Performing Department
VET MED DEAN
Non Technical Summary
Several physiologic conditions drive lipid mobilization (lipolysis) in the adipose tissues (AT) of dairy cows, including periparturient energy deficits and the presence of inflammatory stimuli during infectious disease. Should lipolysis become excessive or dysregulated, AT inflammation results, and cows become susceptible to metabolic and inflammatory diseases, milk yield losses, and reproductive complications. Such challenges come at substantial costs to US producers and compromise the welfare of dairy cows.In addition to energy-dense lipids, cows' fat cells (adipocytes) release endocannabinoids during lipolysis, which are signaling molecules that regulate inflammation and metabolism. We hypothesize that these molecules regulate the inflammatory responses of immune cell populations, and the expression of their receptors vary in AT based on the energy demands faced by the cow. We will test this hypothesis through two aims. Aim 1 will reveal how the stimulation of cannabinoid receptors on immune cells in AT influences their numbers, inflammatory properties, and abilities to regulate lipolysis. For this aim, we will harvest and culture mononuclear cells and adipocytes from dairy cows, expose them to cannabinoid receptor-activating compounds, and evaluate the cells' responses. Aim 2 will investigate the relationships between the expression of cannabinoid receptors and abundance of immune cells in the AT of pre- and post-partum dairy cows. In this aim, we will collect AT from mature dairy cows at -21, +14, and +21 days relative to calving and, using microscopic and molecular biological techniques, assess how the distribution of cannabinoid receptors and abundance of mononuclear cells change relative to one another. Findings from Aims 1 and 2 will be published in peer-reviewed journals, presented at scientific meetings, and shared with industry stakeholders.This project will allow us to elucidate the role of endocannabinoids in the regulation of metabolic and inflammatory responses in the AT of dairy cows. We anticipate that these findings will support the development of targeted nutritional and pharmacological approaches to managing dairy cattle, especially in early lactation. Long-term, these efforts will help improve the health and productivity of dairy cows and, in addition, the sustainability and productivity of the US dairy industry.
Animal Health Component
100%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Goals / Objectives
The primary goal of this project is to determine the molecular mechanisms by which endogenous signaling molecules known as endocannabinoids regulate metabolism and inflammation in the fat of dairy cows. By better understanding these fundamental processes, this project aims to provide insight that may be applied to improve the health, welfare, and productivity of dairy cows and promote dairy industry sustainability and food security. This project will provide comprehensive research training to the PD in advanced molecular biological techniques, data analysis, and scientific communications. The specific objectives of this project consist of: Research objectives1. Investigate the relationships between cannabinoid receptor activation and mononuclear cell proliferation, activation, and regulation of lipolysisDetermine how cannabinoid receptor activation modulates the proliferation and activation of cultured mononuclear cellsAssess the role of cannabinoid receptor signaling on mononuclear cells' capacity to regulate lipolysis in adipocytes2. Characterize the relationships between mononuclear cell abundance, phenotype, and endocannabinoid system component distribution in the adipose tissue of periparturient cowsDetermine how the abundance and phenotype of mononuclear cells change in cows' adipose tissues throughout the periparturient periodEvaluate how the distribution of endocannabinoid system components within adipose tissue relates to mononuclear cell abundance and phenotypeTraining objectives1. Technical skill developmentDevelop expertise in monocyte and adipocyte culture and functional assays to investigate the role of the endocannabinoid system on cell proliferation, inflammatory phenotype, and lipolysisProvide hands-on training in advanced molecular biological techniques including gene expression analysis (RT-qPCR, RNA sequencing, sm-FISH), cell morphology and surface marker expression quantitation (flow cytometry), lipidomic and proteomic analyses, capillary electrophoresis, and immunohistochemistry2. Data analysisGain proficiency in bioinformatics tools and statistical analysis software for analyzing gene expression data, protein expression, image analysis, and cell phenotyping resultsIntegrate and interpret complex datasets to draw accurate and meaningful biological conclusions3. Scientific communicationAdvance the ability to communicate scientific findings through the preparation of research manuscripts in peer-reviewed journals and conference proceedingsProvide opportunities to generate figures and graphics and present research findings at scientific conferences, departmental seminars, and stakeholder outreach eventsPromote mentorship and leadership skills through training of undergraduate and graduate research students and attendance of workshops and seminars
Project Methods
To determine how endocannabinoid (eCB) receptor activation influences immune cell activity and fat breakdown in cows, we will collect blood and fat samples.Aim 1: We will examine how eCB receptor activation alters the behavior of immune cells and mobilization of lipids from fat cells. For the first experiment, we will culture immune cells derived from the blood and expose them to different cannabinoid receptor-targeting agents and observe how they differ in number, type, and function. In the second experiment, we will grow fat cells harvested from cows' fat along with the treated immune cells to observe the effects on fat breakdown and inflammatory responses.Aim 2: We will analyze fat samples from pre- and post-partum cows to understand how the relationships between eCB system components and the characteristics of immune cells change in the tissue throughout this time. Fat samples will be collected from cows prior to and following calving, and we will study the expression and distribution of eCB-associated genes and proteins.The data generated from this project will be shared in peer-reviewed scientific publication and poster and oral presentation format at scientific conferences and stakeholder meetings. The results of this project will help clarify how the eCB system may regulate inflammation and metabolism in the fat of cows, which may improve cow health and productivity.