مورد إلكتروني
Mapping Metabolism: Monitoring Lactate Dehydrogenase Activity Directly in Tissue.
| العنوان: | Mapping Metabolism: Monitoring Lactate Dehydrogenase Activity Directly in Tissue. |
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| المؤلفون: | Jelinek, David |
| المصدر: | Journal of visualized experiments : JoVE; vol 2018, iss 136; 1940-087X |
| بيانات النشر: | eScholarship, University of California 2018-06-21 |
| تفاصيل مُضافة: | Jelinek, David Flores, Aimee Uebelhoer, Melanie Pasque, Vincent Plath, Kathrin Iruela-Arispe, M Luisa Christofk, Heather R Lowry, William E Coller, Hilary A |
| نوع الوثيقة: | Electronic Resource |
| مستخلص: | Mapping enzymatic activity in space and time is critical for understanding the molecular basis of cell behavior in normal tissue and disease. In situ metabolic activity assays can provide information about the spatial distribution of metabolic activity within a tissue. We provide here a detailed protocol for monitoring the activity of the enzyme lactate dehydrogenase directly in tissue samples. Lactate dehydrogenase is an important determinant of whether consumed glucose will be converted to energy via aerobic or anaerobic glycolysis. A solution containing lactate and NAD is provided to a frozen tissue section. Cells with high lactate dehydrogenase activity will convert the provided lactate to pyruvate, while simultaneously converting provided nicotinamide adenine dinucleotide (NAD) to NADH and a proton, which can be detected based on the reduction of nitrotetrazolium blue to formazan, which is visualized as a blue precipitate. We describe a detailed protocol for monitoring lactate dehydrogenase activity in mouse skin. Applying this protocol, we found that lactate dehydrogenase activity is high in the quiescent hair follicle stem cells within the skin. Applying the protocol to cultured mouse embryonic stem cells revealed higher staining in cultured embryonic stem cells than mouse embryonic fibroblasts. Analysis of freshly isolated mouse aorta revealed staining in smooth muscle cells perpendicular to the aorta. The methodology provided can be used to spatially map the activity of enzymes that generate a proton in frozen or fresh tissue. |
| مصطلحات الفهرس: | Animals, Mice, L-Lactate Dehydrogenase, Biochemistry, Issue 136, Lactate dehydrogenase, in situ, enzymatic activity, mouse skin, metabolism, metabolic map, Stem Cell Research, Biochemistry and Cell Biology, Psychology, Cognitive Sciences, article |
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| الإتاحة: | Open access content. Open access content public |
| ملاحظة: | Journal of visualized experiments : JoVE vol 2018, iss 136 1940-087X |
| أرقام أخرى: | CDLER oai:escholarship.org:ark:/13030/qt4kx9787c qt4kx9787c https://escholarship.org/uc/item/4kx9787cTest https://escholarship.orgTest/ 1287292343 |
| المصدر المساهم: | UC MASS DIGITIZATION From OAIster®, provided by the OCLC Cooperative. |
| رقم الانضمام: | edsoai.on1287292343 |
| قاعدة البيانات: | OAIster |
| الوصف غير متاح. |