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Abstract
A neural-specific beta-tubulin mRNA is expressed in the developing central nervous system shown by whole-mount in situ hybridization experiments. Of special interest is the fact that from the late blastula (stage 9; Nieuwkoop and Faber, 1967; Hausen and Ribesell, 1991) until the early neurula (stage 13) the signal can be found not only in the presumptive neural plate but also in the presumptive epidermis. Later in development (from stage 13) the specific mRNA becomes restricted to the presumptive brain and spinal cord area. The results are discussed in the context of predisposition and (pre)determination.
Fig. 1. Localization of a neural-specific mRNA by whole-mount in situ hybridization. (A) Late blastula (stage 9; Nieuwkoop and Faber. 1967). The
signal is found in the dorsal (O) and the ventral (V) ectoderm. AP animal pole, VP vegetal pole. (8) Mid-gastrula (stage 11 1/2, large yolk plug; lateral view)
The signal can be identified on the dorsal side (O) and also in the ventral (V, presumptive epidermis) and lateral parts (YP yolk plug). (C) Late gastrula (stage
12; dorsal view). The presumptive neural plate is intensively stained (left side: cranial, right side: caudal). (DJ Early neurula (stage 13; lateral view). The
signal is now found mainly in the neural plate. (E) Late neural fold stage (stage 17. dorsal view). Only the presumptive brain area and the presumptive
spinal cord are stained. (F, G) Initial neural tube stage (stage 19. F dorsal view. G view on the furure head area). The signal is restricted to the presumptive
brain and spinal cord (SP) zone. A/so the presumptive cement gland (CE) /s stained. (H, J) Larval stage (stage 37; H lareral view, J dorsal view). The
distribution of the transcript can be found In the brain and the spinal cord. Note that also the optic nerve (OP) is stained. (KJ Advanced larva (stage 43,
dorsal view). The staining is detected in the brain. the spinal cord and a/so clearly in the optic nerve (arrow).
Fig. 2 Cross section of an early neurula (stage 14, caudal part). It was
incubated with antisense 35S-RNA of the clone 24-10 (f3-tubulin). The signal
is observed by Epi-Polarization(Grunz. 1990c) in rhe inner layers of the
medullary plate only. e epithelial layer of the neuroectoderm,s sensorial
layer of the neuroectoderm, a archenteron.
Fig. 3. Demonstration of the neural-specific expression of the 13-
tubulin gene (clone 24-1 0). RNA preparations from different adult tissues
were compared. 10,ugof RNA was applied on each lane. (A) The N'rTRANEmembrane
was washed 2 times with 2x SSPE at 6S:C. The film was
exposed for 2 days. IB) The same membrane as In (A) was washed again
with 0.2 x SSPE (high stnngencv) at 6fY'C for 20 min followed by a film
exposure for 3 days. The stringency in Fig. 38 corresponds to that used in
the whole-mount In Situ preparations. B, brain; K, kidney; L. liver; Lu, lung;
M, muscle; S. skin
