Biology Reference
In-Depth Information
the EPO standard curve range. Instead, to measure the ACF
levels of EPO and S100E of rats injected with AAV systemi-
cally it is recommended to assay 2-10
L of ACF.
43. It is important to underline that in control rats the levels of
endogenous EPO (in both serum and ACF) are below the
ELISA sensitivity that corresponds to 0.33 pg/mL.
44. The rat and murine EPO proteins are similarly recognized by
the Quantikine Mouse/Rat Erythropoietin ELISA as reported
by the producer.
45. Before putting the O.C.T. block on the chuck it is important
to mark the block side that corresponds to the temporal part of
the eye. To do this, make a transverse incision in the block
using the trimming blade.
46. Make sure that the eyes are cut along the horizontal meridian.
47. The sections are progressively distributed on slides so that each
slide contains representative sections of either the first (slides
1-15) or the second half (sections 16-30) of the eye.
48. In rats the light damage is typically confined to rods and the
cell loss is mainly restricted to the outer nuclear layer (ONL)
that only contains the PR nuclei. Therefore, the measurement
of the ONL thickness or the count of rows of PR nuclei repre-
sent appropriate methods to quantify the damaging effect of
light. Another method that can be used to estimate PR cell loss
after light damage is based on the quantification of rod-specific
products (e. g. proteins involved in rod phototransduction).
49. It is important to consider that in rodents retinal light damage
results in a graded response with areas containing little or no
damage located close to more severely affected regions. Within
the superior hemisphere, light damage is particularly severe in
a region that is between 1 and 2 mm away from the optic nerve
head (
50
).
μ
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