Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. ytm o ogtime, long a ecological For in system. evolution adaptive investigate to Nassarius contribute would level genetic on communities. TTX resist snails as organisms such how animals, animals. of of range fish kinds various wide puffer genes a in in in appears understood also accumulation well resistance be TTX crab TTX to (2014) and with needed al STX is related et pufferfish, TTX Feroudj of candidates Except mechanism transportation. gene accumulation detailed TTX some the with revealed But related analysis be microarray possibly which DNA fish, puffer in fish proteins puffer in in 2013) found cure were and (2000) transportation (2010 poisoning and al toxin binding TTX and et with related Takashi research possibly external evolution were which the adaptive proteins frog Some defense for fish, critical to in particularly as body such be their insensitivity, would medicine. site in target level STX by genetic toxins or in to TTX resistant accumulate be mullusca affinities channels can could and high possibly sodium animals with they to some since site bind However, environment binding specifically can one animals. (STX) sharing of Saxitoxin by and animals (TTX) of Tetrodotoxin neurotoxin, The potent animals. As other of that for basis from genetic different INTRODUCTION important is provided mutation that species In “L” resistance. Nassarius acid and two detoxification. accumulation in amino to TTX communities one referred of nontoxic which had research and evolution of succinctus acid toxic adaptive some N. delta-aminolevulinic both mainly on, of of 2, communities so analysis member gene non-toxic and transcriptome family channel and X2 protein sodium toxic WAS isoform the subunit, among Sec24D up-regulated level, oxidase protein genes more activity c transport expression produced cytochrome catalytic protein generally different 83-like, and dehydratase, communities mostly protein binding toxic shock The and The non-toxic heat to process, species. and included communities. metabolic due both TTX toxic non-toxic of for and incident both than patterns categories cellular death genetic classification for genes the their and top analysis expression, reveal the poisoning to transcriptome genetic for variciferus) For people accounted conducted gene Nassarius serious to we and resistance. succinctus due caused Here and (Nassarius possibly has accumulation species is body. Nassarius Nassarius resistance two significant their snail TTX in us that in sea communities provide reported accumulation Eating could been (TTX) animals has channels. tetrodotoxin in It sodium resistance communities. and in ecological accumulation mutation in (TTX) evolution adaptive tetrodotoxin into of insights basis genetic the Understanding Abstract 2020 12, August 1 Nassarius in Zou TTX Shanmei of evolution adaptive for patterns Genetic ajn giutrlUniversity Agricultural Nanjing 8 13 lerne octopus blue-ringed , a lc h idn ftxnadtu euti h oi resistance toxin the in result thus and toxin of binding the block can pce-ihgnso asria,i motn nmitiigteblneo aieecological marine of balance the maintaining in important is Nassariinae, of genus species-rich a , 1,2,3,32 1 2,17 nesadn o T sacmltdi nml n o nml eitt TTX to resist animals how and animals in accumulated is TTX how Understanding . eotdtedsrbto n oaiaino T-idn rti n hi homologous their and protein TTX-binding of localization and distribution the reported 34 rtyprfidsm T-idn rtisi ue s.YtuYmsiae al et Yotsu-Yamashita fish. puffer in proteins TTX-binding some purified firstly Nassarius 9 n n frogs and and , scnue spplrfo nAincutisa cnmcspecies economic as countries Asian in food popular as consumed is 3,10-12 thsbe eotdta h uaini oimchannel sodium in mutation the that reported been has It . 1 4,5,6 hc ol edt osnn n death and poisoning to lead could which , 1,2,3,14-16 Understanding . 19 7 employed xanthid , 2,17 19 21 . . . Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. eeec eune pcfi rmr eedsge osqec hmfrconfirmation. for them sequence to of RESULTS designed unigenes were all Then primers specific database. sequences reference as reference NCBI in others and and mouse human, mollusca, genes. from functional 18 genes. Q-PCR of channels target total the tool. sodium A as software 18S. in selected expression gene relative were Mutations the the protein to with toxin-binding normalized determined were were coding changes values with Fold threshold related Cycle closely above. DEGs synthetized cDNA the Dis using Possion with confirmation. 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TTX toxicity the and treatment collection, toxin. Sample for evolution adaptive the understanding METHODS better AND for MATERIALS level mutation and expression genetic in in resistance two resistance in specimens and non-toxic communities. and accumulation succinctus toxic different both TTX related for for possibly evolution involving analysis transcriptional is adaptive basis performed This to genetic lead accumulate. could not the which do Currently live others some they environments but was sea body incident the their poisoning in with food TTX accumulate the can that populations detected one studies within previous in populations Our accumulated TTX China. to in due especially years, several last of nassariids eating However, s .variciferus N. du hnesgnst hc h T id.W onoddavailable downloaded We binds. TTX the which to genes channels odium 28 h eeepeso ee a acltdwt SM h C nlsswspromdwt all with performed was analysis PCA The RSEM. with calculated was level expression gene The . and eeslce o rncitoa nlsssnete nlddbt oi n o-oi specimens. non-toxic and toxic both included they since analysis transcriptional for selected were 29 . asru variciferus Nassarius eempe oterfrnedtbs.Wieteuiee eebs ace othe to matched best were unigenes the While database. reference the to mapped were 24 Nassarius a sdt efr env sebyatrfitrn u a ed.Teassembly The reads. raw out filtering after assembly novo de perform to used was eltm C a efre ihteS atqC atrMx(ihRox) (High Mix Master qPCR Fast SG the with performed was PCR Real-time Nassarius Nassarius pce a different was species ,amn orva hi eei atrsfrTXacmlto and accumulation TTX for patterns genetic their reveal to aiming ), psbd n h oiiyo different of toxicity the and body sp’s aecue udeso odpioigaddahicdnsi the in incidents death and poisoning food of hundreds caused have ae ntetasrp ngnso l ape,w retrieved we samples, all of unigenes transcript the on Based h ucinlantto a efre rmdtbssof databases from performed was annotation functional The 2 22 h ape sdi hssuyhv entse for tested been have study this in used samples The 22 ae ntetxct test, toxicity the on Based . 25-26 hc en htsome that means which , 27 h RadItrrSa5wr sdt get to used were InterProScan5 and NR The . sn h olo o-omlzdraswith reads non-normalized of pool the using fe N xrcintemRNA the extraction RNA After .succinctus N. 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Data may be preliminary. eeslce o PR h PRo hs eetdgnswspromdfrbt oi pcmn (treat specimens toxic both for performed was genes selected genes these functional of potential qPCR as pathways The found the DEGs qPCR. transport, multiple hand, for sequencing, RNA selected other transcriptome were spliceosome, the of like confirmation On transcription, accuracy For about on. mainly so were factors. genes and transcription Por- downregulated endoplas- basal pathways, disease, the in Huntington Metabolic with disease, processing metabolism Parkinson related protein trans- chlorophyll disease, were dehydratase, protein Alzheimer and genes acid (NAFLD), II, phyrin Thermogenesis, upregulated disease delta-aminolevulinic contraction, and main liver muscle 2, Cardiac I fatty the pathways, member Non-alcoholic subunit Metabolic with from phosphorylation, family related oxidase genes Oxidative protein pathways reticulum, upregulated c mic WAS The most cytochrome X2, the 83-like, others. groups, isoform and (control) protein non-toxic Sec24D non-toxic the shock protein and with heat (treatment) port Compared toxic included classification 2). from groups (Table top obtained toxic species the were both genes for for expressed accounted groups different biological function common from molecular most process The from metabolic activity and catalytic cellular S2). (Figure the and categories which binding for and enrichment, Gene process, functional ontologies The process. three of biological (HQ). including and patterns specimens DEGs, component non-toxic coincident for For cellular the performed function, was with molecular enrichment genes. comparison of functional in up-regulated and genes classification more (GO) up-regulated in showed Ontology more and (ZD) specimens (ZL) produced toxic non-toxic also between the specimens with (HL) genes toxic Compared down-regulated 4). each and (Fig. , up-regulated species revealed both for (DEGs) specimens non-toxic genes expressed 3, different Fig. The in different shown generally As were analysis. specimens PCA non-toxic variciferus by and calculated CDS toxic was the the sample succinctus between each for level for difference level expression genes. expression much the gene expressed no the different was unigenes, and to There patterns expression 300-500bp. 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N. rcui suis Trichuris .californica A. .variciferus N. respectively produced N. N. N. Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. opiae o sebywt N-e.I h uuesuis ewudepo opeesv samples and comprehensive toxic various of employ of would genes genes channel we channel sodium studies, sodium complete different the future the cover in explore the not communities that to non-toxic do In technologies or sequences sequence degradation advanced RNA-seq RNA-seq. more the RNA with and that to by assembly be due confirmed may for Whether further reasons regions complicated be the resistance. channel should suggest of TTX sodium We resistance domains possessing RACE-PCR. TTX the or D4 animals with blast other and site reads mutational all D1 blast, potential of the a that Unfortunately, is from “L” electrophysiology. different sodium acid the was amino get “L” new to this site references acid gene channel amino sodium cytochrome one the the against of that clustered genes reasonable were channel TTX. is unigenes accumulating it transcriptome specimens Thus, the disease. in All respiration. upregulated Huntington in path- highly and role is the disease key involve subunit contraction, plays Parkinson oxidase II subunit muscle c disease, and oxidase Cardiac Alzheimer I c pathways, (NAFLD), cytochrome subunit Metabolic The oxidase disease phosphorylation, the c liver Oxidative all Cytochrome fatty like of Non-alcoholic removal mechanism. diseases the various special is of by One which drugs removed ways reticulum. detoxification, or possibly endoplasmic wastes is is in metabolic reticulum body processing as endoplasmic the toxic protein such of groups, from of materials functions genes non-toxic pathway toxic important upregulated the the significant most involved with most the protein Compared the downregulated shock of were and species. subunit heat oxidase upregulated The both c (both for specimens groups. cytochrome in- non-toxic DEGs groups and DEGs of common protein non-toxic of that shock most and heat from analysis the toxic different PCA selected the was we The between specimens Furthermore, genes) toxic activities samples. of mechanism species. toxic patterns special both in expression that for occur gene suggest resistance the may that and This dicated accumulation genes. For TTX up-regulated specimens. with For more non-toxic of showed associated that specimens communities. than toxic more non-toxic two significantly the was , specimens than toxic of genes genes up-regulated regulated more produced munities communities. non-toxic all, evolution and of toxic adaptive First from the patterns species, to mutation TTX-resistant lead and typical expression can the genetic as to environments study, due Different in be this Species could uncertain. In TTX. resistance was with toxin animals animals the to bind in of that but specially reported toxin TTX understanding the been of evolution where animals has adaptive mechanism in their It for genes channel in only medicine. sodium TTX not in of mutation toxin, and cure accumulating TSX food-poisoning animals PSP, for of like also various toxin, in accumulate resistance could resistance and toxin animals produce that thus and pointed body researches more and More sodium found sodium new DISCUSSION of this with of comparison III sequence in Domain same II, and the Domain sequences. obtained in II we ‘L’ of Domain confirmation, acid than unigenes from PCR as amino more By new identified from with one gene 6). was score found channels identify unigene (Fig. we with species matched sites, other database acid this all amino reference of the in acids Among genes amino channels. channels The CL8899.Contig1 sodium (unigene the sequence 80%. to 3425bp of matched total best A groups was control species. general, both and In for treat genes for gene. channel level reference genes. expression 5. the channel actual Fig. as Sodium and the selected -([?][?]Ct) in was 2 as 18S the tendency consistent that The showed indicated group). results (control statistic the specimens non-toxic and group) .succinctus N. .variciferus N. Nassarius. .succinctus N. and ae ntetasrp ngnso ohseis etidt ereetesodium the retrieve to tried we species, both of unigenes transcript the on Based eotie 2adD oan of domains D3 and D2 obtained We .variciferus N. vntog ebatdtecenraso ohseis(5b)t l reference all to (150bp) species both of reads clean the blasted we though even Nassarius notntl,n efc ace oimcanl ee eefound were genes channels sodium matched perfect no Unfortunately, . Nassarius 31,32 ti infiatt eeltegntcbsso oi accumulation toxin of basis genetic the reveal to significant is It . aetxcadnntxccmuiisfo ieetsaareas. sea different from communities non-toxic and toxic have eosrtdsmlrDGpten.Gnrlytetxccom- toxic the Generally patterns. DEG similar demonstrated species. .succinctus N. 35-36 4 hssrnl ugs htteTXin TTX the that suggest strongly This . Nassarius and .succinctus N. .variciferus N. ol o eotie yunigene by obtained be not could 1,13,31-,32 ihlnt f32b where 3425bp of length with eeue orva their reveal to used were .variciferus N. l)from All) u h accumulation the But . Nassarius .succinctus N. .succinctus N. Nassarius h up- the , are ’s Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. 2 i,Y,Bon .&Mse,F erdtxn curnei tlpdfoso ot ia Science rica. costa of frogs atelopid in Official occurrence Toxicon Tetrodotoxin: criniger. F. gobius Mosher, goby & (1975) a G. ,151-152 Brown, from Y., tetrodotoxin Kim, of Toxinology 12. on Isolation potent Society Y. International a Hashimoto, the Tarichatoxin-tetrodotoxin: & of Journal H.G. T. Fischer, Noguchi, & 11. H.D. Buchwald, F.A., Fuhrman, Science H.S., neurotoxin. International Mosher, the of of eggs 10. Journal the in Official compound Toxicon tetrodotoxin-like maculosa). Toxinology a in on (hapalochlaena of toxin Society Occurrence octopus major I. blue-ringed the Spence, Fish venomous & as Sci. M.E.H. the tetrodotoxin Howden, of Soc. D.D., Occurrence Sheumack, a Japanese. al. in 9. Bull. et tetrodotoxin J. floridus. of Maruyama, Atergatis Occurrence K., crab, Fish Koyama, xanthid al. Sci. A., a et Uzu, Soc. K. T., Japanese. Noguchi, Hashimoto, Bull. Y., 8. Ueda, sauliae. J., Charonia Maruyama, “boshubora” T., shell, Noguchi, trumpet the H., Genetics of in Narita, Trends Journal animals. 7. Official in resistance Toxicon tetrodotoxin of axon. evolution Adaptive squid B. Venkatesh, 22 and & T.W. muscle Soong, frog 6. on nortetrodotoxin Toxinology of on Actions Society of International C.Y. blockage Kao, and structure 5. derivatives:chemical Tetrodotoxin R.N. Science replacements Poston, conductance. acid & membrane amino nerve J.W. Interacting Moore, al. Science fugu T., et resistance. of 4.Narahashi, Y. epibatidine tissues Lu, evolve the J.C., to among Santos, frogs W., and poison Sachs, fish allow C.M., puffer puffer Borghese, of R.D., Toxicon to plasma 3.Tarvin, proteins analysis. the homologous blot in western of protein by Distribution examined binding N. pardalis tetrodotoxin Araki, and & resistance saxitoxin N. tetrodotoxin Okoshi, of fish basis H., Genetic Yamaki, M., al. 2.Yotsu-Yamashita, et S. Brenner, Biology S.L., Current See, pufferfishes. N., in Dandona, S.Q., Agri- Lu, Nanjing B., 1.Venkatesh, of Center Bioinformatics CA. Berkeley, the UC by (31600294)” of Reference: China support Platforms of the Foundation Computer thank Science Biological also Natural and We University, National cultural “The Council. from Scholarship grant China a by and supported complete was the study This exploring on focus should Acknowledgments studies Future TTX-binding. the various Nassarius the of with of genes channels blocking related toxin sodium channel mainly for the possessing sodium in are mutation animals found other which was the with “L” groups, be Compared acid non-toxic amino diseases. new and various one toxic and resistance, detoxification the of all mechanism both between metabolic specimens genes non-toxic expression catalytic the and different with binding compared and variciferus genes both process N. upregulated for metabolic two more categories and in showed classification resistance cellular top specimens the and the for accumulation expression, accounted TTX gene activity of For patterns analysis. genetic transcriptome the revealed study This Conclusion ,621-626(2006) . h etsokpoenadctcrm xds uui eefuda h otsignificant most the as found were subunit oxidase c cytochrome and protein shock heat The . 44 10-10(1964) ,1100-1110 22 ,811-81(1984) 15 Nassarius 26-02(2005) ,2069-2072 20 14-00(1982) ,1043-1050 156 9699(1967) ,976-979 pce n ealdmtblcptwy o T eitnein resistance TTX for pathways metabolic detailed and species 11 55 5 3537(1973) ,305-307 11-14(2010) ,1119-1124 357 .succinctus N. 16-26(2017) ,1261-1266 49 18-8 (1983) ,1887-189 .variciferus N. and 47 959 (1981) ,935–94 .variciferus N. hc ol possibly could which , Nassarius .succinctus N. h toxic The . pce by species 189 and Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. 9 ui,S lvre ..Tesgicneo iia eeepeso rfie.Gnm Res Genome profiles. expression gene digital of significance The J.M. for Claverie, tool Methods. (1997) & Nature universal S. 2. a Audic, bowtie with 29. Blast2go: alignment gapped-read al. Fast et S.L. Salzberg, M. (2012) & Talon, Bioinformatics a B. J., research. Langmead, without Terol, genomics 28. or functional J.M., with in Garcia-Gomez, analysis data S., and rna-seq Gotz, visualization from transcript annotation, A., quantification novo Conesa, transcript De accurate 27. Rsem: al. L. Bioinformatics et BMC Bo, genome. P.D. & reference C.N. Blood, Dewey, Nature M., analysis. 26. and Grabherr, generation reference M., for platform Yassour, trinity the A., Protocols using rna-seq Papanicolaou, from transcriptome Full-length reconstruction B.J., sequence Biotechnology al. Haas, Nature et genome. D.A. reference Thompson, 25. a J.Z., without Levin, data survey M., rna-seq Yassour, A from B.J., al. assembly Haas, et Biology M.G., A. Genome Grabherr, Mcpherson, analysis. A., 24. data Cervera, rna-seq D., for Gomez-Cabrero, China practices S., of Tarazona, in best P., University populations of Madrigal, Ocean and A., of species Conesa, toxicity, Journal 23. between barcoding. relationships gene The ex- multiple C. and reveal Wang, (2019) detection & barcoding 1515-1522 toxin C. multigene on Wang, One character–based based J., Plos and community. nassarius Song, dangerous distance S., and Monophyly, Zou, complex L. 22. a Kong, nassarius: & in Q. diversity cryptic Li, traordinary S., Zou, 21. Molecular accel- facilitates genes. loss toxin Domain metalloproteinase Evolution venom al. snake et and duplicate C. Biology Renjifo, of gene R., neofunctionalization Camila, on and R.A., analysis evolution Harrison, microarray erated S.C., Dna Wagstaff, Toxicon N.R., pufferfish. al. Casewell, et in fugu 20. S. accumulation Watabe, tetrodotoxin fish ,G., to Kaneko puffer related Y., Kurosu, possibly of T., candidates of membranes Matsumoto, channel H., Communications properties muscle Feroudj, na+ Research Binding 19. Biophysical skeletal voltage-gated & A. a to Biochemical Sugimoto, of pardalis. and structure M., off. membranes primary cle Isemura, brain the Y., and to Nitanai, pardalis 3h-saxitoxin K., and Nishimori, 3h-pbtx-3 M., Yotsu-Yamashita, pufferfish analyzed pardalis, of takifugu 18. Localization pufferfish, the al. of et Toxicon tissues H. staining. the Yamaki, in immunohistochemical in N., (pstbp) by resistance protein Araki, binding tetrodotoxin K., tetrodotoxin Watanabe, and adaptation: N., saxitoxin complex Okoshi, M., a Yotsu-Yamashita, of sodium 17. history Toxin-resistant Evolutionary Evolution H.H. W.F. and Zakon, Gilly, Evolution Biology & & salamanders. Molecular C.T. H.A. family. Hanifin, Fozzard, gene J.W., 16. complete Kyle, a Y., across Lu, (2008) evolution ,1016-1024 adaptive D.M., of parallel Hillis, diversification Evolutionary M.C., channels: P.C. Jost, Ruben, Nature thamnophis & interaction. 15. E.D. snake predator–prey Brodie, a garter E.D., in the channels Brodie, sodium E., in ttx-resistant Fujimoto, genes S.L., Geffeney, channel (2014) 14. sodium .11 Evolution voltage-gated evolution and Parallel three Biology al. Molecular in et sirtalis. C.R. resistance Feldman, S.V., tetrodotoxin Edwards, D.E., of Janes, J.P., Chuckalovcak, J.W., Mcglothlin, 13. 8 19-52(2013) ,1494-1512 28 69 6724 (2011) 2637-2649 , 2224(2015) ,232-244 12 3333(2011) ,323-323 72 2-8(2013) ,23-28 6 17 181(2016) , α- 434 uui fn1 rmseea mus- skeletal from (fmna1) subunit 267 7973(2005) ,759-763 4342(2000) ,403-412 77 29 6-2(2013) ,68-72 6462(2011) ,644-652 21 37-66(2005) ,3674-3676 7 e77 (2012) ,e47276 7 9 986-995 , ,357-359 18 25 , Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. L 54.266 82 4780.22 73.33 73.99 76.52 94.7 94.59 93.98 94.47 98.27 98.19 97.88 98.09 6.62 6.65 6.65 6.7 44.12 44.31 44.32 44.66 Ratio(%) Reads Clean Q30(%) Reads Clean 60.43 Q20(%) Reads 55 Clean 59.9 Bases(Gb) HD1 Clean Total 58.37 HL3 Reads(Mb) Clean Total HL2 Reads(Mb) HL1 Raw Total Sample 1 Table S1 TABLE S2 FIGURE S1 FIGURE succinctus. 6 performed. FIGURE was resistance and accumulation TTX ZD1. generated with with genes ZL3 comparison the up-regulated in that 5 more (blue) means FIGURE generated down-regulated ZD1-VS-ZL3 than HL3 HD1. (orange) the with genes comparison that up-regulated in means more (orange) HD1-VS-HL3 down-regulated Examples: than group. (blue) treatment as set communities in non-toxic and toxic 4 the FIGURE that indicated patterns. It expression specimens). similar toxic show HL: generally & ZL specimens, non-toxic HD: 3 FIGURE The patterns. annotated 2 FIGURE disease. and health coast. in 1 stress FIGURE reticulum endoplasmic and legends interferons biology, Figure molecular I and Type cell A.D. of review Garg, Pharmacology International toxicity. J., drug-induced tetrodotoxin in Sprooten a stress 36. reticulum of endoplasmic properties to of some related Perspectives. Role Toxicon. & and genes F. Research niphobles. novel Purification Bernard, Takifugu of F., K. kusafugu, Fabienne, Identification of Michiko, 35. plasma F., al. blood Kiyoshi, to the et adaptations Y., from T. protein molecular Kunio, Saito, binding reveals M., S., Takashi, genome Akimoto, Scallop 34. S., al. Sato, Toxicon pufferfish. et H., in J. Kondo, intoxication Zhang, tetrodotoxin J.H., X., Lee, Xun, mutation 33. X., Communications. channel Nature Hu, Sodium neurotoxins. X., and al. Nature Sun, life et semi-sessile psp. Y., T. of Li, Scheuer, risk S.P., increases 32. Macquarrie, clams K., in resistance Konoki, saxitoxin L., to for Connell, leading Systems V.M., Intelligent reconstructing Bricelj, on and Conference evaluating, 31. International detecting, for (1999) Proceedings. program 138-48 sequences. a Biology est Estscan: Molecular P. in Bucher, regions & coding C.V. potential Jongeneel, C., Iseli, 30. N.succinctus h la ed ult erc fe ult control. Quality after metrics quality reads clean The . h ieidctdi elwi el dnie yorstudy. our by identified newly is yellow in indicated site The ossec f2-[]?C)adata xrsinlvlb TPR i oeta Esrelated DEGs potential Six RT-PCR. by level expression actual and -([?][?]Ct) 2 of Consistency C ltfrdffrneo xrsinlvlbtentetxcadnntxccmuiis(D& (ZD communities non-toxic and toxic the between level expression of difference for plot PCA ucinlantto o both for annotation Functional itiuino noae pce o both for species annotated of Distribution Espten ewe oi n o-oi pcmn of specimens non-toxic and toxic between patterns DEGs oi n o-oi omnte of communities non-toxic and Toxic oeta mn cd uainst eae ihTXrssac nsdu hne of channel sodium on resistance TTX with related site mutation acids amino Potential h o lsicto aeoisfrteDG fbt species. both of DEGs the for studied. categories specimens classification all top for The quality and content base of Distribution n Din ZD and 4 pyi california Aplysia N.variciferus 1,021(2016). (1),e00211 nntxcsmls eesta oto ru n LadZ were ZL and HL and group control as set were samples) (non-toxic pce aeacutdfrtetpproportion. top the for accounted take species Nassarius 49 350 9993(2007) ,939-953 .succinctus N. 7 pce rm7databases 7 from species 6-1 (2020). ,63-118 .succinctus N. 8 12 (2017) ,1721 and 434 and .succinctus N. .variciferus N. 7377(2005) ,763-767 .variciferus N. 38 and 463 , olce rmChina from collected hc hwsimilar show which .variciferus. N. ± 6 (2000). 468 HD N. Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. ui-3dw 08NA 1028 NA NA down 484 2844 pathways secretion NA Salivary Metabolic NA pathway fly degradation signaling - Lysine pathway IL-17 signaling Hedgehog 1533-1556 NA 1079 632 NA Spliceosome 1826 NA 823 down down 1315 738 736 cascades 751 coagulation down and Complement down down 861-1061 down NA 4214-7132 down up reticulum endoplasmic in down processing down pathways Protein Metabolic export metabolism down Protein chlorophyll and Porphyrin 2221-3222 Spliceosome Spliceosome cancer Gastric pathways Amoebiasis 1124 Metabolic metabolism up chlorophyll 1507-1399 and down Porphyrin 986 mucin-13 4-like 979 channel NA 376 ion NA 4-like ligand-gated protein NA receptor-related lipoprotein NA low-density 2207 NA NA 2C N-methyltransferase histone-lysine NA X1 isoform disease T Huntington up B 465 disease; troponin 72.16 homolog 453 NA Parkinson disease 40 disease; 953 Huntington factor 1369 Alzheimer cytoskeleton disease; pre-mRNA-processing (NAFLD); up actin Parkinson 74.95 disease of disease; 263 liver Regulation Alzheimer 686 precursor 74.53 fatty junction; (NAFLD); protocadherin-15b 415-678 up Non-alcoholic Adherens disease Thermogenesis; X1 570 liver contraction; isoform fatty [EC:3.4.21.120] up muscle muscle oviductin Non-alcoholic 71.04 Cardiac NA striated Thermogenesis; pathways; reticulum chain, up contraction; endoplasmic Metabolic up heavy muscle pathway in phosphorylation; myosin 70.42 436-611 Cardiac signaling 6-like Oxidative processing pathways; Wnt polypeptide Protein Metabolic light phosphorylation; myosin 70.6 382-852 Oxidative up protein 71.89 uncharacterized 597-737 91.03 mucin-5B up 1855 73.06 up 1906 NA up 91.05 up AGAP011395-PA inhibitor pathway 91.04 673 25 Pathway factor 3044-3563 up up protein tissue RNA-binding up PROTEIN: X2 up QUALITY 90.65 1 isoform LOW protein Sec24D binding Length protein enhancer transport 90.71 I protein type up trend virus Expression immunodeficiency 90.95 human A1b up ribonucleoprotein GH24401-RA 95.53 nuclear transcript heterogeneous from product up 96.85 gene GH24401 95.42 up dehydratase 96.86 up delta-, aminolevulinate, 96.88 protein up recognition up signal dehydratase 96.74 acid delta-aminolevulinic A2/B1 ribonucleoprotein 96.75 nuclear heterogeneous LOC102093901 96.85 protein uncharacterized A-like 98.45 helicase RNA 1-like ATP-dependent cytoskeletal II type 98.42 9 keratin, polypeptide box 6.66 (Asp-Glu-Ala-His) DEAH A helicase 6.74 RNA ATP-dependent 6.7 X1 isoform mucin-2 platelets-like 6.73 for adhesin 2 serine-rich member 6.67 family protein WAS 5 6.69 protein frizzled-related secreted 6.7 H1E-like I histone subunit oxidase 6.64 c II cytochrome subunit 44.41 oxidase c cytochrome 83-like 44.91 protein shock heat 44.65 44.86 Definition 44.47 44.59 species 2 44.67 Table 44.24 61.54 59.92 Ratio(%) Reads Clean 59.92 ZD3 63.16 Q30(%) ZD2 Reads Clean 63.16 ZD1 Q20(%) Reads 63.16 Clean ZL3 Bases(Gb) ZL2 62.14 Clean Total ZL1 60.55 Reads(Mb) Clean Total HD3 Reads(Mb) HD2 Raw Total Sample otdffrn xrse ee mn l oi tetet n o-oi cnrl ruso two of groups (control) non-toxic and (treatment) toxic all among genes expressed different Most 8 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. i.2 Fig. NA NA 3356 255 1 ataxia Spinocerebellar Fig. down NA 1456 down Pathway transport RNA 1396 Length NA NA trend Expression 548 down factors 1035 transcription 780 Basal down 751 down down down [EC:3.4.17.21] dipeptidase down acidic N-acetylated-alpha-linked protein receptor ncharacterized lipoprotein density low very mucus/gel-forming oligomeric 2, mucin homeolog L 3 tektin-3 factor M-like associated member protein 4 binding TATA-box family domain lectin Nup205 C-type protein complex pore nuclear Definition 9 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. i.4 Fig. 3 Fig. 10 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. i.5 Fig. 11 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. i.6 Fig. 12 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. 13 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. 14 Posted on Authorea 12 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159724546.61271404 — This a preprint and has not been peer reviewed. Data may be preliminary. 15