Adipose Tissue Expansion Analysis Service
InquiryOverview of Adipose Tissue Expansion
White adipose tissue serves as the primary repository for energy storage in the form of triacylglycerols and plays a critical role in maintaining whole-body energy homeostasis. Due to its high plasticity, white adipose tissue expands, contracts, and reorganizes throughout adulthood to adapt to fluctuations in energy balance caused by various metabolic cues, including obesity, dietary interventions, and physical activity. The mass of white adipose tissue is regulated by the interplay of adipocyte volumetric changes, mediated by triacylglycerol synthesis and breakdown, as well as processes of adipocyte generation and cell death. Research indicates that the expandability of subcutaneous white adipose tissue is a key determinant of metabolic dysregulation associated with obesity. Protheragen offers a comprehensive exploration into the mechanisms underlying white adipose tissue expansion, employing cutting-edge imaging techniques, molecular biology, and bioinformatics.
Targets for Developing Anti-Obesity Therapeutics
Our analysis is pivotal for pinpointing critical molecular and cellular targets associated with obesity. These identified targets undergo rigorous validation, and we help clients meticulously screen and characterize potential drug candidates.
Anti-Obesity Therapy Development
Subsequently, we conduct an anti-obesity therapy development phase that transforms these validated targets into viable drug candidates through the processes of design, synthesis, and initial pharmacological and toxicity assessments.
Preclinical Studies of Anti-Obesity Therapeutics
Finally, the our preclinical studies of anti-obesity therapeutics phase encompasses extensive animal studies to thoroughly evaluate the efficacy, safety, and pharmacokinetics of the candidate drugs, thereby facilitating the transition to clinical trials.
Unlock Your Body's Potential with Adipose Tissue Expansion Analysis
We utilize advanced stable isotope methodology to precisely measure adipogenesis in adipose tissue expansion analysis. Specifically, we employ thymidine enriched with a high concentration of the rare stable isotope nitrogen, serving as a tracer for cell division. The stable isotopes offer significant advantages: they are non-toxic, ensuring no interference from reagent-induced artifacts, and their safe use in humans allows for seamless translation of findings from model organisms to clinical studies. To measure the incorporation of tracers in adipose tissue components, we utilize isotope ratio mass spectrometry (IRMS). Additionally, for precise and detailed visualization of stable isotopes within histological sections, we employ multi-isotope imaging mass spectrometry (MIMS). MIMS combines precise tracer measurement with imaging resolution comparable to transmission electron microscopy, enabling detailed analysis of adipogenesis dynamics. We analyze these processes during aging and in obesity models, focusing on quantifying adipocyte turnover as a key factor in understanding the pathophysiology of obesity and its expansion mechanisms.
Workflow
Animal Housing and Care
We house the mice at a controlled temperature, with a consistent light and dark cycle, and provide access to food and water. Aging studies and those involving specific genetic models are conducted with a diet that is balanced in protein, fat, and carbohydrates. For diet-induced obesity studies, we use high-fat and control chow diets matched for sucrose content.
Insulin and Glucose Tolerance Tests
We perform insulin tolerance tests by injecting a dose of regular insulin intraperitoneally after a short period of fasting. For glucose tolerance tests, we inject glucose intraperitoneally after an overnight fast. Blood glucose levels are measured using a glucometer, and serum insulin levels are quantified using an ELISA kit.
Stable Isotope Labeling
We dissolve 15N-thymidine in water and administer it to mice by subcutaneous injection. For long-term labeling, we implant subcutaneous osmotic minipumps that deliver a continuous dose. For shorter-term labeling in diet-induced obesity mice, we use subcutaneous injections.
MIMS Analysis
We fix the tissue with paraformaldehyde, embed it in LR white, and section it. The sections are mounted on silicon chips and analyzed using the nanometer-scale secondary ion mass spectrometer. Each tile is acquired at a high resolution, and nuclei are manually traced using a customized plugin. We assign an identity to each nucleus and reimage cells with unclear features at higher resolution for reevaluation.
IRMS Analysis
We mince adipose tissue and digest it with collagenase in a shaking water bath to dissociate cells. The digest is filtered and centrifuged to separate adipocytes, which are then washed. Stromal vascular cells are isolated by straining and washing. For subfraction analysis, antibody-coated microbeads are used for positive or negative selection. Cells are aliquoted, dried, and genomic DNA is isolated. Samples are analyzed using an elemental analyzer coupled to IRMS, with unlabeled mice serving as the natural background, and results are expressed as a percentage above the natural ratio.
Histology Analysis
We fix adipose tissue specimens with paraformaldehyde, embed them in paraffin, section them, and stain them using standard methods. Cross-sectional areas of adipocytes are calculated by tracing the adipocyte periphery. For immunohistochemistry, we perform antigen retrieval by heating samples in a citrate-based buffer. We use a rat monoclonal primary antibody to murine F4/80 and detect it with a biotinylated goat anti-rat IgG secondary antibody. We identify and count "crown-like structures" using a blinded observer.
Statistical Analysis
We perform statistical analyses using JMP 10.0 and Prism 6.0. We use the Student's t-test for comparing two independent, normally distributed groups and the Mann-Whitney test for non-normally distributed groups. For multiple non-normally distributed groups, we use the Kruskal-Wallis test with Dunn's multiple comparison correction. Proportions are compared using the χ2 test. A p-value less than 0.05 on a two-tailed test indicates significance.
Workflow
Applications
- Our service can be used to explore how the expandability of subcutaneous adipose tissue influences metabolic health and the development of obesity-related complications.
- Our service can be used to track changes in adipose tissue volume during weight loss or gain interventions, offering a quantitative measure of treatment effectiveness.
- Our service can be used in studying the physiological and pathological changes that occur in adipose tissue during obesity, such as inflammation and metabolic dysfunction.
Advantages
- We utilize cutting-edge imaging technologies such as MIMS and IRMS for high-resolution visualization and precise measurement of adipogenesis.
- We conduct studies on aging, specific genetic models, and diet-induced obesity, offering insights into various physiological and pathological conditions.
- We integrate molecular biology and bioinformatics to explore the mechanisms underlying white adipose tissue expansion comprehensively.
Publication Data
DOI: 10.1371/journal.pone.0080274
Journal: Frontiers in Cell and Developmental Biology
Published: 2023
IF: 4.6
Result: In this piece, the author delves into the processes by which white adipose tissue grows, which can happen through two main ways: hypertrophy, where the fat cells get larger, and hyperplasia, where the number of fat cells increases. The article emphasizes the crucial role of this tissue expansion in preserving overall metabolic well-being. The article highlights that impaired white adipose tissue expansion in obesity leads to lipid deposition in non-adipose organs, causing metabolic dysfunction. While hyperplasia has been traditionally viewed as essential for healthy white adipose tissue expansion, recent studies question the role of adipogenesis in the transition from impaired white adipose tissue expansion to metabolic health issues. The review summarizes recent findings and emerging concepts on white adipose tissue dynamics, emphasizing their significance in obesity, health, and disease.
Frequently Asked Questions
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What are the functions of Adipose?
As a pivotal element in maintaining whole-body energy homeostasis, adipose tissue has been sculpted by an ancestral recollection of environmental influences and both lifelong and short-term nutritional statuses. Its primary function is to ensure a consistent supply of energy to vital organs, particularly the brain. This tissue possesses the capacity to absorb and store essential nutrients, including glucose, which it converts through lipogenesis, and fatty acids (FAs). Additionally, it serves as a repository for structural molecules such as cholesterol.
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In which other periods can hyperplasia of adipose tissue occur?
- Last half of pregnancy
- The first year of life
- The beginning of puberty
Protheragen is dedicated to delving into the intricate mechanisms of adipose tissue expansion and its interplay with metabolic health. Our scope encompasses foundational research on the expansion of white adipose tissue under diverse physiological conditions and an exploration of sex-based disparities in regional white adipose tissue distribution. contact us to known more about our Obesity Pathobiology Research Service if you want to research how these differences influence the susceptibility to obesity and associated metabolic disorders.
Reference
- White, U. Adipose tissue expansion in obesity, health, and disease. Frontiers in Cell and Developmental Biology. 2023, 11: 1188844. (CC BY)
All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.