Isolation of Intact, Whole Mouse Mammary Glands for Analysis of Extracellular Matrix Expression and Gland Morphology
Autor: | Katherine Keck, Abigail Hielscher, Christopher Thompson |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Physiology General Chemical Engineering Morphogenesis Biology General Biochemistry Genetics and Molecular Biology Extracellular matrix Mice 03 medical and health sciences Mammary Glands Animal stomatognathic system Peritoneum Biopsy medicine Animals medicine.diagnostic_test General Immunology and Microbiology General Neuroscience Tissue Processing Anatomy Immunohistochemistry Extracellular Matrix Entire gland 030104 developmental biology medicine.anatomical_structure Female Subcutaneous tissue |
Zdroj: | Journal of Visualized Experiments. |
ISSN: | 1940-087X |
Popis: | The goal of this procedure was to harvest the #4 abdominal mammary glands from female nulliparous mice in order to assess ECM expression and ductal architecture. Here, a small pocket below the skin was created using Mayo scissors, allowing separation of the glands within the subcutaneous tissue from the underlying peritoneum. Visualization of the glands was aided by the use of 3.5x-R surgical micro loupes. The pelt was inverted and pinned back allowing identification of the intact mammary fat pads. Each of the #4 abdominal glands was bluntly dissected by sliding the scalpel blade laterally between the subcutaneous layer and the glands. Immediately post-harvest, glands were placed in 10% neutral buffered formalin for subsequent tissue processing. Excision of the entire gland is advantageous because it primarily eliminates the risk of excluding important tissue-wide interactions between ductal epithelial cells and other microenvironmental cellular populations that could be missed in a partial biopsy. One drawback of the methodology is the use of serial sections from fixed tissues which limits analyses of ductal morphogenesis and protein expression to discrete locations within the gland. As such, changes in ductal architecture and protein expression in 3 dimensions (3D) is not readily obtainable. Overall, the technique is applicable to studies requiring whole intact murine mammary glands for downstream investigations such as developmental ductal morphogenesis or breast cancer. |
Databáze: | OpenAIRE |
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