Biochemical and behavioral analysis of Ceratitis capitata at the final

Biochemical and behavioral analysis of Ceratitis capitata at the final

Biochemical and behavioral analysis of Ceratitis capitata at the final step of morphogenesis:
imago emergence and wing maturation.
Bochicchio, Pablo A., Martín M. Pérez,; Alejandro Rabossi & Luis A Quesada-Allué.
Instituto de Investigaciones Bioquímicas Buenos Aires- CONICET, Dpto. Química Biológica –FCEyN – UBA, Fundación Instituto Leloir.
Patricias Argentinas 435 (1405), Bs. As., Argentina. e-mail: [email protected]
The final step in morphogenesis of the adult fly is the acquisition of the definitive body shape and the wing maturation.
Newly eclosed flies have wings that are highly folded and compact.
We focused our interest in the extrication behavior, the acquisition of the final body shape and wing maturation of Ceratitis capitata. These were correlated with behavioral
performance with few metabolic parameters as well as with the sclerotization machinery.
To be able to analyze in detail a significant number of flies and parameters, a novel imaging recording system were set up.
EXPERIMENTAL SETUP
1
A
CAM 1
POST- ECLOSION BEHAVIOR
2
CAM 2
ADULT EMERGENCE EVENTS
3
A
tf
t0
B
B
Pharate adult
D
Exarate adult
P
A
10 mm
V
Extrication; Walking; Stationary
Wild-type C. capitata adults were reared in a chamber, at 23 + 1°C, 50–60%
relative humidity, with a photoperiod of 16:8 h light: dark cycle.
A- Scheme of the film system and scenario. Flies were recorded using two cameras
(day and night). Each camera recorded simultaneously twelve arenas. B- In each
arena a pharate adult of 264 h was glued in the center.
Example of behavioral patterns registered in petri-dish arenas. (A) Record of fly
pathways, starting at extrication point (t0) and ending when the ultimate size and
shape of the adult was attained (tf). (B) Diagram of C. capitata adult locomotor
activity during the extrication (E) and the walking phase (W) and the motionless
phase (S).
Time duration displayed by C. capitata during ecdysis and postemergence period. WSP%: percentage of wing spreading period.
ENERGETIC RESERVES DURING LATE PHARATE AND EXARATE ADULT
1,4
0,8
0,6
0,4
0,2
0
2,5
2
1,5
1
0,5
2,5
-1
Em
1
OXYGEN CONSUMPTION DURING
PHARATE AND EXARATE ADULT
Pharate adult
-1
Em
1 día
WSP
%
Extrication behavior
-
Post-emergence behavior
1- Walking phase
1,5
1
0,5
0
-3
5
C. capitata
2
0
-3
5 días
-3
-1
Em
1
5
6.2
2- Motionless phase:
Wing expansion
Wing tanning
93.8
Total time (Extrication + Post-emergence)
100
LATE
NBAD SYNTHASE ACTIVITY IN THORAX AND ABDOMEN OF C. capitata
5
Exarate adult
0.4
300
BALA
264
0
0.25
2
24
144
480 hs
200
150
0.2
0.1
100
14
12
NBAD
10
8
0.0
-3
-1
0
5
1
*
6
4
2
0
0
10
20
30
40
50
Síntesis NBAD (pmol/Thorax +
Abdomen)
3000
240
250
0.3
cpm.10-3
Oxygen consumption / fly (l/min.)
3
Total Lipids (mg /fly)
1
0.5
3
3,5
1,2
glycogen (ug /mg tissue)
Trehalose content (ug/ fly)
4
2500
2000
1500
1000
500
0
60
m48
Retention time (min)
m24
0
1
2
24
48
horas
NBAD SYNTHASE ACTIVITY IN WINGS OF C. capitata
6
1,2
264
0
0.25
1
2
24
144
480 hs
1
Síntesis NBAD (pmol/ala)
240
0,8
0,6
0,4
0,2
0
48m
24m
0
1
2
24
48
144
480
tiem po de s de la e m ergencia (hs )
7
CONCLUSIONS
• We identified two clearly defined periods just after ecdysis where the fly acquires the final body shape and expands the wings. These periods were denominated: walking period
and motionless periods.
• During the motionless period occurs the expansion, the sclerotization and the tannification of the wings. After which, the imago is able to fly.
• The oxygen consumption in just emerged exarate adult of C. capitata, was four fold higher with respect to pharate adult.
• A rapid recovery of glycogen and lipid reserves occur immediately after emergence and were associated with an important increase in the metabolic rate.
• In thorax and abdomen the maximal N-B-alanyldopamine synthesis was observed during the first two hours after the imago emergence and then decreased to scarce levels.
• Strikingly, N-B-alanyldopamine synthase activity in wings remained high from the emergence until the 6th day.
• The correlation of behavior with final sclerotization of wings is reported here for the first time.
144
480