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A factor influencing sodium regulation in crustaceans
Life Sciences Vol . 6, pp . 2171-2175, 1967 . Printed in Great Britain .
Pergamon Press Ltd.
A FACTOR IIIFI1JFaiCING SODIUM REGULATION IN CRUSTACEAIPS ; R . R ~~^=^^~+-thi*a sad B . T . Scheer Department of Biology, University of Oregon, Eugene, Oregon 9îu03
(Received 14 June 1967 ; in final form 2 August 1967) The well-known ability of euryhaline enietaceana to regulate the ionic and osmotic concentration of their body fluids is n variety of environmental situations suggests that the regulation mny involve endocrine controls, but the evidence for such controls remains limited sad equivocal (1) .
Control
of water permeability by a neurosecretory factor in the eyestalk vas demonstrated in Carcinus maenes (2), is HemiArapsus nudes (3) and in Procambarus clarkü (4) .
Heller and Smith (5) showed that extract of cregfish eyestalk
affects the water balance of frogs .
Bliss et, al . (6) reported the presence
of a hormone caacerning the voter balance in the central nervws system, but not in the eyestalks of lead crab, Cecnrcinua lateralis .
Since ion transport
sad ion permeability are of hindnmental importance to euryhaliae cruatacesas, we should expect saax controls of these phenameae .
The present report deal
with the demonstration that a factor in the extract of the cephalothorax of the prawn Pandalua
ordeai decreases the outflux of Na+ ions from the body of
the shore crab Aemigrapeus nudes . Materials sad Methods Mete crabs in the iatermolt stage srcre used throughout the course of Ns+ outflux ezperlments .
For the measurement of Na+ autfluz the crabs were
injected with 22Na+ and placed in a bath of known volume .
The contents of the
bath were circulated coatiauoualy through s vial placed in the well of e
~ Supported by NSF Grant GB 3181 On leave from Sri Venkateawara University, Tirupati, India 2171
SODIUM REGULATION
2172
acintillatioa coaster.
Vol . 6, No . 20
The counter xas connected xith n rate meter whose
output xaa recorded caatiauoual3r xith a recording potentiaaeeter (Fiß . 1) . lla+ outflux xas computed from the following equation : 4 ~ Qe Qb Q
ln
AB 1Q - QbA AB 2Q - QbA
xhare 4 is the efflux during a time period (30 or 60mia), Qe is the total
amount of 23Fa+ in the hemolymph, Qb the Na+ in the bath, 0. the total Ha+ in the system (Qe + Qb), A the total 22A8+ is the system, and AB l sad AB2 the amounts of ~Na+ at the beginning end end of the time period respectively . ~e equatiaa was derived as the asaumptiens that uaidirectionnl flux rates are proportiensl to total Aa+ in the hemolymph sad bath respectively and that the eaimal is is a steady at ate. The eyestalk sad cephalothorax extracts of the pram Paadnlus
ordani
(in the intermolt stage) were tested for their effect on Ba+ outflux of the crabs .
Caatrol animals xere administered xith aeliae solution . Results sad Diacusaim
Injections of eyestalk extract had no effect on Pa+ flux .
This is in
agreement xith Bryan's (6) report that eyestalk removal does nox affect the ability of the crayfish to alter the rate of Na+ uptake in response to changes in blood Aa+ level,
When va.rioua fractions of cephalothorax extract obtained
after chromatography (î) on G-10 Sephadex (Fig . 2) xere tested, it xes found that fraction III contains an active factor that markedly decreases the outfluz of Aa+ ions from the body of the crab .
The effect of this active factor
vas tested is txo xeys . In one aeries of experiments, after an initial period of Na+ efflux measurement the crabs xere injected xith the extract . decreased immediately.
The efflux rate
Injection of saline had no such effect (Table 1) .
Vol . 6, No . 20
SODIUM REGULATION
.
2173
.fi .
.
.
.
.e
:-: .i R=' ".
;i
,~Yf!
-
"
..Y
:
~r :: :3ss+-,s ;
'":(".TP :.r.
°i'~~i' _." i°.~t: .'. .f ''3s
...
"h
' .:ï'Wr
-i :.~ " iaf'~f ." Y : : .;ti71. " i . . :e
~
_
_
FIG, 1 Typical recorda of Na+ outflux . Middle :
Saline injected .
III injected . vea 2Q ul .
Top:
Bottom :
Eyestnlk extract injected,
Cephalothorax extract fraction
Ia all cases the volume of extract or saline injected
Arrwa indicate time of injection.
SODIUM REGULATION
2174
Vol .
6, No .
20
TABLE 1 Effect of Cephalothorax Extract (Fraction III) of Pandalus
ordaai
an fta+ Outflux of the Crab Hemigrapsus audus . Aa+ Outflux uM/2 hours Control
Before
After
Difference
1
1333
1133
200
2
1958
602
3
1352
1356 1131
241
Injected
1685
523
1162
2
25T3
1163
1410
3
2383
594
1789
1
Variance ratio of differences 33 .8 ; P
BEPHADE% G-10 COLUMN 100 X 2 S CY . B.4 YL / TUBE FLOW RATE : 9.33 YL / HR
2h
I N
1 2
TUBE NUMBER
FIO . 2 Chromatography of cephalathorax extract oa Sephadex G-10 In another type of experiment the extract or esline vas injected at the
beginning of the experiment end then the Hn+ outflux was measured for 3 hours, In the animals injected xith .frnetioa III the Aa+ outflux was considerably lox
when compared to control animals (Table 2) .
Vol . 6, No . 20
2175
SODIUM REGULATION
TABLE 2 Effect of Cephalothoraz Eztract (Fraction III) of Paadalua ~ordaai oa Aa+ Outflux of the Crnb Hemigrapaus audua Na+ Outflux LM first 3 hours Control 5011 S . D.
t
Injected
P
2935
1864
S . D . = 851
0.06
These results suggest the presence of a "factor" is the extracts of the cephalothoraz, controlling Ae+ regulation in crustaceans,
If the factor is
neurosecretory in origin, it probably nriaea in the cells of the central nervous system proper rather than in the neurosecretory complex of the eyestalk .
The active factor probably decreases the permeability of the gills
to Ae+ ions . Further atudiea to eaelyze the pathvaya of Aa+ fluxes in Hemiarapaua nudue , to know hw rates change in reaponae to altered environmental salinity and to determine the site for possible control are in progress . References 1.
B . T . SC~ER, Arch . Aaat . Micr, Morph, Exp , ~4, 531 (1965) .
2.
D . B . CARLISLE, Pubbl . Staz . Zool . Napoli ~, 22~ (1956) .
3,
R . L . LEHMAA and B . T . SCNEER, Phyaiol . Comp . Oecol . 4, 164 (1956) .
4,
F, I, KAMEMOTO, IQ{ANETfi N, KATO, sad LOIS E . TUCKER, Am, Zool . 6, 213 (1966) .
5.
R . RELLER and B, SMITH, J . Exp . Hiol . 2~, 388 (1948) .
6.
DOROTHY E, BLISS, STEFANIE M . E, WANG and EDWIA A MARTINEZ, Am, Zool . 6, 197 (1966) .
7.
G . W . BRYAA, J, Ezp, Biol . ~, 100 (1960b) .
8.
L . H . KLEIAHOLZ and FRAACFS KIMBALL, Gen . Comp . Eadocrinol . ~, 336 (1965) .
Pergamon Press Ltd.
A FACTOR IIIFI1JFaiCING SODIUM REGULATION IN CRUSTACEAIPS ; R . R ~~^=^^~+-thi*a sad B . T . Scheer Department of Biology, University of Oregon, Eugene, Oregon 9îu03
(Received 14 June 1967 ; in final form 2 August 1967) The well-known ability of euryhaline enietaceana to regulate the ionic and osmotic concentration of their body fluids is n variety of environmental situations suggests that the regulation mny involve endocrine controls, but the evidence for such controls remains limited sad equivocal (1) .
Control
of water permeability by a neurosecretory factor in the eyestalk vas demonstrated in Carcinus maenes (2), is HemiArapsus nudes (3) and in Procambarus clarkü (4) .
Heller and Smith (5) showed that extract of cregfish eyestalk
affects the water balance of frogs .
Bliss et, al . (6) reported the presence
of a hormone caacerning the voter balance in the central nervws system, but not in the eyestalks of lead crab, Cecnrcinua lateralis .
Since ion transport
sad ion permeability are of hindnmental importance to euryhaliae cruatacesas, we should expect saax controls of these phenameae .
The present report deal
with the demonstration that a factor in the extract of the cephalothorax of the prawn Pandalua
ordeai decreases the outflux of Na+ ions from the body of
the shore crab Aemigrapeus nudes . Materials sad Methods Mete crabs in the iatermolt stage srcre used throughout the course of Ns+ outflux ezperlments .
For the measurement of Na+ autfluz the crabs were
injected with 22Na+ and placed in a bath of known volume .
The contents of the
bath were circulated coatiauoualy through s vial placed in the well of e
~ Supported by NSF Grant GB 3181 On leave from Sri Venkateawara University, Tirupati, India 2171
SODIUM REGULATION
2172
acintillatioa coaster.
Vol . 6, No . 20
The counter xas connected xith n rate meter whose
output xaa recorded caatiauoual3r xith a recording potentiaaeeter (Fiß . 1) . lla+ outflux xas computed from the following equation : 4 ~ Qe Qb Q
ln
AB 1Q - QbA AB 2Q - QbA
xhare 4 is the efflux during a time period (30 or 60mia), Qe is the total
amount of 23Fa+ in the hemolymph, Qb the Na+ in the bath, 0. the total Ha+ in the system (Qe + Qb), A the total 22A8+ is the system, and AB l sad AB2 the amounts of ~Na+ at the beginning end end of the time period respectively . ~e equatiaa was derived as the asaumptiens that uaidirectionnl flux rates are proportiensl to total Aa+ in the hemolymph sad bath respectively and that the eaimal is is a steady at ate. The eyestalk sad cephalothorax extracts of the pram Paadnlus
ordani
(in the intermolt stage) were tested for their effect on Ba+ outflux of the crabs .
Caatrol animals xere administered xith aeliae solution . Results sad Diacusaim
Injections of eyestalk extract had no effect on Pa+ flux .
This is in
agreement xith Bryan's (6) report that eyestalk removal does nox affect the ability of the crayfish to alter the rate of Na+ uptake in response to changes in blood Aa+ level,
When va.rioua fractions of cephalothorax extract obtained
after chromatography (î) on G-10 Sephadex (Fig . 2) xere tested, it xes found that fraction III contains an active factor that markedly decreases the outfluz of Aa+ ions from the body of the crab .
The effect of this active factor
vas tested is txo xeys . In one aeries of experiments, after an initial period of Na+ efflux measurement the crabs xere injected xith the extract . decreased immediately.
The efflux rate
Injection of saline had no such effect (Table 1) .
Vol . 6, No . 20
SODIUM REGULATION
.
2173
.fi .
.
.
.
.e
:-: .i R=' ".
;i
,~Yf!
-
"
..Y
:
~r :: :3ss+-,s ;
'":(".TP :.r.
°i'~~i' _." i°.~t: .'. .f ''3s
...
"h
' .:ï'Wr
-i :.~ " iaf'~f ." Y : : .;ti71. " i . . :e
~
_
_
FIG, 1 Typical recorda of Na+ outflux . Middle :
Saline injected .
III injected . vea 2Q ul .
Top:
Bottom :
Eyestnlk extract injected,
Cephalothorax extract fraction
Ia all cases the volume of extract or saline injected
Arrwa indicate time of injection.
SODIUM REGULATION
2174
Vol .
6, No .
20
TABLE 1 Effect of Cephalothorax Extract (Fraction III) of Pandalus
ordaai
an fta+ Outflux of the Crab Hemigrapsus audus . Aa+ Outflux uM/2 hours Control
Before
After
Difference
1
1333
1133
200
2
1958
602
3
1352
1356 1131
241
Injected
1685
523
1162
2
25T3
1163
1410
3
2383
594
1789
1
Variance ratio of differences 33 .8 ; P
BEPHADE% G-10 COLUMN 100 X 2 S CY . B.4 YL / TUBE FLOW RATE : 9.33 YL / HR
2h
I N
1 2
TUBE NUMBER
FIO . 2 Chromatography of cephalathorax extract oa Sephadex G-10 In another type of experiment the extract or esline vas injected at the
beginning of the experiment end then the Hn+ outflux was measured for 3 hours, In the animals injected xith .frnetioa III the Aa+ outflux was considerably lox
when compared to control animals (Table 2) .
Vol . 6, No . 20
2175
SODIUM REGULATION
TABLE 2 Effect of Cephalothoraz Eztract (Fraction III) of Paadalua ~ordaai oa Aa+ Outflux of the Crnb Hemigrapaus audua Na+ Outflux LM first 3 hours Control 5011 S . D.
t
Injected
P
2935
1864
S . D . = 851
0.06
These results suggest the presence of a "factor" is the extracts of the cephalothoraz, controlling Ae+ regulation in crustaceans,
If the factor is
neurosecretory in origin, it probably nriaea in the cells of the central nervous system proper rather than in the neurosecretory complex of the eyestalk .
The active factor probably decreases the permeability of the gills
to Ae+ ions . Further atudiea to eaelyze the pathvaya of Aa+ fluxes in Hemiarapaua nudue , to know hw rates change in reaponae to altered environmental salinity and to determine the site for possible control are in progress . References 1.
B . T . SC~ER, Arch . Aaat . Micr, Morph, Exp , ~4, 531 (1965) .
2.
D . B . CARLISLE, Pubbl . Staz . Zool . Napoli ~, 22~ (1956) .
3,
R . L . LEHMAA and B . T . SCNEER, Phyaiol . Comp . Oecol . 4, 164 (1956) .
4,
F, I, KAMEMOTO, IQ{ANETfi N, KATO, sad LOIS E . TUCKER, Am, Zool . 6, 213 (1966) .
5.
R . RELLER and B, SMITH, J . Exp . Hiol . 2~, 388 (1948) .
6.
DOROTHY E, BLISS, STEFANIE M . E, WANG and EDWIA A MARTINEZ, Am, Zool . 6, 197 (1966) .
7.
G . W . BRYAA, J, Ezp, Biol . ~, 100 (1960b) .
8.
L . H . KLEIAHOLZ and FRAACFS KIMBALL, Gen . Comp . Eadocrinol . ~, 336 (1965) .