from notes posted ....
The Blood-Brain Barrier
Evolutionarily selective pressure and advantage to developing a barrier came from the need to preserve chemical homeostasis from any fluctuations around the synaptic active zones.
BBB acts as a physical, metabolic, and transport barrier restricting traffic of nutrients and other molecules (through epithelial cells, astrocytes)
Posses challenges to drug therapy, as to how to transport drugs past this barrier
-> but lipids can still pass-through (like 1st gen antihistamines for allergies, they pass-through so it makes you dizzy)
Posses challenges to drug therapy, as to how to transport drugs past this barrier
-> but lipids can still pass-through (like 1st gen antihistamines for allergies, they pass-through so it makes you dizzy)
stroke and inflammation may cause BBB to break open and cause harm
In situ hybridization
The cDNA can also be used to determine which cells, produce a particular mRNA using in situ hybridization. Labeled nucleic acid is incubated with fixed tissue or cells under conditions where only specifically bound hybrid is stable. Auto radiography reveals the position of endogenous RNA. Controls with RNase help prove that hybridization is to RNA not genetic material.
Immunochemistry
The visualization of antigens in their normal cellular and tissue
environment.The primary antibody, which detects the antigen of interest
is generally detected by a secondary antibody, which is linked to an
enzyme (e.g., horseradish peroxidase or alkaline phosphatase), a
chromogen (e.g., FITC, Fast Red, etc.), or, in the case of electron
microscopy, an electron dense material (a gold particle).
Knock-out, Knock-in, Transgenics
Knock-out: a procedure to disrupt the sequence of a gene and
interfere with its expression.
Knock-in: a variation of gene targeting that uses homologous
recombination but allows expression of altered genetic sequences
in place of the endogenous gene. This approach allows the test of
more subtle mutations than is allowed by a simple knock out.
Transgenics: Introduction of a gene under a particular promoter
into the germ line allows propagation of an organism that will
express or even conditionally express a particular gene. The
transgene is expressed in addition to the normal gene.
Molecular Biology of the Cell. Alberts et
al.
Chapter:
Manipulating Proteins, DNA, and RNA
(Chapter 8 in 4th or 5th Ed.)
or
Recombinant DNA Technology
(Chapter 7 in 3rd Ed.)
In situ hybridization
The cDNA can also be used to determine which cells, produce a particular mRNA using in situ hybridization. Labeled nucleic acid is incubated with fixed tissue or cells under conditions where only specifically bound hybrid is stable. Auto radiography reveals the position of endogenous RNA. Controls with RNase help prove that hybridization is to RNA not genetic material.
Immunochemistry
The visualization of antigens in their normal cellular and tissue
environment.The primary antibody, which detects the antigen of interest
is generally detected by a secondary antibody, which is linked to an
enzyme (e.g., horseradish peroxidase or alkaline phosphatase), a
chromogen (e.g., FITC, Fast Red, etc.), or, in the case of electron
microscopy, an electron dense material (a gold particle).
Knock-out, Knock-in, Transgenics
Knock-out: a procedure to disrupt the sequence of a gene and
interfere with its expression.
Knock-in: a variation of gene targeting that uses homologous
recombination but allows expression of altered genetic sequences
in place of the endogenous gene. This approach allows the test of
more subtle mutations than is allowed by a simple knock out.
Transgenics: Introduction of a gene under a particular promoter
into the germ line allows propagation of an organism that will
express or even conditionally express a particular gene. The
transgene is expressed in addition to the normal gene.
Molecular Biology of the Cell. Alberts et
al.
Chapter:
Manipulating Proteins, DNA, and RNA
(Chapter 8 in 4th or 5th Ed.)
or
Recombinant DNA Technology
(Chapter 7 in 3rd Ed.)
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