Abstract

Rhogocytes are pore cells scattered among the connective tissue of different body parts of gastropods and other molluscs, with great variation in their number, shape and size. They are enveloped by a lamina of extracellular matrix. Their most characteristic feature is the “slit apparatus”, local invaginations of the plasma membrane bridged by cytoplasmic bars, forming slits of ca. 20 nm width. A slit diaphragm creates a molecular sieve with permeation holes of 20×20 nm. In blue-blooded gastropods, rhogocytes synthesize and secrete the respiratory protein hemocyanin, and it has been proposed–though not proven–that in the rare red-blooded snail species they might synthesize and secrete the hemoglobin. However, the cellular secretion pathway for respiratory proteins, and the functional role(s) of the enigmatic rhogocyte slit apparatus are still unclear. Additional functions for rhogocytes have been proposed, notably a role in protein uptake and degradation, and in heavy metal detoxification. Here we provide new structural and functional information on the rhogocytes of the red-blooded freshwater snail Biomphalaria glabrata. By in situ hybridization of mantle tissues, we prove that rhogocytes indeed synthesize hemoglobin. By electron tomography, the first three dimensional (3D) reconstructions of the slit apparatus are provided, showing detail of highly dense material in the cytoplasmic bars close to the slits. By immunogold labelling, we collected evidence that a major component of this material is actin. By genome databank mining, the complete sequence of a B. glabrata nephrin was obtained, and localized to the rhogocytes by immunofluorescence microscopy. The presence of both proteins fit the ultrastructure-based hypothesis that rhogocytes are related to mammalian podocytes and insect nephrocytes. Reactions of the rhogocytes to deprivation of food and cadmium toxification are also documented, and a possible secretion pathway of newly synthesized respiratory proteins through the slit apparatus is discussed.

Highlights

  • Rhogocytes are prominent cells embedded in the connective tissue or floating in the hemolymph of gastropods and other molluscan taxa [1]

  • To-date this has only been proved for podocytes and nephrocytes by the identification of common slit diaphragm proteins [7]

  • Tissue Distribution and Abundance of Rhogocytes A single B. glabrata individual was entirely cut into sections of 3

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Summary

Introduction

Rhogocytes are prominent cells embedded in the connective tissue or floating in the hemolymph of gastropods and other molluscan taxa [1] They are completely surrounded by a lamina of extracellular matrix that suggests a mesenchymal origin of this cell type. Their plasma membrane shows many invaginations that usually appear to be empty but occasionally contain granular material. These extracellular lacunae ( referred to as ‘‘subsurface cisternae’’) are covered by finger-like cytoplasmic bars separated by distinct slits of 20 nm width. To-date this has only been proved for podocytes and nephrocytes by the identification of common slit diaphragm proteins [7]

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