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Figure 3: In situ hybridization analysis of NHE2 mRNA expression and histology. (a), (c) and (b), (d) are paired serial light micrographs of WT and Nhe2-/- pituitary, H&E-stained and in situ hybridization using the antisense probe (resp.). Hybridization with the sense probe yielded no specific signal in either WT or Nhe2-/- samples (data not shown). Three distinct areas of the pituitary are visible in both genotypes (white arrows) at 2x magnification. (e), (g) and (f), (h) are similar pairs at 10x showing that the most dense grain deposits were over the pars distalis (PD), less grain density over the pars intermedia (PI), and much lower grain density over the pars nervosa (PN). Black arrows in (e) and (g) indicate projections from the PI into the PN. The grain deposition over the PI demonstrated a distinct glandular (tubular) arrangement. (i) and (j) are paired WT and Nhe2-/- H&E stained sections at 40x where the difference in thickness of the PI (double headed white arrows) is clear. Cystic dilations (open spaces) of the FS cell canaliculi are clearly apparent in the PD of the Nhe2-/- mice ((b), (d), (f), (h), and (j)); bars : 750 μm in (a)–(d), 40 μm in (e)–(h), and 10 μm in (i) and (j).