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The effect of crustacean eye stalk extract on the endogenous respiration of selected mammalian tissues
Camp. Biochem. Physiaf., 1974, Vol. #A, pp. 803 to 805. Pergamon Press. Printed in Great Britain
SHORT COMMUNICATION THE EFFECT OF CRUSTACEAN EYE STALK EXTRACT ON THE ENDO~ENOUS RESP~~TION OF SELECTED MAMMALIAN TISSUES WILLIAM
L. MENGEBIER”
Department of Biology, Bridgewater College, Bridgewater, Virginia 22812, U.S.A. (Received 9 February 1973)
Abstract-l. Eyestalk extract obtained from crayfish and lobster significantly increased endogenous respiration of brain and kidney homogenates of the rat and mouse respectively. 2. Liver and heart homogenates were not similarly stimulated.
INTRODUCTION
THE UBIQUITOUS effect of crustacean eyestalk extract on a donor species is well documented (Fingerman, 1966, 1968; Rao et al. 1967; &year, 1969). That at least some components of the extract are capable of modifying interspecific, intergeneric and even interphylogenic function has been reported by Thornborough & Mengebier (1967), Adiyodi & Adiyodi (1968) and Keller (1969), but in all these instances, not only the donor but the effected species have been invertebrates. With these results in mind, it was deemed of interest to investigate the possible efforts of these invertebrate derivatives on vertebrate function. This report deals with the effects of crustacean eyestalk extract obtained from two sources, the crayfish Cambarus bartoni and the lobster Homarus americanus, on the endogenous respiration of liver, brain, heart and kidney homogenates of the rat and mouse respectively. MATERIALS
AND METHODS
Specimens of Cumbarus bartoni were collected in the vicinity of Bridgewater, Virginia; eyestalks of Homarus americanus were obtained from animals purchased commercially in Portsmouth, New Hampshire. Five per cent w/v eyestalk homogenates were prepared in O-1 M PO4 buffer, pH 7.4. These homogenates were centrifuged for 180 min at 0°C and 10,000 g. The rest&ant supernatants were utilized in all experiments. * Supported by grants from the Virginia Academy of Science and the National Science Foundation, CU 4022. 803
804
WILLIAM L. MENGEBIER
The endogenous respiration of 10 per cent homogenates of selected tissues from the rat and mouse was determined mannometricaliy. All homogenates were prepared in O-1 M PO* buffer, pH 7.4, and all experiments were carried out at 26°C. Reaction vessels contained l-0 ml of the homogenate, 1.0 ml of PO, buffer, pH 7*4,05 ml of eyestalk extract when applicable and H,O to make. a total volume of 3.0 ml. Center wells contained O-2 ml of 2 N NaOH. Reaction vessels containing only eyestalk extract in equivalent dilutions were prepared for each experiment. All experiments involving lobster extract were performed in the facilities of the Department of Zoology, the University of New Hampshire, Durham, New Hampshire.
RESULTS
The effects of eyestalk extract obtained from the crayfish and lobster on the endogenous respiration of the rat and mouse respectively are shown in Fig. 1. When converted to total oxygen uptake, the increases measured for brain and kidney tissue were statistically significant, with P < O-05 in all instances using the
Control Mouse Rat
-10 -20 Liver
Brain
Kidney
HSWt
FIG. 1. Mean percentage change in Oa uptake over two consecutive IS-min intervals of 10 per cent homogenates of the rat and mouse to which 0.5 ml of crayfish and lobster eyestalk extract respectively were added. ‘N’ refers to the number of experiments.
Student’s t-test. There was no significant difference between control values for the liver and heart when compared with homogenates to which eyestalk extract had been added. No endogenous respiratory activity could be measured from flasks containing only extract at appropriate dilutions. Both crayfish and lobster extracts were inactivated by boiling. Crayfish eyestalk extracts were dialyzed against glass distilled water at 18°C for 24 hr, but no change in activity was found.
EFFECTOF CRUSTACEAN EYE STALKEXTRACTON MAMMALIAN TISSUES
80.5
DISCUSSION
The concept of invertebrate substances affecting vertebrate tissues may be considered as an example of biochemical evolution. If one accepts Florey’s (1966) comparison of the Crustacean eyestalk with both the hypothalmus and anterior pituitary of mammals, the results obtained in this series of experiments could point toward a hormonal involvement. At the present time, one may only conjecture as to the selectivity shown by both crayfish and lobster extracts for mammalian brain and kidney tissue. It is possible that in the case of liver and heart, membrane permeability may have been altered or even abolished thus furthering a selectivity of some evolutionary significance. The active component or components of the extracts are unknown. There is some resemblance of the crayfish extract to that of the diabetogenic hormone described by Kleinholz (1966), in that it is inactivated by boiling, but not by dialysis. REFERENCES ADIYODI R. & Adiyodi K. (1968) Ovary-inhibiting effect of extract of crab eyestalk on female American cockroaches. Gen. Sz compar. Endow. 10, 14-20. FINGERMANM. (1966) Neurosecretory control of pigmentary effecters in crustaceans. Am. Zoologist 6, 169-180. FINGERMANM. (1968) Crustacean color changes with emphasis on the fiddler crab. Scientiu 103, 1-16. FLO~XY E. (1966) General and Comparative Physiology, p. 267. Saunders W. B., Philadelphia. KEIIIIR R. (1969) Unte~uchungen zur Artspezifitat eines crustaceen Hormons, Z. vergl. Physiol. 63, 137-145. KLEINHOLZ L. H. (1966) Separation and purification of crustacean eyestalk hormones. Am. Zoologist 6, 161-168. PUYEARR. (1969) Molt cycle regulation of nucleotide pyrosphosphate in the hepatopancreas of the blue crab, Callinectes sapidus Rathbun. Camp. Biochem. Physiol. 28, 159-168. RAO K. R., BARTELLC. K. & FINGERMANM. (1967) Relationship between the response of melanophores in the fiddler crab Uca pugilator, and the concentration of eyestalk extract, 2. vergl. Physiol. 56, 232-236. THORNBOROUGH J. & MENGEBKERW. L. (1967) Direct stimulation of respiratory enzyme activity by crustacean eyestalk extract. Camp. Biochem. Physiol. 20, 351-353. Key Word Index-Endogenous americanus.
respiration; eyestalk extract; Cambarus bartoni; bonus
SHORT COMMUNICATION THE EFFECT OF CRUSTACEAN EYE STALK EXTRACT ON THE ENDO~ENOUS RESP~~TION OF SELECTED MAMMALIAN TISSUES WILLIAM
L. MENGEBIER”
Department of Biology, Bridgewater College, Bridgewater, Virginia 22812, U.S.A. (Received 9 February 1973)
Abstract-l. Eyestalk extract obtained from crayfish and lobster significantly increased endogenous respiration of brain and kidney homogenates of the rat and mouse respectively. 2. Liver and heart homogenates were not similarly stimulated.
INTRODUCTION
THE UBIQUITOUS effect of crustacean eyestalk extract on a donor species is well documented (Fingerman, 1966, 1968; Rao et al. 1967; &year, 1969). That at least some components of the extract are capable of modifying interspecific, intergeneric and even interphylogenic function has been reported by Thornborough & Mengebier (1967), Adiyodi & Adiyodi (1968) and Keller (1969), but in all these instances, not only the donor but the effected species have been invertebrates. With these results in mind, it was deemed of interest to investigate the possible efforts of these invertebrate derivatives on vertebrate function. This report deals with the effects of crustacean eyestalk extract obtained from two sources, the crayfish Cambarus bartoni and the lobster Homarus americanus, on the endogenous respiration of liver, brain, heart and kidney homogenates of the rat and mouse respectively. MATERIALS
AND METHODS
Specimens of Cumbarus bartoni were collected in the vicinity of Bridgewater, Virginia; eyestalks of Homarus americanus were obtained from animals purchased commercially in Portsmouth, New Hampshire. Five per cent w/v eyestalk homogenates were prepared in O-1 M PO4 buffer, pH 7.4. These homogenates were centrifuged for 180 min at 0°C and 10,000 g. The rest&ant supernatants were utilized in all experiments. * Supported by grants from the Virginia Academy of Science and the National Science Foundation, CU 4022. 803
804
WILLIAM L. MENGEBIER
The endogenous respiration of 10 per cent homogenates of selected tissues from the rat and mouse was determined mannometricaliy. All homogenates were prepared in O-1 M PO* buffer, pH 7.4, and all experiments were carried out at 26°C. Reaction vessels contained l-0 ml of the homogenate, 1.0 ml of PO, buffer, pH 7*4,05 ml of eyestalk extract when applicable and H,O to make. a total volume of 3.0 ml. Center wells contained O-2 ml of 2 N NaOH. Reaction vessels containing only eyestalk extract in equivalent dilutions were prepared for each experiment. All experiments involving lobster extract were performed in the facilities of the Department of Zoology, the University of New Hampshire, Durham, New Hampshire.
RESULTS
The effects of eyestalk extract obtained from the crayfish and lobster on the endogenous respiration of the rat and mouse respectively are shown in Fig. 1. When converted to total oxygen uptake, the increases measured for brain and kidney tissue were statistically significant, with P < O-05 in all instances using the
Control Mouse Rat
-10 -20 Liver
Brain
Kidney
HSWt
FIG. 1. Mean percentage change in Oa uptake over two consecutive IS-min intervals of 10 per cent homogenates of the rat and mouse to which 0.5 ml of crayfish and lobster eyestalk extract respectively were added. ‘N’ refers to the number of experiments.
Student’s t-test. There was no significant difference between control values for the liver and heart when compared with homogenates to which eyestalk extract had been added. No endogenous respiratory activity could be measured from flasks containing only extract at appropriate dilutions. Both crayfish and lobster extracts were inactivated by boiling. Crayfish eyestalk extracts were dialyzed against glass distilled water at 18°C for 24 hr, but no change in activity was found.
EFFECTOF CRUSTACEAN EYE STALKEXTRACTON MAMMALIAN TISSUES
80.5
DISCUSSION
The concept of invertebrate substances affecting vertebrate tissues may be considered as an example of biochemical evolution. If one accepts Florey’s (1966) comparison of the Crustacean eyestalk with both the hypothalmus and anterior pituitary of mammals, the results obtained in this series of experiments could point toward a hormonal involvement. At the present time, one may only conjecture as to the selectivity shown by both crayfish and lobster extracts for mammalian brain and kidney tissue. It is possible that in the case of liver and heart, membrane permeability may have been altered or even abolished thus furthering a selectivity of some evolutionary significance. The active component or components of the extracts are unknown. There is some resemblance of the crayfish extract to that of the diabetogenic hormone described by Kleinholz (1966), in that it is inactivated by boiling, but not by dialysis. REFERENCES ADIYODI R. & Adiyodi K. (1968) Ovary-inhibiting effect of extract of crab eyestalk on female American cockroaches. Gen. Sz compar. Endow. 10, 14-20. FINGERMANM. (1966) Neurosecretory control of pigmentary effecters in crustaceans. Am. Zoologist 6, 169-180. FINGERMANM. (1968) Crustacean color changes with emphasis on the fiddler crab. Scientiu 103, 1-16. FLO~XY E. (1966) General and Comparative Physiology, p. 267. Saunders W. B., Philadelphia. KEIIIIR R. (1969) Unte~uchungen zur Artspezifitat eines crustaceen Hormons, Z. vergl. Physiol. 63, 137-145. KLEINHOLZ L. H. (1966) Separation and purification of crustacean eyestalk hormones. Am. Zoologist 6, 161-168. PUYEARR. (1969) Molt cycle regulation of nucleotide pyrosphosphate in the hepatopancreas of the blue crab, Callinectes sapidus Rathbun. Camp. Biochem. Physiol. 28, 159-168. RAO K. R., BARTELLC. K. & FINGERMANM. (1967) Relationship between the response of melanophores in the fiddler crab Uca pugilator, and the concentration of eyestalk extract, 2. vergl. Physiol. 56, 232-236. THORNBOROUGH J. & MENGEBKERW. L. (1967) Direct stimulation of respiratory enzyme activity by crustacean eyestalk extract. Camp. Biochem. Physiol. 20, 351-353. Key Word Index-Endogenous americanus.
respiration; eyestalk extract; Cambarus bartoni; bonus