No abstract

Recent years have led to increased effort to describe and understand the peripheral nervous system and its influence on central mechanisms and behavior in gastropod molluscs. This study revealed that an antibody raised against keyhole limpet hemocyanin (KLH) cross-reacts with an antigen(s) found extensively in both the central and the peripheral nervous systems of Biomphalaria alexandrina. The results revealed KLH-like immunoreactive (LIR) neurons in the cerebral, pedal, buccal, left pleural, right parietal, and visceral ganglion within the CNS with fibers projecting throughout all the peripheral nerves. Numerous KLH-LIR peripheral sensory neurons located in the foot, lips, tentacles, mantle, esophagus, and penis exhibited a bipolar morphology with long tortuous dendrites. KLH-LIR cells were also present in the eye and statocyst, thus suggesting the labeling of multiple sensory modalities/cell types. KLH-LIR cells did not co-localize with tyrosine hydroxylase (TH)-LIR cells, which have previously been described in this and other gastropods. The results thus provide descriptions of thousands of peripheral sensory neurons, not previously described in detail.Future research should seek to pair sensory modalities with peripheral cell type and attempt to further elucidate the nature of KLH-like reactivity. These findings also emphasize the need for caution when analyzing results obtained through use of antibodies raised against haptens conjugated to carrier proteins, suggesting the need for stringent controls to help limit potential confounds caused by cross-reactivity.In addition, this study is the first to describe neuronal cross-reactivity with KLH in Biomphalaria, which could provide a substrate for host-parasite interactions with a parasitic trematode, Schistosoma.

Catecholaminergic neurons are abundant in molluscs and are involved in a variety of behaviors such as feeding, respiration, learning, and locomotion. However, previous identification of these neurons has relied almost exclusively on immunohistochemistry using antibodies, which have not been fully validated for use in molluscs. We employed tissue‐specific quantitative PCR in adults of Lymnaea stagnalis (a pulmonate gastropod) and whole‐mount in situ hybridization in larvae to both quantify and visualize messenger RNA of the catecholamine synthesis enzyme, tyrosine hydroxylase (TH). TH messenger RNA was found to localize primarily in the foot and the central nervous system, with smaller quantities present in the cephalic sensory organs. Additionally, we performed western blots that validated a popular antibody used as a marker for catecholaminergic neurons in molluscs. Taken together, these data indicate that TH messenger RNA is present in the central and peripheral nervous system of L. stagnalis and support the specificity of past immunohistochemical labeling of the TH protein. These findings have potentially broad implications, given the wide range of biological processes that have been studied in L. stagnalis.