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Fig. 1. Neuropilin 1 is transiently expressed in the developing mouse SMG.
(A) Semi-quantitative RT-PCR analysis of neuropilins on the cDNA
samples prepared from SMGs at the indicated developmental stage.
Npn1, but not Npn2, was transiently detected in the
embryonic SMG. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh)
transcript is shown as an internal control. The amount of Npn1
transcript was normalized against the internal control at each stage for
comparison. (B) RNA in situ hybridization analysis of Npn1
transcript on tissue sections. Npn1 was detectable in the rudimentary
SMG at E12.5 (a). The expression of Npn1 transcript peaked at
E15.5 (b), and was dramatically decreased at E17.5 (c), P1
(d) and adult mouse (e). No signals were detectable in the SMG
by using antisense Npn2 or sense Npn1 probes. The boxed areas in a and b are
magnified and shown in a' and b',b'',
(original magnification 400x), respectively. Areas within dashed lines:
salivary gland. (C) Colocalization of Npn1 transcript with
E-cadherin, but not with fibronectin, in E15.5 SMG cultured ex vivo.
(D) Semi-quantitative RT-PCR analysis of the cDNA samples prepared from
either SMG epithelium or the surrounding mesenchyme. Npn1 was mainly
expressed in the SMG epithelium. Fgfr2 and Fgf7 were
controls for specific expression in Epi and in M, respectively. Scale bars: 50
µm in B; 100 µm in C. DA, descending aorta; DRG, dorsal root ganglion;
Epi, epithelium; GAPDH, Gapdh internal control; IF,
immunofluorescence staining; ISH, RNA in situ hybridization; M, mesenchyme;
Nc, nasal cavity; PC, positive control; SC, spinal cord; SCG, superior
cervical ganglion; SMG, submandibular gland.
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