Lipoprotein-induced cell growth and hemocyanin biosynthesis in rhogocytes

Abstract

Rhogocyte is a unique molluscan cell that synthesises a supramolecular respiratory protein known as hemocyanin. Its ability to synthesise the protein has eluded the scientists despite hemocyanin’s importance as a carrier protein and complex molecule with anti-viral activity. Although a hypothetical model of hemocyanin release from the rhogocytes lacunae was proposed based on colloid-osmotic pressure mechanism, lack of in vitro studies limits further validation of this model. In this study, we aim to investigate the impact of cell culture conditions and nature of hemocyanin biosynthesis of rhogocyte cells dissociated from Haliotis laevigata mantle tissue. Population of cells with different hemocyanin expression levels was profiled using flow cytometry, while hemocyanin concentrations in the media were elucidated by ELISA assay. We demonstrated that addition of lipoprotein supplement into the media resulted in a burst secretion of hemocyanin into the culture media. Over 7 days of culture, the population of cells tagged with hemocyanin antibody increased steadily while hemocyanin release in the media decreased significantly. Variation of culture medium, temperature, growth supplement type and concentration also impacted the cell growth and hemocyanin biosynthesis. These results indicated the possibility of an active process triggered by the addition of supplement to synthesise the protein at the highest amount during the first hour. The current study provides a glimpse of the hemocyanin biosynthesis by rhogocyte that may be significant to understand the cell ability to synthesise supramolecular protein and secretion through lacunae.

Gill histology and ultrastructure in Aplysia depilans (Mollusca, Euopisthobranchia)

“…Taking into consideration that these cells were found bordering the hemolymph space of the gill, they could produce hemolymph proteins. In gastropods, rhogocytes are responsible for the synthesis and secretion of the respiratory protein hemocyanin (Martin et al, 2011; Sairi et al, 2022; Sminia & Boer, 1973), but these connective tissue cells were rarely reported in gills (Cajaraville et al, 1990; Haszprunar, 1996). In rhogocytes of the gastropods Lymnaea stagnalis and Haliotis tuberculata , particles that resemble hemocyanin particles in shape and size were observed within dilated rough endoplasmic reticulum cisternae (Albrecht et al, 2001; Sminia & Boer, 1973).…”