In Vivo Effects of Dopamine and Dopaminergic Antagonists on Testicular Maturation in the Red Swamp Crayfish, Procambarus clarkii

7 Pages·1995·3.3 MB·English

by  R Sarojini

Reference: Bin/ Bull 189: 340-346. (December, 1995)
In Vivo Effects of Dopamine and Dopaminergic
on Testicular Maturation in the
Antagonists
Red Swamp Procambams clarkii
Crayfish,
RACHAKONDA SAROJINI, RACHAKONDA NAGABHUSHANAM,
AND MILTON FINGERMAN*
Department ofEcology. Evolution, andOrganismal Biology.
Tulane University. New Orleans. Louisiana 70118
Abstract. In vivo, dopamine (DA) inhibits testicular of crustaceans, including crayfishes, is well established.
maturation in the red swampcrayfish, Procambamsclar- 5HT-like immunoreactivity in the central nervous system
kii. Crayfish given DA injections had a smallertesticular ofthe red swampcrayfish Procambamsclarkii. the species
index, smaller testicular lobes, fewer mature sperm, and used in the present study, was demonstrated by several
less-well-developedandrogenic glandsthan didthecontrol investigators (Fujii and Takeda, 1988; Arechiga et al..
crayfishgiven physiological saline. Malesadministered 5- 1990; Real and Czternasty, 1990). In addition, 5-HT has
hydroxytryptamine(5-HT)oraDAreceptorblocker, spi- been identified and quantitatively measured by high per-
perone or pimozide, showed enhanced testicular matu- formanceliquidchromatography (HPLC)inProcambams
rationand morehighlydevelopedandrogenicglandsthan clarkiiby Kulkarni and Fingerman (1992). Usingthecrab
did the control crayfish. When equimolar amounts of5- Carcimis macnas. Kerkut et al. (1966) provided the first
HT and DA were co-injected, the actions ofDA and 5- convincingevidencefortheexistenceofDA inthenervous
HT were found to be antagonistic. These results can be system ofa crustacean. Neurons with DA-like immuno-
explainedbyassuming notonlythat 5-HTtriggersrelease reactivity have been visualized in the crayfish Orconectes
of the gonad-stimulating hormone (GSH) but that DA limosiis (Elekes et al.. 1988), the lobster Homarus gam-
(a)triggersreleaseofthegonad-inhibitinghormone(GIH), mams (Barthe el al.. 1989). and Procambams clarkii
(b)inhibitsGSH release, or(c)doesboth (a) and (b), with (Mercier et al.. 1991). By use of HPLC, Elofsson et al.
GSH and GIH affecting the androgenic glands directly, (1982) showed the presence ofDA in the nervous system
thereby regulating release of the androgenic gland hor- ofthe crayfish Pacifastacus leniusculus.
monethat hasthe well-established roleofstimulatingtes- In decapod crustaceans the major neuroendocrine
ticular maturation and spermatogenesis. component ofthe eyestalk, the medulla terminalis X-or-
gan-sinus gland complex, is the source ofthe gonad-in-
Introduction hibiting hormone (GIH) (Panouse, 1943). In contrast, a
Biogenic amines function as neurotransmitters in a gonad-stimulatinghormone(GSH)ispresentinthebrain
wide array of animals (Werman, 1966; Gerschenfeld. and thoracic ganglia (Otsu. 1960, 1963; Eastman-Reks
1973; Fingerman, 1985). Among the demonstrated roles and Fingerman, 1984). Data from thislaboratory provide
ofat least some ofthe biogenic amines in crustaceans is the basis for the hypothesis that 5-HT triggers release of
regulation ofrelease ofneurohormones (Fingerman and GSH in bothsexesofthefiddlercrab Ucapugilator(Rich-
Nagabhushanam, 1992; Fingerman et ai. 1994). ardson et al.. 1991; Sarojini et al.. 1993) and in Procam-
The presence of the biogenic amines 5-hydroxytrypt- bamsclarkii (Kulkarni and Fingerman, 1992; Sarojini et
amine (5-HT)and dopamine (DA) in the nervoussystems al. 1994). On the other hand, DA has so far been found
to antagonize the gonad-stimulating action of 5-HT in
Received 8 May 1995;accepted 14 September 1995. females of Procambams clarkii (Sarojini et al.. 1995a)
*To whom correspondenceshould be sent. and in males of Uca pitgilator(Sarojini el al.. 1995b).
340
CRAYFISH TESTICULAR MATURATION 341
In male crustaceans, in addition to the two neurohor- Company (St. Louis, MO). The drugs were dissolved in
mones, GSH and GIH, the androgenic gland hormone crayfish physiological saline (Van Harreveld, 1936). To
(AGH) hasa majorrole in thecontrol ofspermatogenesis. prepare the stock solution of spiperone a few drops of
The function ofthe androgenic gland in controlling de- acetic acid were added to facilitate solubilization. When
velopmentand maturation ofthe reproductivesystem and DA was used 1 X 10~6 mol, 1 X 10~7 mol and I
secondary sexual characteristics in male crustaceans was X 10~s mol percrayfish were injected. Theamountsof5-
first described by Charniaux-Cotton ( 1954). Initiation of HT, si ' and pimozide injected were 1 X 10~6 mol
AGH
spermatogenesis isdue to circulating (Payen, 1973). per crayfish. The volume injected into each crayfish was
Spermatogenesis stops when the androgenic glands are 100 pi.
removed (Charniaux-Cotton, 1964; Puckett, 1964; Na- Thetesticular index(TI)wasdetermined foreachcray-
gamine el til.. 1980). Removal ofboth eyestalks, thereby fish used in these experiments according to the standard
removing the source of GIH, results in hypertrophy of formula:
the androgenic glands and precocious spermatogenesis
Weight ofthe testes
(Meusy, 1965; Demeusy, 1967; Payen ct a/.. 1971). Thus. ^
GIH appears to exert its effect on the testes indirectly, by Weight ofthe crayfish
GinShiHbiting the androgenic glands. On the other hand, a Thetestesand androgenicglandswere removed from each
is required to activate the androgenic glands for ofthecrayfish used in theseexperimentsafterthecrayfish
spermatogenesis to occur (Juchault and Legrand. 1965), were weighed at the time ofsacrifice. When these organs
aprocessthat Payen(1980)referredtoasapositivecontrol were removed the testes were weighed. The testes and
ofthe androgenic glands by a neurohormone. Gupta el androgenic glands were then fixed for 24 h in Bouin's
ul. (1989) suggested from their studies ofthe crab Para- fluid, dehydrated in an alcoholic series, and embedded in
tiscldpu/eiustaohayndirnocdrreaosmeuisntthhaetthheemionlacytmipvehplhiatesreooffGtIheHtewsittehs spatraaifnfeidnw(imt.hp.De5l6af-i5el8dC's).heSmeacttoixoynlsin(7f^omll)owweedrebycuctoaunnd-
concomitant decreases in the liters ofGSH and AGH. terstaining with alcoholic eosin (Bancroft and Stevens,
In view ofthe facts that 5-HT stimulates gonadal mat- 1982). The diameters of50 testicular follicles (^m) in the
DurAatiaonntaignobnoitzhesmatlheisaancdtifoenmaolfe5P-roHcTamibnafreumsalcelasrkoifiathnids tmeisctresosocfoepaechfitmtaedlewiwtehraenmoecausluarremdicbryomuesteeorf.aThceomnpuomubnedr
swclphaeerctkiiheise,,r(tDbh)iAswhiiennhtvihebseitrtisg5ta-etsHitoTincuawlnaadrsmDadAetsuiragacntteiadonnttaoignodnPeirtsoetciracmamilbnlaeyr(ouans) doliifkaemmwaeittseuerrmsee(asusrpuner)reodmf.p5Te0hrecefleollxslpieicnlreiemwaecanhstsaanwldseroroedgepetneeirrcfmogirlnmaeenddd.twwTeihrceee
gonadal maturation and spermatogenesis in the male andtheaveragedresultsarepresented inthefigureswhere
carfafyefcitsehd,wahnedn(Dc)Awhoertahedroptahmeinaenrdrgoigcerneiccepgtloarndbslowciklelrbies ebaarcsh1vCalaunedreSpCreisnenFtisgutrhee m7ewahinchfo,ra2s0wec'rlalyfeixsph,laeixncebpetlofwo.r
iDnAjected. This is the first report that shows injection of represent the means for 40 crayfish. The data were ana-
affects the androgenic glands ofany crustacean. lyzed by means ofStudent's /-test with significance set at
the 95% confidence interval. Standard errors ofthe means
Materials and Methods were also calculated.
Experimental animals
Results
Specimens of the red swamp crayfish, Procambarus
clarkii, were purchased from a local seafood dealer. In Effect of D ion the testes
the laboratory they were maintained in freshwater tanks
wherethe waterwas recirculated constantly through sand To determine the response of the testes to DA. each
filtration units. Male intermolt crayfish with a carapace time the experiment was done 50 male crayfish were di-
ulesnegdthfoorft3he0s-e35expmemrimaenndtsa.bTohdeycwreaiygfhitshowfe1r1e-m12aigntmaiwneerde avsidtehdeiinntiotifalivecognrtoroulp,saonfd10theiaschg.roTuhpeoffircstragyrfiosuhp, swehrivcehd
at a room temperature of 24 2C. with 12 h of light received no treatment, was sacrificed on the first day of
daily, from 8:00 A.M. to 8:00 P.M., and were fed com- theexperiment. The initial control crayfishwereweighed,
mercial crayfish food daily. and then theirtestesand androgenic glands weredissected
out.Then,asstatedabove,thepairedtesteswereweighed,
and thetestesand androgenicglandswere fixed in Bouin's
Drugs fluid. A simultaneous control group received only phys-
5-HT creatinine sulfate, DA hydrochloride. spiperone, iological saline in 100 /ul doses. The last three groups ran
and pimozide were purchased from the Sigma Chemical concurrently with the simultaneous control group and
342 R. SAROJINI ET AL.
0.10 100
X
_ -
-
z
a<i Q
O
C-/5
t-/3
0.00
1C sc DA DA 10- 10 10'c
DA DA DA
TREATMENT TREATMENT
X F1i0g~u6rmeol1.perEcfrfeacytfisohf)odiffdfoerpeanmtidnoese(sDA()1oxnthIeCTm8e,a1nxtest1i0c"u'l,arainnddex1 > F1i0g~u6rmeol2.perEfcfreacytfisohf)doifffdeorpenatmidnoese(sDA(1)oxn t1h0e~8m,e1anxtes1t0i~c7u,laarnldobe1
tcoofhnattnhreoblca.rrasEyfr1irCso.hr,1PbCanTru6swDanrAhe,aSrauEnsdMda1rB(kaTir7i.DSA1CC..isBinasiirtiganl1i0fcioc*natnDrtolAly;i(SsPCs.i<gsni0im.fu0icl5a)tnatlnlaeyrogu(esPr d(siPiam<muel0tt.ea0rn5e)oofutshtahcneonbctarrraosylf.i1sCEh.r.rloPOr'rb6oaiDrwAsn.ahraaernuSdxEM1d0.a~r7kBiaDirA..S1CC.isisniitginailficcoannttrloyl;larSgCe.r
<0.05)largerthan bar 10"" DA.
Itisevident from Figures 1-3 that DA inhibitedtesticular
received 1 X 10~6 mol, 1 X 1(T7 mol, and 1 X 1(T8 mol maturation. The responses to the three concentrations of
DA per crayfish respectively in lOO/ul doses. Injections DA used strongly suggest that this inhibition is dose-re-
were administered on the 1st, 5th, and 10th days. The lated, asin Figures 1 and 3 wherethe inhibition produced
simultaneouscontrolgroupandthosegiven DAweresac- by 1 X 10~6 mol DA per crayfish is significantly greater
rificedon the 15thdayand processed in the same manner than that produced by 1 X 10~8 mol DA per crayfish.
as the initial control group.
The TI and mean testicular lobe diameter of the si-
multaneous control group were significantly larger than
the corresponding values of the initial control group, S
showing that during the 15 days ofthe experiments the a-t
testes were undergoing maturation (Figs. 1,2). Further- C/3
more, the simultaneous control testes contained mature a
sperm whereas the initial control testes had none (Fig. 3). at
The TI and mean testicular lobe diameter ofthe crayfish
that received 1 X 1(T6 mol DA injections were signifi-
cantly smaller than the corresponding values for the si- O
multaneous control crayfish that received only physio- a
logical saline. Furthermore, there were no mature sperm S
in the testicular lobes of the crayfish that received the Z3 2 -
injections of 1 X 1CT6 mol DA in contrast to the simul-
taneouscontrol crayfish. The crayfish that received injec-
tions ofthe two lower doses ofDA (1 X 1(T7 mol and 1 1C SC
X 10~8 mol) also had a smaller TI and mean testicular DA
TREATMENT
lobediameterthan thesimultaneouscontrol crayfish, but
only the difference between the testicular lobe diameter Figure 3. Effect of different doses (1 X 10~8, 1 X 10"', and 1
ofthe simultaneouscontrolsand thecrayfish that received 10~6mol per crayfish) ofdopamine (DA) on the mean number of
itTnwhjoeeclttoiewsotneesrsodofofs1etXsheo1fc0rDa7yAfmiocslhonDtthaAaitnweradescmesiatvatteuidrsteiicnsajlpelecytrismio,gnnsbifuoitfcastnihtg.e- Sa1mCnaC.dtuiisrn1ie0st~iisag8lpniDecfrAoim.cnatpnrBetoalrlry;fsolSlalC1ri0,cg~lees7riiD(nmPuAtlh<teaatn0ned.es0to5eu1s)0so~tfc8hotaDnhnteArocblraaa.rrysefEir1ssrChi,,ognrPi1rfb0ioa'ccr"aisnutDanlArhye.a(nSPiIExO<M"d.a70r.DkB0iAa5i..)r
nificantly fewer than in the simultaneous control group. largerthan bar Ifr6 DA.
CRAYFISH TESTICULAR MATURATION 343
0.30 200
0.00
5-HT COMB
TREATMENT
TREATMENT
Figure 5. Effect ofdifferent treatmentson the mean testicularlobe
Figure4. Effectofdifferenttreatmentsonthemeantesticularindex diameter ofthe crayfish. Pnicninhariis clarkii. 1C. initial control; SC.
cboooizfnnittadhrteeoilpoc;ernraSyoPcff,riasyh11,fiXxPshr;1o100c5~~a-66mHmbmToa,olnlt1ssDXpicApl1ae0rrp"koei6inr-meco1prlCae,yr5fici-nrsiHtahiyTafl+ipscehor1:ntcXPrrIoa.yl1f;10i"sSxh6C;,m1Co0s~Olim6Mu5mBl-ot,HlaTncpeoiopmmue--sr ccsxrioammy1ubf0lii~stn6haa.mntoeiElorournspoiorcmfobonaIztrixrsdoela1;;r0e~5S6P-S,mHEoTM.l1.XD1BAaxIrCp1eTS0r'C~mc6orimaslyosflisigpsn5hiip-f+eiHrcTao1nnXtpeleyr1pCe(cTrPr6acy<mrfoaiy0slf.hi;05s5h-C);HOlTPMaIr,Bpgee,rr1
crayfish. Error bars are SEM. Bar SC is significantly (P < 0.05) larger than bar1C.but barSCissignificantly(P<0.05)smallerthan barsSP,
than bar 1C. butbarSCissignificantly(P<0.05)smallerthan barsSP. PI. 5-HT. and COMB. Bar 5-HT issignificantly (P < 0.05) largerthan
PI. 5-HT. and COMB. Bar 5-HT issignificantly (P< 0.05) largerthan barCOMB.
barCOMB.
Effects oftheDA receptor Mockers spiperone ami
pimozide. 5-HTalone, and5-HTin combination with
DA on the testes
For each replicate ofthis set ofexperiments, 6 groups 100
of 10 crayfish were selected from the stock. One group 90 -
served as the initial control: the crayfish of this group 80 -
weretreatedin thesamewayasthe initial control crayfish
ofthe DA dose-response experiment. The crayfish in the
simultaneous control group received physiological saline P 60 -
in 100 jul doses. Two groups received 1 X 10"" mol ofthe 50 -
DA receptorblockersspiperoneand pimozide respectively
in 100 n\ doses. Anothergroup received 1 X 10 6 mol of O 40 -
5-HT per crayfish in 100-^1 doses and the last group re- W 30 -
ceived 1 X 10~6 mol DA in 50-^1 doses + 1 X 10 6 mol 20 -
5-HT in 50-jil doses per crayfish, respectively. Injections
were administered on the 1st, 5th, and 10th days. All the 10 -
crayfish thatreceived injectionsweresacrificedon the 15th
day, and their testes were processed in the same manner 1C SC SP PI 5-HT COMB
as those ofthe initial control group. TREATMENT
As in the previous experiment, the Tl and testicular Figure6. Effectofdifferenttreatmentsonthemeannumberofma-
lobe diameter of the simultaneous control crayfish were turesperm perfollicleinthetestesofthecrayfish.Procambarusclarkii,
significantly larger than the corresponding values for the 1C.initialcontrol;SC.simultaneouscontrol;SP. 1 X 10'6molspiperone
tcirnoiontltiratloelsctoteenssttdreiosdlhgharadovnueopsm(oaFmitgeusr.mea4,stpu5er)rema,nsdpt,eheramlsti(hmFouiulgg.tha6n)te.hoeFuosirncitotinha-el p+n5ie-frH1iccTraXanpytefl1riy0s~ch(r6:PamyPfoI<i,lsh0I:.5x0-C5H)O1T0Ml~aBp6re,gmreorcclortmaphybaifinminsoahbz.taiirdEoern1rCpo,oerfrbb1ucartXrasybfIaai0rsr~he6S;SmC5Eo-liMHsT.Ds.iABg1anirpXfeiSrc1Ca0cn~rtai6lsyyfmsioi(sglhP-
crayfishthateach received 100-/ulinjectionsof1 X 10~6 mol <0.05) smaller than bars SP. PI. 5-HT, and COMB. Bar 5-HT issig-
of either of the DA receptor blockers (spiperone or nificantly(P<0.05)largerthan barCOMB.
344 R. SAROJINI ET AL
pimozide). theTI and mean testicularlobediameterwere
significantly larger than the corresponding values for the
simultaneous control crayfish that received physiological
saline alone. Furthermore, there was a statistically signif-
icant greater number of mature sperm in the testicular
follicles ofthe crayfish that received either spiperone or
pimozide than in the simultaneous control crayfish. It is
clear from these results that spiperone and pimozide in-
duced testicular maturation.
TheTIand meantesticularlobediameterofthecrayfish
that received 100 n\ of 1 X 10 h mol 5-HT were signifi-
cantly larger than the corresponding values for the si-
multaneous control crayfish (Figs. 4. 5), and the number
ofmature sperm in the testes ofthe crayfish given 5-HT 5-HT COMB
was also significantly greater than for the simultaneous
control crayfish (Fig. 6). The combination ofequimolar TREATMENT
amounts ofDA and 5-HT produced significant increases Figure 7. Effect ofdifferenttreatmentsonthe mean cell size in the
in the TI, testicular lobe diameter, and sperm count but androgenicglandsofthecrayfish,Pnvambamsclarkii. 1C,initialcontrol;
tsaihbgalnteitfDoicAiannhatilnbyditle5cs-soHmtpThlaeancttdeiladynt5tah-geoHnsTitsiatmlioucnlaeal.tloyT,rhyebausctetirDoeAnsuolwtfsa5ss-hHnoTo.wt SocDrzCAai,,ydfseiis1pmhe;uxrlStcP1ra.0any'e1f7oi*usmhso:1lc05o'-nD6tHrAmToo,llp;1esr1Xp(icTrp16ae0Dy~rfAo6in,smeho;1pleX1r50-~c1rH8(aTTyD6fAipms,eohr;lc1PrDIaX,yAf1ilpsx(heT;r81Cc(mrOTao6yMlfmBiosD,hl:Acp1oip0mme~--r7
bination of 1 > ICT'mol DA per crayfish + 1 X 10~6mol 5-HT per
EinffcecotmsbionfaDtAi,onDwAitahntDaAgonoinsttsh,e5a-ndHrTogaelonniec,galnadnd5-HT (ctrPhaayn<fibs0ah.r.0s5E)1rCr,somr1a0lb~la6errDsAta,hraen10S"bEa7MrD.sAS,BPaa,rndPSI,C105i~s-8HsDTiAg,n,iafbniucdtanCbtlaOyrMS(BPC.<isBs0ai.gr0n5i5)f-iclHaaTnrtgleiyrs
The androgenic glands of the initial control crayfish significantly (P< 0.05) largerthan barCOMB.
consisted of only a few cords ofcells closely associated
with the vas deferens. These cells had a thin rim of ho-
mogeneous cytoplasm around the nucleus. The cells of granular than in either control group. As with the testes,
thesimultaneouscontrol crayfish weresignificantlylarger while the combination of5-HT and DA produced signif-
than thoseintheinitialcontrolglands(Fig. 7). The means icant growth of the androgenic glands, this growth was
in Figure 7 forthe initial and simultaneous controls rep- significantly less than that produced by 5-HT alone, ad-
resentdata from 40crayfish versus 20crayfish fortherest ditional evidenceofantagonisticactionsofDA and 5-HT
of the groups because the means for the initial and si- on the reproductive system ofmale red swamp crayfish.
multaneous controls are based on the averages of these
controls from the crayfish used in the two sets ofexperi- Discussion
ments that provided the data for Figures 1-3 and 4-6.
The cells of the androgenic glands of the crayfish that The present study demonstrates for the first time in a
rDeAceipveerdcirnajyefcitsihonsweorfe1 nXot10s~i6g,ni1fiXca1n0tl~y7,doirff1erXent10~in8 msiozle ctrhaeyrfmiosrhe,anthiinshiisbitthoeryfirascttrieopnorotfoDfAtheoneftfehcettoefstDesA. Faunrd-
from those ofthe initial control group. The cells in the any of its antagonists on the androgenic glands of any
androgenic glands ofall the crayfish that received injec- crustacean. DA alone inhibited testicularand androgenic
tionsofDA,regardlessofthedose used, weresignificantly gland maturation (Figs. 1-3, 7). On the other hand, 5-
smaller than the cells in the concurrent control glands. HTand the DA receptorblockersspiperone and pimozide
The inhibitory effects ofthe three concentrations ofDA induced testicular and androgenic gland maturation
on the androgenic glands do not provide clear evidence (Figs. 4-7).
ofa dose-related response, although the highest concen- In Pmcambantx clarkii, as stated above, gonadal mat-
tration produced somewhat more inhibition than did the uration is regulated by both stimulatory and inhibitory
GSH
two lesser doses. The androgenic glands ofcrayfish that neurohormones. maturation being stimulated by
received a DA receptor blocker, spiperone or pimozide, from the brain andthoracicgangliaand inhibited byGIH
5-HT alone or 5-HT in combination with DA showed from the eyestalk neuroendocrine system. Our previous
significantly greater development of their androgenic studies with Procambarus clarkii (Sarojini ct a/. 1993,
glands over the initial and simultaneous controls. The 1994) showed that 5-HT stimulates gonadal maturation
GSH
cytoplasm in these enlarged glands was more dense and in males and females, presumably by stimulating
CRAYFISH TESTICULAR MATURATION 345
release and that DA inhibits ovarian development. The reduced AGH in the blood, resulting in at least some in-
evidence for 5-HT and DA presence in the nervous sys- hibition of testicular maturation and spermatogenesis.
tems ofcrayfish (Fujii and Takeda, 1988; Arechiga el al. Experiments are currently in progress to evaluate these
1990; Real and Czternasty. 1990; Mercier et al., 1991; suggested modes ofaction of DA. That DA can have a
Kulkarni and Fingerman, 1992) was already demon- stimulatory role in the release of a neurohormone was
strated. shown forthe red pigment-concentrating hormone, asre-
The roles of DA and 5-HT in regulation of gonadal ported by Fingerman and Fingerman (1977) and Quack-
maturation in vertebrates is documented. Goldfish, Car- enbushand Fingerman (1984)whoperformed in vivoand
assius auratus, fed the DA agonist apomorphine had el- //; vitroexperimentson release ofthis neurohormone with
evated plasma levels ofgrowth hormone whereas the cir- thefiddlercrab, Ucapugilator. Theconcentrationsofbio-
culating levels of gonadotropic hormone were reduced genieaminesused intheseexperimentsarequite likethose
(Wong et al.. 1993). Long-term feeding ofgoldfish with injected by other investigators while studying the same
apomorphine induced significant increases in both the species, Procambarusclarkii. Livingstone etal. (1980) in-
body weight and length. 5-HT stimulates gonadotropic jected 5.7 X 10"6 mol 5-HT and 6.5 X 10~6 mol octo-
hormone release in the goldfish (Somoza et al.. 1988; So- pamine per crayfish, and Arechiga et al. (1990) injected
moza and Peter, 1991). This effect of5-HT may be due 1 X 10~9 to 1 X 10~3 mol 5-HT per crayfish.
to direct action on the gonadotrophs or to inhibition of Because DA inhibited testicular maturation in Pro-
DA release from nerve terminals in the pars distalis. DA cambarusclarkii. it isworth mentioningthepotential ap-
inhibits release of this gonadotropic hormone (Yu and plication of DA analogues in crayfish farming. Supple-
Peter, 1992). Similarly, DA appears to inhibit luteinizing menting the crayfish diet with long-lasting DA agonists
hormone release in the frog, Rana temporaria(Sotowska- may slow reproductive activity ofcrayfish and simulta-
Brochocka el al.. 1994). neously lead to enhanced somatic growth.
The crayfish that received 5-HT alone had a larger TI
and mean testicularlobediameterandalso had more ma- Literature Cited
ture sperm in their testicular lobes than did the simulta-
neous control group (Figs. 4-6) which is consistent with Arechiga, H., E. Banuelos, E. Frixione, A. Piconcs, and L. Rodriguez-
tthhaetearrelcieeirverdesu5l-tsHTofinSacroojmibniinaettialo.n(w1i99t4h).DAThehacdraayfsiisgh- BancSdrorosofaxt.y,tJr1.y9p9Dt0.a.,miannMedo.dA/u.lSaEt.t\eipvo.ennBsi.oofl19c18r5a20y.:fis1hT23hr-ee1ot4ri3ny.alansdenPsritaicvtiitcyeboyfHi5s-thoy--
nificantly larger TI and mean testicular lobe diameter, logical Techniques, 2nd Ed. Churchill Livingstone, New York. Pp.
and also a greater number ofmature sperm, when com- 109-121.
pared with the simultaneous controls (Figs. 4-6), but all Bart1h9e8,9.J. DY.o,paNm.inMeonasn,d mDo.toCratatcateirvti,tyMin.thGeeflfoabrsdt,eraHnodmaFn.isClgaraamc-.
three values were significantly smaller than the corre- nitini*. Brain Re*.. 497: 368-373.
sponding values of the crayfish given 5-HT alone. The Charniaux-Cotton, H. 195-1. Decouvertechez unCrustace Amphipode
DA in the mixture was not able to antagonize fully the (Orchestia gammarella) d'une glande endocrine responsable de la
stimulatoryactionofthe 5-HT. Thisantagonism between differenciationdescaracteressexuelsprimairesetsecondairesmales.
the effects produced by 5-HT and DA on the testes is C. R Acad. Sci. Paris. 239: 780-782.
Charniaux-Cotton, H. 1964. Endocrinologieetgenetiquedusexechez
reminiscent of that seen with the erythrophores of Uca lesCrustacessuperieurs.Ann. Endocrinol. 25: 36-42.
pugilatorwhere the pigment-dispersingeffect of5-HTand Demeusy, N. 1967. Modalitesd'action du controle inhibiteurpedon-
the pigment-concentrating effect of DA were reduced culaireexercesurlescaracteressexuelsexternesmalesdu Decapode
when mixtures of 5-HT and DA were co-injected (Fin- BrachyoureCareinut maenusL. C. R .lead. Sci. Paris. 265D: 628-
gerTmhaendaatnadoFbitnagineerdmawni,th1t9h7e7)D.A antagonistsDuAsed in the Eastt6i3ms0as.nu-eRaenkds,cySc.l.iacnAdMMP. oFinngoevarrmiaann.g1r9o8w4.th iEnffveicvtosaofndneiunrovietnrdooicnritnhee
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Hungarica36: 129-146.
leaTdhiengitnohipbrietcoorcyiaocutsiotnesotifcDulAaromnattuhreatainodnr.ogenic glands ElofIsdseonnt,ificR.a,tioLn.anLdaxqumaynrt,itatEi.veRmoesacsngurreenm,entasndofCb.iogHeannicssaomni.nes19a8n2d.
and testes in Procambarusclarkiicanbeexplained as fol- DOPAinthecentralnervoussystemandhamolymphofthecrayfish.
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greelneiacsgeloafndGs.1HWefrhoympotthheeseiyezsettahlaktnDeAuroeietnhdeorc(rai)nsetismyusltaetme,s FFiinnggceehrrrmmoaamnna.,toMMp..h,o1ra9en8s5d..SA.mT\hV.e/.Fopoihlnygs2ie5or:lmoa2gn3y.3a-1n29d5727p..harAmnatcaogloongisytoifccarcutsitoancseaonf
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