Nerve injury induces a rapid efflux of nitric oxide (NO) detected with a novel NO microsensor.
An early step in repair of the leech CNS is the appearance of endothelial nitric oxide synthase (eNOS) immunoreactivity and NOS activity, but coincident generation of NO at the lesion after injury has not been shown. This is important because NO can regulate microglial cell motility and axon growth. Indirect measurement of NO with the standard citrulline assay demonstrated that NO was generated within 30 min after nerve cord injury. A polarographic NO-selective self-referencing microelectrode that measures NO flux noninvasively was developed to obtain higher spatial and temporal resolution. With this probe, it was possible to demonstrate that immediately after the leech CNS was injured, NO left the lesion with a mean peak efflux of 803 +/- 99 fmol NO cm(-2) sec(-1). NO efflux exponentially declined to a constant value, as described through the equation f(t) = y(o) + ae(-t/tau), with tau = 117 +/- 30 sec. The constant y(o) = 15.8 +/- 4.5 fmol cm(-2) represents a sustained efflux of NO. Approximately 200 pmol NO cm(-2) is produced at the lesion (n = 8). Thus, injury activates eNOS already present in the CNS and precedes the accumulation of microglia at the lesion, consistent with the hypothesis that NO acts to stop the migrating microglia at the lesion site.
Kumar
S M
SM
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
Porterfield
D M
DM
Muller
K J
KJ
Smith
P J
PJ
Sahley
C L
CL
D000818:Animals; D002490:Central Nervous System / Q000737:chemistry / Q000502:physiology*; D002956:Citrulline / Q000378:metabolism; D007865:Leeches; D008839:Microelectrodes; D017628:Microglia / Q000166:cytology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009409:Nerve Crush; D009416:Nerve Regeneration / Q000502:physiology*; D009474:Neurons / Q000378:metabolism*; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D011048:Polarography / Q000295:instrumentation
D002956:Citrulline; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D019331:NG-Nitroarginine Methyl Ester