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Nidogen plays a role in the regenerative axon growth of adult sensory neurons through Schwann cells.

Lee HK, Seo IA, Suh DJ, Park HT - J. Korean Med. Sci. (2009)

Bottom Line: Since Schwann cells play a critical role in peripheral nerve regeneration, nidogen may play a role in it via regulation of Schwann cells.Here, we demonstrate direct evidence that nidogen induces elongation of regenerative axon growth of adult sensory neurons, and that the effect is Schwann cell dependent.Taken together, it is likely that nidogen is required for proper regeneration of peripheral nerves after injury.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, College of Medicine, Dong-A University, Busan, Korea.

ABSTRACT
We previously reported that nidogen is an extracellular matrix protein regulating Schwann cell proliferation and migration. Since Schwann cells play a critical role in peripheral nerve regeneration, nidogen may play a role in it via regulation of Schwann cells. Here, we demonstrate direct evidence that nidogen induces elongation of regenerative axon growth of adult sensory neurons, and that the effect is Schwann cell dependent. Continuous infusion of recombinant ectodomain of tumor endothelial marker 7, which specifically blocks nidogen function in Schwann cells, suppressed regenerative neurite growth in a sciatic nerve axotomy model. Taken together, it is likely that nidogen is required for proper regeneration of peripheral nerves after injury.

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Related in: MedlinePlus

The effects of Fc and nidogen-Fc on the neurite outgrowth of adult DRG explants. (A) DRG explants from adult rats were cultured in collagen matrix, and the culture medium was supplemented with Fc (20 µg/mL) or nidogen-Fc (20 µg/mL) for 40 hr. Explants are immunostained with anti-Tuj1 to label axons. Scale bar; 100 µm. (B) Quantification of mean length of neurites. Values shown are mean lengths±standard deviation.
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Figure 1: The effects of Fc and nidogen-Fc on the neurite outgrowth of adult DRG explants. (A) DRG explants from adult rats were cultured in collagen matrix, and the culture medium was supplemented with Fc (20 µg/mL) or nidogen-Fc (20 µg/mL) for 40 hr. Explants are immunostained with anti-Tuj1 to label axons. Scale bar; 100 µm. (B) Quantification of mean length of neurites. Values shown are mean lengths±standard deviation.

Mentions: For explant cultures in collagen gel, collagens were prepared as previously described (20). Thoracic and lumbar DRGs were removed from 40-50 days old male Sprague-Dawley rats (Samtako, Osan, Korea). Our college ethics committee approved that our protocol fulfilled the guide of animal experiments established by The Korean Academy of Medical Sciences (DIACUC-07-11). Following the dissection of a DRG into 2-3 explants under a stereomicroscope, the explants were implanted into collagen gel (Fig. 1) or placed on a 6 well-Millicell culture membrane (Millipore, Billerica, MA, U.S.A., 10-12 explants/membrane, Fig. 2) as described before (19). Then, Fc or nidogen-Fc (21) containing Dulbecco's modified Eagle's medium with 5% fetal bovine serum was added. After 2 or 3 days of culture, the explants were fixed with 4% paraformaldehyde and immunostained with an anti-Tuj1 antibody (1/2,000, Covance, Princeton, NJ, U.S.A.). The neurite outgrowth was examined under a Laser confocal microscope (Carl Zeiss, Hamburg, Germany). Quantification of the axonal elongation from the DRG explants (20 explants in each of three independent experiments) in collagen gel (Fig. 1) was carried out with LSM-510 software (Carl Zeiss) after photographing the explants under the Laser confocal microscope. Line draw tools were employed to automatically measure the length of the TuJ1-positive neurites (180-200 neurites per group), and the difference of mean length between groups was analyzed using the Student's t test. Parameters with values P<0.05 were considered to be different significantly.


Nidogen plays a role in the regenerative axon growth of adult sensory neurons through Schwann cells.

Lee HK, Seo IA, Suh DJ, Park HT - J. Korean Med. Sci. (2009)

The effects of Fc and nidogen-Fc on the neurite outgrowth of adult DRG explants. (A) DRG explants from adult rats were cultured in collagen matrix, and the culture medium was supplemented with Fc (20 µg/mL) or nidogen-Fc (20 µg/mL) for 40 hr. Explants are immunostained with anti-Tuj1 to label axons. Scale bar; 100 µm. (B) Quantification of mean length of neurites. Values shown are mean lengths±standard deviation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2719197&req=5

Figure 1: The effects of Fc and nidogen-Fc on the neurite outgrowth of adult DRG explants. (A) DRG explants from adult rats were cultured in collagen matrix, and the culture medium was supplemented with Fc (20 µg/mL) or nidogen-Fc (20 µg/mL) for 40 hr. Explants are immunostained with anti-Tuj1 to label axons. Scale bar; 100 µm. (B) Quantification of mean length of neurites. Values shown are mean lengths±standard deviation.
Mentions: For explant cultures in collagen gel, collagens were prepared as previously described (20). Thoracic and lumbar DRGs were removed from 40-50 days old male Sprague-Dawley rats (Samtako, Osan, Korea). Our college ethics committee approved that our protocol fulfilled the guide of animal experiments established by The Korean Academy of Medical Sciences (DIACUC-07-11). Following the dissection of a DRG into 2-3 explants under a stereomicroscope, the explants were implanted into collagen gel (Fig. 1) or placed on a 6 well-Millicell culture membrane (Millipore, Billerica, MA, U.S.A., 10-12 explants/membrane, Fig. 2) as described before (19). Then, Fc or nidogen-Fc (21) containing Dulbecco's modified Eagle's medium with 5% fetal bovine serum was added. After 2 or 3 days of culture, the explants were fixed with 4% paraformaldehyde and immunostained with an anti-Tuj1 antibody (1/2,000, Covance, Princeton, NJ, U.S.A.). The neurite outgrowth was examined under a Laser confocal microscope (Carl Zeiss, Hamburg, Germany). Quantification of the axonal elongation from the DRG explants (20 explants in each of three independent experiments) in collagen gel (Fig. 1) was carried out with LSM-510 software (Carl Zeiss) after photographing the explants under the Laser confocal microscope. Line draw tools were employed to automatically measure the length of the TuJ1-positive neurites (180-200 neurites per group), and the difference of mean length between groups was analyzed using the Student's t test. Parameters with values P<0.05 were considered to be different significantly.

Bottom Line: Since Schwann cells play a critical role in peripheral nerve regeneration, nidogen may play a role in it via regulation of Schwann cells.Here, we demonstrate direct evidence that nidogen induces elongation of regenerative axon growth of adult sensory neurons, and that the effect is Schwann cell dependent.Taken together, it is likely that nidogen is required for proper regeneration of peripheral nerves after injury.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, College of Medicine, Dong-A University, Busan, Korea.

ABSTRACT
We previously reported that nidogen is an extracellular matrix protein regulating Schwann cell proliferation and migration. Since Schwann cells play a critical role in peripheral nerve regeneration, nidogen may play a role in it via regulation of Schwann cells. Here, we demonstrate direct evidence that nidogen induces elongation of regenerative axon growth of adult sensory neurons, and that the effect is Schwann cell dependent. Continuous infusion of recombinant ectodomain of tumor endothelial marker 7, which specifically blocks nidogen function in Schwann cells, suppressed regenerative neurite growth in a sciatic nerve axotomy model. Taken together, it is likely that nidogen is required for proper regeneration of peripheral nerves after injury.

Show MeSH
Related in: MedlinePlus