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FIG. 6. Indirect immunofluorescence of MAb-A5 in the mesencephalic region in development. (a) A section of the rostra1 part of the optic
tectum at stage 40 (X90). (b) A section caudal to the section in (a). (c) A section of the rostra1 part of the optic tectum at stage 45 (X70). (d)
A section of the caudal part of the optic tectum at stage 45. Arrows indicate the stained regions. |
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FIG. 4. Whole mount of the diencephalon and mesencephalon of tadpoles
(stage 51). (a) Retinal central pathways in a Xenopus tadpole.
All retina1 axons from the right eye were labeled with HRP to show
the projection to the visual centers, i.e., the optic tectum (OT), nucleus
Belonci (NB), corpus geniculatum thalamicum (CGT), and basal optic
nucleus (BON). OC: optic chiasm. For the method of HRP labeling,
see Fujisawa et ul. (1981). (b) The tadpole brain stained with MAb-A5
by the ABC method. The optic tectum, the nucleus Belonci, and the
corpus geniculatum thalamicum are positively stained. The staining
of the basal optic nucleus is obscure. (c) The tadpole brain stained
with MAb-B2 by the ABC method. All visual centers are negative
(X40). |
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FIG. 5. Indirect immunofluorescent analysis of the expression of A5-antigen in the optic tectum of eye-enucleated tadpoles (stage 51) in
which the right eye was enucleated at stage 29. The left tecta (*) was not innervated by optic nerves (X175). |
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Fig.1 Indirect immunofluorescence of MAb-45 and MAb-B2 on cryostat sections of the optic tectum of Xenopus tadpoles (stage 53). (a) A coronal section of the mesencephalon stained with MAb-A5. Only the outer layers of the optic tectum (OT) show the fluorescence (arrow),
whereas the tegmentum (TG) is negative (X100). (b) A section similar to (a) stained with MAb-BX. Both the optic tectum and tegmentum are
stained. (c) Immunofluorescence with MAb-A5 in the optic tectum is restricted to the 8th and 9th plexiform layers (X160). (d) The section next
to (c) is stained with MAb-B2. The 3rd, 5th, and 7th plexiform layers of the optic tectum show immunofluorescence with MAb-B2, but the 8th
and 9th layers are practically negative. (e) The same section as (d) viewed with bright-field optics. The numbers indicate the 3rd, 5th, 7th, 8th,
and 9th plexiform layers of the optic tectum, respectively. |
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FIG. 2. Indirect immunofluorescence of MAb-A5 and MAb-B2 on cryostat sections of the diencephalon of Xenopus tadpoles (stage 53). (a) A coronal section stained with MAb-A5. The nucleus Belonci shows immunofluorescence (arrow) (X130). (b) The section next to (a) stained with
MAb-B2. The nucleus Belonci is negative. (c) A coronal section of the diencephalic region that is located rostra1 to the region shown in (a).
The corpus geniculatum thalamicum is stained with MAb-A5 (arrow). (d) The section next to (c) stained with MAb-B2. The corpus geniculatum
thalamicum is negative. |
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FIG. 3. Indirect immunofluorescence of MAb-A5 and MAb-B2 on cryostat sections of the mesencephalic region, located rostrally to the
sections shown in Fig. 1. (a) A coronal section stained with MAb-A5. The pretectum shows immunofluorescence (arrow). A small region
corresponding to the basal optic nucleus shows the faint fluorescence at the ventrolateral part (*) (X110). (b) The section next to (a) stained
with MAb-B2. The pretectum and the basal optic nucleus are negative. (c-e) Enhanced image showing localization of MAb-A5 binding in a
ventrolateral part of the mesencephalon processed by a color image analyzer: (c) integrated image (128 times) (X250), (d) specific object image
extracted as the highest area which corresponds to the basal optic nucleus, and (e) image in (d) superimposed on (c). |
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FIG. 7. Immunofluorescent staining of viable tadpole brains with MAb-A5 (a) and MAb-B2 (b) (stage 53). Numbers indicate the 3rd, 5th, 7th,
8th. and 9th plexiform layers of the optic tectum (X140) |
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FIG. 8. MAb-A5 indirect immunofluorescent staining of trypsin-digested optic tectum (stage 53). (a) Intact optic tectum was treated with
0.05% trgpsin for 2 hr and then fixed and stained with MAb-A5 (b) Control nontrypsinized optic tectum stained with MAb-A5 (X100). |