Immunohistochemical study of cholecystokinin peptide in rat spinal motoneurons.
With the aid of indirect immunofluorescence histochemistry and sequence specific antibodies a possible localization of cholecystokinin (CCK) peptide in spinal motoneurons has been analyzed. To increase peptide levels, the sciatic nerve was ligated, and the area around the ligation was studied 24 hours later. For comparison, antisera raised against calcitonin gene-related peptide (CGRP) and substance P were employed. With CCK specific antisera (directed to the N-terminal portion of CCK-8 or the midportion of CCK-33) accumulation of peptide-like immunoreactivity (LI) was observed in large, dilated axonal swellings proximal to, but at some distance from, the ligature. Such accumulations were also observed with C-terminally directed CCK antiserum, but in addition numerous axons of smaller diameter extending up to the ligation contained this type of immunoreactivity. The latter antiserum is thought to cross-react with CGRP. In fact, this staining pattern was indistinguishable from the one seen after incubation with CGRP antiserum. In contrast substance P-LI could not be seen in the larger dilated axons but only in large numbers of thinner fibers close to the ligation. Double staining experiments revealed that the large dilations contained both CGRP- and CCK-specific LI. Distal to the ligation CGRP- and substance P- but no specific CCK-LI could be observed. The present findings support the view that CCK mRNA in spinal motoneurons is translated into CCK peptide, at least after axotomy, and that the peptide is transported into the motoneuron axon. However, compared to CGRP the CCK levels are presumably low, and the functional role of CCK peptide in motoneurons remains to be established.
Cortés
R
R
Department of Histology and Neurobiology, Karolinska Institutet, Stockholm, Sweden.
Aman
K
K
Arvidsson
U
U
Terenius
L
L
Frey
P
P
Rehfeld
J F
JF
Walsh
J H
JH
Hökfelt
T
T
D000818:Animals; D001369:Axons / Q000378:metabolism; D015740:Calcitonin Gene-Related Peptide / Q000276:immunology; D002766:Cholecystokinin / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D007106:Immune Sera / Q000276:immunology; D008026:Ligation; D008297:Male; D009046:Motor Neurons / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012584:Sciatic Nerve; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism*; D013373:Substance P / Q000276:immunology
D007106:Immune Sera; D012333:RNA, Messenger; D013373:Substance P; D002766:Cholecystokinin; D015740:Calcitonin Gene-Related Peptide