Induction of Autophagy by Amino Acid Starvation in Fish Cells

Autophagy is well established as a starvation-induced process in yeast and mammalian cells and tissues. To elucidate the cellular mechanisms induced by starvation in fish, we characterized the induction of autophagy in cultured zebrafish cells under starvation conditions. As an autophagic marker protein, the microtubule-associated protein 1-light chain 3B protein (MAP1-LC3B) was cloned from the fish cells, and its expression and localization were characterized. In zebrafish embryonic (ZE) cells, posttranslational modifications produced two distinct forms of MAP1-LC3B, i.e., a cytosolic form and a membrane-bound form (types I and II, respectively). Immunofluorescence microscopy revealed fluorescently labeled autophagosomes in cells stably transfected with a green fluorescent protein (GFP)–MAP1-LC3B fusion protein and showed that this protein accumulated in punctate dots in a time-dependent manner in response to amino acid starvation. Starvation also induced the degradation of long-lived proteins. Treatment with 3-methyladenine and wortmannin, two class-III inhibitors of phosphoinositide 3-kinase (PI3K), repressed autophagy under starvation conditions, indicating that the PI3K class-III pathway regulates starvation-induced autophagy in fish

A novel silk-like shell matrix gene is expressed in the mantle edge of the Pacific oyster prior to shell regeneration.

During shell formation, little is known about the functions of organic matrices, especially about the biomineralization of shell prismatic layer. We identified a novel gene, shelk2, from the Pacific oyster presumed to be involved in the shell biosynthesis. The Pacific oyster has multiple copies of shelk2. Shelk2 mRNA is specifically expressed on the mantle edge and is induced during shell regeneration, thereby suggesting that Shelk2 is involved in shell biosynthesis. To our surprise, the database search revealed that it encodes a spider silk-like alanine-rich protein. Interestingly, most of the Shelk2 primary structure is composed of two kinds of poly-alanine motifs-GXNA(n)(S) and GSA(n)(S)-where X denotes Gln, Arg or no amino acid. Occurrence of common motifs of Shelk2 and spider silk led us to the assumption that shell and silk are constructed under similar strategies despite of their living environments

Quantification of the Flavor and Taste of Gonads from the Sea Urchin Mesocentrotus nudus Using GC–MS and a Taste-Sensing System

Sea urchin gonads are a delicious seafood item of high commercial value. Our past studies have revealed that the gonads of the sea urchin Mesocentrotus nudus fed the basal frond portion of fresh Saccharina kelp (BS) or the sporophylls of fresh Undaria (SU) during May–July are of high-quality. The present study investigated the flavor and taste of BS and SU gonads in comparison with those from non-fed M. nudus (NF) using gas chromatography–mass spectrometry (GC–MS) and gas chromatography (GC)-sniffing techniques, and a taste-sensing system. Data of the estimated intensity of taste (EIT) were compared with assessment of gonads from M. nudus collected from an Eisenia bed (fishing ground) and a barren in July. Gonads from both BS and SU released pleasant green, sour, and fruity aromas characteristic of butyl acetate, which are here recognized essential flavor components of high-quality gonads. The gonads of BS and SU had a strong umami taste compared to those of NF, and the Eisenia bed and the barren. The most marketable M. nudus gonads were assessed to be those with green and fruity aromas from butyl acetate, sweet aroma from benzaldehyde, umami EIT > 13.8, bitterness EIT < 3.1, and without any unpleasant sulfurous odor from sulfur-containing compounds