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Fig. 3. Absence of most skeletal muscles in
Six1–/–Six4–/–
E18.5 fetuses (A-L) and in
Six1–/–Six4–/–
E13.5 embryos (M-V). (A,C,E,G,I,K,M,O,Q,S,U)
Six1–/+Six4–/+ animals;
(B,D,F,H,J,L,N,P,R,T,V)
Six1–/–Six4–/–
animals. General view of E18.5 heterozygous (A) and double dKO (B) fetuses, as
revealed by X-gal staining. Skeletal muscles are absent from the limbs,
abdominal ventral muscles are missing and, at the shoulder and hip levels,
most muscles are lacking. X-gal Eosin staining on transversal sections (C-H)
or immunostaining performed with fast-type myosin heavy chain antibody My32
(I-L) confirmed the general axial muscle hypoplasia and that limbs have no
muscle. (C,D) Shoulder level, with the humerus (hu), thymus (thy) and vertebra
(v) indicated. Most muscles are lacking in the dKO fetuses, except epaxial
muscles, which appear incorrectly shaped, near the vertebral column. (E,F) Hip
level revealing deep back muscles in the control and dKO fetus. (G,H) Hip
level revealing that proximal muscles around the femur (fe) are lacking in the
dKO fetus, and that ventral-most muscles are also lacking (see enlargement of
the boxed ventral region. int, intestine. (I,J) Forelimb and (K,L) hindlimb
levels, revealing the absence of muscles in the dKO fetuses. r, radius; u,
ulna; t, tibia; f, fibula. General view of E13.5 heterozygous (M) and dKO (N)
fetuses, as revealed by X-gal staining. Abdominal ventral muscles are missing
and, at the shoulder level, most muscles are lacking. Transverse sections
(O-V) revealing desmin expression after immunostaining. (O,P) Eye level,
showing that most muscles are absent in the dKO embryo. Eo, extraocular
muscles; pt, pterigoid muscles; ca, capitalis muscles; nt, neural tube (Q,R)
Tongue level, revealing absence of tongue muscle (to) in the dKO embryo. Ma,
masseter. (S,T) Forelimb level, revealing absence of any desmin-positive cells
in the forelimb of dKO embryo. fl, forelimb; he, heart; db, deep back muscles;
st, spinotrapezius; cm, cutaneus maximus; ii, intercostals interni; bb, biceps
brachii, pe, pectoralis; tt, transversus thoracis. (U,V) Diaphragm level,
showing that ventral muscles and diaphragm are lacking in dKO embryos. li,
liver; dia, diaphragm; sm, scalenus medius; ie, intercostales externi; ra,
rectus abdominis; ta, transversus abdominis.
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