Experimental Methods in Endocrinology Flashcards
What should students be able to explain by the end of the lecture?
The different endocrine methodologies that enable quantitative and qualitative measurement of hormones and/or receptors
What are the basic procedures in endocrinology methods?
Understand the value of functional assays in endocrine research and diagnostics
What information can be gained from histological studies?
Cytological characteristics of endocrine tissue
What are the characteristics of hypertrophic cells?
Enlarged cells
What does atrophic refer to in cell characteristics?
Diminished cells
What can significant changes in organ or cell size indicate?
Clues to an underlying pathophysiology
What does histological staining reveal about cells?
Basic components of the cell
What does Hematoxylin stain interact with?
Acidic parts (phosphoric acid of DNA + RNA)
What does Eosin stain interact with?
Basic parts (cytoplasm)
What is the purpose of immunohistochemistry?
Detect and label specific antigens using specific antibodies
What type of molecules can be characterized using immunohistochemistry?
- Hormones
- Receptors
- Cell signalling molecules
- Transcription factors
- Other proteins
How are antibodies prepared for immunohistochemistry?
By injecting hormones or receptors obtained from one species into another species
What is the principle behind radioimmunoassay (RIA)?
Detection and quantification of low concentrations of hormones in blood or other fluids
What does the radioimmunoassay procedure depend on?
Binding of the specific antibody to the radiolabeled antigen (hormone)
What is gene expression?
The process by which a gene gets turned on in a cell to make RNA and protein
What can hormones induce in target tissue?
Changes in levels of mRNA or protein
What is the role of Polymerase Chain Reaction (PCR)?
Measure gene expression by amplifying specific DNA sequences
What does In Situ Hybridization localize?
Nucleotides (DNA and RNA) within cells/tissues
What do functional assays measure?
Dynamic measurements of hormone secretion
What do stimulation tests determine?
The capacity of the target gland to respond to its control mechanism
What do suppression tests determine?
Whether the negative feedback mechanisms that control a hormone’s release are intact
What is the definition of transgenic animals?
Animals that contain exogenous genes or gene modifications
What are the potential benefits of genetic engineering in endocrinology?
- Modify hormone responses
- Affect performance, growth, or health of production animals
- Disease-resistance genes
- Genes for better feed utilization
What is the effect of inserting GH genes from rats into mice?
Production of an exceptionally large phenotype
True or False: Upregulation of RNA always means upregulation of protein production.
False
Fill in the blank: The principle of radioimmunoassay involves the competition between unlabeled hormone and __________ for binding to the antibody.
[radiolabeled hormone]
