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Korean J Parasitol > Volume 33(3):1995 > Article

Original Article
Korean J Parasitol. 1995 Sep;33(3):155-164. English.
Published online Sep 20, 1995.  http://dx.doi.org/10.3347/kjp.1995.33.3.155
Copyright © 1995 by The Korean Society for Parasitology
Localization of actin and myosin in Cryptosporidium parvum using immunogold staining
J R Yu,*1 and J Y Chai2
1Department of Parasitology, College of Medicine, Kon-Kuk University, Choongju 380-701, Korea.
Received August 04, 1995; Accepted August 26, 1995.

Abstract

The location of actin and myosin of the several stages of Cryptosporidium parvum was observed. The tissue antigen of C. parvum was prepared through immunosuppression of ICR mice with Depomedrol®. The thin sectioned specimens, which were incubated with the IgG fraction of the rabbit polyclonal antibodies raised against chicken back muscle actin and bovine uterus myosin, were treated with 10 nm gold-conjugated goat anti-rabbit IgG. Electrodense particles were located mainly on the pellicles of all observed developmental stages of the parasites. The number of actin gold particles in the cytoplasm increased when the parasite was dividing actively as in case of meronts. Especially in macrogametocytes, a lot of actin and myosin particles were synthesized and storaged as amilopectin-like bodies. There were many actin gold particles along the microspikes of cytoplasmic membranes in various developmental stages. The actin and myosin observed in this study may play important roles to control the shape of the parasites and movements of cytoplasmic membranes as cytoskeletal proteins.

Figures


Figs. 1-4
Positive control of immunogold staining with primary antibodies against action (Fig. 1) and myosin (Fig. 2) in the rat abdominal muscle. Negative control of immunogold staining missing primary antibodies against actin(Fig. 3) and myosin (Fig. 4). Bar; 200 nm.


Figs. 5-8
The localities of actin shown by immunogold staining. In the trophozoite, gold particles are seen on the pellicle and cytoplasm around the feeder organelle (Fig. 5). In the uninucleated meront, gold particles ard dispersed all over the cytoplasm and pellicle except the nucleus and dense bobies (Fig. 6). In the type II meront, the pellicle and membranes between th merozoites have gold particles (Fig. 7). In the macrogametocyte, amylopectin-like bodies and cytoplasm around the feeder organelle show many gold particles, whereas wall forming bodies do not (Fig. 8). ap; amyolpectin-like body, d; dense body, fo; feeder organelle, mv; microvilli, nu; nucleus, p; pellicle, r; rhoptry, rb; residual body, w; wall forming body. Bar; 200 nm.


Figs. 9-13
The localities of actin shown by immunogold staining. In the developing oocyst, gold particles are located along the oocyst wall and cytoplasmic membrane and the actin detected at the center of the cytoplasm (asterisk) is thought to be involved in mitosis of the cell (Fig. 9). In the mature meront including one unescaped merozoite, the pellicle, merozoites membrane and cytoplasmic residuum show many gold particles (Fig. 10). Mature oocysts containing one sporozoite show the same patterm as mature meronts (Fig. 11). Some vacant parasitophorous vacuoles reveal long projectons like filopodia (arrow head) and microspikes (arrow) and they have many action gold particles (Fig. 12,Fig. 13). cr; cytoplasmic residuum, mz; merozoite, ow; oocyst wall, p; pellicle, sp; sporozoite. Bar; 200 nm.


Figs. 14-17
The localities of myosin shown by immunogold staining. In the trophozoite (Fig. 14) and uninucleated meront (Fig. 15). gold particles are detected along the pellicles, but they are very few in the nucleus and cytoplasm. In the macrogametocyte, amylopectin-like bodies are stained well with gold particles (Fig. 16). In the type II meront, gold particles are distributed along the pellicle and merozoites membranes also (Fig. 17). ap; amylopectin-like body, mv; microvilli, nu; nucleus, p;pellicle, r; rhoptry, w; wall forming body, Bar; 200 nm.


Figs. 18-19
The localities of myosin shown by immunogold staining. In the developing oocysts a small number of gold particles were detected along the oocyst wall (Fig. 18). In an unidentified developmental stage. gold particles showed well the localization of myosin along the pellicle (arrow heal; Fig. 19). Bar; 200nm.

References
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