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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

Distribution of chondroitin 4-sulphate (C-4-S) in the salmon spine. In the longitudinal sections a, b, and the transverse section, c collected at 700 d°, staining could hardly be seen. d, e Longitudinal sections at 900 d°, staining in developing IVRs appeared, very strongly in the central part (arrows), enlarged in e. f Transverse section showing that staining appeared in graded levels towards the lumen. g At 1100 d°, staining showed two–three distinct globes in the IVRs, enlarged in h and indicated by arrows. In addition, staining was seen in the layer of IVR towards lumen. i Transverse section showing that C-4-S was present in the notochord. j Longitudinal section at 1400 d°, the pattern of distinct globes and wings became more evident (arrows), enlarged in k. l Transverse section showed the distinct lamellar pattern of the notochordal sheath. No staining was seen in the lumen of the notochord at any stages of development. Scale bar 100 µm; nl notochord lumen
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Fig5: Distribution of chondroitin 4-sulphate (C-4-S) in the salmon spine. In the longitudinal sections a, b, and the transverse section, c collected at 700 d°, staining could hardly be seen. d, e Longitudinal sections at 900 d°, staining in developing IVRs appeared, very strongly in the central part (arrows), enlarged in e. f Transverse section showing that staining appeared in graded levels towards the lumen. g At 1100 d°, staining showed two–three distinct globes in the IVRs, enlarged in h and indicated by arrows. In addition, staining was seen in the layer of IVR towards lumen. i Transverse section showing that C-4-S was present in the notochord. j Longitudinal section at 1400 d°, the pattern of distinct globes and wings became more evident (arrows), enlarged in k. l Transverse section showed the distinct lamellar pattern of the notochordal sheath. No staining was seen in the lumen of the notochord at any stages of development. Scale bar 100 µm; nl notochord lumen

Mentions: Replacing the enzyme cABC lyase with cACII produced a different labelling pattern (Fig. 5). With this enzyme, only C-4-S creates the immunostaining. At 700 d°, staining could hardly be seen in the notochordal sheath and no staining was found in chondrogenic tissue (Fig. 5a–c). This indicates that the dominating GAG in the notochordal sheath at 700 d° was DS. However, at 900 d°, an increase in the expression of C-4-S was detected, mainly restricted to the developing IVRs of the notochordal sheath, appearing as dark brown circular structures placed centrally in the IVR (Fig. 5d–f). At 1100 d°, the staining clearly showed two distinct spots in the IVRs (arrows, Fig. 5g, h), which persisted at 1400 d° (arrows, Fig. 5j, k). Furthermore, transverse sections from 1100 and 1400 d° revealed that C-4-S was arranged in the NS in concentric lamellae (Fig. 5i, l). The results indicate that C-4-S was arranged as a gradient, with strongest staining towards lumen and confined to more well-outlined structures, whereas DS showed a more widespread distribution. No staining for C-4-S was seen in the lumen of the notochord at any stages of development and in contrast to DS only weak staining could be detected in the chondrogenic zones.Fig. 5


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)

Distribution of chondroitin 4-sulphate (C-4-S) in the salmon spine. In the longitudinal sections a, b, and the transverse section, c collected at 700 d°, staining could hardly be seen. d, e Longitudinal sections at 900 d°, staining in developing IVRs appeared, very strongly in the central part (arrows), enlarged in e. f Transverse section showing that staining appeared in graded levels towards the lumen. g At 1100 d°, staining showed two–three distinct globes in the IVRs, enlarged in h and indicated by arrows. In addition, staining was seen in the layer of IVR towards lumen. i Transverse section showing that C-4-S was present in the notochord. j Longitudinal section at 1400 d°, the pattern of distinct globes and wings became more evident (arrows), enlarged in k. l Transverse section showed the distinct lamellar pattern of the notochordal sheath. No staining was seen in the lumen of the notochord at any stages of development. Scale bar 100 µm; nl notochord lumen
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4495713&req=5

Fig5: Distribution of chondroitin 4-sulphate (C-4-S) in the salmon spine. In the longitudinal sections a, b, and the transverse section, c collected at 700 d°, staining could hardly be seen. d, e Longitudinal sections at 900 d°, staining in developing IVRs appeared, very strongly in the central part (arrows), enlarged in e. f Transverse section showing that staining appeared in graded levels towards the lumen. g At 1100 d°, staining showed two–three distinct globes in the IVRs, enlarged in h and indicated by arrows. In addition, staining was seen in the layer of IVR towards lumen. i Transverse section showing that C-4-S was present in the notochord. j Longitudinal section at 1400 d°, the pattern of distinct globes and wings became more evident (arrows), enlarged in k. l Transverse section showed the distinct lamellar pattern of the notochordal sheath. No staining was seen in the lumen of the notochord at any stages of development. Scale bar 100 µm; nl notochord lumen
Mentions: Replacing the enzyme cABC lyase with cACII produced a different labelling pattern (Fig. 5). With this enzyme, only C-4-S creates the immunostaining. At 700 d°, staining could hardly be seen in the notochordal sheath and no staining was found in chondrogenic tissue (Fig. 5a–c). This indicates that the dominating GAG in the notochordal sheath at 700 d° was DS. However, at 900 d°, an increase in the expression of C-4-S was detected, mainly restricted to the developing IVRs of the notochordal sheath, appearing as dark brown circular structures placed centrally in the IVR (Fig. 5d–f). At 1100 d°, the staining clearly showed two distinct spots in the IVRs (arrows, Fig. 5g, h), which persisted at 1400 d° (arrows, Fig. 5j, k). Furthermore, transverse sections from 1100 and 1400 d° revealed that C-4-S was arranged in the NS in concentric lamellae (Fig. 5i, l). The results indicate that C-4-S was arranged as a gradient, with strongest staining towards lumen and confined to more well-outlined structures, whereas DS showed a more widespread distribution. No staining for C-4-S was seen in the lumen of the notochord at any stages of development and in contrast to DS only weak staining could be detected in the chondrogenic zones.Fig. 5

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