Functional morphology of Helix pomatia salivary gland cells was studied at light microscopic level by using different histochemical methods. Three cell types could be demonstrated in the salivary gland: mucocytes, granular and vacuolated cells. The distribution and the number of the different cell types were different in active and inactive snails. In active feeding animals, dilatated interlobular salivary ducts were observed, which were never present in inactive ones. In active animals an additional cell type, the cystic cell could also be observed. Periodic acid Schiff staining revealed both mucuos and serous elements in the salivary gland. Furthermore, hematoxyline-eosin staining indicated the occurrence of a cell layer with high mitotic activity in the acini. Applying immunohistochemical methods with monoclonal mouse anti-human Ki-67 clone, B56 and polyclonal rabbit anti-human Ki-67 antibodies, we also were able to demonstrate the occurrence of dividing cells in the salivary gland. Analysis of 1-2 µm semi-thin Araldite sections stained with toluidine-blue showed that the saliva can be released, in addition to possible exocytosis, by the lysis of cystic cells. Using an apoptosis kit, we could also establish that this process was due to rather an apoptotic than a necrotic mechanism. In the salivary gland of active snails, where an intensive salivation takes place, significantly more apoptotic cells occurred, if compared to that of inactive animals. It is suggested that programmed cell death may also be involved in the saliva release.
The ultrastructure, neuroanatomy and central projection patterns, including the intercellular connections of the statocyst hair cells of the pond snail, Lymnaea stagnalis, were studied, applying different intra- and extracellular cellular staining techniques combined with correlative light- and electron microscopy. Based on the ultrastructure different hair cells could be distinguished according to their vesicle and granule content, meanwhile the general organization of the sensory neurons was rather uniform, showing clearly separated perinuclear and “vesicular” cytoplasmic regions. Following intra- and extracellular labeling with fluorescence dyes or HRP a typical, local arborization of the hair cells was demonstrated in the cerebral ganglion neuropil, indicating a limited input-output system connected to the process of gravireception. Correlative light- and electron microscopy of HRP-labeled hair cells revealed both axo-somatic and axo-axonic output contacts of hair cell varicosities, and input on sensory axons located far from the terminal arborizations. Our findings suggest (i) a versatile ultrastructural background of hair cells corresponding possibly to processing different gravireceptive information, and (ii) the synaptic (or non-synaptic) influence of gravireception at different anatomical (terminal, axonal and cell body) levels when processed centrally. The results may also serve as a functional morphological background for previously obtained physiological and behavioral observations.