• Applied Microscopy
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• v.23 no.2
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• pp.53-77
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• 1993

In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

• Applied Microscopy
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• v.27 no.3
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• pp.295-308
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• 1997

Liver tissues were taken from rats 20 min after administration of dehydrocholic acid and biligrafin, fasted for 8 days, and bile duct ligation for 48 hr. And the zonulae occludentes around the bile canaliculi were investigated by the freeze fracture method. In the normal rats, the zonulae occludentes showed networks consisting of one to four strands in many regions. After administration of dehydrocholic acid, the depth of zonulae occludentes were increased and networks consist of six to nine strands. The zonulae occludentes, however, were disorganized, and strands were reduced after administration of biligrafin. In the fasting rats, the zonulae occludentes showed one or two strands which were loosely linked. After ligation of bile duct, the zonulae occludentes were disorganized, and strands were reduced or still in a state not to be reduced. The strands were interrupted by gaps or linear particles in all the groups, especially in the biligrafin and the ligation. The free ends were remarkable in the fasting group and contact or surround the lateral gap junctions in the dehydrocholic acid group. The discontinuities and free ends of strands were also encountered in the normal group, thus suggesting that the barrier is not complete even in the normal state. The aggregation of particles appeared within the meshwork of zonulae occludentes in all the group, except for the ligation. From the evidence, it is assumed that zonulae occludentes are labile, differ in the structure according to the situation of bile canaliculi and show regional alterations even in the same bile canaliculi, that zonulae occludentes are affected by mechanical stress or certain other factors, and that there is a correlation between the strands and particle aggregations in association with the alteration of zonulae occludentes.