Whole mounted hindbrains (A-D”) and transverse hindbrain sections
(A’-C’, E-H) from e11.5 embryos of indicated genotypes. Discrete
sensory afferent projections were visualized by labeling distinct cranial nerves
(indicated by Roman numbers) with lipophilic dyes. V - trigeminal nerve, dV -
descending trigeminal tract, VII - facial nerve, VIII – inner ear
vestibular nerve, IX - glossopharyngeal nerve, X - vagus nerve, XII -
hypoglossal nerve. (A-C, A’-C’) In control, Lmx1a
KO, and Lmx1b KO embryos, different afferent projections run
parallel to the anterior-posterior axis of the hindbrain, do not significantly
overlap with each other, and do not cross the dorsal midline occupied by the
wide choroid plexus epithelium derived from the IVth ventricle roof
plate.
(D-D”) Dorsal whole mount view showing that in
Lmx1a/b DKO embryos, inner ear vestibular (VIII),
trigeminal (V), and solitary tract (ST) afferents extend aberrantly close to or
cross the dorsal midline. Arrow (D-D”) shows location of the dorsal
midline, which is heavily populated by inner ear vestibular (VIII) afferent
fibers. D’ and D” are high power views of D.
(A’-C’, E-H) Transverse hindbrain sections showing that
similar to control, Lmx1a KO and Lmx1b KO
embryos, in Lmx1a/b DKO hindbrain, the descending trigeminal
tract (dV) was located ventral to inner ear vestibular (VIII) fibers. However,
in contrast to control, Lmx1a KO and Lmx1b KO
embryos (A’-C’), inner ear vestibular (VIII) fibers extended to
and populated the dorsal midline roof plate (RP) covering the central canal in
Lmx1a/b DKO littermates (E-H). Note the near normal
location of facial branchial motor neurons (FBM) and ventral motor neurons (Vm)
in the Lmx1a/b DKO (E, F) near the floor plate (FP), and
solitary tract afferents crossing the roof plate (G). G is a high power view of
E, without the magenta channel; panels F and H show two different sections from
another embryo.
Scale bar: 100 μm.