Article
Full Access
Lumbar dorsal root ganglia of the cat: A quantitative study of peptide immunoreactivity and cell size
M. G. Garry,
K. E. Miller,
V. S. Seybold,
M. G. Garry
Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis, Minnesota 55455
Search for more papers by this authorK. E. Miller
Department of Neurological Surgery, University of Miami, School of Medicine, Miami, Florida 33176
Search for more papers by this authorV. S. Seybold
Graduate Program in Neuroscience, University of Minnesota, Minneapolis, Minnesota 55455
Search for more papers by this authorM. G. Garry,
K. E. Miller,
V. S. Seybold,
M. G. Garry
Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis, Minnesota 55455
Search for more papers by this authorK. E. Miller
Department of Neurological Surgery, University of Miami, School of Medicine, Miami, Florida 33176
Search for more papers by this authorV. S. Seybold
Graduate Program in Neuroscience, University of Minnesota, Minneapolis, Minnesota 55455
Search for more papers by this authorAbstract
The purpose of the present study was to quantify the extent to which several peptides and serotonin coexist with substance P or somatostatin in selected lumbar dorsal root ganglia of the cat. The technique for the simultaneous visualization of two antigens by immunofluorescence was used to investigate the coexistence of neuropeptides in the lumbar dorsal root ganglia of colchicine-treated cats. Perikarya immunoreactive for calcitonin gene-related peptide, galanin, leu-enkephalin, somatostatin, and substance P were visualized in both the lumbar 5 and 6 dorsal root ganglia. In contrast, no immunoreactivity was observed for adipokinetic hormone, bombesin, dynorphin A, met-enkephalin, oxytocin, tyrosine hydroxylase, thyrotropin-releasing hormone, vasopressin, vasoactive intestinal peptide, or serotonin in either ganglion examined. Substance P coexisted with calcitonin-gene-related peptide, somatostatin, and leu-enkephalin. Somatostatin was colocalized with calcitonin gene-related peptide, leu-enkephalin, and substance P but coexisted with galanin minimally. The cell area of immunoreactive perikarya was also examined. Data concerning the cross-sectional area of immunoreactive cells indicated that somatostatin-immunoreactive perikarya were generally the largest population observed (up to ∼6,000 μm2). Somatostatin and calcitonin gene-related peptide, as well as substance P and calcitonin gene-related peptide, coexisted in populations of cell bodies that had a smaller size (less than 2,000 μm2). These results suggest that certain peptides which coexist in the dorsal root ganglia may provide histochemical markers for functional groups of primary afferent neurons.
Literature Cited
- Appel, N. M., and R. P. Elde (1988) The intermediolateral cell column of the thoracic spinal cord is comprised of target-specific subnuclei: Evidence from retrograde transport studies and immunohistochemistry. J. Neurosci. 8: 1767–1775.
- Appel, N. M., M. W., Wessendorf, and R. P. Elde (1987) Thyrotropin-releasing hormone in spinal cord: Coexistence with serotonin and with substance P in fibers and terminals apposing identified preganglionic sympathetic neurons. Brain Res. 415: 137–143.
- Basbaum, A. I., L., Cruz, and E. Weber (1986) Immunoreactive dynorphin B in sacral primary afferent fibers of the cat. J. Neurosci. 6: 127–133.
- Basbaum, A. I., and E. J. Glazer (1983) Immunoreactive vasoactive intestinal polypeptide is concentrated in the sacral spinal cord: A possible marker for pelvic visceral afferent fibers. Somatosens. Res. 1: 69–82.
- Brown, P. B., and J. L. Fuchs (1975) Somatotopic representation of hindlimb skin in cat dorsal horn. J. Neurophysiol. 38: 1–9.
- Cameron, A. A., J. D., Leah, and P. J. Snow (1989) The coexistence of neuropeptides in feline sensory neurons. Neuroscience 27: 969–980.
- Cameron, A. A., J. D., Leah, and P. J. Snow (1986) Electrophysiological and morphological characteristics of feline dorsal root ganglion cells. Brain Res. 363: 1–11.
- Costa, M., J. B., Furness, and I. L. Gibbins (1986) Chemical coding of enteric neurons. In T. H. Ökfelt, K. Fuxe, and B. Pernow (eds): Progress in Brain Research, Vol. 68. Amsterdam: Elsevier Science Publishers, pp. 217–239.
- Cuello, A. C., G., Galfrè, and C. Milstein (1979) Detection of substance P in the central nervous system by a monoclonal antibody. Proc. Natl. Acad. Sci. USA 7: 3532–3536.
- Dalsgaard, C.-J., S. R., Vincent, T. Hökfelt, J. M. Lundberg, A. Dahlström, M. Schultzberg, G. J. Dockray, and A. C. Cuello (1982) Coexistence of chole cystokinin- and substance P-like peptides in neurons of the dorsal root ganglia of the rat. Neurosci. Lett. 33: 159–163.
- Elde, R., T., Hökfelt, O. Johansson, and L. Terenius (1976) Immunohistochemical studies using antibodies to leucine-enkephalin: Initial observations on the nervous system of the rat. Neuroscience 1: 349–351.
- Erichsen, J. T., A., Reiner, and H. J. Karten (1982) Co-occurrence of substance P-like and leu-enkephalin-like immunoreactivities in neurons and fibres of avian nervous system. Nature 295: 407–410.
- Franco-Cereceda, A., H., Henke, J. M. Lundberg, J. B. Petermann, T. Hökfelt, and J. A. Fischer (1987) Calcitonin gene-related peptide (CGRP) in capsaicin-sensitive substance P-immunoreactive sensory neurons in animals and man: Distribution and release by capsaicin. Peptides 8: 399–410.
- Fuxe, K., L. F., Agnati, T. McDonald, V. Locatelli, T. Hökfelt, C.-J. Dalsgaard, N. Battistini, N. Yanaihara, V. Mutt, and A. C. Cuello (1983) Immunohistochemical indications of gastrin releasing peptide-bombesin-like immunoreactivity in the nervous system of the rat Codistribution with substance P-like immunoreactive nerve terminal systems and coexistence with substance P-like immunoreactivity in dorsal root ganglion cell bodies. Neurosci. Lett. 37: 17–22.
- Gibbins, I. L., J. B., Furness, and M. Costa (1987) Pathway-specific patterns of the co-existence of substance P, calcitonin gene-related peptide, cholecystokinin and dynorphin in neurons of the dorsal root ganglia of the guinea pig. Cell Tissue Res. 248: 417–437.
- Gibbins, I. L., J. B., Furness, M. Costa, I. MacIntyre, C. J. Hillyard, and S. Girgis (1985) Co-localization of calcitonin gene-related peptide-like immunoreactivity with substance P in cutaneous, vascular and visceral sensory neurons of guinea pigs. Neurosci. Lett. 57: 125–130.
- Gibson, S. J., J. M., Polak, S. R. Bloom, I. M. Sabate, P. M. Mulderry, M. A. Ghatei, G. P. McGregor, J. F. B. Morrison, J. S. Kelley, R. M. Evans, and M. G. Rosenfeld (1984) Calcitonin gene-related peptide immunoreactivity in the spinal cord of man and of eight other species. J. Neurosci. 4: 3101–3111.
- Harper, A. A., and S. N. Lawson (1985) Conduction velocity is related to morphological cell type in rat dorsal root ganglion neurons. J. Physiol. (Lond.) 359: 31–46.
- Hökfelt, T., Z., Wiesenfeld-Hallin, M. Villar, and T. Melander (1987) Increase of galanin-like immunoreactivity in rat dorsal root ganglion cells after peripheral axotomy. Neurosci. Lett. 83: 217–220.
- Hökfelt, T., R., Elde, O. Johansson, R. Luft, G. Nilsson, and A. Arimura (1976) Immunohistochemical evidence for separate populations of somatostatin-containing and substance P-containing primary afferent neurons in the rat. Neuroscience 1: 131–136.
- Johnson, D. E., T. J., Kulik, J. E. Lock, R. P. Elde, and T. R. Thompson (1982) Bombesin, calcitonin, and serotonin immunoreactive pulmonary neuroendocrine cells in acute and chronic neonatal lung disease. Pediatr. Res. 16: 446–454.
- Ju, G., T., Hökfelt, E. Brodin, J. Fahrenkrug, J. A. Fischer, P. Frey, R. P. Elde, and J. C. Brown (1987) Primary sensory neurons of the rat showing calcitonin gene-related peptide immunoreactivity and their relation to substance P-, somatostatin-, galanin-, vasoactive intestinal polypeptide- and cholecystokinin-immunoreactive ganglion cells. Cell Tissue Res. 247: 417–431.
- Kawatani, M., I. P., Lowe, I. Nadelhaft, C. Morgan, and W. C. DeGroat (1983) Vasoactive intestinal peptide in visceral afferent pathways to the sacral spinal cord of the cat. Neurosci. Lett. 42: 311–316.
- Kayahara, T., T., Takimoto, and S. Sakashita (1981) Synaptic junctions in the cat spinal ganglion. Brain Res. 216: 277–290.
- Kuo, D. C., J. J., Oravitz, R. Eskay, and W. C. DeGroat (1984) Substance P in renal afferent perikarya identified by retrograde transport of fluorescent dye. Brain Res. 323: 168–171.
- Kuo, D. C., M., Kawatani, and W. C. DeGroat (1985) Vasoactive intestinal peptide identified in the thoracic dorsal root ganglia of the cat. Brain Res. 330: 178–182.
- Larsson, L. I. (1981) A novel immunocytochemical model system for specificity and sensitivity screening of antiserum against multiple antigens. J. Histochem. Cytochem. 29: 408–410.
- Leah, J. D., A. A., Cameron, W. L. Kelly, and P. J. Snow (1985) Coexistence of peptide immunoreactivity in sensory neurons of the cat. Neuroscience 16: 683–690.
- Lee, K. H., K., Chung, J. M. Chung, and R. E. Coggeshall (1986) Correlation of cell body size, axon size, and signal conduction velocity for individually labelled dorsal root ganglion cells in the cat. J. Comp. Neurol. 243: 335–346.
- Lee, Y., N., Hayashi, C. J. Hillyard, S. I. Girgis, I. MacIntyre, P. C. Emson, and M. Tohyama (1987) Calcitonin gene-related peptide-like immunoreactive sensory fibers form synaptic contact with sympathetic neurons in the rat celiac ganglion. Brain Res. 407: 149–151.
- Lee, Y., K., Takami, Y. Kawai, S. Girgis, C. J. Hillyard, I. MacIntyre, P. C. Emson, and M. Tohyama (1985) Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P. Neuroscience 15: 1227–1237.
- Lu, G. W., and V. Miletic (1988) EPSP-like activity in cat dorsal root ganglion neurons. Soc. Neurosci. Abstr. 14: 695.
- Lundberg, J. M., A., Franco-Cereceda, X. Hau, T. Hökfelt, and J. A. Fischer (1985) Co-existence of substance P and calciton in gene-related peptide-like immunoreactivities in sensory nerves in relation to cardiovascular and bronchoconstrictor effects of capsaicin. Eur. J. Pharmacol., 108: 315–319.
- Maley, B., and R. P. Elde (1982) Immunohistochemical localization of putative neurotransmitters within the feline nucleus tractus solitarii. Neuroscience, 7: 2469–2490.
- McCarthy, P. W., and S. N. Lawson (1987) Calcitonin gene-related peptide- like immunoreactivity (CGRP-LI) is found in primary afferent neurones with A- as well as those with C-fibres, in the rat. Soc. Neurosci. Abstr. 13: 296.
- McGregor, G. P., S. J., Gibson, I. M. Sabate, M. A. Blank, N. D. Christofides, P. D. Wall, J. M. Polak, and S. R. Bloom (1984) Effect of peripheral nerve section and nerve crush on spinal cord neuropeptides in the rat; increased VIP and PHI in the dorsal horn. Neuroscience 13: 207–216.
- Miletic, V., and H. Tan (1988) Iontophoretic application of calcitonin generelated peptide produces a slow and prolonged excitation of neurons in the cat lumbar dorsal horn. Brain Res. 446: 169–172.
- Miller, K. E., and V. S. Seybold (1987) Comparison of met-enkephalin, dynorphin A-, and neurotensin-immunoreactive neurons in the cat and rat spinal cords: I Lumbar cord. J. Comp. Neurol. 255: 293–304.
- Molander, C., and G. Grant (1985) Cutaneous projections from the rat hindlimb foot to the substantia gelatinosa of the spinal cord studied by transganglionic transport of WGA-HRP conjugate. J. Comp. Neurol. 237: 476–484.
- Morgan, C., I., Nadelhaft, and W. C. DeGroat (1981) The distribution of visceral primary afferents from the pelvic nerve to Lissauer's tract and the spinal gray matter and its relationship to the sacral parasympathetic nucleus. J. Comp. Neurol. 201: 415–440.
- Nadelhaft, I., and A. Booth (1984) The location and morphology of preganglionic neurons and the distribution of visceral afferents from the rat pelvic nerve: A horseradish peroxidase study. J. Comp. Neurol. 226: 238–245.
- O'Dononue, T. L., V. J., Massari, C. J. Pazoles, B. M. Chronwall, C. W. Schults, R. Quirion, T. N. Chase, and T. W. Moody (1984) A role for bombesin in sensory processing in the spinal cord. J. Neurosci. 4: 2956–2962.
- Panula, P., M., Hadjiconstantinou, H.-Y. T. Yang, and E. Costa (1983) Immunohistochemical localization of bombesin/gastrin-releasing peptide and substance P in primary sensory neurons. J. Neurosci. 3: 2021–2029.
- Platt, J. L., and A. F. Michael (1983) Retardation of fading and enhancement of intensity of immunofluorescence by p-phenylenediamine. J. Histochem. Cytochem. 31: 840–842.
- Price, J. (1985) An immunohistochemical and quantitative examination of dorsal root ganglion neuronal subpopulations. J. Neurosci. 5: 2015–2059.
- Price, J., and A. W. Mudge (1983) A subpopulation of rat dorsal root ganglion neurones is catecholaminergic. Nature 301: 241–243.
-
Randic, M.,
P. D., Ryu, and
L. Urban
(1986)
Effects of polyclonal and monoclonal antibodies to substance P on slow excitatory transmission in rat spinal dorsal horn.
Brain Res.
383: 15–27.
10.1016/0006-8993(86)90003-X Google Scholar
- Randic, M., G., Gerber, P. D. Ryu, and I. Kangrga (1987) Inhibitory actions of galanin and somatostatin 28 on rat spinal dorsal horn neurons. Soc. Neurosci. Abstr. 13: 1308.
- Randic, M., and V. Miletic (1977) Effects of substance P in cat dorsal horn neurones activated by noxious mechanical stimuli. Brain Res. 128: 164–169.
- Ryu, P. D., G., Gerber, and M. Randic (1987) Multiple actions of calcitonin gene-related peptide on rat spinal dorsal horn neurons. Soc. Neurosci. Abstr. 13: 1312.
- Sasek, C. A., V. S., Seybold, and R. P. Elde (1984) The immunohistochemical localization of nine peptides in the sacral parasympathetic nucleus and the dorsal gray commissure in rat spinal cord. Neuroscience 12: 855–873.
- Sasek, C. A., and R. P. Elde (1986) Coexistence of enkephalin and dynorphin immunoreactivities in neurons in the dorsal gray commissure of the sixth lumbar and first sacral spinal cord segments in rat. Brain Res. 381: 8–14.
- Sasek, C. A., P. A., Schueler, W. S. Herman, and R. P. Elde (1985) An antiserum to locust adipokinetic hormone reveals a novel peptiderigic system in the rat central nervous system. Brain Res. 343: 172–175.
- Schalmbruch, H. (1986) Fiber composition of the rat sciatic nerve. Anat. Rec. 251: 71–81.
- Schueler, P. A., and R. P. Elde (1988) Differential expression of ubiquitin within the rat brain: Discretely localized increases following salt loading. J. Chem. Neuroanat. 1: 165–173.
- Skofitsch, G., and D. M. Jacobowitz (1985) Calcitonin gene-related peptide coexists with substance P in capsaicin sensitive neurons and sensory ganglia of the rat. Peptides 6: 747–754.
- Stefanini, M., C. deMartino, and L. Zamboni (1967) Fixation of ejaculated spermatozoa for electron microscopy. Nature 216: 173–174.
- Sternberger, L. A., P. H. Hardy, Jr., J. J. Cuculis, and H. G. Meyer (1970) The unlabelled antibody-enzyme method of immunohistochemistry Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in the identification of spirochetes. J. Histochem. Cytochem. 18: 315–333.
- Tuchscherer, M. M., C., Knox, and V. S. Seybold (1987) Substance P and cholecystokinin-like immunoreactive varicosities in somatosensory and autonomic regions of the rat spinal cord: A quantitative study of coexistence. J. Neurosci. 7: 3984–3995.
- Tuchscherer, M. M., and V. S. Seybold (1985) Immunohistochemical studies of substance P, cholecystokinin-octapeptide and somatostatin in dorsal root ganglia of the rat. Neuroscience 14: 593–605.
- Tuchscherer, M. M., and V. S. Seybold (1989) A quantitative study of the coexistence of peptides in varicosities within the superifical laminae of the dorsal horn of the rat spinal cord. J. Neurosci. 9: 195–205.
- Wessendorf, M. W., and R. P. Elde (1985) Characterization of an immunofluorescence technique for the demonstration of coexisting neurotransmitters within nerve fibers and terminals. J. Histochem. Cytochem. 33: 984–994.
- Wiesenfeld-Hallin, Z., T., Hökfelt, J. M. Lundberg, W. G. Forssmann, M. Reinecke, F. A. Tschopp, and J. A. Fischer (1984) Immunoreactive calcitonin gene-related peptide and substance P coexist in sensory neurons to the spinal cord and interact in spinal behavioral responses of the rat. Neurosci. Lett. 52: 199–204.
- Willard, A. L., and R. Nishi (1987) Neuropeptides mark functionally distinguishable cholinergic enteric neurons. Brain Res. 422: 163–167.
- Yaksh, T. L., and T. A. Rudy (1976) Chronic catheterization of the spinal subarachnoid space. Physiol. Behav. 17: 1031–1036.
Citing Literature
