|
|
|
|
 |
|||
|
|||
Ultrastructural localization of neuropeptides and GABA in rat dorsal horn: a comparison of different immunogold labeling techniques
A Merighi, JM Polak, G Fumagalli and DT Theodosis
Department of Histochemistry, Royal Postgraduate Medical School, London, UK.
Several immunogold techniques were used to determine the ultrastructural localization of calcitonin gene-related peptide (CGRP), tachykinin, somatostatin, and gamma-amino-butyric acid (GABA) immunoreactivity in the dorsal horn of rat spinal cord. The immunocytochemical reactions were carried out directly on ultrathin sections from non-osmicated frozen tissue, non-osmicated low temperature-embedded (Lowicryl K4M) tissue, and osmicated epoxy- embedded material. Preservation of ultrastructural morphology and immuno-labeling efficiency were compared. Morphology of subcellular organelles was relatively good in ultra-thin frozen sections, which showed the highest immunoreactivity. However, only very small samples of tissue could be examined. Although there was relatively good immunolabeling in the Lowicryl K4M-embedded tissue, the ultrastructure of the neuropil, and particularly that of synapses, was poorly maintained. In contrast, the osmicated epoxy-embedded material offered optimal morphological preservation together with accurate subcellular localization of all antigens under study. The latter approach thus enabled clear visualization of CGRP, tachykinin, and somatostatin immunoreactivity restricted to large dense-cored vesicles (90-150 nm diameter) in many axonal and synaptic profiles in the superficial layers of the dorsal horn. CGRP- and tachykinin-positive profiles were also present in the tract of Lissauer. GABA immunoreactivity was present mainly in axons and terminals, and less frequently in somatic and dendritic profiles. In terminals, which often formed symmetrical synapses on immunonegative dendritic profiles, it was associated with small (30-60 nm diameter) clear vesicles and mitochondria. Double immunolabeling was possible on all preparations, but the osmicated, epoxy-embedded material clearly showed co-localization of peptides, especially of CGRP and tachykinins, within the same dense-cored vesicles in axonal fibers and/or terminals. On the other hand, peptide and GABA immunoreactivity were consistently seen in different nerve profiles. In a few cases, GABAnergic terminals were seen to synapse on tachykinin-positive fibers.
Volume 37,
Issue 4,
pp. 529-540,
04/01/1989
Copyright © 1989 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. V.T. Lobo, F. J. M. Alonso, M. I. Arenas, E. Caso, B. Fraile, and R. M. del Rio Ultrastructural Staining with Sodium Metaperiodate and Sodium Borohydride J. Histochem. Cytochem., January 1, 2002; 50(1): 11 - 20. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. S. Vilim, E. C. Cropper, D. A. Price, I. Kupfermann, and K. R. Weiss Peptide Cotransmitter Release from Motorneuron B16 in Aplysia californica: Costorage, Corelease, and Functional Implications J. Neurosci., March 1, 2000; 20(5): 2036 - 2042. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. R. Koerber, K. Mirnics, A. M. Kavookjian, and A. R. Light Ultrastructural Analysis of Ectopic Synaptic Boutons Arising From Peripherally Regenerated Primary Afferent Fibers J Neurophysiol, April 1, 1999; 81(4): 1636 - 1644. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Aimar, L. Pasti, G. Carmignoto, and A. Merighi Nitric Oxide-Producing Islet Cells Modulate the Release of Sensory Neuropeptides in the Rat Substantia Gelatinosa J. Neurosci., December 15, 1998; 18(24): 10375 - 10388. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. S. Vilim, D. A. Price, W. Lesser, I. Kupfermann, and K. R. Weiss Costorage and Corelease of Modulatory Peptide Cotransmitters with Partially Antagonistic Actions on the Accessory Radula Closer Muscle of Aplysia californica J. Neurosci., December 15, 1996; 16(24): 8092 - 8104. [Abstract] [Full Text] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |