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Catecholamines in social wasps
Comp. Biochem. Physiol., 1974,Vol. 48A, pp. 369 to 373. Pergamon Press. Printed in Great Blitnin
CATEC~OLAMI~ES
IN SOCIAL WASPS
JACOB ISHAY,l ZALMAN, ABRAHAMa YONA GRUNFELD2 and SIMON GITTER IDepartment of Physiology and Pharmacology, Saclder School of Medicine, Tel-Aviv University; and *Israel Institute for Biological Research, Ness-Ziona, Israel
(Received 6 June 1973) Abstract-l. Various tissues originating from several species of social wasps showed significant amounts of catecholamines (noradrenaline, adrenaline, dopamine and S-hydrox~~t~ne). 2. Dopamine proved to be the most abundant among the catecholamines examined. 3. The data are discussed in regard to possible functions of catecholamines in wasps.
INTRODUCTION IN EARLIER studies,
Owen (1971) and Edery et al. (1972) brought forward evidence about the presence of considerable amounts of catecholamines in the venom of Apis mell@ra, Vespda arena&a and Vespa OrientaEis.We became interested in investigating further the source of these amines and whether they are to be found solely in the venom or are localized as well in other organs of both adults and larvae.
MATERIALS
AND METHODS
Wasps of the following species and origin were studied: Vespa orientalis (V.O.) and Polistes gal&us (P.gu.), collected in Israel; V. crabro (V,c.), Paraves$ula germanica (F.g.), P. vulgaris (P.v.), Dolichovespula sax&a (D.s.) and D. media (D.m.), all collected in Germany. The wasps were gathered alive with their nests, according to the method of Ishay (1964) and then segregated with respect to age groups and castes, depending on availability of the material. For the biochemical determinations of catecholamines in the wasp tissues, the hemolymph, fat bodies and midgut of larvae; hemolymph of prepupae; venom sacs and whole bodies of workers and queens-the latter freed previously of the venom sacsand those of males, were lyophilized. Catecholamines (CA) levels in all mentioned tissues were assayed on the lyophilized samples according to the fluorimetric methods of Chang (1964) and Maickel et al. (1968). RESULTS
The distribution of ~techol~ines summarized in Table 1.
(CA) in the various tissues of wasps is 369
Worker’s venom sacs 48.0 Queen’s whole body 0.4 Male’s whole body
44.6
1.50
Imagines Worker’s whole body
1 +O
41.6 0 0
V.C.
4.1
O-4
1.7
1.2 0.7
V.O.
Pupae Hemolymph
Hemolymph
Prepupae
Midgut
Larvae Saliva Hemolymph Fat body
Material
70.0
0
0.3
2.2
0 0
P.g.
21.4
0
0
2.4
0 0
P.v.
33.2 0 O-8
0
1.8
0 0
D.s.
Noradrenaline (nslmg dry wt.1
0 0
0.4
0 0
D.m.
0
2-l
0
P.ga.
58.0 0.9
1.6
2.5
2.2 0.8
V.O.
29.1
0
5.0
0.7
36.6 0 0
V.C.
TABLE l-CATECHOLAMINE LEVELSIN WASPTISSUES
24.3
0
0
0.2
0.5
0 0
P.g.
17.2
0
0
1.1
0 0
P.v.
Adrenaline (ng/mg dry wt.)
15.0 0.8 o-2
0.4
0
0 0
D.s.
0.7 0.3
0.8
0‘3
0 0
D.m.
0
0.7
O
P.ga.
E % (J z
E $
B
k
z
-*
r E &
icr E E z
“!
F &
8
IN SOCIAL
CA~CHO~~
WASPS
371 0
0
oorj
0
64 M
-I-
005
0
00
372
JACOB ISHAY, ZALMAN ABRAHAM, YONA GRUNFELD AND SIMON GITTER
As can be seen from the Table, the amounts of CA present in the different tissues of the species examined vary considerably. In general it appears that the levels of CA are relatively higher in adult wasps than in larvae. Among the tissues of adults the greatest concentrations of CA were observed in venom sacs, which in turn exhibited a particular abundance of serotonin (5-HT). Although venom sacs still contained significant amounts of dopamine (DA), noradrenaline (NA) and adrenaline (A), the cumulative amounts of these three amines did not constitute more than 30 per cent of DA in any one of the species. Contrary to this finding in adults DA is the main amine found in larvae and in the pupal hemolymph of those species which were examined. An exception to the rule is the even higher content of 5-HT in the saliva of larvae in V. crabro. DISCUSSION
The findings of this study clearly indicate the presence of considerable amounts In adults, the highest amounts were of various catecholamines in wasp tissues. found in venom sacs whereas in larvae the greatest concentration was found in saliva and midgut. Considering the physiological role of the above three mentioned systems one can certainly regard their respective functions as part of the excretory system. Thus, the contents of the midgut are emptied immediately upon pupation (Imms, 1946) ; larval saliva is usually transferred to the adults as part of the phenom“trophallaxis” (Wheeler, 1928), and the contents of enon of food exchange, venom sacs serve as the source of poisonous substances to be injected into prey. In the latter, the high levels of catecholamines are most probably related to the venom’s defensive role directed towards other arthropods. Particularly with respect to DA this has been clearly shown. Thus, DA was found to cause enhancement of heart rate in insects (Miller & Metcalf, 1968; Owen, 1971) and also bring about increased activity of the insect central nervous system (Gahery & Boistel, 1965), both of which could contribute to the toxic action of the venom which as suggested by Owen (1971) is achieved through accelerationof hemolymph circulation, with a resulting rapid distribution of the venomous components their sites of action and the ultimate production of excitation and pain. Regarding 5-HT this amine is known to occur in large amounts in invertebrates (Welsh, 1964) as well as in vertebrate venoms (Welsh, 1966) and most likely fulfills a similar function as DA. In spite of DA and 5-HT constituting the dominant catecholamines found, in the aforementioned tissues small amounts of NA and It should be pointed out, however, that A could be detected in the remaining. these amines were also found in tissues not necessarily associated with venom sacs or other excretory organs. Thus, the presence of NA and A, although only in minute amounts in hemolymph of V.O. larvae and V.C. pupae, P.g. prepupae and in D.s. and Vm. male whole-body, leads one to consider the possibility of neurotransmitter functions of these endogenous catecholamines in invertebrates. Although Welsh & King (1970) in their study of catecholamines in Planaria sp. presented evidence for the presence of NA, these authors discuss the questions as to the adequacy of the methods used in the identification of hormones. In this
CATECHOLAMINES IN SOCIAL. WASPS
373
respect we feel that the methods used by us leave no doubt as to the reliability of our findings. If for any reason one might suspect that the presence of NA and A in the hemolymph of larvae or pupae might be considered as components which are to be transferred at a later stage into the venom sacs, certainly the presence of NA in males (imagines) cannot be attributed to that same function, as wasp males do not produce venom. The finding of the relatively high levels of DA in the hemolymph of all the species examined and its well-known function as precursor of biogenic amines, lends further support to our assumption that these catecholamines are indeed endogenous transmitters in insects. REFERENCES CHANG C. C. (1964) A sensitive method for spectrophotofluorometric assay of catecholamines. Int.J. Neuropharm. 3, 643-649. EDERY H., ISHAY J., LASS Y. & GITTER S. (1972) Pharmacological activity of Oriental hornet (Vespa orientalis) venom. Toticon 10, 13-23. GAHERYY. & BOISTELJ. (1965) Study of some pharmacological substances which modify the electrical activity of the sixth abdominal ganglion of the cockroach Periplaneta americana. In The Physiology of the Insect Central Nervous System (Edited by TREHERNEJ. E. & BFAMENTJ. W.), pp. 73-78. Academic Press, New York. IMMSA. D. (1946) Textbook of Entomology. Methuen, London. MAICKELR. P., Cox R. H., JR., SILLANTJ. & MILLER F. P. (1968) A method for the determination of serotonin and norepinephrine in discrete areas of rat brain. Int. J. Neuropharn. 7, 275-281. MILLER T. & -CALF R. L. (1968) Site of action of pharmacologically active compounds on the heart of Periplaneta americana L. J. insect Physiol. 14, 383-394. OWEN M. D. (1971) Insect venoms: identification of dopamine and noradrenaline in wasp and bee stings. Experientia 27, 544-545. WELSH J. H. (1964) Composition and mode of action of some invertebrate venoms. Ann. Rev. Pharmacol. 4, 293-304. WELSH J. H. (1966) Serotonin and related tryptamine derivatives in snake venoms. Mem. Inst. Butantan Symp. Int., 33, 509-518. WELSH J. H. & KING E. C. (1970) Catecholamines in planarians. Comp. Biochem. Physiol. 36, 683-688. WHEELER W. M. (1928) The Social Insects. Kegan Paul, Trench & Trubner, New York. Key Word Index-Wasps;
catecholamines; noradrenaline; adrenaline; dopamine; 5-HT.
CATEC~OLAMI~ES
IN SOCIAL WASPS
JACOB ISHAY,l ZALMAN, ABRAHAMa YONA GRUNFELD2 and SIMON GITTER IDepartment of Physiology and Pharmacology, Saclder School of Medicine, Tel-Aviv University; and *Israel Institute for Biological Research, Ness-Ziona, Israel
(Received 6 June 1973) Abstract-l. Various tissues originating from several species of social wasps showed significant amounts of catecholamines (noradrenaline, adrenaline, dopamine and S-hydrox~~t~ne). 2. Dopamine proved to be the most abundant among the catecholamines examined. 3. The data are discussed in regard to possible functions of catecholamines in wasps.
INTRODUCTION IN EARLIER studies,
Owen (1971) and Edery et al. (1972) brought forward evidence about the presence of considerable amounts of catecholamines in the venom of Apis mell@ra, Vespda arena&a and Vespa OrientaEis.We became interested in investigating further the source of these amines and whether they are to be found solely in the venom or are localized as well in other organs of both adults and larvae.
MATERIALS
AND METHODS
Wasps of the following species and origin were studied: Vespa orientalis (V.O.) and Polistes gal&us (P.gu.), collected in Israel; V. crabro (V,c.), Paraves$ula germanica (F.g.), P. vulgaris (P.v.), Dolichovespula sax&a (D.s.) and D. media (D.m.), all collected in Germany. The wasps were gathered alive with their nests, according to the method of Ishay (1964) and then segregated with respect to age groups and castes, depending on availability of the material. For the biochemical determinations of catecholamines in the wasp tissues, the hemolymph, fat bodies and midgut of larvae; hemolymph of prepupae; venom sacs and whole bodies of workers and queens-the latter freed previously of the venom sacsand those of males, were lyophilized. Catecholamines (CA) levels in all mentioned tissues were assayed on the lyophilized samples according to the fluorimetric methods of Chang (1964) and Maickel et al. (1968). RESULTS
The distribution of ~techol~ines summarized in Table 1.
(CA) in the various tissues of wasps is 369
Worker’s venom sacs 48.0 Queen’s whole body 0.4 Male’s whole body
44.6
1.50
Imagines Worker’s whole body
1 +O
41.6 0 0
V.C.
4.1
O-4
1.7
1.2 0.7
V.O.
Pupae Hemolymph
Hemolymph
Prepupae
Midgut
Larvae Saliva Hemolymph Fat body
Material
70.0
0
0.3
2.2
0 0
P.g.
21.4
0
0
2.4
0 0
P.v.
33.2 0 O-8
0
1.8
0 0
D.s.
Noradrenaline (nslmg dry wt.1
0 0
0.4
0 0
D.m.
0
2-l
0
P.ga.
58.0 0.9
1.6
2.5
2.2 0.8
V.O.
29.1
0
5.0
0.7
36.6 0 0
V.C.
TABLE l-CATECHOLAMINE LEVELSIN WASPTISSUES
24.3
0
0
0.2
0.5
0 0
P.g.
17.2
0
0
1.1
0 0
P.v.
Adrenaline (ng/mg dry wt.)
15.0 0.8 o-2
0.4
0
0 0
D.s.
0.7 0.3
0.8
0‘3
0 0
D.m.
0
0.7
O
P.ga.
E % (J z
E $
B
k
z
-*
r E &
icr E E z
“!
F &
8
IN SOCIAL
CA~CHO~~
WASPS
371 0
0
oorj
0
64 M
-I-
005
0
00
372
JACOB ISHAY, ZALMAN ABRAHAM, YONA GRUNFELD AND SIMON GITTER
As can be seen from the Table, the amounts of CA present in the different tissues of the species examined vary considerably. In general it appears that the levels of CA are relatively higher in adult wasps than in larvae. Among the tissues of adults the greatest concentrations of CA were observed in venom sacs, which in turn exhibited a particular abundance of serotonin (5-HT). Although venom sacs still contained significant amounts of dopamine (DA), noradrenaline (NA) and adrenaline (A), the cumulative amounts of these three amines did not constitute more than 30 per cent of DA in any one of the species. Contrary to this finding in adults DA is the main amine found in larvae and in the pupal hemolymph of those species which were examined. An exception to the rule is the even higher content of 5-HT in the saliva of larvae in V. crabro. DISCUSSION
The findings of this study clearly indicate the presence of considerable amounts In adults, the highest amounts were of various catecholamines in wasp tissues. found in venom sacs whereas in larvae the greatest concentration was found in saliva and midgut. Considering the physiological role of the above three mentioned systems one can certainly regard their respective functions as part of the excretory system. Thus, the contents of the midgut are emptied immediately upon pupation (Imms, 1946) ; larval saliva is usually transferred to the adults as part of the phenom“trophallaxis” (Wheeler, 1928), and the contents of enon of food exchange, venom sacs serve as the source of poisonous substances to be injected into prey. In the latter, the high levels of catecholamines are most probably related to the venom’s defensive role directed towards other arthropods. Particularly with respect to DA this has been clearly shown. Thus, DA was found to cause enhancement of heart rate in insects (Miller & Metcalf, 1968; Owen, 1971) and also bring about increased activity of the insect central nervous system (Gahery & Boistel, 1965), both of which could contribute to the toxic action of the venom which as suggested by Owen (1971) is achieved through accelerationof hemolymph circulation, with a resulting rapid distribution of the venomous components their sites of action and the ultimate production of excitation and pain. Regarding 5-HT this amine is known to occur in large amounts in invertebrates (Welsh, 1964) as well as in vertebrate venoms (Welsh, 1966) and most likely fulfills a similar function as DA. In spite of DA and 5-HT constituting the dominant catecholamines found, in the aforementioned tissues small amounts of NA and It should be pointed out, however, that A could be detected in the remaining. these amines were also found in tissues not necessarily associated with venom sacs or other excretory organs. Thus, the presence of NA and A, although only in minute amounts in hemolymph of V.O. larvae and V.C. pupae, P.g. prepupae and in D.s. and Vm. male whole-body, leads one to consider the possibility of neurotransmitter functions of these endogenous catecholamines in invertebrates. Although Welsh & King (1970) in their study of catecholamines in Planaria sp. presented evidence for the presence of NA, these authors discuss the questions as to the adequacy of the methods used in the identification of hormones. In this
CATECHOLAMINES IN SOCIAL. WASPS
373
respect we feel that the methods used by us leave no doubt as to the reliability of our findings. If for any reason one might suspect that the presence of NA and A in the hemolymph of larvae or pupae might be considered as components which are to be transferred at a later stage into the venom sacs, certainly the presence of NA in males (imagines) cannot be attributed to that same function, as wasp males do not produce venom. The finding of the relatively high levels of DA in the hemolymph of all the species examined and its well-known function as precursor of biogenic amines, lends further support to our assumption that these catecholamines are indeed endogenous transmitters in insects. REFERENCES CHANG C. C. (1964) A sensitive method for spectrophotofluorometric assay of catecholamines. Int.J. Neuropharm. 3, 643-649. EDERY H., ISHAY J., LASS Y. & GITTER S. (1972) Pharmacological activity of Oriental hornet (Vespa orientalis) venom. Toticon 10, 13-23. GAHERYY. & BOISTELJ. (1965) Study of some pharmacological substances which modify the electrical activity of the sixth abdominal ganglion of the cockroach Periplaneta americana. In The Physiology of the Insect Central Nervous System (Edited by TREHERNEJ. E. & BFAMENTJ. W.), pp. 73-78. Academic Press, New York. IMMSA. D. (1946) Textbook of Entomology. Methuen, London. MAICKELR. P., Cox R. H., JR., SILLANTJ. & MILLER F. P. (1968) A method for the determination of serotonin and norepinephrine in discrete areas of rat brain. Int. J. Neuropharn. 7, 275-281. MILLER T. & -CALF R. L. (1968) Site of action of pharmacologically active compounds on the heart of Periplaneta americana L. J. insect Physiol. 14, 383-394. OWEN M. D. (1971) Insect venoms: identification of dopamine and noradrenaline in wasp and bee stings. Experientia 27, 544-545. WELSH J. H. (1964) Composition and mode of action of some invertebrate venoms. Ann. Rev. Pharmacol. 4, 293-304. WELSH J. H. (1966) Serotonin and related tryptamine derivatives in snake venoms. Mem. Inst. Butantan Symp. Int., 33, 509-518. WELSH J. H. & KING E. C. (1970) Catecholamines in planarians. Comp. Biochem. Physiol. 36, 683-688. WHEELER W. M. (1928) The Social Insects. Kegan Paul, Trench & Trubner, New York. Key Word Index-Wasps;
catecholamines; noradrenaline; adrenaline; dopamine; 5-HT.