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Fig. 7. The mutant ENCCs form aggregates on a 2D substrate. Rings of E13.5
control and mutant distal midgut were plated on a mixture of ECM gel and
fibronectin. After 2 days in culture, neuron cell bodies were detected with
HuD (red), neuronal processes with NF160 (red), glial cells with B-FABP
(green) and nuclei with DAPI. (A-G) Control culture, at low (A) and
high (B-G) magnification. Neurons and glial cells form scattered networks on
SMC (B,C) or directly on ECM (D-G). (H-N) Mutant culture, at low (H)
and high (I-N) magnification. Green arrows indicate aggregates of neurons and
glial cells that adhere directly to ECM. White arrows show spheroid aggregates
that do not adhere to SMC or ECM. (I,J) Aggregates on the SMC layer. (K,L) An
aggregate adhering to ECM. (M,N) High magnifications of spheroid aggregates
stained for HuD and NF160 (red) and B-FABP (green) (M) and for X-Gal (N). Red
arrows indicate neuronal processes linking aggregates to the explants.
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