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Sulphated glycosaminoglycans and proteoglycans in the developing vertebral column of juvenile Atlantic salmon (Salmo salar).

Hannesson KO, Ytteborg E, Takle H, Enersen G, Bæverfjord G, Pedersen ME - Fish Physiol. Biochem. (2015)

Bottom Line: In addition, the distribution of the different GAG types in normal and malformed vertebral columns from 15 g salmon was compared.A changed expression pattern of GAGs was found in the malformed vertebrae, indicating the involvement of these molecules during the pathogenesis.Our study reveals the importance of GAGs in development of vertebral column also in Atlantic salmon and indicates that a more comprehensive approach is necessary to completely understand the processes involved.

View Article: PubMed Central - PubMed

Affiliation: Nofima AS, 1430, Ås, Norway.

ABSTRACT
In the present study, the distribution of sulphated glycosaminoglycans (GAGs) in the developing vertebral column of Atlantic salmon (Salmo salar) at 700, 900, 1100 and 1400 d° was examined by light microscopy. The mineralization pattern was outlined by Alizarin red S and soft structures by Alcian blue. The temporal and spatial distribution patterns of different types of GAGs: chondroitin-4-sulphate/dermatan sulphate, chondroitin-6-sulphate, chondroitin-0-sulphate and keratan sulphate were addressed by immunohistochemistry using monoclonal antibodies against the different GAGs. The specific pattern obtained with the different antibodies suggests a unique role of the different GAG types in pattern formation and mineralization. In addition, the distribution of the different GAG types in normal and malformed vertebral columns from 15 g salmon was compared. A changed expression pattern of GAGs was found in the malformed vertebrae, indicating the involvement of these molecules during the pathogenesis. The molecular size of proteoglycans (PGs) in the vertebrae carrying GAGs was analysed with western blotting, and mRNA transcription of the PGs aggrecan, decorin, biglycan, fibromodulin and lumican by real-time qPCR. Our study reveals the importance of GAGs in development of vertebral column also in Atlantic salmon and indicates that a more comprehensive approach is necessary to completely understand the processes involved.

No MeSH data available.


Related in: MedlinePlus

Western blotting of proteins isolated from normal and malformed vertebrae of 15 g salmon showing that C-0-S was primarily detected in molecules of higher sizes, whereas C-4-S showed additional bands in the region <60 kDa. Also the main band, appearing by mAb 5D4 against KS, exhibited a molecular size just below the 60-kDa marker. Some broad, weak stain was furthermore seen in the region for higher molecular sizes against KS, most clear in samples from malformed vertebra. mAb against C-6-S was only found this epitope in samples obtained from malformed vertebrae, appearing as a band in the region above the 250-kDa molecular marker. N normal, MF malformed
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Fig11: Western blotting of proteins isolated from normal and malformed vertebrae of 15 g salmon showing that C-0-S was primarily detected in molecules of higher sizes, whereas C-4-S showed additional bands in the region <60 kDa. Also the main band, appearing by mAb 5D4 against KS, exhibited a molecular size just below the 60-kDa marker. Some broad, weak stain was furthermore seen in the region for higher molecular sizes against KS, most clear in samples from malformed vertebra. mAb against C-6-S was only found this epitope in samples obtained from malformed vertebrae, appearing as a band in the region above the 250-kDa molecular marker. N normal, MF malformed

Mentions: Proteoglycans represent a large family of molecules of different sizes which may carry same GAG chains. The results showed that C-0-S was primarily detected in molecules of higher sizes, whereas C-4-S showed additional bands in the region <60 kDa (Fig. 11). Also the main band, appearing by mAb 5D4 against KS, exhibited a molecular size just below the 60-kDa marker in accordance with the sizes for the small proteoglycans as decorin, biglycan, lumican and fibromodulin. Some broad, weak stain was furthermore seen in the region for higher molecular sizes, most clear in samples from malformed vertebrae. Regarding the mAb against C-6-S, we only found this epitope in samples obtained from malformed vertebrae, appearing as a band in the region above the 250-kDa molecular marker. The result suggests that this epitope is associated with PG with high molecular size, which changed into lower molecular size able to enter the gel during the pathological development.Fig. 11


Sulphated glycosaminoglycans and proteoglycans in the developing vertebral column of juvenile Atlantic salmon (Salmo salar).

Hannesson KO, Ytteborg E, Takle H, Enersen G, Bæverfjord G, Pedersen ME - Fish Physiol. Biochem. (2015)

Western blotting of proteins isolated from normal and malformed vertebrae of 15 g salmon showing that C-0-S was primarily detected in molecules of higher sizes, whereas C-4-S showed additional bands in the region <60 kDa. Also the main band, appearing by mAb 5D4 against KS, exhibited a molecular size just below the 60-kDa marker. Some broad, weak stain was furthermore seen in the region for higher molecular sizes against KS, most clear in samples from malformed vertebra. mAb against C-6-S was only found this epitope in samples obtained from malformed vertebrae, appearing as a band in the region above the 250-kDa molecular marker. N normal, MF malformed
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4495713&req=5

Fig11: Western blotting of proteins isolated from normal and malformed vertebrae of 15 g salmon showing that C-0-S was primarily detected in molecules of higher sizes, whereas C-4-S showed additional bands in the region <60 kDa. Also the main band, appearing by mAb 5D4 against KS, exhibited a molecular size just below the 60-kDa marker. Some broad, weak stain was furthermore seen in the region for higher molecular sizes against KS, most clear in samples from malformed vertebra. mAb against C-6-S was only found this epitope in samples obtained from malformed vertebrae, appearing as a band in the region above the 250-kDa molecular marker. N normal, MF malformed
Mentions: Proteoglycans represent a large family of molecules of different sizes which may carry same GAG chains. The results showed that C-0-S was primarily detected in molecules of higher sizes, whereas C-4-S showed additional bands in the region <60 kDa (Fig. 11). Also the main band, appearing by mAb 5D4 against KS, exhibited a molecular size just below the 60-kDa marker in accordance with the sizes for the small proteoglycans as decorin, biglycan, lumican and fibromodulin. Some broad, weak stain was furthermore seen in the region for higher molecular sizes, most clear in samples from malformed vertebrae. Regarding the mAb against C-6-S, we only found this epitope in samples obtained from malformed vertebrae, appearing as a band in the region above the 250-kDa molecular marker. The result suggests that this epitope is associated with PG with high molecular size, which changed into lower molecular size able to enter the gel during the pathological development.Fig. 11

Bottom Line: In addition, the distribution of the different GAG types in normal and malformed vertebral columns from 15 g salmon was compared.A changed expression pattern of GAGs was found in the malformed vertebrae, indicating the involvement of these molecules during the pathogenesis.Our study reveals the importance of GAGs in development of vertebral column also in Atlantic salmon and indicates that a more comprehensive approach is necessary to completely understand the processes involved.

View Article: PubMed Central - PubMed

Affiliation: Nofima AS, 1430, Ås, Norway.

ABSTRACT
In the present study, the distribution of sulphated glycosaminoglycans (GAGs) in the developing vertebral column of Atlantic salmon (Salmo salar) at 700, 900, 1100 and 1400 d° was examined by light microscopy. The mineralization pattern was outlined by Alizarin red S and soft structures by Alcian blue. The temporal and spatial distribution patterns of different types of GAGs: chondroitin-4-sulphate/dermatan sulphate, chondroitin-6-sulphate, chondroitin-0-sulphate and keratan sulphate were addressed by immunohistochemistry using monoclonal antibodies against the different GAGs. The specific pattern obtained with the different antibodies suggests a unique role of the different GAG types in pattern formation and mineralization. In addition, the distribution of the different GAG types in normal and malformed vertebral columns from 15 g salmon was compared. A changed expression pattern of GAGs was found in the malformed vertebrae, indicating the involvement of these molecules during the pathogenesis. The molecular size of proteoglycans (PGs) in the vertebrae carrying GAGs was analysed with western blotting, and mRNA transcription of the PGs aggrecan, decorin, biglycan, fibromodulin and lumican by real-time qPCR. Our study reveals the importance of GAGs in development of vertebral column also in Atlantic salmon and indicates that a more comprehensive approach is necessary to completely understand the processes involved.

No MeSH data available.


Related in: MedlinePlus