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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
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?
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)
TA cloning (A- over hang)
incorporation of restriction sites in PCR oligoprimers
Classical methods to detect and analyze gene transcription products
RNA in situ hybridization
+ Quantitative. Size. Integrity. Amount
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.
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
2.DNA or RNA labeling. DNA-oligo, Use
RNA- Use dioxigenin-UTP.
3.Put on the brain slice and let it hybridize over night at 60
C. Then wash away probe.
4. Observe with radioactive x-ray film.
P 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
S 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..."