Abstract
Axonal transport of neprilysin, a putative neuropeptide degrading-enzyme, was examined in the proximal, middle, and distal segments of rat sciatic nerves using a double ligation technique. Neprilysin activity was significantly increased not only in the proximal segment but also in the distal segment 12–120 h after ligation, and the maximal neprilysin activity was found in the proximal and distal segments at 96 and 72 h, respectively. Western blot analysis of neprilysin showed that its immunoreactivities in the proximal and distal segments were 2.8- and 2.4-fold higher than that in the middle segment, indicating that neprilysin is transported by anterograde and retrograde axonal flow. These observations suggest that neprilysin may be involved in the metabolism of neuropeptides in nerve terminals or synaptic clefts.
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Abbreviations
- Aβ:
-
Amyloid-β
- CCK:
-
Cholecystokinin
- CPE:
-
Carboxypeptidase E
- DFP:
-
Diisopropylfluorophosphate
- N-DNS-AG:
-
N-Dansyl-d-alanylglycine
- N-DNS-AGNPG:
-
N-Dansyl-d-alanylglycyl-p-nitrophenylalanylglycine
- N-DNS-R:
-
N-Dansyl-arginine
- HRP:
-
Horseradish peroxidase
- IAA:
-
Iodoacetic acid
- NEP:
-
Neprilysin
- OD:
-
Optical density
- pAb:
-
Polyclonal antibody
- PAM:
-
Peptidylglycine α-amidating monooxygenase
- PBS:
-
Phosphate-buffered saline
- PCMS:
-
p-Chloromercuriphenylsulfonic acid
- SP:
-
Substance P
- TPBS:
-
Tween-PBS
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Ohkushi, G., Suzuki, N., Kobayashi, S. et al. Axonal Transport of Neprilysin in Rat Sciatic Nerves. J Mol Neurosci 53, 96–102 (2014). https://doi.org/10.1007/s12031-013-0202-6
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DOI: https://doi.org/10.1007/s12031-013-0202-6