Search Results (7)

5-Hydroxytryptamine (5-HT) plays a vital role in the reproductive process of vertebrates and is also present in many invertebrates. The cDNA of the Sepiella japonica 5-HT₆ receptor (Sj5-HT₆r) was first cloned by RACE (Rapid Amplification of cDNA Ends). The length was 1450 bp, and the predicted open reading frame (ORF) was 1116 bp, which encoded 371 amino acids. Sequence characteristics analysis showed that Sj5-HT₆r shares a high degree of identity with 5-HT₆r from other cephalopods and forms a sister branch to bivalves. Subcellular localization showed that Sj5-HT₆r protein was localized on the HEK293T cell membrane surface. Quantitative Real-time PCR (qPCR) analysis demonstrated that Sj5-HT₆r was highly expressed in reproductive organs of both sexes. In particular, transcripts with significant expression were observed at stage III of female gonadal development in tissues of the ovary and nidamental gland, and at stage IV in tissues of the accessory nidamental gland. In situ hybridization (ISH) experiment results indicated that Sj5-HT₆r mRNA was primarily distributed in all regions of the optic lobes except the plexiform zone. These results may provide a basis for the future exploration of the reproductive regulation of 5-HT and 5-HT₆ receptors in S. japonica. Full article

Due to overfishing and marine pollution, the status of fishery resources and the ecological environment in the sea areas have considerably changed. The proliferation and release of fishery resources have become the main means of human intervention to maintain and conserve the resource populations. The annual output of Sepiella japonica (S. japonica), once one of the four major seafood species in the East China Sea, has now recovered to a level of 4000 t. However, do the traditional spawning grounds of S. japonica still exist? Have the spawning grounds shifted? These scientific questions are worthy of attention. Based on the survey data of fishery resources and environment at 120 stations in coastal Zhejiang waters from 2015 to 2022, the spatial and temporal distribution and resource center of gravity of S. japonica in springtime were analyzed. Random forest (RF) was used to explain the importance of six environmental variables, including water depth, sea surface temperature, sea surface salinity, chlorophyll a, pH, and dissolved oxygen. The generalized additive model (GAM) nonparametric smoothing function was used to analyze the relationship between environmental factors and the distribution of S. japonica inhabiting the offshore areas of Zhejiang province, and the effects of environmental factors on spawning habitat selection of S. japonica were revealed. This study found that there was a significant interannual variation in S. japonica resources, with an overall increasing trend in the resource. The spawning grounds were mainly distributed in the Jiushan Islands Marine Reserve, the Dachen Islands Marine Reserve, and their nearby sea areas. The resource peaked at latitude 28.3° N. Additionally, the most important variables affecting the distribution of S. japonica were depth of water, followed by sea surface salinity, pH, dissolved oxygen, sea surface temperature, and chlorophyll a. S. japonica mainly inhabited sea areas with a depth of 15~25 m and a sea surface salinity of 26~32. When the pH ranged from 7.6 to 8.3, dissolved oxygen ranged from 6 to 9 mg/L, sea surface temperature ranged from 14 to 17 °C, and chlorophyll a ranged from 2.5 to 5 µg/L, S. japonica was more likely to be present. This study provides insights into the spatial distribution of S. japonica in offshore Zhejiang province, offering a reference for the rational utilization and scientific protection of this resource. Full article

Temperature is a crucial environmental factor that affects embryonic development, particularly for marine organisms with long embryonic development periods. However, the sensitive period of embryonic development and the role of autophagy/apoptosis in temperature regulation in cephalopods remain unclear. In this study, we cultured embryos of Sepiella japonica, a typical species in the local area of the East China Sea, at different incubation temperatures (18 °C, 23 °C, and 28 °C) to investigate various developmental aspects, including morphological and histological characteristics, mortality rates, the duration of embryonic development, and expression patterns of autophagy-related genes (LC3, BECN1, Inx4) and apoptosis marker genes (Cas3, p53) at 25 developmental stages. Our findings indicate that embryos in the high-temperature (28 °C) group had significantly higher mortality and embryonic malformation rates than those in the low-temperature (18 °C) group. Furthermore, high temperature (28 °C) shortened the duration of embryonic development by 7 days compared to the optimal temperature (23 °C), while low temperature (18 °C) caused a delay of 9 days. Therefore, embryos of S. japonica were more intolerant to high temperatures (28 °C), emphasizing the critical importance of maintaining an appropriate incubation temperature (approximately 23 °C). Additionally, our study observed, for the first time, that the Early blastula, Blastopore closure, and Optic vesicle to Caudal end stages were the most sensitive stages. During these periods, abnormalities in the expression of autophagy-related and apoptosis-related genes were associated with higher rates of mortality and malformations, highlighting the strong correlation and potential interaction between autophagy and apoptosis in embryonic development under varying temperature conditions. Full article

Insulin-like peptide receptor (ILPR) can effectively regulate ovarian development in invertebrates, but its effect in cuttlefish has not been reported. We isolated and characterized a ILPR gene from Sepiella japonica, referred to as SjILPR. This gene displayed significant homologies to Octopus bimaculoides ILPR, and contained all typical features of insulin receptors and tyrosine kinase domain structure. SjILPR is expressed in all detected tissues, with the highest expression in the ovary. During ovarian development stages, its expression levels in the ovary, pancreas, and liver were correlated to the female reproductive cycle. After the silencing of SjILPR in vivo, comparative transcriptome analysis identified 4314 differentially expressed genes (DEGs) in the injected group, including 2586 down-regulated genes and 1728 up-regulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed that 832 DEGs were assigned to 222 pathways, many pathways of which were related to gonadal development. Four down-regulated genes relevant to ovarian development (Vitellogenin 1, Vitellogenin 2, Cathepsin L1-like, and Follistatin) were selected to confirm the accuracy of RNA-seq data by qRT-PCR. These results showed that SjILPR might regulate ovarian development to control reproduction by affecting the expression of the relevant genes in female S. japonica. Full article

The insulin-like peptide (ILP) family is well known for regulating reproduction in invertebrates, while its role in mollusks remains largely unknown. In this study, we first isolated and characterized the ILP gene in the cuttlefish Sepiella japonica. The full-length SjILP cDNA obtained was 926 bp and encoded a precursor protein of 161 amino acids. The precursor protein consisted of a signal peptide, a B chain, a C-peptide, and an A chain. It possessed the typical features of ILP proteins, including two cleavage sites (KR) and eight conserved cysteines. To define the function of SjILP, the expression of SjILP in different tissues and ovarian development stages were analyzed using qRT-PCR. SjILP was mainly expressed in the ovary, and its gene expression correlated with ovarian development. Furthermore, silencing SjILP using RNA interference (RNAi) dramatically decreased the expression levels of four ovarian-development-related genes (vitellogenin1, vitellogenin2, cathepsin L1-like, and follistatin). These data suggest the critical role of SjILP in the regulation of ovarian development in S. japonica. Full article

Neuropeptides are released by neurons that are involved in a wide range of brain functions, such as food intake, metabolism, reproduction, and learning and memory. A full-length cDNA sequence of an FMRFamide gene isolated from the cuttlefish Sepia pharaonis (designated as SpFMRFamide) was cloned. The predicted precursor protein contains one putative signal peptide and four FMRFamide-related peptides. Multiple amino acid and nucleotide sequence alignments showed that it shares 97% similarity with the precursor FMRFamides of Sepiella japonica and Sepia officinalis and shares 93% and 92% similarity with the SpFMRFamide gene of the two cuttlefish species, respectively. Moreover, the phylogenetic analysis also suggested that SpFMRFamide and FMRFamides from S. japonica and S. officinalis belong to the same sub-branch. Tissue expression analysis confirmed that SpFMRFamide was widely distributed among tissues and predominantly expressed in the brain at the three development stages. The combined effects of SpFMRFamide+SpGnRH and SpFLRFamide+SpGnRH showed a marked decrease in the level of the total proteins released in the CHO-K1 cells. This is the first report of SpFMRFamide in S. pharaonis and the results may contribute to future studies of neuropeptide evolution or may prove useful for the development of aquaculture methods for this cuttlefish species. Full article

The peptide FMRFamide is one of the well-known peptides involved in multiple physiological processes in the phylum Mollusca. In this study, a FMRFamide gene (GenBank accession No. KJ933411) was identified in a cuttlefish species called Sepiella japonica and was designated as SjFMRFamide. The total length of the SjFMRFamide sequence was found to be 1880 bp while the open reading frame contained 996 bp encoding a protein of 331 amino acid residues with a predicted isoelectric point (pI) and molecular weight (MW) of 9.18 and 38.8 kDa along with a 333 bp 5′-untranslated region (UTR) and 551 bp 3′-UTR. The deduced SjFMRFamide precursor protein contains one signal peptide and expresses four kinds FMRFamide-related peptides including a single copy of FLRFamide, ALSGDAFLRFamide, and FIRFamide and multiple copies of FMRFamide. Results of phylogenetic relation analysis strongly indicated that the sequence of this gene shares high identity with the genes of known FMRFamides. Spatial expression analysis indicated the highest mRNA expression of SjFMRFamide in the brain of male and female cuttlefishes among the eight tissues analyzed. An in situ hybridization assay of the brain indicated that SjFMRFamide was transcribed in several functional lobes, which suggests that it might be related to many physiological regulatory mechanisms. This is the first study describing FMRFamide in S. japonica and the results may contribute to future studies of neuropeptide evolution or may prove useful for the development of aquaculture methods for this cuttlefish species. Full article