Abstract
In humans, injuries and diseases can result in irreversible tissue or organ loss. This well-known fact has prompted several basic studies on organisms capable of adult regeneration, such as amphibians, bony fish, and invertebrates. These studies have provided important biological information and helped to develop regenerative medicine therapies, but important gaps concerning the regulation of tissue and organ regeneration remain to be elucidated. To this aim, new models for studying regenerative biology could prove helpful. Here, the description of the cephalic tentacle regeneration in the adult of the freshwater snail Pomacea canaliculata is presented. In this invasive mollusk, the whole tentacle is reconstructed within 3 months. Regenerating epithelial, connective, muscular and neural components are already recognizable 72 h post-amputation (hpa). Only in the early phases of regeneration, several hemocytes are retrieved in the forming blastema. In view of quantifying the hemocytes retrieved in regenerating organs, granular hemocytes present in the tentacle blastema at 12 hpa were counted, with a new and specific computer-assisted image analysis protocol. Since it can be applied in absence of specific cell markers and after a common hematoxylin-eosin staining, this protocol could prove helpful to evidence and count the hemocytes interspersed among regenerating tissues, helping to unveil the role of immune-related cells in sensory organ regeneration.
Highlights
Invertebrates have provided important contributions to the understanding of human biology, and their usage in biological research has been increasing in the last few decades [1,2,3]
Studies in the freshwater snail Lymnaea stagnalis demonstrated the crucial role played by resident phagocytes for the regeneration of neural components, a feature shared by several invertebrate species, suggesting an ancient origin and conserved relationships between phagocytic cell functions and neural regeneration [16]
Cephalic tentacles play a major role in P. canaliculata biology, as they are utilized for food search, co-specific recognition, and orienting [23,24]
Summary
Invertebrates have provided important contributions to the understanding of human biology, and their usage in biological research has been increasing in the last few decades [1,2,3]. Cephalic tentacles play a major role in P. canaliculata biology, as they are utilized for food search, co-specific recognition, and orienting [23,24] These long tentacles present a well-defined organization, with muscular and connective tissue representing a sort of scaffold for the nervous component that serves the olfactory epithelium [23]. No specific molecular markers are available for tracking tissue and circulating hemocytes in P. canaliculata [25,30,31], hampering hemocyte quantification outside the hemolymph On these basis, histological techniques and a new protocol of computer-assisted image analysis were combined to document and count the hemocytes in the regenerating cephalic tentacles. Our morphological observations and software-based quantifications indicate that in the immediacy of amputation, Group II granular hemocytes increase significantly in the forming blastema
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