Abstract
BackgroundThe lipid signaling molecule, ceramide, is a key component of the vertebrate stress response, however, there is limited information concerning its role in invertebrate species. In order to identify genes involved in ceramide metabolism in bivalve molluscs, Pacific oyster genomic resources were examined for genes associated with ceramide metabolism and signaling.ResultsSeveral genes were identified including full-length sequences characterized for serine palmitoyltransferase-1, 3-ketodihydrosphingosine reductase, acid ceramidase, and ceramide glucosyltransferase. Genes involved in ceramide synthesis and metabolism are conserved across taxa in both form and function. Expression analysis as assessed by quantitative PCR indicated all genes were expressed at high levels in gill tissue. The role of the ceramide pathway genes in the invertebrate stress response was also explored by measuring expression levels in adult oysters exposed to Vibrio vulnificus. Two genes demonstrated increased expression during the bacterial challenge: a gene involved in hydrolytic breakdown of ceramide (acid ceramidase) and a gene involved in de novo generation of ceramide (3-ketodihydrosphingosine reductase), suggesting a possible role of ceramide in the invertebrate stress and immune responses.ConclusionsIn silico and laboratory results support that Pacific oysters have the basic components of the ceramide metabolism pathway. These results also indicate that ceramide may have analogous functions in vertebrates and invertebrates. The gene expression pattern of acid ceramidase and 3-kethodihydrosphingosine reductase in response to bacterial exposure especially supports that ceramide and sphingolipid metabolism may be involved in the oyster’s stress and/or immune responses.
Highlights
The lipid signaling molecule, ceramide, is a key component of the vertebrate stress response, there is limited information concerning its role in invertebrate species
A total of 23 sequences associated with ceramide metabolism were identified by analyzing publicly available Crassostrea gigas sequences (Table 1)
Gene identification This study identified a suite of genes in the Pacific oyster homologous to vertebrate genes associated with ceramide metabolism, including the sequencing and characterization of serine palmitoyltransferase-1 (Cg-sptlc1), acid ceramidase (Cg-Acid ceramidase (AC)), 3-ketodihydrosphingosine reductase (Cg-3-ketodihydrosphingonsine reductase (3KDSR)), Figure 4 Amino acid alignment of translated Cg-3KDSR with protein sequence from H. sapiens [GenBank: Q06136], M. musculus [GenBank: NP_081810], and D. rerio [GenBank: NP_957433]
Summary
The lipid signaling molecule, ceramide, is a key component of the vertebrate stress response, there is limited information concerning its role in invertebrate species. Ceramide is a sphingolipid that serves as an important signaling molecule for a variety of cellular processes including differentiation, proliferation, inflammation, and apoptosis [reviewed in 1 and 2]. The diversity of processes in which ceramide plays a role as a signaling molecule indicates its importance across a variety of life stages and environmental conditions. The accumulation of ceramide can halt embryonic development [3], inhibit insulin signaling [4], and promote apoptosis during cellular stress [5]. Leukemia cells exposed to various exogenous stressors (ionizing radiation, hydrogen peroxide, UV radiation, and heat shock) showed elevated levels of ceramide and increased apoptosis [8]
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