Assessments of Reactive Astrogliosis Following CNS Injuries - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 53

In other words, the volume and complexity of each astrocyte is
signifi cantly underestimated by the visualization of intermediate
fi laments. Techniques to determine the expression of intermedi-
ate fi laments in CNS injury models typically utilize standardized
techniques for IHC using commercially available antibodies against
GFAP. Table 1 lists sources for the most commonly used GFAP
antibodies and detailed protocols for carrying out IHC in brain or
spinal cord tissue are described in the Materials, Methods, and
Notes sections below.
It recently has been discovered that in healthy tissue, cortical
and hippocampal astrocytes are organized into adjacent, but non-
overlapping domains and that one domain can contact greater than
100,000 synapses in rodent brain and over 1,000,000 in human
brain ( 9, 10 ). Additionally, recent studies demonstrate that under
some conditions of CNS brain injury, this “tiling” of astrocyte
processes can be lost in reactive astrogliosis ( 11 ). For example,
Nedergaard and colleagues recently utilized diolistic labeling of
fi xed brain tissue and subsequent analysis in reactive astrocytes to
demonstrate that seizure/epilepsy caused a loss of astrocytic
domain organization. Moreover, they reported that this loss of
astrocyte domains was not a universal feature of astrogliosis, as it
was not found in a mouse model of neurodegeneration even
though extensive reactive astrogliosis was observed ( 11 ). Thus, a
current hypothesis of the alteration of domain organization in
reactive astrogliosis is that in mild-moderate gliosis these distinct
domains remain, but in severe gliosis and scar formation, particu-
larly when proliferation is involved, astrocyte processes interdigi-
tate and overlaps resulting in a loss of distinct domains ( 2 ).
Techniques for diolistic labeling using a Helios Gene Gun System

Table 1
Antibodies commonly used to detect intermediate GFAP
in CNS astrocytes

Protein Antibody type Vendor

Z0334 Dako, Inc., Carpinteria, CA
http://www.dakousa.com
MAB360 Milipore, Inc., Billerica, MA
http://www.milipore.com
Ab4674 Abcam, Inc., Cambridge, MA
http://www.abcam.com
SC-6171 Santa Cruz Biotechnology, Inc.,
Santa Cruz, CA
http://www.scbt.com
A list of commonly used, commercially available antibodies against GFAP
protein is shown. Both monoclonal and polyclonal antibodies are available
that are derived from several species. The antibody number and Web URL are
provided for each vendor

GFAP Rabbit polyclonal

Mouse monoclonal

Chicken polyclonal

Goat polyclonal