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Ultrastructural antigenic localization in Paragonimus iloktsuenensis during developmental stage by immunogold labeling method
Hoon Sik Kim,1 and Ok Ran Lee*2
1Choongang Internal Medicine Clinic, Taejeon City 303-301, Korea.
2Department of Parasitology, Soonchunhyang University College of Medicine, Chonan 330-090, Korea.
Received October 06, 1995; Accepted November 09, 1995.
Abstract
Antigenic localization in Paragonimus iloktsuenensis worm tissues (tegument, intestine and vitelline gland) in different developmental stages of 2 weeks, 3 weeks, 4 weeks, 5 weeks and 33 weeks from albino rats (Sprague-Dawley) infected with P. iloktsuenensis was observed by electron microscopy. These worm tissues of different developmental stage of P. iloktsuenensis was observed on electromicrograph by immunogold labeling method using P. iloktsuenensis infected rat serum of 10 weeks. Antigenic localization was demonstrated as labeling of gold particles in tissues on electronmicrograph. In tegument, gold particles were labeled on tegumental tissue, generally more numerous on secretory granules in tegumental syncytium 2 weeks than those on the other elder developmental stages, but there was a little variation in antigenicity according to individual worm tissue. In general, antigenicity in tegumental tissue was not strong (gold particles: 0.1- 5/1 µm2). In intestine, a large number of gold particles (15-18/1 µm2) were labeled in intestinal epithelium. Gold particles were concentrated especially on secretory granules in cytoplasm, and gold particles were labeled not only in cytoplasmic protrusions, but also in intestinal luminal contents. Intensity of labeling of gold particles was not correlated with developmental stage of worms. In vitelline gland, a large number of gold particles were labeled on vitelline globules. The gold particles in vitelline globules (8-11/1 µm2) were concentrated in protoplasm among segmental globules.
Figures
Fig. 1 Tegument of Paragonimus iloktsuenensis on electronmicrograph. a: Showing tegumental surface (S), tegumental syncytium (TS), basal lamana (BL), muscle (MU), tegumental cell (TC), in 33 weeks developmental worm. Bar = 0.5 µm (× 2,400) b: Magnification of Fig. 1a. M: mitochondria bar = 0.5 µm (× 8,400) c: Tegumental syncytium of 5 weeks developmental worm. Showing the numerous secretory granules (SG), mitochondria (M), and rough endoplasmic reticulum (RER). Bar = 2 µm (× 48,000).
Figs. 2-3 Fig. 2. Intestinal epithelium of P. iloktsuenensis on electronmicrograph. Numberous secretory granules of several size are located in the cytoplasm of intestinal epithelial cell (IEC). and long slender cytoplasmic protrusion (CP) is projected toward intestinal lumen, the RER is well developed. 5 weeks developmental worm. Bar = 1 µm (× 39,000). Fig. 3. Vitellin gland of P. iloktsuenensis on electronmicrograph. a: Vitelline lobules (VL) of 33 weeks developmental worm. Each vitelline lobules is composed of vitelline cells (VC) of various developmental stage. Bar = 0.5 µm (× 6,000). b: Vitelline cell (VC) of 33 weeks developmental worm. In well developed vitelline cell, numerous vitelline globules (VG) are located in cytoplasm, and a large amount of RER are surrounded nucleus (N). NU: nucleolus. Bar = 1 µm (× 12,000). c: Vitelline globules (VG) of 33 weeks developmental worm. Showing the cleavaged vitelline globules. Bar = 1 µm (× 24,000).
Figs. 4-8 Tegumental syncytium in several developmental stages of P. iloktsuenensis on lectronmicrograph by immunogold labeling method P. iloktsuenensis infected rat serum. Gold particles (GP) are labeled mostly on the secretory granules (SG) in cytoplasm. Bar = 2 µm. Fig. 4. 2 weeks worm (× 48,000). Fig. 5. 3 weeks worm (× 48,000). Fig. 6. 4 weeks worm (× 39,000). Fig. 7. 5 weeks worm (× 60,000). Fig. 8a. 33 weeks worm (× 54,000). Fig. 8b. 33 weeks worm (× 24,000). *a and b are different worms. In the b worm, a number of gold particles are labeled in similar desity with those in the 2 weeks worm.
Figs. 9-14 Intestinal epithelium in different developmental stages of P. iloktsuenensis on electronmicrograph by immumogold labeling method using P. iloktsuenensis infected rat serum. Gold particles are abundantly labeled on secretory granules, and on cytoplasmic protrusions (CP). Bar 2 µm. Fig. 9. 2 weeks worm (× 30,000). Fig. 10. 3 weeks worm (× 39,0000. Fig. 11. 4 weeks worm (× 39,000). Fig. 12. 5 weeks worm (× 39,000). Fig. 13. 33 weeks worm (× 33,000). Fig. 14. Intestinal luminal contents of 33 weeks developmental worm. Gold particles in vitelline globules were concentrated in protoplasm among cleavaged globules.
Figs. 15-18 Vitelline globules in defferent developmental stages of P. iloktsuenensis on electronmicrograph by immunogold labeling method using P. iloktsuenensis infected rat serum. Gold particles are labeled on the protoplasm between vitelline globules. Bar = 3 µm. Fig. 15. 3 weeks worm (× 84,000). Fig. 16. 4 weeks worm (× 48,000). Fig. 17. 5 weeks worm (× 60,000). Fig. 18. 33 weeks worm (× 84,000).
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