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Fig. 6. Cre-mediated recombination leads to enlarged optic nerves. X-gal staining
of the proximal optic nerve immediately adjacent to the retina (A) and distal
optic nerve near the chiasm (asterisk) (B). Optic nerves were dissected from
2-month-old mice with the genotype of hGFAP-cre+;
Rosa26-lacZ/+ (n=3). (C,D) Adjacent sections to A,B were
subjected to double-labeling immunofluorescence with anti-lacZ (red)
and anti-GFAP (green). Most of the lacZ-positive cells are
GFAP-expressing astrocytes. (E) hGFAP-cre-mediated recombination in
the optic nerve was revealed by PCR analysis. Upper panel: a PCR assay
identifying the floxed NF1 allele (x1) and recombined floxed allele
(
) indicated that a significant number of the floxed NF1 alleles in the
optic nerve (ON), cerebral cortex (Ctx) and hippocampus (Hp) of the
Nf1flox/+;hGFAP-cre+ mice transformed into the
recombined alleles. Bottom panel: a PCR assay that identifies the wild type
(+) and the floxed (x1) NF1 allele confirmed the genotype of the tissues
analyzed. (F) A representative of control (left) and mutant (right) eyes with
the optic nerves and chiasm (arrowheads). High-magnification of view of
control (G) and mutant (H) optic nerves with chiasms (asterisk). The mutant
nerve has a conspicuous enlargement (indicated by broken lines and
arrowheads). Scale bars: 100 µm in A-D; 1 mm in F-H.
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