Faculty Mentor Information
Dr. Bethaney Fehrenkamp (Mentor), University of Idaho and Idaho WWAMI Medical Education, University of Washington
Additional Funding Sources
This research was made possible through the University of Idaho College of Agricultural and Life Sciences and the American Society for Parenteral and Enteral Nutrition (ASPEN), Idaho WWAMI Medical Education, an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant #P20GM103408, and the University of Idaho Office of Undergraduate Research - Summer Undergraduate Research Fellowship.
Abstract
Introduction: While numerous correlational studies suggest that human milk (HM) optimizes infant gut development, little is known about the physiological process. To investigate HM's protective mechanisms for intestinal development, we used the neonatal piglet model. The recommended time to exclusively breastfeed an infant is six months, which is equivalent to one month in piglets. We hypothesized that HM-fed piglets would show increased paracellular junctions and reduced inflammatory markers in the intestines compared to infant formula (IF)-fed piglets.
Methods: We reared littermate Yorkshire-duroc piglets for 28 days, assigning them to HM-fed (n=6) and IF-fed (n=6) groups, alongside controls reared on sow’s milk (n=2) at the farm. Following euthanasia and tissue collection, we extracted RNA from intestinal tissues, purified them of DNA contaminants, synthesized complementary DNA (cDNA), and performed quantitative-PCR (qPCR). We examined paracellular junction proteins (Claudin 1, Tight Junction Protein 1, Tight Junction Protein 2, Occludin, and Cadherin 1), immunomodulatory cytokines (Interleukin (IL)-2, IL-4, IL-8, IL-10, IL-1B, and TNFa), and lymphocyte markers (CD3e, CD8a, CD20, CD79b). Results were normalized to the reference gene beta actin within individuals and averaged across feeding groups.
Results: We used ANOVA, analysis of variance, with multiple comparisons to determine differences between feeding groups.
Discussion:Contrary to our expectations, there were no significant differences in the expression of inflammatory markers or paracellular junctions. However, as part of a larger project, it is still clear HM plays a role in optimizing gut development. Future research will involve a global transcriptomic analysis of the ileum using next-generation sequencing data.
Transcriptional Investigations into Neonatal Gut Development Related to Early Feeding
Introduction: While numerous correlational studies suggest that human milk (HM) optimizes infant gut development, little is known about the physiological process. To investigate HM's protective mechanisms for intestinal development, we used the neonatal piglet model. The recommended time to exclusively breastfeed an infant is six months, which is equivalent to one month in piglets. We hypothesized that HM-fed piglets would show increased paracellular junctions and reduced inflammatory markers in the intestines compared to infant formula (IF)-fed piglets.
Methods: We reared littermate Yorkshire-duroc piglets for 28 days, assigning them to HM-fed (n=6) and IF-fed (n=6) groups, alongside controls reared on sow’s milk (n=2) at the farm. Following euthanasia and tissue collection, we extracted RNA from intestinal tissues, purified them of DNA contaminants, synthesized complementary DNA (cDNA), and performed quantitative-PCR (qPCR). We examined paracellular junction proteins (Claudin 1, Tight Junction Protein 1, Tight Junction Protein 2, Occludin, and Cadherin 1), immunomodulatory cytokines (Interleukin (IL)-2, IL-4, IL-8, IL-10, IL-1B, and TNFa), and lymphocyte markers (CD3e, CD8a, CD20, CD79b). Results were normalized to the reference gene beta actin within individuals and averaged across feeding groups.
Results: We used ANOVA, analysis of variance, with multiple comparisons to determine differences between feeding groups.
Discussion:Contrary to our expectations, there were no significant differences in the expression of inflammatory markers or paracellular junctions. However, as part of a larger project, it is still clear HM plays a role in optimizing gut development. Future research will involve a global transcriptomic analysis of the ileum using next-generation sequencing data.