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Behavioral and Physiological Responses of Sea Slug (onchidium reevesii) to Low-Frequency Noise

Journal of Ocean University of China 2022年第6期21卷 1607-1614页

作者： TU Zhihan LI Chenqi JIA Jingjing SHEN Heding National Demonstration Center for Experimental Fisheries Science Education Shanghai Ocean UniversityShanghai 201306China International Research Center for Marine Biosciences Ministry of Science and TechnologyShanghai Ocean UniversityShanghai 201306China Shanghai Collaborative Innovation Center for Cultivating Elite Breeds and Green-Culture of Aquaculture Animals Shanghai 201306China

A large amount of marine noise pollution from coastal industrial construction and the water transportation industry affects the growth and development of marine life,which is a big issue threatening marine organisms.However,most studies have focused on vertebrates,such as marine mammals and fishes,and little is known about the effects of noise on invertebrates.Therefore,the impacts of low-frequency noise(100,300,and 500 Hz)on behavioral and physiological responsesof the sea slug(onchidium reevesii)were investigated.Under laboratory conditions,sea slugs were stimulated with low-frequency noise at 100,300,and 500 Hz for 1 h.Then,hemolymph enzyme activities(glucose,albumin,triglycerides,superoxide dismutase,catalase,and malondialdehyde)were measured,and mRNA expression of the hsp70 gene was detected in hemolymph and the nervous system by quantitative realtime polymerase chain reaction(qRT-PCR)analysis,while expression of the hsp70 protein was analyzed by immunohistochemistry.The results demonstrated that sea slugs were away from the stimulus source under the influence of low-frequency noise compared to the control group.Enzyme activities,as well as hsp70 gene mRNA and protein expression levels,were significantly higher in the noiseexposed groups than those in the control group.Overall,these changes indicate that low-frequency noise caused oxidative stress in sea slugs in vivo,and the oxidative damage gradually increased when the noise frequency was increased from 100 to 500 Hz.

关键词： ocean noise onchidium reevesii low-frequency noise biochemical stress mollusks

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Expression of mapk1 and egr1 genes in onchidium reevesii under tidal stimulation

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Aquaculture and Fisheries 2023年第1期8卷 42-49页

作者： Rongyu Wu Rongcheng Rao Xiaoming Zhang Heding Shen Key Laboratory of Aquatic Germplasm Resources Exploration and Utilization of Ministry of Education Shanghai Ocean UniversityShanghai201306China National Experimental Teaching Demonstration Center of Aquaculture Science Shanghai Ocean UniversityShanghai201306China Classification and Evolution of Marine Animal System Shanghai Ocean University Key Laboratory of Shanghai UniversitiesShanghai201306China

Previous studies have speculated that tidal rhythm of onchidium reevesii was linked to its memory ability, which has not been well explained. Mapk1 and egr1 genes are closely related to memory formation in the MAPK signaling pathway that have been extensively studied in vertebrates. These two genes are involved in Long-term potentiation (LTP), which is generally regarded as one of the main molecular mechanisms underlying learning and memory. To investigate the relationship between tidal rhythm of O. reevesii and its memory ability, we studied the molecular mechanism of mapk1 and egr1 genes cooperating with tidal rhythm. The full-length cDNA sequence of mapk1 gene was cloned by RACE method and analyzed by bioinformatics, then qRT-PCR was used to analyze its expression levels in each tissue. A homology analysis and phylogenetic tree reconstruction revealed that O. reevesii is most closely related to the Biomphalaria glabrata. The qRT-PCR showed that mapk1 was expressed in all the tissues, but the highest expression was in the ganglion. We speculated that mapk1 is related to memory formation of O. reevesii. Then we used qRT-PCR to detect the expression of mapk1 and egr1 in ganglion of O. reevesi under tidal stimulation. The expression of mapk1 and egr1 at the rising tide points were significantly higher (P reevesii was affected by the tide for a long time and formed the tide memory.

关键词： onchidium reevesii Tide rhythm mapk1 egr1

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Analyses of the differential expression of cloned Caspase-3 and Caspae-9 genes,of onchidium reevesii,exposed to extreme hot and cold conditions

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Aquaculture and Fisheries 2021年第6期6卷 583-592页

作者： Fengshen Zhao Heding Shen Key Laboratory of Aquatic Germplasm Resources Exploration and Utilization of Ministry of Education Shanghai Ocean UniversityShanghai201306China National Experimental Teaching Demonstration Center of Aquaculture Science Shanghai Ocean UniversityShanghai201306China Classification and Evolution of Marine Animal System Shanghai Ocean University Key Laboratory of Shanghai UniversitiesShanghai201306China

During low tide intertidal organisms face severe temperature fluctuations.This situation may be more pronounced in the context of climate change.To investigate the molecular response mechanism of onchidium reevesii when stressed by hot and cold temperatures,we cloned its Caspase-3 and Caspase-9 genes and detected their expression characteristics using RT-qPCR.The results showed that the full-length of the cDNA of Caspase-3 consist of 1831 bp,and included an open reading frame(ORF)of 894 bp,a 50UTR of 155 bp and a 30 UTR of 300 bp,and encoding for 297 amino acids.The full-length of the cDNA of Caspase-9 consisted of 2425 bp,and included an ORF of 1373 bp,a 50 UTR of 290 bp,and a 30 UTR of 759 bp,and encoding for 457 amino acids.During the evolution of species the two genes have been greatly conserved.A homology analysis and phylogenetic tree reconstruction revealed that O.reevesii is most closely related to Aplysia californica.The RT-qPCR showed that both genes were expressed in all of the tissues and that expression was highest in the hepatopancreas.It is speculated that these genes play an essential role in the immunity of O.reevesii to certain diseases.Conditions of temperature stress(40�C and 10�C)showed that both heat and cold stresses could induce significant changes in both genes.Compared to the control group,under cold stress conditions the levels of expression of Caspase-3 and Caspase-9 were significantly upregulated(P<0.05)in each experimental group.When exposed to a constant temperature of 10�C,their expression peaked after 1 h(4.32-fold,P<0.05;and 5.58-fold,P<0.05,respectively)and was at its lowest after 12 h(1.93-fold,P<0.05;and 2.35-fold,P<0.05,respectively).Their expression was upregulated after 24 h(2.68-fold,P<0.05;and 4.21-fold,P<0.05,respectively).The expression of both genes showed similar trends.Under heat stress,the expressions of the two genes showed a wave pattern.When exposed to a constant temperature of 40�C,their expression was up-regulated after 1,6,and 24 h,and down-regulated after 3 a

关键词： onchidium reevesii Caspase-3 Caspase-9 Temperature Stress

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