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Fig. 3. MHB abnormalities in En1cre/+;
Otx2flox/flox embryos. (A-W) In situ hybridisation on E9.5
and E10.5 En1cre/+ and En1cre/+;
Otx2flox/flox embryos with Otx2 (A,I,Q),
Fgf8 (B,J,R), Gbx2 (C,K,S), Wnt1 (D,L,T),
Otx1 (E,M,U), En1 (F,N,V), Otx2-5' (G,O,W)
and Pax6 (H,P) probes. (X) Schematic representation summarising MHB
abnormalities detected in En1cre/+;
Otx2flox/flox embryos. The parasagittal section in Q-W is
focussed on the ventral midbrain of conditional mutants and approximately
corresponds to the area included in the dotted square in J. In conditional
mutant embryos, the arrows in A,C-H,I,K-Q,S-W indicate the corresponding
position of the ventralmost expression of Fgf8, while the arrows in
B,J,R indicate the corresponding position of the ventral and posterior border
of the functional Otx2 domain (Otx2). The curved
arrows in J,K,N,X indicates the dorsal and anterior rotation of the MHB whose
ventral site remains in a fairly normal position; the arrowheads in
A,B,E,F,I,J,M,N indicate the posterior border of Pax6; and the
arrowheads in Q,U,W indicate the ventral midbrain area lacking functional
Otx2 transcripts (Otx2) but still transcribing
Otx1 and Otx2 (Otx2-5'). (X) Red, green,
orange and yellow correspond to the hindbrain, posterior midbrain, anterior
midbrain and forebrain, respectively. MHB, midbrain-hindbrain boundary; Pt,
pretectum; Fb, forebrain; Mb, midbrain; Hb, hindbrain.
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and 5-HT (L,R,ß) antibodies and with Otx2
)
Schematic representation summarising the expression pattern of Shh, Nkx6.1 and
Nkx2.2 and neuronal cell types detected in control and mutant embryos. The
broken lines in A indicate the approximate position of frontal (G-L) and
horizontal (S-V, M-R) sections; the position of frontal sections of the triple
mutant (W-ß) are at a level similar to that of the conditional mutant,
but in the triple mutant this position is posterior to Fgf8
expression; the arrowheads in B-E and the arrows in S,U,V indicate the
corresponding position of Fgf8 expression and the arrows in T
indicate the posterior border of functional Otx2 domain; the inset in
H,N,X highlights the ventral expression of Nkx2.2; the bracket demarcates the
neuroepithelium expressing Nkx2.2 and generating Ser neurons in the midbrain
of mutant embryos (H,L,X,ß) and in the rostral hindbrain of control
embryos (N,R). OM oculomotor nucleus; RN, red nucleus; Fb, forebrain; Mb,
midbrain; Hb, hindbrain; TN, trochlear nucleus; OMn, oculomotor nerve; e, eye;
DA, dopaminergic cells; Ser, serotonergic cells.