PSYCH 4644: FINAL REVIEW Flashcards
Berthold’s Experiment
Transplanted testes developed blood connections and became functional (i.e. produced sperm )
Concluded that the testes produced a blood-born product that affects anatomy and behavior (since nerve connections severed)
1849: A. A. Berthold performed first formal endocrinology experiment
Demonstrated non-neural contribution by the testes required for normal development of a rooster
Prepubertal castration of male chick was used to make capons for more tender meat.
Berthold wanted to know if these extensive effects were dependent on neural connection to the testes.
• Transplanted testes were fully functional
• Birds with transplanted testes were entirely normal
– Normal appearance
• Combs, wattles, plumage
– Normal behavior
• Normal vocalizing
• Normal aggression
• Normal mating

• Hypothesized secretory product carried by blood to target tissues
– Learned later to be testosterone
Techniques in
Behavioral Endocrinology
Assays: measure hormone levels.
Bioassays
RIA, EIA, ELISA
Ablation and replacement
Lesion
Electrical stimulation
Optogenetics
Electrophysiological recording
Pharmacological Techniques: agonists & antagonists
Immunocytochemistry
Immediate early genes
Autoradiography
Western blot
In situ hybridization
Genetic manipulations
Histology
Brain Imaging
Endocrinology Technique #1:
Ablation & Replacement
This method is like that employed by Berthold.
It is the removal (ablation) of a suspected hormone source or by replacement via implantation or by injection of suspected hormonal chemicals
Technique #2:
Bioassays
The use of a biological response to determine the PRESENCE or AMOUNT of a particular substance in a sample.
Identifies the chemical processes in a hormone’s actions.
Involves testing the hormone on a living animal or cell culture.
Bioassays are often usefully conducted on alternate species from which a hormone has been derived.
Bioassay for Prolactin:
In this case, the cell height of the crop sac is going to differentiate dependent upon the amount of prolactin injected.
Technique #3: Immunoassays
The most famous form is the radioimmunoassay (RIA) and it is a technique that was first able to measure hormones PRECISELY.
Rosalyn Yalow received the Nobel Prize in for her work on this technique.
Radioimmunoassays
Type of Immunoassay.
- Antibody to a Hormone and the Labeled Hormone are mixed in solution.
- The Labeled Hormone binds with the Antibody.
- Unlabeled Hormone is added.
- The Unlabeled Hormone and Labeled Hormone compete for binding sites.
- The percentage of bound Labeled Hormone is measured and a standard curve is developed.
- The standard curve is used to determine the amount of Unlabeled Hormone in a given sample.
Technique #4: Immunocytochemistry
A technique that it uses antibodies to determine the LOCATION of hormones or hormones receptors.
Often, these antibodies are attached to a fluorescent dye for viewing under a fluorescent microscope.
Technique #5
Autoradiography
Radiolabeled hormones are injected into a tissue or animal and the sections taken will reveal sites of absorption and radioactivity.
A technique used to detect a radiolabeled substance, such as a hormone, in a cell or organism, by placing a thin slice of the material in contact with a photographic emulsion, which displays darkened silver grains in response to the radioactive emissions.
Technique #6
Blot Tests
A cellular technique that is used to discern if a particular protein or nucleic acid is found in a given tissue.
The various forms of these techniques involve electrophoresis of homogenates of tissues from which proteins are extracted.
Electrophoresis will separate these proteins on the basis of electrical charge.
Technique #7
“in-Situ” Hybridization
A technique that will allow examination of gene expression in cells, tissues, or organs are identified for a specific protein.
Technique #8
Stimulation Followed With Recordings
Direct electrical stimulation of neuronal pools (brain, spinal cord, or peripheral nerves) can elicit electrical activity in cells and tissues of neurall relevant sites.
Rat with electrodes implanted in the brain to allow stimulation of specific regions of the hypothalamus (a major endocrine secreting region).
Technique #9
Pharmacological Techniques
The use of chemical agents that may alter the action or function of a hormone.
The 2 major groups are:
AGONISTS – hormone mimics
ANTAGONISTS – hormone blockers
These compounds can be administered to an animal in a variety of ways, but the two most common forms are:
CANNULATION – a form of permanent form of injection tube is implanted in a target region. Used typically for multiple administrations of a pharmacological agent.
SIMPLE INJECTION – using syringes to administer a single dose of a pharmacological agent
Technique #10
Anastomosis
An older technique whereby the blood systems of two different animals is interconnected to study the endocrine systems of the two animals.
Technique #11
Microdialysis
Much like kidney dialysis, it is a technique that allows the administration of very minute quantities of neurotransmitters, hormones and/or other pharmacological agents into a conscious animal.
But the experimental benefit is that samples (chemical or electrical) from the site are also possible.
Technique #12
Brain Imaging
Scanning techiques of various forms that are used to remotely monitor and test the activity of body structure function, esp. that of the brain.
There are several types of these devices including:
Positron Emission Tomography: (PET): will show functioning of brain regions in real time.
Computer Assisted Tomography (CT) – uses x-rays to give a 3-D image of the brain within a particular plane of space.
Magnetic Resonance Imaging (MRI) - using non-ionizing radiation energy to see images similar to CT scans.
Technique #13
Genetic Manipulations
Engineering knockout mice
Technique #13
Microarrays
This tool is capable of showing the expression (activity) of genes within an animal’s genome.
AKA, “Gene Arrays”
Leptin Example:
Both mice have a defective “ob” gene, resulting in development of obesity.
The normal-weight mouse was given daily injections of the leptin protein, the protein that the “ob” gene encodes, to rescue the effects of its mutation.
Histology!!!
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Optogenetics!!!
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WHAT ARE HORMONES ?
Chemicals produced and released in very small amount by ENDOCRINE glands into the bloodstream
Hormones are not released constantly but instead are released in spurts (pulsatile secretion)
Travel some distance to target organs or tissues
Target cells have specific receptors
Regulates cellular events that lead to activation of enzymatic pathways or to effects on gene expression and protein synthesis
Principal actions of hormones:
Metabolism
Growth
Developmental Processes
Behavior
Hormonal Communication
- Uses hormones
- Released into circulatory system
- Travels long distances (1mm-2m)
- Can travel anywhere in the body
- Slow, graded
- Mediate long-term processes
- Little voluntary control
Think blood
Neural Communication
- Uses neurotransmitters
- Released locally into synapse
- Travels short distances (20-30 nm)
- Can travel only along neural tracts
- Fast, all or none
- Mediates fast changes
- Some voluntary control
Think action potentials..
Endocrine system
Consists of endocrine glands which have cells that release chemical messages (i.e. hormones) into the blood stream
Endocrine effects as contrasted with:
- Autocrine
- Paracrine
Not mutually exclusive:
A hormone can act in an autocrine, paracrine, and endocrine fashion.