This track provides a link into the
Allen Brain Atlas (ABA)
images for this probe. The ABA is an extensive
database of high resolution in-situ hybridization images of adult
male mouse brains covering the majority of genes.
The ABA created a platform for high-throughput in situ hybridization
(ISH) that allows a highly systematic approach to analyzing gene expression in
the brain. ISH is a technique that allows the cellular localization of mRNA
transcripts for specific genes. Labeled antisense probes, specific to a
particular gene, are hybridized to cellular (sense) transcripts and subsequent
detection of the bound probe produces specific labeling in those cells
expressing the particular gene. This method involves tagged nucleotides
detected by colorimetric methods.
The platform used for the ABA utilizes this non-isotopic approach, with
digoxigenin-labeled nucleotides incorporated into a riboprobe produced by in
vitro transcription. This method produces a label that fills the cell body,
in contrast to autoradiography that produces scattered silver grains surrounding
each labeled cell. To enhance the ability to detect low level expression, the
ABA has incorporated a tyramide signal amplification step into the protocol that
greatly increases sensitivity. The specific methodology is described in detail
within the ABA Data Production Processes document.
Thanks to the Allen
Institute for Brain Science in general, and Susan
Sunkin in particular, for coordinating with UCSC on this annotation.