Summary
Morphological changes in the motor and sensory neurons in the lumbar spinal cord and the dorsal root ganglia were investigated at different survival times following the injection of the B subunit of cholera toxin (CTB) into the medial gastrocnemius muscle. Unconjugated CTB, visualized immunohistochemically, was found to be retrogradely transported through ventral and dorsal roots to motor neurons in the anterior horn, each lamina in the posterior horn, and ganglion cells in the dorsal root ganglia at L3–L6. The largest numbers of labeled motor neurons and ganglion cells were observed 72 h after the injection of CTB. Thereafter, labeled ganglion cells were significantly decreased in number, whereas the amount of labeled motor neurons showed a slight reduction. Motor neurons had extensive dendritic trees filled with CTB, reaching lamina VII and even the pia mater of the lateral funiculus. Labeling was also seen in the posterior horn, but the central and medial parts of laminae II and III had the most extensively labeled varicose fibers, the origin of which was the dorsal root ganglion cells. The results indicate that CTB is taken up by nerve terminals and can serve as a sensitive retrogradely transported marker for identifying neurons that innervate a specific muscle.
Similar content being viewed by others
References
Brunner R, Zimmermann P, Klußmann FW (1980) Localization and neurophysiological properties of motoneurons of the m. triceps surae of the rat after retrograde labelling with Evans blue. Cell Tissue Res 212:73–81
Brushart TM, Mesulam M-M (1980) Transganglionic demonstration of central sensory projections from skin and muscle with HRP-lectin conjugates. Neurosci Lett 17:1–6
Burke RE, Strick PL, Kanda K, Kim CC, Walmsley B (1977) Anatomy of medial gastrocnemius and soleus motor nuclei in cat spinal cord. J Neurophysiol 40:667–680
Carpenter MB, Stein BM, Shriver JE (1968) Central projections of spinal dorsal roots in the monkey II. Lower thoracic, lumbosacral and coccygeal dorsal roots. Am J Anat 123:75–118
Craig AD, Mense S (1983) The distribution of afferent fibers from the gastrocnemius-soleus muscle in the dorsal horn of the cat, as revealed by the transport of horseradish peroxidase. Neurosci Lett 41:233–238
Eccles JC, Fatt P, Landgren S, Winsbuly GJ (1954) Spinal cord potentials generated by volley in the large muscle afferents. J Physiol 125:590–606
Egger MD, Egger LD (1982) Quantitative morphological analysis of spinal motoneurons. Brain Res 253:19–30
Eringer C, Ericksen JT (1986) Transsynaptic retrograde transport of fragment C of tetanus toxin demonstrated by immunohistochemical localization. Brain Res 380:383–388
Grant G, Arvidson J, Rovertson B, Ygge J (1979) Transganglionic transport of horseradish peroxidase in primary sensory neurons. Neurosci Lett 12:23–28
Ha H, Kao T, Tan EC (1980) Muscle sensory neurons in the spinal ganglia in the rat determined by the retrograde transport of horseradish peroxidase. Exp Neurol 70:438–445
Hamano K, Mannen H, Ishizuka N (1978) Reconstruction of trajectory of primary afferent collaterals in the dorsal horn of the cat spinal cord, using Golgi-stained serial sections. J Comp Neurol 181:1–6
Holliday M (1980) Organization of motor pools in the chick lumber lateral motor column. J Comp Neurol 194:143–170
Homma S, Kohno K, Okado N (1989) Localization of serotonin positive fibers in specific motor neuron pools of the lumbosacral spinal cord of the chicken. Biomed Res 10 [Suppl 3]:233–239
Kristensson K (1970) Transport of fluorescent protein tracer in peripheral nerves. Acta Neuropathol (Berl) 16:293–300
Kristensson K, Olsson Y (1971) Retrograde axonal transport of protein. Brain Res 29:263–265
LaMotte C (1977) Distribution of the tract of Lissauer and the dorsal root fibers in the primate spinal cord. J Comp Neurol 172:529–562
Lasek R, Joseph BS, Whitlock DG (1969) Evaluation of a radioautographic neuroanatomical tracing method. Brain Res 8:319–336
LaVail JH, LaVail MM (1972) Retrograde axonal transport in the central nervous system. Science 176:1416–1417
Light AR, Perl ER (1979) Reexamination of the dorsal root projection to the spinal dorsal horn including observations on the differential termination of coarse and fine fibers. J Comp Neurol 186:117–132
Meckler RL, Baron R, McLachlan EM (1990) Selective uptake of C-fragment of tetanus toxin by sympathetic preganglionic nerve terminals. Neuroscience 36:823–829
Melian CR, Grant G (1990) Distribution of lumbar dorsal root fibers in the lower thoracic and lumbosacral spinal cord of the rat studied with choleragenoid horseradish peroxidase conjugate. J Comp Neurol 299:470–481
Mesulam M-M, Brushart TM (1979) Transganglionic and anterograde transport of horseradish peroxidase across dorsal root ganglia: a tetramethylbenzidine method for tracing central sensory connections of muscles and peripheral nerves. Neuroscience 4:1107–1117
Molander C, Grant G (1986) Laminar distribution and somatotopic organization of primary afferent fibers from hindlimb nerves in the dorsal horn. A study by transganglionic transport of horseradish peroxidase in the rat. Neuroscience 19:297–312
Molander C, Xu Q, Grant G (1984) The cytoarchitectonic organization of the spinal cord of the rat. I. The lower thoracic and lumbosacral cord. J Comp Neurol 230:133–141
Mong FSF (1990) Dendritic distributions of motor neurons innervating fast and slow muscles of the hind limb of rats. J Hirnforsch 31:259–267
Nishino H, Ono T, Sakai K (1977) Transposition of horseradish peroxidase in rat sciatic nerve. J Physiol Soc Jpn 39:331
Okado N, Homma S, Ishihara R, Sako H, Kohno K (1988) Differential innervation of specific motor neuron pools by serotonergic fibers in the chick spinal cord. Neurosci Lett 94:29–32
Oldfors A (1986) Cholera toxin B-subunit incorporation into synaptic vesicles of the neuromuscular junction of the rat. Experientia 42:415–417
Ralston HJ III (1968) Dorsal root projections to dorsal horn neurons in the cat spinal cord. J Comp Neurol 132:303–330
Ramón y Cajal S (1909) Histologie du système nerveux de l'homme et des vertébrés. Instituto Ramón y Cajal, Madrid
Rexed B (1952) The cytoarchitectonic organization of the spinal cord in the cat. J Comp Neurol 96:415–496
Rexed B (1954) A cytoarchitectonic atlas of the spinal cord in the cat. J Comp Neurol 100:297–380
Rodin BE, Sampogna SL, Kruger L (1983) An examination of intraspinal sprouting in dorsal root axons with the tracer horseradish peroxidase. J Comp Neurol 215:187–198
Roigrok TJH, Crowe A, Donkelaar HJT (1982) The distribution of motoneurons innervating hindlimb muscles in the terrapin Pseudemys scripta elegans. Neurosci Lett 28:157–162
Ruigrok TJH, Crowe A, Donkelaar HJT (1985) Dendrite distribution of identified motoneurons in the lumbar spinal cord of the turtle Pseudemys scripta elegans. J Comp Neurol 238:275–285
Scheibel ME, Scheibel AB (1969) Terminal patterns in cat spinal cord-III. Primary afferent collaterals. Brain Res 13:417–443
Sickles DW, Oblak TG (1984) Quantitative differences in horseradish peroxidase-labeling of α-motoneurons. Neurosci Lett 49:69–75
Strick PL, Bruke RE, Kanda K, Kim CC, Walmsley B (1976) Differences between alpha and gamma motoneurons labeled with horseradish peroxidase by retrograde transport. Brain Res 113:582–588
Swett JE, Woolf CJ (1985) The somatotopic organization of primary afferent terminals in the superficial laminae of the dorsal horn of the rat spinal cord. J Comp Neurol 231:66–77
Szentágothai J (1964) Neuronal and synaptic arrangement in the substantia gelatinosa Rolandi. J Comp Neurol 122:219–240
Ulfhake B, Kellerth J-O (1983) A quantitative morphological study of HRP labeled cat α-motoneurons supplying different hindlimb muscles. Brain Res 264:1–19
Wan XST, Trojanowski JQ, Gonatas JO (1982) Cholera toxin and wheat germ agglutinin conjugates as neuroanatomical probes their uptake and clearance, transganglionic and retrograde transport and sensitivity. Brain Res 243:215–224
Zenker W, Mysicka A, Neuhuber W (1980) Dynamics of the trans-ganglionic movement of horseradish peroxidase in primary sensory neurons. Cell Tissue Res 207:479–489
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hirakawa, M., McCabe, J.T. & Kawata, M. Time-related changes in the labeling pattern of motor and sensory neurons innervating the gastrocnemius muscle, as revealed by the retrograde transport of the cholera toxin B subunit. Cell Tissue Res 267, 419–427 (1992). https://doi.org/10.1007/BF00319364
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00319364