Reference: Bio!. Bull. 195: 112-114. (October. 1998) Effect of Methyl Farnesoate on Late Larval Development and Metamorphosis in the Prawn Macrobrachium rosenbergii (Decapoda, A Palaemonidae): Juvenoid-like Effect? URI ABDU'. PETER TAKAC2 HANS LAUFER3 AND AMIR SAGI1 * , , 1 Department ofLife Sciences, Ben-Gnrion University ofthe Negev, P. O. Box 653, Beer-Sheva 84105, Israel; 2Institute ofZoology, Slovak Academy ofSciences, Bratislava 84206, Slovak Republic; ami 'Department ofMolecular and Cell Biology, University ofConnecticut, Storrs, Connecticut 06269-3125 Abstract. Methyl farnesoate (ME), the unepoxidated al., 1985), but its physiological significance is not clear. formofinsectjuvenile hormone III. wasdetected in larvae The first report of the presence of MF in Crustacea was of the freshwater prawn Macrobrachium rosenbergii, that of Laufer et al. ( 1987), who demonstrated that MF which metamorphose to post-larvae following 1 1 larval is secreted by the mandibular organ of the spider crab stages. The possible role of ME as a morphogen was Libiniu emarginata and is found in its hemolymph. Since studied by administering the compound toM. rosenbergii then, MFhasbeen found in avariety ofcrustaceans (Borst larvae via an Anemia vector. Higher ME levels caused i-tat.. 1987; Laufer and Borst, 1988; Landau etal., 1989), earlier retardation of late larval growth, and the highest including the freshwater prawn Macrobrachium rosen- ME dose retarded larval development. Furthermore. sig- bergii (Sagi et al., 1991 ). nificantly affected the patterns of metamorphosis and the JHs are involved in the regulation ofa substantial num- appearance of intermediate individuals exhibiting both berofinsect functions, including development, reproduc- larval and post-larval morphology and behavior. Three tion, and behavior. JHs are probably best known for their intermediate types weredefined, two ofwhich were found effect on the growth and differentiation of insect epider- only at the MF-treated groups and one that was exclusive mal cells. When JHs are present during larval instars, the to the higher dose treatments. The relative abundance of cuticle that is secreted has larval characteristics; in the intermediate specimens increased from 2% in the control absence of JHs. the cuticle exhibits pupal characteristics to 32% in the high MEconcentration, which suggests that (see Laufer and Borst, 1983, for a review). Exogenous ME has ajuvenoid-like effect in this decapod crustacean. application of JHs may cause the appearance of various intermediate forms, and in some cases, of supernumerary Introduction larvae, as a result of differences in the growth rates of Juvenile hormones (JHs) a family ofepoxidated ses- various parts of the body (allometric growth) (reviewed quiterpenoids act as growth regulators in insects (Wig- in Sehnal, 1983). It has been suggested that exogenous glesworth, 1970). Methyl farnesoate (ME), the unepoxi- JH applied during a critical period may function at the dated form of JH III, has been detected in some insect cellular level, resulting in mosaic animals with character- species (Slama. 1971; Lanzrein el ai. 1984; Bruning et istics of two different stages (see Riddiford, 1994, for a review). Thus, it is likely that the effect of a particular juvenoid on insect development is determined by the de- R*eAcuetihvoerd 6toNowvheommbercor1r9e9s7p;oancdceenpcteedshIoIIulJdulyhe19a98d.dressed. E-mail: velopmental timing and the effective concentration ofthe juvenoid (Slama. 1995). sagia@bgumail.bgu.ac.il Ahhreviatiims: MF, methyl farnesoate; JH,juvenile hormone. The regulation oflarval development and metamorpho- 112 MF AND METAMORPHOSIS IN MACROBRACH1UM 113 sis in crustaceans is not fully understood. In cyprids of washed with4% NaCl and acetonitrile, and extracted with barnacles, JH I and JH analogs caused premature meta- hexane. The hexane fraction was evaporated by means of morphosis without attachment of the cyprid to the sub- a rotary evaporator. The dry matter was dissolved in hex- strate (Gomez et cil., 1973; Ramenofsky et al., 1974; aneandpassedthrough aSep-Paksilicacartridge (Waters, Tighe-Ford, 1977). In decapods, JH analogs retarded the Milford, MA), and the cartridge was rinsed with methy- development of larvae ofthe mud crab Rhithropanopeus lene chloride (Abdu etal., 1998). The methylene chloride htirrixii (Christiansen et al., 1977a, b) and the growth of was evaporated, and the residue was dissolved in 100 /j\ early larval stages and post-larvae ofthe estuarine shrimp of hexane, loaded into a Beckman HPLC and analyzed Palaemonetes pugio. In contrast, growth was enhanced according to Sagi etal. ( 1991 ). The nonbiological isomer, in the premetamorphic stages of P. pugio (McKenney cis-trans MF, was used as the internal standard, and the and Celestial, 1993). Treatment of Homants americanus retention times were compared to an external standard MF larvae with JH III increased the duration of larval devel- containing both the biological (all-trans ) and nonbio- MF opment and influenced the size of the carapace and ap- logical isomers. The putative trans-trans peaks of pendages (Hertz and Chang. 1986). Injections ofJH I into larvae at stages 4, 6, 9, 10, and 1 1 were collected from H. americanus larvae gave rise to a number ofintermedi- the HPLC and loaded onto a Hewlett Packard gas chro- ate stages (Charmantier et al.. 1988). matograph with a 30-rn cross-linked and surface-bound MF is the only JH-like compound found thus far in dimethylpolysiloxane column (0.5 min to 70C. 70- crustaceans. A few studies have targeted its regulatory 280C. 20C/min) connected to a mass spectrometer. role in development and metamorphosis in decapods. Ap- plication of MF to H. americanus larvae prolonged meta- Effect ofmethylfarnesoate on late larval growth and morphosis by a small but significant amount of time development (Borst et al.. 1987). In M. rosenbergii, MF retarded Three experimental groups, each containing 15 M. ro- growth and development in larval stages 5 to 9 (Abdu et senbergii stage-9 larvae, were fed with Anemia enriched acmtiaae.sbnIeontl1fo9saou9rnos8dmM).e.imnesrcetorcsautemssnotbra(ecpSrehgnaioynisds,eirswthhaaeirncedphrsCoeicxhmheiailsnbasgire,tstso1oa9tf8"h6olc)aso.revmaSimlnuochdnheevmtieisylmpoteeph"--e cawwSlaiiE.tttLe(hhsC1.O09t.T9)h28h.e1)e..EevA0axe.cp3hc5eiho,crnlittoermrroeelna0att.glam5orle9noneust/pyjgsw(wteaMatesFshmy/lfpcmeeoldnrasfwliaoicscrtotcmhheooedArlddnoifeinanmgn1i2dttahodraSeeArenUbkrPdirpeEculphRalesie-d--t of larval development (Sollaud. 1923). encompassing 11 tic containers (300 ml) that were constantly aerated. The satnadgeSso,of,ol1l9o69w)e.dLbayrvmaeltadmeovreplhoopsmiesntinitnoMp.ostr-olsaernvbaeerg(iUinios twaaitneerdwaats27replac1eCd.daTihley,laanrdvaethweetreempfeerdataudreliwbiatsummawiint-h characterized by significant growth of the carapace from Anemia nauplii that were enriched daily with MF. Five stage 1 through stage 11. During metamorphosis (from larvae perreplicate were randomly sampledtwice a week; stage 1 1 to post-larva), growth is allometric, with in- larval stage wasdetermined andthe length ofthe carapace creases in size being limited to certain parts of the body was measured with an ocularmicrometer; the larvae were (excluding the carapace) (Abdu. 1996). then returned to the growth chamber. In the calculation In this study, M. rosenberMgiFi was used as a model spe- of the average larval stage for each sampling, all post- cmieenstfoarnsdtumdeytianmgorthpehorsoliesooff crusitnatcheeanlsa.teMlaFrvawlasdevienldoepe-d i1n1disvtiadgueasls)(inwcelruedidnegsibgontahtepdosats-la1r2.vaeThaendavienrtaegremeldairavtael odeftemcotrepdhoinlotghieclaalrvafee,ataunrdesitsduerfifencgt omnettahmeormapnhiofseisstatwiaosn bsteatgweeeanndtrtehaetmeanvtesr,agaendcatrhaepascteatilsteincgatlhswigenrieficcaonmcpeawraesd studied. ANOVA tested using one-way followed by the least significant difference (LSD) test P = 0.05. The experiment Materials and Methods wasrepeated three times with similarresults; each experi- Detection ofmethylfarnesoate in Macrobrachium ment contained 180 larvae. rosenbergii lan-ae Assessment ofmetamorphosis in Macrobrachium M. rosenbergii larvae were hatched and reared ac- rosenbergii cording to Daniels etal. (1992). Larval stages were determined according to Uno and Soo (1969). To detect the Among the many changes occurring during metamor- presence of MF, about 1 g of larvae at each ofthe stages phosis (Ling, 1969; Uno and Soo, 1969), the following 4, 6, 9, 10, and 1 1, and post-larvae were collected and six predominant changes were selected to distinguish be- homogenized in 2 ml of4% NaCl and 5 ml ofacetonitrile. tween larvae and post-larvae of M. rosenbergii in this The homogenates were filtered through a-cellulose. study; ( 1) In larvae, the abdomen is bent such that the 114 U. ABDU ET AL. cis-transMF the pereiopods. During the metamorphic process these 4.00 -I 4.88 long exopodites degenerate and become reduced to knobs at the postmetamorphic molt (Ling, 1969). (3) In stage 1 1 larvae, the endopodite of the second pereiopod has three segments, the second and third segments being at 3,00- an angle to the first, which is not fully developed. In contrast, in post-larvae there are four segments, all of which are straight, and the first segment is fully developed, as in the adult. (4) The carapace median tooth is evident from larval stage 4 onward, but disappears during metamorphosis and is absent in the post-larva. (5) In stage I 1 larvae, the entire dorsal margin of the rostrum is equipped with numerous small teeth, but no ventral teeth 1.00 are present, whereas the rostrum of the post-larva has I 1 large dorsal teeth and 5 ventral teeth. (6) Larvae are planktonic. swimming tail up. and in most cases tail first, ventral side up; during metamorphosis, they settle to the 5.00 5.50 6.00 bottom. Post-larvae are benthic, swimming and behaving as do the juveniles. Retention time(min ) The relative abundance of the different intermediate Figure 1. HPLC detection ofmethyl farnesoate in Macrdbnicliiuin types were compared between treatments, and the statisti- rosenbergii larvae at stage 4. Cis-trans methyl farnesoate was used as cal significance was tested using the Mann-Whitney U the internal standard. test. Results pleopods and the walking legs are not in the same plane, Presence ofmethylfarnesoate in Macrobrachium whereas in post-larvae, the abdomen is straight, with the rosenbergii larvae pleopods and the walking legs in the same plane. (2) MF was found in hexane extracts of M. rosenbergii Larvae swim mainly by beating the long exopodites of larvae at stages 4, 6, 9, 10, and 1 1, and in post-larvae. A 50,000 1 69 - i1n12c3r-us1t1a2c7e.an reproduction and development. Insect Biochem. 17: senbergii larvae were within the physiological range (Ta- Kruning. K., R. R. Shivers, and B. Lanzrrin. 1985. Methyl tarne- kac, pers. comm. and Abdu et cii, 1998). the possibility soateandjuvenilehormone III in normal and precocene-treatedem- of nonspecific toxicity could not be entirely ruled out. In bryos of the ovoviviparous cockroach Nauphaeta cinereu. Int. ./. crustaceans, intermediate larval forms may be produced Im-cru-hr. RcproJ. Ocv. 8: 269-278. by eyestalk removal during a "critical period," as has C'hacnrgu,stEa.ceSa.n, Mmo.ltJi.ngB:ruacem,ulatnid-hSo.rmLo.naTlamsoynstee.m.19A9m3..ZoRo/e.gu3l3a:ti3on24o-f been shown for Rhithropanopeus harrisii (Costlow. 329. 1966a), Sesartnci reticiiliittini (Costlow, 1966b). H. anieri- Charmantier, G., and D. E. Aiken. 1987. Intermediate larval and canus (Charmantierand Aiken, 1987), Pahu'immetes vnr- postlarval stages ofHomarus americanus H. Milne Edwards. 1837 ians (Le Roux, 1984). andAlphensheterochaelis (Knowl- (Crustacea: Decapoda). J. Crust. Biol. 7: 525-535. Charmantier, G., M. Charmantier-Daures, and D. E. Aiken. 1988. ton, 1994). SnyderandChang ( 1986) suggested that inter- Larval development and metamorphosis of the American lobster mediate larval forms in H. umericuinis were probably a Homarusnmericamis(Crustacea: Decapoda):effectofeyestalkabla- manifestation of inadequate nutrition or other stresses. tion and juvenile hormone injection. Gen. Comp. Endocrinol. 70: oHfowienvteerrm,edKinaotewlftoornms( 1i9n94A). shehtoewreodchatehaltisthlearavpaep,eawrhaincche Chri3s1t9i-a3n3s3en., M.E., J.D. Costlow, Jr., and R.J. Monroe. 1977a. have a Iccithotropic mode of nutrition, was due to direct Edfefveecltospomfenttheojfuvtehneilmeudh-ocrrmaobneRhmiithmriocpaZnRo-p5e1u5s (liAalrtrio.ssiiid)inonvalrairovuasl or indirect hormonal control from the eyestalk. The re- salinities and cyclic temperatures. Mar. Biol. 39: 269-27. ports of the isolation and characterization of mandibular Christiansen, M.E., J.D. Costlow, Jr., and R.J. Monroe. 1977h. MF AND METAMORPHOSIS IN MACROBRACHIUM 19 Effects of the juvenile hormone mimic ZR-512 (Altozar) on larval of the juvenile female spider crab Libinia emarginata. Invertebr. development of the mud-crab Rhithropanopeus harrisii in various Reprod. Dcvel. 31: 63-68. salinities and cyclic temperatures. Mar. Biol. 39: 281-288. Le Roux, A. 1984. Quelques effetsde 1'ablationdes pedonculesocu- Costlow,J.D.,Jr. 1966a. Theeffectofeyestalkextirpationonlarval lairs surles larves et les premiers stadesjuveniles de Palaemonetes development of the mud crab. Rhithropanopeus harrisii (Gould). various(Leach)(Decapoda.Palaemonidae).Bull. Soc.Zool. Fr. 109: Gen. Comp. Endocrinol. 3: 120130. 43-60. Costlow,J.D.,Jr. 1966b. Theeffectofeyestalkextirpationon larval Ling,S.W.1969. ThegeneralbiologyanddevelopmentofMacmbra- development of the crab Sesarma reticiilanim Say. Pp. 209-224 chium rosenbergii. De Man. FAO Fish. Rep. 57: 589-606. in Sonit' Contemporary Sunlit'* in Marine Science. H. Barnes, ed. Liu, L.. and H. Laufer. 1996. Isolation and characterization ofsinus Appleton-Century-Crofts. New York. gland neuropeptides with both mandibularorgan inhibiting and hy- Daniels,W.H., L. R. D'Abramo,and L.D. Parseval. 1992. Design perglycemic effects from the spidercrab. Libinia emarginata. Arch. saynsdtemmanfaorgefmreesnhtwaotferaprcalowsnesd,,Mraeccrirocburlaatcihngiu"mclreosaernwbaetregri"i DhaetcMhaenr,y McKIennsencetyB,ioCc.hLe.m,.JrP.h,yasinodl.D.32M:.3C7e5l-es3t8i5a.l. 1993. Variationsinlarval GomJ1eu8zv7,e9n.iEl.J.eDS.hh,oerlDlm.foiJsn.heFRameuisl.mkinc1es1r::,6eV5fVf-.e7cA3t.s.oNnewcmirarnip,edancrdusCt.acIeraenlamnde.ta1m9o7r3-. PresgdPthurwyrosiiwicntohghl,.taGon1x.d0i5cDmoC.es,:itsaH2b.3bo9ylV-iV2osa4jmn5ta.oisfnesaken,cteAs.gtruJoa.rwiLtnehentsrzeh,gruilamanptdorP.aJ.lCaMoe.mmopRn.aebtBieinsoocpvhtiiecgmhi..o Hertpzh,osi\sV..SA.c,ieannced1E7.9:S.8C1h3a-n8g1.4.1986. Juvenile hormone effects on h1o99r6m.one.BiAorcchhe.miIsntsreyctofBiporcohteemin.sPthhyastiobli.nd32a:nd40m7e-t4a1b9o.lizejuvenile metamorphosisoflobsterlarva.Int. J. Invertebr. Reprod. Devel. 10: Ramenofsky, M., D.J. Faulkner, and C. Ireland. 1974. Effects of 71-77. juvenile hormone on cirriped metamorphosis. Biochem. Biophys. Homola. E., and E.S. Chang. 1997a. Distribution and regulation of Res. Com/mm. 60: 172-178. esterases that hydrolyze methyl farnesoate in Homarus americam/s Riddiford. L.M. 1994. Cellular and molecular actions of juvenile and other crustaceans. Gen. Comp. Endocrinol. 106: 62-72. hormone I. General considerations and premetamorphic actions. Pp. Homola, E., and E.S. Chang. 1997b. Methyl farnesoate: crustacean 213-274 inAdvances in Insect Physiology vol24. P. D. Evans, ed. juvenile hormone in search offunctions. Comp. Biochem. Physiol. Academic Press, London. 117B: 347-356. Sagi, A., E. Homola, and H. Laufer. 1991. Methyl farnesoate in King, L.E.. Q. Ping, G.D. Prestwich, and S.S. Tobe. 1993. The theprawnMacrobrachium rosenbergii: synthesis bythe mandibular characterization of haemolymph methyl farnesoate binding protein organinvitro, andlitersinthehemolymph. Comp. Biochem. Physio/. and the assessment ofmethyl farnesoate metabolism by the haemo- 99*8: 879-882. lymph andothertissues from Procambarusclarkii. Insect Binchem. Sehnal, F. 1983. Juvenile hormone analogues. Pp. 657-672 in Endo- Mul. Biol. 25: 405-501. crinology ofInsects, R. G. H. Downer and H. Laufer. eds. Alan R. Knowlton, R.E. 1994. Effects oflarval eyestalkextirpation on mor- Liss. New York. phogenesis and molting in the snapping shrimp Alplteus hetero- si.mi,i. K. 1971. Insectjuvenile hormone and analogues. Annu. Rev. cliaelis Say. J. E.\p. Zool. 270: 162-174. Biochem. 40: 1079-1102. Landau, M., H. Laufer, and E. Homola. 1989. Control of methyl Slama, K. 1995. The present status of the mode of action of insect farnesoate synthesis in the mandibular organ of the crayfish Pro- juvenile hormone. Netli. J. Zool. 45: 71-78. cambarus clarkii: evidence for peptide neurohormones with dual Snyder,M.J.,andE.S.Chang. 1986. Effectsofeyestalkablationon functions. Invertebr. Reprotl. Devel. 16: 165-168. larvalmoltingratesandmorphologicaldevelopmentoftheAmerican Lanzrein, B., H. Imboden, C. Burgin, E. Bruning, and H. Gf'eller. lobster, Homarus americanus. Biol. Bull. 170: 232-243. m1e9t84h.yl fOanrnetsiolaertse,,oarnigdine,cdaynsdterfouindcstiionnsemobfryjouvneincildeevheolrompomneentIIoIf, Sollnaaued",. MI..PEa.rti1e92de3s.cripLteived.evLealocpopnedmeennstatiloanrvapirroegrdeesssiv"ePadleae1m'oonntio-- Btiheosyonvtohveisviis,parMuestacbooclkirsomacahndNMauopdheoeotfaAcctiinoenreoaf.InPvpe.rt4e5br4a-t4e6H5ori-n Takagecn,esPe.., BJu.lSl..BB.iolA.hl5,7:an5d09H-.60L3a.ufer. 1997. Seasonal differences mones. J.HoffmanandM. Porchet,eds.Springer-Verlag.NewYork. Linibmineitahyelmafragrinneastoaa.teInevsetretreabsre.aRcteipvriotdy.inDetviesls.ue3s1:of2t1h1e-2s1pi6d.er crab Laufer, H., and K. H. Albrecht. 1990. Metabolism ofmethyl farne- Tamone,S.L.,G.D. Prestwich, and E.S. Chang. 1997. Identifica- soatein vitroby peripheral tissuesofthespidercrabLibiniiiI'liiargi- tion andcharacterizationofmethyl farnesoate bindingproteinsfrom nata (Decapoda). Pp. 217-222 inAdvances in Invertebrate Repro- the crab. Cancermagister. Gen. Comp. Endocrinol. 105: 168-175. duction 5, M. Hoshi and O. Yamashita. eds. Elsevier, Amsterdam. Tighe-Ford, J.D. 1977. Effects ofjuvenile hormone analogues on Laufer,H.,andD.W.Borst. 1983. Juvenilehormoneanditsmecha- larval metamorphosis in the barnacle Eliminus modestus Darwin. nism ofaction. Pp. 203-216 in Endocrinology ofInsects. R.G. H. (Crustacea: Cirripedia). J. E.\p. Mar. Biol. Ecol. 25: 163-176. Downer and H. Laufer. eds. Alan R. Liss, New York. Uno,Y.,andK.C.Soo. 1969. LarvaldevelopmentofMacrobrachium Laufer, H., and D.W. Borst. 1988. Juvenile hormone in Crustacea. rosenbergii De Man, reared in the laboratory. J. Tokyo Univ. Fish. Pp. 305-313 in Endocrinology of Selected Invertebrate Types. 55: 179-190. R. G. H. Downer and H. Laufer, eds. Alan R. Liss, New York. Wainwright, G., S.G. Webster, M.C. Wilkinson, J.S. Chung, and Laufer,H..D. W. Borst, F.C. Baker,C. Carrasco,M.Sinkus, C.C. H.H.Rees. 1996. Structureandsignificanceofmandibularorgan- Reuter, L. Tsai, and D.A. Schooley. 1987. The identification of inhibitinghormone inthecrab. Cancerpagurus. J. Biol. Client. 271: a crustaceanjuvenile hormone. Science 235: 202-205. 12749-12754. Laufer, H., P. Takac,J.S.B. Ahl,and M. R. Laufer. 1997. Methyl Wigglesworth, V. B. 1970. Insect Hormones. University Reviews in farnesoate and the effect ofeyestalk ablation on the morphogenesis Biology, p. 159. Oliver and Boyd, Edinburgh.

Effect of Methyl Farnesoate on Late Larval Development and Metamorphosis in the Prawn Macrobrachium rosenbergii (Decapoda, Palaemonidae): A Juvenoid-like Effect?

Similar Effect of Methyl Farnesoate on Late Larval Development and Metamorphosis in the Prawn Macrobrachium rosenbergii (Decapoda, Palaemonidae): A Juvenoid-like Effect?