Dietary-induced modification of calcium transport in mitochondria isolated from flight-muscle of developing sheep blowfly Lucilia cuprina

Vol. 79, No. 1, 1977

BIOCHEMICAL

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

DIETARY-INDUCED MODIFICATION

OF CALCIUM TRANSPORT

IN MITOCHONDRIA ISOLATED FROM FLIGHT-MUSCLE OF DEVELOPING SHEEP BLOWFLY LUCILIA Fyfe

L. Bygrave

and Robyn L.

CUPRINA

Smith

Department of Biochemistry, Faculty of Science, National University, Canberra, A.C.T. 2600, Australia

Australian Received

September

16,1977

SUMMARY: The activity of mitochondrial calcium transport in fZight-muscZe of the sheep blowfly Lucilia cuprina is shown to depend on the nature of the diet upon which the adult fly is maintained. The finding raises the possibility that calcium transport in mitochondria from other tissues and species also might be influenced by the dietary intake. Numerous biochemical of

cell

species

and it

and tissues

ionized

transport plays

Ca.

by the

located

role

in the

Particularly

compelling

that

mitochondrial

Ca transport

opment in insect goes transient

certain

tumour In extending

possibility

that

influenced

also

system attributes

Copyright All rights

described

the

cell

these

(Ca)

events

view

that

often

the

inner

Ca

membrane in refs.

stems from observations

changes markedly

during

devel-

and mammalian tissue(43), hormonal

ions

concentration

Ca (reviewed

evidence

flight-muscle changes after

in vivo(6)

that

mitochondrial

in controlling

in a range

by calcium

intracellular

supports

l-3).

liver

some time

Much evidence

system

a major

for

in part

events

are influenced

has been advocated

may be controlled of

and physiological

perturbation

and may have undergone

of

permanent

underrat

changes

in

lines('7,8). this

line

of

mitochondrial by the

to enable

these

0 1977 by Academic Press, Inc. of reproduction in any form reserved.

we considered

Ca transport

dietary

previously(4)

research,

activity

the might

intake.

The developing

appeared

to have particular

ideas

to be tested.

be

insect

We reDort

here

154 ISSN

0006-291X

BIOCHEMICAL

Vol. 79, No. 1, 1977

results

of experiments

flight-muscle flies. the

in which

mitochondria

Ca transport

isolated

One group was maintained other

on a diet

show that flies

maintained

maintained

EXPERIMENTAL: Division

flies feed

on Diets

duct

of

hydrates,

Ltd.)

75.2%;

and moisture, each diet.

flight-muscle at the

Ca transport nique(9).

A consisted

the

approximately

indicated

chondria

periods

transport

activity

6 hours

after

on Diets between

emergence

la).

155

(a product :

of

carbo-

minerals,

4.0%

quantities all

described(4)

in legends

to

figures.

Red-quench

legends

(phosphate)

influence

flight-muscle

had been maintained A large

tech-

to figures,

on Ca transport

the two groups

(Fig.

cubes (a pro-

as previously

1 show the

A and B.

to

from the

phosphate

that

and allowed

were isolated

by LuciZia

from adults

adult

at 28OC throughout

effects

on Ca transport

different

3%;

in the

Inorganic

Data in Fig.

isolated

that

and

sugar

the Ruthenium

are described

marked concentration-dependent

concentration

than

composition

80 flies

was measured using

mitochondria(l0).

from

were reared

in liberal

Mitochondria

stages

of

fat,

was provided

RESULTS AND DISCUSSION:

We

by the

jars

following

life-cycle.

Details

B).

provided

baby food,

were maintained

developmental

kindly

glass

13.8%;

Water

A) and

Upon emergence,

Flies

of

(Diet

Canberra,

B was "Farex"

protein,

4%.

of the

Diet

with

(Diet

greater

previously(4).

Diet

Australia

stages

C.S.I.R.O.,

A or B.

of adult

mitochondria

and pupae,

in muslin-covered

C.S.R.).

protein

in

B.

larvae

as described

were placed

Glaxo

on Diet

LuciZia

alone

A is significantly

of Entomology,

maintained

with

in flight-muscle

on Diet

was compared

from two groups on sugar

supplemented

Ca transport

in flies

with

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

induces in LuciZia

of phosphate mitofor

difference

in Ca

is seen as early

A similar

pattern

of

as response

BIOCHEMICAL

Vol. 79, No. 1, 1977

(A)

30

01

AND BIOPHYSICAL

RESEARCH COMMUNICATIONS

(6)

0

15

30

0

15

30

02

mM Phosphate

0

24 48 72 Hrs. after emergence

Fig.1. Ca transport measured as a function of phosphate concentration in flight-muscle mitochondria of Lucilia maintained on different diets. Measurement of Ca transport was made in reaction media containing in a final vol. of 1OOmMKCl, 25mM @-glycerol phosphate and varying l.Oml: concentrations of inorganic phosphate as indicated. The final pH and temp. were 7.4 and 25OC, respectively. Mitochondria (1 mg. protein) obtained from flight-muscle of flies maintained on diet A (0) or diet B (0) for the . . were added and after a 1 min. f,"~~~~~i~~d'5"o"",~d~a2+ (containing approx. 0.3~Ci~~Z+) was added to initiate the reaction. At 30 sec. intervals, 100~1 of reaction medium was transferred to Eppendorf tubes containing 0.5mM ethanedioxybis-(ethylamine) tetraacetic acid (EGTA) plus 5uM Ruthenium Red. The tubes were centrifuged and portions of the supernatant counted for radioThe ages of the flies from which activity (see ref. 9). mitochondria were isolated were (a) 6 hr; (b) 15 hr.; (c) 24 hr.; (d) 48 hr. and (e) 72 hr. Fig.2. Changes during development of transport measured in the presence of Data are derived from Fig. 1 and show with 2.5mM phosphate at the different in mitochondria from flies maintained B CO>.

of Ca transport transport isolated emergence

to phosphate

activity from the

flies

is

depressed

maintained

response

concentration

pattern

about

mitochondrial Ca 2.5mM phosphate. Ca transport activity developmental stages on diet A (0) or

is obtained

50% in those B.

is

similar

156

(Fig,

Ca

mitochondria

At 15 hours

on Diet also

but

lb)

after but

Vol. 79, No. 1, 1977

high

BIOCHEMICAL

phosphate

previously

concentrations

apparent

by phosphate

by 24 hours

Ca transport phosphate feature 72 hours

is

accentuated

of

maintained

depressed of

at

flies

Ca transport

that

flies

greater

than

on Diet

A (Fig.

all

lc); of

on Diet.

Id)

Ca transport

maintained

2.5mM is

concentrations

(Fig.

as

of mitochondrial

maintained

by 48 hours

emergence

by mitochondria (Fig.

in flies

activity

after

to inhibit

concentrations

in mitochondria is

begin

The marked inhibition

reported(l0).

Ca transport

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

and it

This

B.

is not

until

at 2.5mM phosphate

on Diet

A, begins

to decline

lc). Data

in Fig.

transport, during

2 derived

maintained

in flies

on Diet

maximal

activity

Other

of both

for

that

the

sexes.

those

at least

experiments,

indicated

exhibit

while

in the energy transport

data

and occur

of which

that

the dietary

port

activity.

be drawn about changes, provide

we feel

which

in both not

the

systems

data

that

such information.

further

types

of the

differences

for

driving

the

first

of the

Ca

fly.

they

mitochondrial

mitochondrial

157

for

in that

exploitation Since

of mitochondria.

sex of the

do not permit

details

in flies

in the presence

responsible

can influence

the molecular

activity A retain

occurred

to reflect

shows, we believe

While

Those

are not presented,

are significant

regimen

on Diet

activities

independently

The above findings

transport

maintained

seen appear

transducing

evidence

reduced

the respiratory

differences

changes

on the two diets.

changes to Ca transport

Also

1 show how Ca

48 hours.

or absence of ADP were similar Thus the

in Fig.

of 2.5mM phosphate,

maintained

B clearly

in development

clear

those

measured in the presence development

early

from

provide time, Ca trans-

conclusions

to

dietary-induced of this

system will

Ca transport

BIOCHEMICAL

Vol. 79, No, 1, 1977

in insect

flight-muscle

properties(l,4) general

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

and mammalian tissues

we consider

these

findings

share

many

may have quite

relevance.

ACKNOWLEDGEMENT: We thank the Australian Wool Trust Fund for financial support of this work and Drs A.J. Howells and K.L. Williams for helpful discussions. REFERENCES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10.

Current Topics in Bioenergetics, 6, 259-318 F.L., (1977). Baker, P.F., Sot. Exp. Biol. Symp., 30, 67-88 (1976). E. and Crompton, M., Sot. Exp. Biol. Symp., Carafoli, 30, 89-115 (1976). Bygrave, F.L., Daday, A.A. and Doy, F.A., Biochem. J., 146, 601-608 (1975). Bygrave, F.L. and Ash, G.R., FEBS Letters, 80, 271-274 (1977). Dorman, D.M., Barritt, G.J. and Bygrave, F.L., Biochem. J. 150, 389-395 (1975). F.L., Biochem. J., 144, 551Thorne, R.F.W. and Bygrave, 558 (1974). in Control Mechanisms in Cancer (W.E. Criss, Bygrave, F.L., T. Ono and J.R. Sabine, eds.) pp. 411-423, Raven Press, New York (1976). Reed, K.C. and Bygrave, F.L., Biochem. J. 140, 143-155 (1974). F.L., Biochem. J. in press. Smith, R.L. and Bygrave, Bygrave,

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