Amino acids modulate ion transport and fluid secretion by insect Malpighian tubules
Matthew H. Hazel1,
Juan P. Ianowski1,
Robert J. Christensen1,
Simon H. P. Maddrell2 and
Michael J. O'Donnell1,*
1
Dept of Biology, McMaster University, 1280 Main Street West, Hamilton,
Ontario, L8S 4KI, Canada
2
Dept of Zoology, Downing Street, Cambridge, CB2 3EJ, UK

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Fig. 1. Effects of amino acids on secretion rates of Drosophila
melanogaster tubules. The continuous-exposure protocol was used (see
Materials and methods). Amino acids were tested at 20 mmol l-1
except for threonine and tyrosine, which were tested at 10 mmol l-1
and 0.5 mmol l-1, respectively, due to their lower solubility in
Drosophila saline. For each amino acid, a corresponding set of
controls was run in amino-acid-free saline. Each bar represents the mean
secretion rate (+S.E.M.) as a percentage of the corresponding control mean.
N=8-16 tubules for each amino acid and corresponding control group.
Asterisks represent significant (P<0.05) increases or decreases in
secretion rate relative to the controls. The inset shows a doseresponse
curve (mean ± S.E.M.) for glutamine.
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Fig. 2. Time course of the effects of glutamine and cysteine on secretion rates of
Drosophila tubules. Values are means ± S.E.M. Filled circles
represent control tubules in amino-acid-free Drosophila saline
(N=7), filled squares represent tubules exposed to 20 mmol
l-1 cysteine at t=40 min (N=12), and open circles
represent tubules exposed to cysteine at t=40 min, and then to both
20 mmol l-1 cysteine and 20 mmol l-1 glutamine at
t=80 min (N=3).
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Fig. 3. Time course of the effects of glutamine on secretion rates of 4th instar
Rhodnius Malpighian tubules. The experiment represents a typical
rescue protocol. Values are means ± S.E.M. All tubules were started in
amino-acid-free control saline and were stimulated with 5-HT at t=0
min. The time of glutamine (20 mmol l-1) addition is indicated by
the arrow. Filled circles indicate tubules exposed to the glutamine
(N=8), and open circles indicate control tubules (N=8).
Asterisks indicate significant differences from the value for the same group
at t=75 min.
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Fig. 4. Graph summarizing the results of rescue experiments for Rhodnius
prolixus. All amino acids were tested at 20 mmol l-1 except
threonine, which was tested at 10 mmol l-1 due to its solubility.
Each bar represents the mean secretion rate (+ S.E.M.) as a percentage of the
corresponding control mean. N=7-9 tubules for each amino acid and
corresponding control group. The broken line indicates the control rate
(100%). Asterisks represent significant (P<0.05) increases or
decreases in secretion rate relative to the controls. Tyrosine was not tested
because its low solubility did not permit preparation of an appropriate stock
solution at 10x the final concentration. The insets show
doseresponse curves for stimulation by glutamine and inhibition by
arginine and lysine. Values are means ± S.E.M. (N=17-13
tubules for each amino acid and corresponding control group).
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Fig. 5. Secretion rates of 4th instar Rhodnius Malpighian tubules set up
using the pre-incubation protocol. Values are means ± S.E.M.
(N=8 for both groups). The experimental tubules (filled circles) were
preincubated for 2.5 h in 20 mmol l-1 glutamine, whereas the
control tubules (open circles) were pre-incubated for 2.5 h in amino-acid-free
control saline. All tubules were then transferred to amino-acid-free control
saline and stimulated with 5-hydroxytryptamine (5-HT) at the time indicated by
the arrow.
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Fig. 6. Graph summarizing the results of pre-incubation experiments for
Rhodnius tubules. All amino acids were tested at 20 mmol
l-1 except for threonine (10 mmol l-1) and tyrosine (0.5
mmol l-1). Each bar represents the mean secretion rate (+ S.E.M.)
as a percentage of the corresponding control mean. The broken line indicates
the control rate (100%). N=7-16 tubules for each amino acid and
corresponding control group. Asterisks indicate significant
(P<0.05) increases or decreases in secretion rate relative to
controls. The inset shows the extent to which the inhibition produced by
cysteine could be reversed by subsequent addition of 20 mmol l-1
glutamine.
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Fig. 7. Graph summarizing the results of pre-incubation experiments for
Drosophila tubules. All amino acids were used at 20 mmol
l-1 except for threonine (10 mmol l-1) and tyrosine (0.5
mmol l-1). Each bar represents the mean secretion rate (+ S.E.M.)
as a percentage of the corresponding control mean. The broken line indicates
the control rate (100%). N=8-16 tubules for each amino acid and
corresponding control group. Asterisks represent significant
(P<0.05) increases or decreases in secretion rate relative to
controls.
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Fig. 8. Secretion rate and secreted fluid Na+ and K+
concentrations for 4th instar Rhodnius Malpighian tubules set up in a
rescue experiment. Values are means ± S.E.M. (N=8 tubules for
each group). Filled circles represent tubules rescued with 20 mmol
l-1 glutamine at t=60 min (indicated by the arrow), and
open circles represent control tubules. Asterisks represent significant
(P<0.05) increases or decreases relative to controls.
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Fig. 9. Secretion rate and secreted fluid Na+ and K+
concentrations for 4th instar Rhodnius Malpighian tubules set up in a
pre-incubation experiment. Values are means ± S.E.M. (N=8
tubules for each group). Filled circles represent tubules pre-incubated with
20 mmol l-1 glutamine, and open circles represent control tubules
pre-incubated in amino-acid-free saline. Asterisks represent significant
(P<0.05) increases or decreases relative to controls. Tubules were
stimulated with 5-hydroxytryptamine (5-HT) at t=0 min.
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Fig. 10. Secretion rate and secreted fluid pH for 4th instar Rhodnius
tubules pre-incubated for 2 h in 20 mol l-1 glutamine (filled
circles), 20 mol l-1 arginine (open circles) or amino-acid-free
saline (filled squares). Values are means ± S.E.M. (N=8
tubules for each group). Asterisks represent significant (P<0.05)
increases or decreases relative to controls.
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© The Company of Biologists Ltd 2003