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Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model.

Leichsenring A, Bäcker I, Wendt W, Andriske M, Schmitz B, Stichel CC, Lübbert H - BMC Neurosci (2008)

Bottom Line: While for a long time, proteases were mainly considered as protein degrading enzymes, they are now receiving growing interest as signalling molecules in the pain pathology.Moreover, we succeeded in measuring the activity of CATX, which was substantially increased after L5T.The differential expression of these proteins exhibited the same spatial distribution and temporal progression in the spinal cord, progressing up to the medulla oblongata in the late phase of chronic pain.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Animal Physiology, Ruhr-University of Bochum, Bochum, Germany. anna.leichsenring@rub.de

ABSTRACT

Background: Ample evidence suggests a substantial contribution of cellular and molecular changes in the spinal cord to the induction and persistence of chronic neuropathic pain conditions. While for a long time, proteases were mainly considered as protein degrading enzymes, they are now receiving growing interest as signalling molecules in the pain pathology. In the present study we focused on two cathepsins, CATS and CATX, and studied their spatiotemporal expression and activity during the development and progression of neuropathic pain in the CNS of the rat 5th lumbar spinal nerve transection model (L5T).

Results: Immediately after the lesion, both cathepsins, CATS and CATX, were upregulated in the spinal cord. Moreover, we succeeded in measuring the activity of CATX, which was substantially increased after L5T. The differential expression of these proteins exhibited the same spatial distribution and temporal progression in the spinal cord, progressing up to the medulla oblongata in the late phase of chronic pain. The cellular distribution of CATS and CATX was, however, considerably different.

Conclusion: The cellular distribution and the spatio-temporal development of the altered expression of CATS and CATX suggest that these proteins are important players in the spinal mechanisms involved in chronic pain induction and maintenance.

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Related in: MedlinePlus

Upregulation of cathepsin protein levels and activities after L5T. A: Western blot analysis of CATX and CATS proform expression in the spinal cord of L5T (n = 5) and sham operated animals (n = 5) at 8 d after injury. Expression levels were normalized relative to the corresponding α-tubulin band. At this time point the L5 transection induced an upregulation of both proteins in all SC segments. The expression level of CATX was substantially higher than that of CATS. Data are means ± SD. B: CATX activities in the lumbar SC 8 d after transection in L5T, sham (n = 7, for each group) and naive animals (n = 4). Each symbol represents the value of a single animal, the bar indicates the mean for the group. CATX activity was significantly higher in L5T than in sham or naive SC. *** p ≤ 0.001. L, lumbar; C, cervical; T, thoracic.
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Figure 4: Upregulation of cathepsin protein levels and activities after L5T. A: Western blot analysis of CATX and CATS proform expression in the spinal cord of L5T (n = 5) and sham operated animals (n = 5) at 8 d after injury. Expression levels were normalized relative to the corresponding α-tubulin band. At this time point the L5 transection induced an upregulation of both proteins in all SC segments. The expression level of CATX was substantially higher than that of CATS. Data are means ± SD. B: CATX activities in the lumbar SC 8 d after transection in L5T, sham (n = 7, for each group) and naive animals (n = 4). Each symbol represents the value of a single animal, the bar indicates the mean for the group. CATX activity was significantly higher in L5T than in sham or naive SC. *** p ≤ 0.001. L, lumbar; C, cervical; T, thoracic.

Mentions: Western blot analysis (n = 5 per group) revealed the proforms of CATS (37 kD) and CATX (34 kD), as the most prominent bands, while the prepro- and mature forms were below the detection level. The proforms of both enzymes were detected in all segments analyzed (L, lumbar; T, thoracic; C, cervical) of the adult rat spinal cord (Fig. 4A). The L5 nerve transsection produced an upregulation in all SC segments for CATS as well as CATX (Fig. 4A). The strongest increase in protein content was found in the T segment for both enzymes (CATS 63.6%; CATX 87.4%), while the increase in the L and C-segments was substantial (34–61.5%) but lower than in the T segment (Fig. 4A). Moreover, our Western blot analysis showed that in all SC segments the level of CATX is more than twice as high as the level of CATS. These results were confirmed in a second experiment with 4 animals per group (see Additional files 1 and 2).


Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model.

Leichsenring A, Bäcker I, Wendt W, Andriske M, Schmitz B, Stichel CC, Lübbert H - BMC Neurosci (2008)

Upregulation of cathepsin protein levels and activities after L5T. A: Western blot analysis of CATX and CATS proform expression in the spinal cord of L5T (n = 5) and sham operated animals (n = 5) at 8 d after injury. Expression levels were normalized relative to the corresponding α-tubulin band. At this time point the L5 transection induced an upregulation of both proteins in all SC segments. The expression level of CATX was substantially higher than that of CATS. Data are means ± SD. B: CATX activities in the lumbar SC 8 d after transection in L5T, sham (n = 7, for each group) and naive animals (n = 4). Each symbol represents the value of a single animal, the bar indicates the mean for the group. CATX activity was significantly higher in L5T than in sham or naive SC. *** p ≤ 0.001. L, lumbar; C, cervical; T, thoracic.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2527007&req=5

Figure 4: Upregulation of cathepsin protein levels and activities after L5T. A: Western blot analysis of CATX and CATS proform expression in the spinal cord of L5T (n = 5) and sham operated animals (n = 5) at 8 d after injury. Expression levels were normalized relative to the corresponding α-tubulin band. At this time point the L5 transection induced an upregulation of both proteins in all SC segments. The expression level of CATX was substantially higher than that of CATS. Data are means ± SD. B: CATX activities in the lumbar SC 8 d after transection in L5T, sham (n = 7, for each group) and naive animals (n = 4). Each symbol represents the value of a single animal, the bar indicates the mean for the group. CATX activity was significantly higher in L5T than in sham or naive SC. *** p ≤ 0.001. L, lumbar; C, cervical; T, thoracic.
Mentions: Western blot analysis (n = 5 per group) revealed the proforms of CATS (37 kD) and CATX (34 kD), as the most prominent bands, while the prepro- and mature forms were below the detection level. The proforms of both enzymes were detected in all segments analyzed (L, lumbar; T, thoracic; C, cervical) of the adult rat spinal cord (Fig. 4A). The L5 nerve transsection produced an upregulation in all SC segments for CATS as well as CATX (Fig. 4A). The strongest increase in protein content was found in the T segment for both enzymes (CATS 63.6%; CATX 87.4%), while the increase in the L and C-segments was substantial (34–61.5%) but lower than in the T segment (Fig. 4A). Moreover, our Western blot analysis showed that in all SC segments the level of CATX is more than twice as high as the level of CATS. These results were confirmed in a second experiment with 4 animals per group (see Additional files 1 and 2).

Bottom Line: While for a long time, proteases were mainly considered as protein degrading enzymes, they are now receiving growing interest as signalling molecules in the pain pathology.Moreover, we succeeded in measuring the activity of CATX, which was substantially increased after L5T.The differential expression of these proteins exhibited the same spatial distribution and temporal progression in the spinal cord, progressing up to the medulla oblongata in the late phase of chronic pain.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Animal Physiology, Ruhr-University of Bochum, Bochum, Germany. anna.leichsenring@rub.de

ABSTRACT

Background: Ample evidence suggests a substantial contribution of cellular and molecular changes in the spinal cord to the induction and persistence of chronic neuropathic pain conditions. While for a long time, proteases were mainly considered as protein degrading enzymes, they are now receiving growing interest as signalling molecules in the pain pathology. In the present study we focused on two cathepsins, CATS and CATX, and studied their spatiotemporal expression and activity during the development and progression of neuropathic pain in the CNS of the rat 5th lumbar spinal nerve transection model (L5T).

Results: Immediately after the lesion, both cathepsins, CATS and CATX, were upregulated in the spinal cord. Moreover, we succeeded in measuring the activity of CATX, which was substantially increased after L5T. The differential expression of these proteins exhibited the same spatial distribution and temporal progression in the spinal cord, progressing up to the medulla oblongata in the late phase of chronic pain. The cellular distribution of CATS and CATX was, however, considerably different.

Conclusion: The cellular distribution and the spatio-temporal development of the altered expression of CATS and CATX suggest that these proteins are important players in the spinal mechanisms involved in chronic pain induction and maintenance.

Show MeSH
Related in: MedlinePlus