Quantitative X-ray microanalysis of P, Ca, and S in the mucus secretory granules of the cryofixed frog palate epithelium.
In respiratory epithelium, the mucus is densely packed inside the secretory granules (SG) of secretory cells (SC) before being released by exocytosis in the airway lumen. We have previously shown that the frog palate is a representative model of respiratory epithelium and that rapid cryofixation is a very effective technique in preserving the integrity of the mucus SG. The concentration of phosphorus (P), sulphur (S), and calcium (Ca) were analysed inside the SG of the SC of frog palate after quick freezing, cryosubstitution, and embedding in Lowicryl resin at low temperature. The experiments were carried out using X-ray microanalysis conducted with energy dispersive spectrometry (EDS) at 100 kV. The quantitation was carried out using the continuum method with reference to Agar standards. The cryofixation permitted us to distinguish two types of SG depending on whether they were electron dense (serous cells) or electron-lucent (mucous cells). A significant (P < 0.001) difference in the S concentration was observed between the individual serous (239 +/- 79 mmol.kg-1) and the mucous SG (161 +/- 48 mmol.kg-1). No significant difference could be identified in the Ca concentration between the two SG phenotypes. In the serous SG, the P content was high (41 +/- 17 mmol.kg-1) compared with the mucous SG where it was not measurable. The comparison of the three element concentrations in each type of secretory cells showed that significant differences in concentration of S and Ca concentration could be observed from one SC cell to another.(ABSTRACT TRUNCATED AT 250 WORDS)
Wagner
D
D
Unité INSERM U 314, Université de Reims, France.
Puchelle
E
E
Hinnrasky
J
J
Girard
P
P
Balossier
G
G
D000818:Animals; D002118:Calcium / Q000032:analysis*; D015925:Cryopreservation; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000648:ultrastructure; D004577:Electron Probe Microanalysis; D008297:Male; D010159:Palate / Q000737:chemistry*; D010758:Phosphorus / Q000032:analysis*; D011893:Rana esculenta; D012137:Respiratory System / Q000737:chemistry / Q000648:ultrastructure; D013455:Sulfur / Q000032:analysis*
D010758:Phosphorus; D013455:Sulfur; D002118:Calcium