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20th century was a century of __________ in Biology.
21st century is a century of ________ in Biology.
Describe Neural Circuits.
- They underlie all aspects of behavior.
- A wide variety of structures and regulatory proteins are required for a single circuit.
- Simple organisms have simple neural circuits and vice versa.
- Even subtle changes in expression of a single gene can affect behavior.
Subtle changes in the expression of a single gene can affect behavior, such as:
- Absence of expression
- changes in degree of expression
- expression of splice isoforms
- changes in expression pattern
- can overlap and take care of several functions
What is Phenylketonuria? What's it an example of?
- It is the inability to break down the AA phenylalaine.
- It is a recessive autosomal inherited trait.
- It's an example of a diseas caused by the change of a single gene.
What are some model organisms in Neural Science?
Mice, Elegans, Fruit flies, monkeys, frogs.
Journals that publish neuroscience:
- Journal of Neuroscience
- Nature Neuroscience
- Behavioral Neuroscience
Where is genetic information stored?
How many chromosomes do humans have? What kind?
46. 22 pairs of autosomes and a 2 sex chromosomes.
What do genes do?
- Regulate behavior
- Emotional regulation of behavior
Human brain and _____ brain react ______ to ________ stimuli in the ______ (region of the brain).
Mice ; Similar ; fear ; amygdala
Two major genetic approaches for studying specific DNA sequences
- 1. Specific amplification (DNA cloning)
- 2. Specific detection
Cloning of PCR products. (Types)
- Blunt-end cloning
- TA cloning (A- over hang)
- incorporation of restriction sites in PCR oligoprimers
Classical methods to detect and analyze gene transcription products
- Northern blotting
- cDNA library
- RNA in situ hybridization
- + Quantitative. Size. Integrity. Amount
- -Not sensitive
- Beneficial because you can isolate small amounts of RNA.
- +Quantitative. Very Sensitive.
- -Need to know sequence by designing primers.
pros and cons of RNA in situ hybridization
- +Allows to analyze where a gene of interest is expressed in the brain. Can be semi-quantitive if radioactive.
- -Not always possible.
- Makes proteins.
- Use antisense probe.
- Doesn't make protein.
- Use sense probe.
Steps of RNA in situ hybridization
- 1. Cut brain sections and place on glass slides using cryostat at -25oC
- 2.DNA or RNA labeling. DNA-oligo, Use 33P or 35S. RNA- Use dioxigenin-UTP. 33P or 35S.
- 3.Put on the brain slice and let it hybridize over night at 60oC. Then wash away probe.
- 4. Observe with radioactive x-ray film.
Why can't 32P be used for in situ?
- It is a very high energy isotope, too much for in situ.
- Can use for Northern and Southern blotting.
Radioactive oligo in situ hybridization
- DNA is labeled using 33P or 35S radioactive dNTP transferase enzyme.
- Oligo is applied to the brain sections in a hybridized buffer over night.
NeuN is a ___________ in ____ neurons.
- protein ; all.
- Serves as a marker.
Visualization of gene expression in vitro and in vivo uses
- GFP (green fluorescent protein)
start at KO mice "methods to create a genetically modified animal..."