meetings in July, 2017 and are being prepared for publication. For Objective 2, we are collaborating with Drs. Jeff Vallet and Clay Lents at the USDA Meat Animal Research Center in Clay Center, Nebraska to evaluate the long-term effects of nursing on uterine development at pregnancy day 13, the time of implantation in adult pigs. Dr. Vallet has identified low and high colostrum consuming gilts at postnatal day (PND) 1 using an immunocrit assay. Low and high immunocrit gilts were allowed to mature and bred in order to collect tissues at pregnancy (Pxd) 13 (around the time of attachment/implantation). We have isolated RNA from uterine tissues for both RNAseq and miRNAseq analysis. Results from these studies are currently being prepared for publication. This collaboration will enable us to address the question of long-term effects of low and high colostrum ingestion on reproductive performance in pigs.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Bagnell CA, Ho TY, George AF, Wiley AA, Miller DJ, Bartol FF. 2017. Maternal lactocrine programming of porcine reproductive tract development. Molec Reprod and Development, doi: 10.1002/mrd.22815.
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Bartol FF, Wiley AA, George AF, Miller DJ, Bagnell CA. 2017. Postnatal reproductive development and the lactocrine hypothesis. J Anim Science 95(5): 2200-2210.
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
George AF, Rahman KM, Camp ME, Prasad N, Bartol FF, Bagnell CA. 2017. Defining age- and lactocrine-sensitive elements of the neonatal porcine uterine microRNA-mRNA interactome. Biol Reprod 96(2): 327-340.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Ho TY, Rahman KF, Camp ME, Wiley AA, Bartol FF, Bagnell CA. 2016. Timing and duration of nursing from birth affect neonatal porcine uterine matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1. Domestic Animal Endocrinology 59: 1-10.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Rahman KM, Camp ME, Prasad N, McNeel AK, Levy SE, Bartol FF, Bagnell CA. 2016. Age and nursing affect the neonatal porcine uterine transcriptome. Biol Reprod 94(2):46, 1-13.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2017
Citation:
Lactocrine signaling and developmental programming - Plenary Lecture. 10th International Conference on Pig Reproduction, Columbia, MI, June 14, 2017
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Maternal lactocrine programming of reproductive tract development, Physiology/Endocrinology Seminar Series, Cornell University, Ithaca, NY, Nov 2, 2016.
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Progress 07/21/16 to 09/30/16
Outputs
Target Audience:The target audience for this research includes agricultural producers who are interested in increasing reproductive efficiency in livestock and others interested in the impact of nursing on development and the importance of milk as a means of continuing maternal/neonatal communication after birth. This research project serves as the basis for training graduate and undergraduate students in hypothesis testing, experimental design, research methodologies and data analysis. These studies also involve additional informal teaching about lactation, maternal programming of development and endocrine disruption. Students also gain the opportunity for experiential learning about pregnancy, parturition and lactation in swine. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research project has provided research training opportunities from July 21, 2016 through Sept 30, 2016 for the following graduate students: Ashley George, Chelsee Holloway. In addition, Dr. Teh Ho, Research Associate in my laboratory has participated in the animal and laboratory work involved in these research studies. In addition, hard-working undergraduate students worked on this project including: Dorothy Knipple and Julie Wang and gained valuable hands-on research experience in conducting these studies. In addition, since I teach a course entitled Animal Reproduction each Fall semester, in Fall 2016 I provided undergraduate research training to undergraduates by presenting this research project to my class of ~ 125 undergraduate students majoring in Animal Sciences, Biotechnology and Biology. 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?In the next reporting period we plan to complete Objective 1, the miRNAseq analyses of uterine tissues with respect to age and nursing, and publish the research. We also will begin the long-term studies described in Objective 2 in collaboration with Dr. Jeff Vallet at the USDA Meat Animal Research Center in Clay Center, Nebraska. Here we will evaluate the long-term effects of nursing on uterine development at PND 14 and at pregnancy day 13, the time of implantation in adult pigs. We are in the early stages of this study, measuring PND 1 immunocrit in neonatal gilts as a measure of colostrum consumption. Next, gilts with either low or high immunocrit levels will be allowed to mature, monitored for estrous cycling then bred to induce pregnancy. The plan is to collect uterine tissues from PxD 13 gilts that had either low and high immunocrit levels at PND 1 to determine whether colostrum availability (measured by immunocrit at PND 1) influences uterine development at the time of implantation.
Impacts
What was accomplished under these goals?
Impact Statement For newborn mammals, both human and animal, consumption of 'first milk' (colostrum) soon after birth delivers nutrients and milk-borne factors (MbFs) to infants that can affect the way that reproductive tissues develop and function in adults. Little is known about MbFs and how they affect development. Colostrum and milk are produced during lactation and delivery of MbFs from mother to infant via nursing is called lactocrine transmission. Proposed research, using a pig model system, will test the lactocrine hypothesis for maternal programming of reproductive system development. The lactocrine hypothesis predicts that neonates deprived of colostrum will have reduced uterine capacity to support conceptus development as adults due to disruption of the uterine developmental program shortly after birth. Understanding mechanisms through which lactocrine signaling, enabled by colostrum consumption shortly after birth, affects development and function of reproductive system tissues will permit refinement of recommendations for care of human infants. This will also be important for improvements in husbandry guidelines designed to optimize reproductive performance in the pig, an economically important domestic animal. The goal of these studies is to improve reproductive efficiency in domestic animals by learning how nursing impacts programming of reproductive tract tissues that may ultimately have consequences for reproductive health and performance.With respect to Objective 1, efforts were directed to complete experiments outlined to define the age-sensitive neonatal porcine uterine transcriptome using microRNA sequencing (miRNAseq). In our previous work, RNAseq was conducted on the same tissues and this research was recently accepted for publication (Nov 2015). Now analyses focus on miRNAseq on uteri (n = 4/group) obtained at birth [postnatal day (PND) 0] and from gilts that were either nursed ad libitum for two days from birth (PND 2N) or fed a commercial milk replacer formula (PND 2R) in lieu of nursing over this same period of time. Results from these miRNAseq studies are currently being prepared for publication. These data were presented at the Annual Meeting of the Society for the Study of Reproduction in July 2016. For Objective 2, we have been collaborating with Dr. Jeff Vallet at the USDA Meat Animal Research Center in Clay Center, Nebraska to evaluate the long-term effects of nursing on uterine development at pregnancy day 13, the time of implantation in adult pigs. Dr. Vallet has identified low and high colostrum consuming gilts at postnatal day (PND) 1 using an immunocrit assay. Uterine tissues from both groups at PND 14 (n= 8/group) have been collected and are ready for analysis. In another set of animals low and high immunocrit gilts were allowed to mature and will be bred in order for us to collect tissues at Pxd 13 (around the time of attachment/implantation). This collaboration will enable us to address the question of long-term effects of low and high colostrum ingestion on reproductive performance in pigs. Analysis of these samples will be conducted as part of this current Hatch Project.
Publications
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