Localization of keyhole limpet hemocyanin‐like immunoreactivity in the nervous system ofBiomphalaria alexandrina

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.

FMRF‐NH2‐related neuropeptides in Biomphalaria spp., intermediate hosts for schistosomiasis: Precursor organization and immunohistochemical localization

“…Standard whole‐mount immunohistochemical protocols were followed (see Beach et al, 2019; Rosa‐Casillas et al, 2021; Vaasjo et al, 2018). Dissections were performed in normal saline (in mmol L −1 : NaCl 51.3, KCl 1.7, MgCl 2 1.5, CaCl 2 4.1, HEPES 5, pH 7.8.…”

Tyrosine hydroxylase messenger RNA corroborates protein localization in the nervous system of the pond snail, Lymnaea stagnalis

“…Much of this past work has used antibodies raised against vertebrate antigens but has not validated primary antibodies by use of western blots. Recent work has identified that unexpected cross‐reactivity can occur with certain antibodies in the molluscan nervous system (Beach et al, 2019). Therefore, we suggest that future work employing IHC on molluscs should be modeled on our approach to validating primary antibodies.…”

Identification and localization of a gonadotropin‐releasing hormone‐related neuropeptide in Biomphalaria, an intermediate host for schistosomiasis

“…Standard whole‐mount immunohistochemical protocols were followed (see Beach et al, 2019; Vaasjo et al, 2018). Tissues were dissected in normal saline (in mmol L −1 : NaCl 51.3, KCl 1.7, MgCl2 1.5, CaCl2 4.1, HEPES 5, pH 7.8.…”