Association of serotonin-like immunoreactive axons with nociceptive projection neurons in the caudal spinal trigeminal nucleus of the rat.
Serotoninergic projections to the spinal dorsal horn are implicated in the modulation of nociceptive transmission. However, morphological evidence indicating that serotoninergic projection fibers make synapses on nociceptive neurons in the medullary dorsal horn is still meager. Thus, we examined whether axonal varicosities with serotonin (5-HT)-like immunoreactivity (5-HT-LI) might make synapses on nociceptive projection neurons in the caudal spinal trigeminal nucleus (Vc) of the rat. Projection neurons were retrogradely labeled with tetramethylrhodamine-dextran amine (TMR-DA) or wheat germ agglutinin-horseradish peroxidase (WGA-HRP) that was injected into the parabrachial or thalamic region. Vc neurons in which c-fos protein-like immunoreactivity (Fos-LI) was induced by subcutaneous injection of formalin into the lip were considered nociceptive. Vc neurons in direct contact with axonal varicosities that bind isolectin I-B4 were also considered nociceptive. Triple labeling for 5-HT, TMR-DA, and Fos as well as that for 5-HT, TMR-DA, and I-B4 were done by using the immunofluorescence and fluorescence histochemical techniques. Confocal laser-scanning microscopy revealed that axonal varicosities with 5-HT-LI were in close apposition to TMR-DA-labeled neurons showing Fos-LI in lamina I and the outer part of lamina II (lamina IIo), and that both axonal varicosities with 5-HT-LI and those binding I-B4 were in close apposition to single neuronal profiles labeled with TMR-DA. The presumed nociceptive neuronal profiles in close apposition to axon terminals with 5-HT-LI were mainly those of laminae I and II neurons as well as dendrites of lamina III neurons. Electron microscopy confirmed that axon terminals with 5-HT-LI and those with I-B4 binding activity in laminae I and II made synapses on somatic and dendritic profiles that were labeled with WGA-HRP. The results indicate that serotoninergic neurons project directly on nociceptive projection neurons in the Vc.
Li
J L
JL
Department of Morphological Brain Science, Faculty of Medicine, Kyoto University, Japan.
Kaneko
T
T
Nomura
S
S
Li
Y Q
YQ
Mizuno
N
N
D000818:Animals; D000918:Antibody Specificity; D001369:Axons / Q000737:chemistry / Q000276:immunology* / Q000648:ultrastructure; D004396:Coloring Agents; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde; D037102:Lectins; D008297:Male; D008854:Microscopy, Electron; D009474:Neurons / Q000737:chemistry / Q000648:ultrastructure; D009619:Nociceptors / Q000737:chemistry / Q000502:physiology*; D010146:Pain / Q000503:physiopathology; D016760:Proto-Oncogene Proteins c-fos / Q000032:analysis / Q000276:immunology; D051381:Rats; D017208:Rats, Wistar / Q000502:physiology*; D012235:Rhodamines; D012701:Serotonin / Q000032:analysis / Q000276:immunology*; D014279:Trigeminal Nucleus, Spinal / Q000166:cytology*; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate
D004396:Coloring Agents; D037102:Lectins; D016760:Proto-Oncogene Proteins c-fos; D012235:Rhodamines; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate; D005557:Formaldehyde; D012701:Serotonin; C005358:tetramethylrhodamine