DEVELOPMENT OF THE DIGESTIVE FUNCTIONS DURING LARVAL STAGE OF JAPANESE FLOUNDER PARALICHTHYS OLIVACEUS

Kurokawa, T., Suzuki, T.

Abstract:

Flounder is a commercially important species in aquaculture of Japan.

Zooplanktons of rotifer and brine-shrimp nauplii are essential to rear

flounder larvae, because complete artificial diet for early larvae has

not yet been established. It is thought that larvae can not utilize the

artificial diet since their digestive organs have not fully developed. To

develop a complete diet, it is important to reveal the digestive ability

at the larval stage. In this study, to elucidate the developmental

process of digestive functions of the pancreas and the intestine during

the larval stage of flounder, distribution of trypsinogen and

aminopeptidase was traced by immunohistochemical methods;

trypsinogen is a proform of major protease synthesized in the

pancreas; aminopeptidase is one of the brush border digestive

enzymes of the intestine. In addition, the developmental process of

diffuse pancreas was followed from primordia stage at larva.

The larvae were kept in a 100 L tank supplied with running sea water

at 17 C, and fed on rotifers from 3 to 30 days post-hatching (dph), on

brine shrimp nauplii from 15 to 45 dph and on artificial diet from 20

dph.

From the pancreas, weak signals to antibody against trypsinogen

(anti-eTrg) were first detected at 2 dph. The signals increased

markedly from 2 to 3 dph. Zymogen granules containing trypsinogen

began to be secreted into the intestinal lumen at 3 dph. Thus, the

pancreas of flounder larva acquires an exocrine function by the time

of the first feeding at 3 dph. The pancreas was a small compact organ

at 3 dph. The pancreas started to elongate along the vein on the

intestine at 20 dph. After metamorphosis (45 dph), the pancreas had

branched out along the veins running to the porta hepatis from the

stomach, pyloric appendages, spleen and intestine. Thus, the pancreas

develops from compact-type organ of larvae to the diffuse-type organ

of the adult at the metamorphosis stage.

From the intestine, weak signals to antibody against aminopeptidase

(anti-ramp) were detected from the brush border of the posterior part

of the intestine at hatching. At 1 dph, the anterior part as well as the

posterior part of the intestine exhibited weak signals to anti-ramp of

intestine. At 2 dph, the rectum had differentiated morphologically,

and both intestine and rectum gave strong signals to anti-ramp. At 3

dph, the brush border of the intestine showed strong signals to

anti-ramp, but the signals of the rectum had been reduced. Thus, the

intestinal epithelial cells have already started to synthesize digestive

enzymes onto the brush border at hatch, and the functional

differentiation of the rectum from the intestine occurs at first feeding

at 3 dph.

It is known that the gastric glands start to synthesize digestive

enzyme (pepsinogen) at metamorphosis in flounder. Therefore, the

digestive organs of flounder larvae, including the pancreas and

intestine, other than the stomach, have acquired the ability to

synthesize digestive enzymes at first feeding.

(National Research Institute of Aquaculture Nansei, Mie, 516-01,

Japan)

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