Destabilizing Protein Polymorphisms in the Genetic Background Direct Phenotypic Expression of Mutant SOD1 Toxicity

Genetic background exerts a strong modulatory effect on the toxicity of aggregation-prone proteins in conformational diseases. In addition to influencing the misfolding and aggregation behavior of the mutant proteins, polymorphisms in putative modifier genes may affect the molecular processes leading to the disease phenotype. Mutations in SOD1 in a subset of familial amyotrophic lateral sclerosis (ALS) cases confer dominant but clinically variable toxicity, thought to be mediated by misfolding and aggregation of mutant SOD1 protein. While the mechanism of toxicity remains unknown, both the nature of the SOD1 mutation and the genetic background in which it is expressed appear important. To address this, we established a Caenorhabditis elegans model to systematically examine the aggregation behavior and genetic interactions of mutant forms of SOD1. Expression of three structurally distinct SOD1 mutants in C. elegans muscle cells resulted in the appearance of heterogeneous populations of aggregates and was associated with only mild cellular dysfunction. However, introduction of destabilizing temperature-sensitive mutations into the genetic background strongly enhanced the toxicity of SOD1 mutants, resulting in exposure of several deleterious phenotypes at permissive conditions in a manner dependent on the specific SOD1 mutation. The nature of the observed phenotype was dependent on the temperature-sensitive mutation present, while its penetrance reflected the specific combination of temperature-sensitive and SOD1 mutations. Thus, the specific toxic phenotypes of conformational disease may not be simply due to misfolding/aggregation toxicity of the causative mutant proteins, but may be defined by their genetic interactions with cellular pathways harboring mildly destabilizing missense alleles

New Insights into the Apoptotic Process in Mollusks: Characterization of Caspase Genes in Mytilus galloprovincialis

Apoptosis is an essential biological process in the development and maintenance of immune system homeostasis. Caspase proteins constitute the core of the apoptotic machinery and can be categorized as either initiators or effectors of apoptosis. Although the genes encoding caspase proteins have been described in vertebrates and in almost all invertebrate phyla, there are few reports describing the initiator and executioner caspases or the modulation of their expression by different stimuli in different apoptotic pathways in bivalves. In the present work, we characterized two initiator and four executioner caspases in the mussel Mytilus galloprovincialis. Both initiators and executioners showed structural features that make them different from other caspase proteins already described. Evaluation of the genes’ tissue expression patterns revealed extremely high expression levels within the gland and gills, where the apoptotic process is highly active due to the clearance of damaged cells. Hemocytes also showed high expression values, probably due to of the role of apoptosis in the defense against pathogens. To understand the mechanisms of caspase gene regulation, hemocytes were treated with UV-light, environmental pollutants and pathogen-associated molecular patterns (PAMPs) and apoptosis was evaluated by microscopy, flow cytometry and qPCR techniques. Our results suggest that the apoptotic process could be tightly regulated in bivalve mollusks by overexpression/suppression of caspase genes; additionally, there is evidence of caspase-specific responses to pathogens and pollutants. The apoptotic process in mollusks has a similar complexity to that of vertebrates, but presents unique features that may be related to recurrent exposure to environmental changes, pollutants and pathogens imposed by their sedentary nature

India ink induces apoptosis in the yellow clam Mesodesma mactroides (Deshayes, 1854). Optical and ultrastructural study

Este artigo reporta o processo inflamatório agudo e celular no marisco brancoMesodesma mactroides, induzido através de injeção de tinta nanquim no pé muscular. Obervações histológicas com microscopia óptica e eletrônica foram realizadas 24 e 48 h após a injeção. A resposta celular inflamatória induzida consistiu de infiltração hemocitária geral sem ocorrência de necrose e atividade apoptótica. Migração de fagócitos carregados de tinta nanquim através do epitélio intestinal foi registrada. Parece que o marisco branco excretou partículas de nanquim pelas brânquias e rim. A tinção positiva para apoptose foi observada na glândula digestiva. Microscopia eletrônica da glândula digestiva revelou mudanças ultraestruturais de estresse do retículo endoplasmático e corpos apoptóticos. O mecanismo pelo qual as partículas de tinta nanquim induzem apoptose continua desconhecido, possivelmente associado ao estresse do retículo endoplasmático. Este trabalho destacou características que exigem discussões na área restrita das respostas inflamatórias de moluscos.This paper reports on the acute inflammatory and cellular process in the yellow clam, Mesodesma mactroides, induced by injection of India ink into the muscular foot. Histological observations with optical and electronic microscopy were made at 24 and 48 h after injection. The induced cellular inflammatory response consisted of a general hemocyte infiltration without necrosis and apoptotic activity. Migration of ink-laden phagocytes across the intestinal epithelium was recorded. It appeared that the yellow clam "excreted" ink particles through the gill and kidney. The positive staining for apoptosis was observed in the digestive gland. Electronic microscopy revealed ultrastructural changes of endoplasmic reticulum stress and apoptotic bodies in the digestive gland. The mechanism by which the India ink particles induce apoptosis remains unknown, but might possibly be associated with the endoplasmic reticulum stress. This work has highlighted features that require further discussion in the restricted field the inflammatory responses of mollusks