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Fig. 3. The esophagi of Sulf1-/-;Sulf2-/- mice
have normal skeletal muscle function, but impaired smooth muscle
contractility. (A) Postnatal development of the esophageal skeletal
muscles in control and Sulf1-/-;Sulf2-/- pups.
The cross-sections of Sulf1+/-;Sulf2+/- control
and Sulf1-/-;Sulf2-/- mice were immunostained
with antibodies against skeletal muscle markers including myosin heavy chain
(MHC) and MYF5 or incubated with Cy3-conjugated alpha-bungarotoxin (alpha-BTx)
to identify acetylcholine (ACh) receptors on muscle. The completion of the
esophageal skeletal muscle formation at P15 was assayed by immunostaining of
the abdominal segments with an alkaline phosphatase-conjugated mouse antibody
against fast skeletal myosin (sk-Myosin). The antigen-antibody complex was
visualized using the substrate BM purple. Arrowheads mark the junction between
the esophagus and the stomach. The
Sulf1-/-;Sulf2-/- esophagi have completed
skeletal muscle formation in the esophagus at P15. Scale bars: 100 µm.
(B) Physiological measurements of the esophageal skeletal muscles. The
lower-half thoracic segments of the esophagi of the adult
Sulf1+/-;Sulf2+/- control and
Sulf1-/-;Sulf2-/- mice were subject to twitch
and tetanus stimuli. Muscle contractility was measured and compared with those
in the presence of selective ion-channel blockers (n=3). The
Sulf1-/-;Sulf2-/- esophagi exhibited comparable
skeletal muscle contractility in response to the electrical stimuli and
ion-channel blocks as the control esophagi. (C) Physiological tests of
esophageal smooth muscle contractility. The smooth muscle of the control and
the Sulf1-/-;Sulf2-/- mutant esophagi were
dissected and their contractile forces in response to various stimuli measured
(n=3). The Sulf1-/-;Sulf2-/- esophagi
showed diminished smooth muscle contractility in response to carbachol, and
partially reduced contractility in response to other chemicals. (D)
Quantification of the esophageal smooth muscle contractility induced by
various stimuli as shown in C.
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