Expression of apoptosis-related factors in muscles infected with Trichinella spiralis.

We found that the expression of mitochondrial apoptosis related genes (Bcl-2 associated protein X, BAX; apoptotic protease activating factor 1, Apaf-1; Caspase 9 and serine/threonine protein kinase, PKB) is elevated in Trichinella spiralis-infected muscles during encapsulation. Micro-dissection of the capsule and subsequent reverse transcription polymerase chain reaction (RT-PCR) confirmed that the expressions of these genes are restricted to the nurse cell. Immunocytochemistry revealed that pro-apoptosis factor (BAX, Apaf-1 and Caspase 9) are predominantly expressed in the basophilic cytoplasm (infected muscle cell origin) and anti-apoptosis factor (PKB) in the eosinophilic cytoplasm (satellite cell origin) of the nurse cell. Electron microscopy revealed that the pre-existing mitochondria in the muscle cells became swollen and disappeared immediately after newborn larva invasion, but new mitochondria of smaller size appeared in the cytoplasm. Nuclear fragmentation and condensation were observed in basophilic cytoplasm which is known to die. Together, the results suggest that the infected muscle cells transform but die through the process of apoptosis which is triggered by factors from the newly formed mitochondria. The anti-apoptosis factor may help the eosinophilic cytoplasm with its survival to ensure nurse cell function.

Adaptive changes in muscle fibers infected with Trichinella spiralis.

Ultrastructural changes which occurred in infected skeletal muscle fibers after infection with larvae of Trichinella spiralis were followed on a daily basis utilizing synchronous infections. No changes were observed in muscle fiber architecture during the first 2 days of intracellular infection. However, on Day 3, a space containing various sarcoplasmic elements developed between the plasma membrane and myofilaments. Widening near the regions of triads was also observed at this time. On Day 4 the space at the outer edge had increased, as did the ones at the triads. In addition, the myofilaments throughout the infected fiber were in a state of partial disarray. Finally, the nuclei were enlarged and had migrated to the central portion of the infected cytoplasm. On Day 5 day, sarcomeres were highly disorganized, and an increase in sarcoplasmic reticulum (SR) was noted. By Day 8, only the extreme periphery of the infected fiber contained Z bands with actin filaments attached. Proliferation of the t tuble system was also evident. At Day 10, myofilaments were completely replaced with SR. Further, the plasma membrane became hyperinvoluted and was associated with a 36-fold increase in the thickness of the glycocalyx. No further enlargement of nuclei occurred after Day 10. Finally, a host-derived double membrane completely surrounded the larva.