Accumulation of α-2,6-sialyoglycoproteins in the muscle sarcoplasm due to Trichinella sp. invasion

Abstract

The sialylation of the glycoproteins in skeletal muscle tissue is not well investigated, even though the essential role of the sialic acids for the proper muscular function has been proven by many researchers. The invasion of the parasitic nematode Trichinella spiralis in the muscles with subsequent formation of Nurse cell-parasite complex initiates increased accumulation of sialylated glycoproteins within the affected area of the muscle fiber. The aim of this study is to describe some details of the α-2,6-sialylation in invaded muscle cells. Asynchronous invasion with infectious T. spiralis larvae was experimentally induced in mice. The areas of the occupied sarcoplasm were reactive towards α-2,6-sialic acid specific Sambucus nigra agglutinin during the whole process of transformation to a Nurse cell.The cytoplasm of the developing Nurse cell reacted with Helix pomatia agglutinin, Arachis hypogea agglutinin and Vicia villosa lectin-B4 after neuraminidase pretreatment.Up-regulation of the enzyme ST6GalNAc1 and down-regulation of the enzyme ST6GalNAc3 were detected throughout the course of this study. The results from our study assumed accumulation of sialyl-Tn-Ag, 6`-sialyl lactosamine, SiA-α-2,6-Gal-β-1,3-GalNAc-α-Ser/Thr and Gal-β-1,3-GalNAc(SiA-α-2,6-)-α-1-Ser/Thr oligosaccharide structures into the occupied sarcoplasm. Further investigations in this domain will develop the understanding about the amazing adaptive capabilities of skeletal muscle tissue.

The synthesis of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE), α-dystroglycan, and β-galactoside α-2,3-sialyltransferase 6 (ST3Gal6) by skeletal muscle cell as a response to infection with Trichinella spiralis

“…The up-regulation of Gne described in this work was in a very good agreement with our previous findings concerning the increased sialic acid biosynthesis in the Nurse cell (Milcheva et al, 2015(Milcheva et al, , 2019(Milcheva et al, , 2020. Considering the up-regulation of Gne however, we set our attention on the sialyltransferase activity.…”

“…In skeletal muscles, the sialic acids are important for the functional maintenance of glycoproteins involved in fi ber structure and neuromuscular junctions (McDearmon et al, 2003;Combs & Ervasti, 2005), development and regeneration (Broccolini et al, 2008), muscle excitability (Johnson et al, 2004;Schwetz et al, 2011) and exercise performance (Hanish et al, 2013). They obviously play some role in the process of development of the skeletal muscle cell into a nurse cell, as already reported (Milcheva et al, 2015(Milcheva et al, , 2019(Milcheva et al, , 2020.…”

“…Infective Trichinella spiralis larvae (code ISS03) were isolated from previously invaded laboratory albino mice ( Mus musculus musculus ), between 30 th and 40 th day post infection (d.p.i.) according to a routine protocol, as already described ( Milcheva et al, 2019 ). Fifteen male mice, 6 – 8 weeks old, were inoculated with 500 infective T. spiralis larvae per os .…”

“…The yield and purity of the collected gDNA were measured using S-300 Spectrophotometer (Boeco, Hamburg, Germany). Hot start PCR was designed on approximately 100 ng gDNA as a template by using Veriti thermoblock (Applied Biosystems of Thermo Fisher Scientific), as already described in details ( Milcheva et al, 2019 ). The products of amplification were visualized on 2.5 % agarose gel supplemented with Simply Safe nucleic acid stain (EurX®, Gdansk, Poland) versus 100-1000 bp DNA Ladder (EurX) and the gels were photographed with a gel documentation system Vision (Scie-Plas Ltd, Cambridge, UK).…”

“…The yield and purity of the collected RNA were measured using S-300 Spectrophotometer (Boeco). Approximately 2 μg total RNA from each sample were used for first strand cDNA synthesis, as already described ( Milcheva et al, 2019 ). The generated cDNA was quantified and the samples were stored at -80°C.…”