Intracellular and extracellular elemental composition of the endolymphatic sac studied by X-ray microanalysis

X-ray microanalysis was performed along with light microscopy (LM) on rapidly frozen and cryo-sectioned endolymphatic sac tissues of adult guinea pigs, to determine the elemental composition of the different cell types in this tissue as well as the content of the sac lumen. The morphological preservation and spatial resolution of cryo-sectioned endolymphatic sac was found adequate for the identification of the different cell types of the sac in the transmission electron microscope. Further cell type identification was performed by comparing scanning transmission electron microscopy images with LM images on adjacent serial sections. X-ray microanalysis demonstrated differences between epithelial and sub-epithelial cells in the intracellular concentrations of sodium, chlorine, potassium, calcium, phosphorus and sulphur. Measurements performed in the lumen of the endolymphatic sac showed elevated sodium and decreased potassium levels as compared with the known levels of these elements in cochlear or vestibular endolymph. High phosphorus and sulphur levels were also found in the endolymph of the sac. Other morphological and analytical findings on the luminal content point out that otoconial destruction and cleaning of the endolymph from the cell debris and other products such as lipids and proteins take place in the endolymphatic sac. Our results suggest that the endolymphatic sac participates in fluid absorption (osmoregulation), ion transport and otoconial destruction. The data support the longitudinal flow theory of the endolymph.