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Figure 7.2 Assay of mitochondrial function in skeletal muscle. (A) ATP is produced within
the mitochondria as a result of the activity of four major complexes (I-IV). Complexes I, II,
and IV (NADH dehydrogenase, succinate dehydrogenase, and cytochrome c oxidase) are
most often dysregulated in mitochondrial disease. (B) Mitochondrial enzymatic function
is commonly assayed on skeletal muscle sections via colorimetric reactions, which
are imaged in bright field. Images are reproduced with permission from the following
sources; NADH-dehydrogenase (
http://csip.cornell.edu/Curriculum_Resources/CEIRP/
cellular_resp.html
,
Cornell University), succinate dehydrogenase (
Kostrominova, 2010
),
and cytochrome
photographed by Dr. Mark Cohen, Case Western University).
c
muscle fibers. Additionally,methodsweredeveloped inwhich the samemuscle
section could be sequentially stained for multiple enzymatic activities. Exam-
ples include staining a muscle section for both SDHand “reversed” ATP activ-
ities (
Horak, 1983
) and successive histochemical staining of the same section of
muscle for mATPase, SDH, and hematoxylin (
Solomon and Dunn, 1988
).
Relatedly,
Hawcroft et al. (1992)
described techniques of SDH staining of
tissue prior paraffin embedding and sectioning. The sequential evaluation of
SDH/COX activities in the same muscle section is currently a predominant
method for histological evaluation of mitochondrial dysfunction (discussed
in
Section 7.1
)(
Barron et al., 2005; Greaves et al., 2010; Sundaram et al.,
2011; Taivassalo et al., 2012; Vladutiu and Heffner, 2000
). Histochemical
staining for SDH was also used to correlate capillary architecture in slow and
fast muscles with SDH activity (
Murakami et al., 2010
).
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