Skip to main content
SpringerLink
Log in

Carbonic anhydrase and horseradish peroxidase: double labelling of rat dorsal root ganglion neurons innervating motor and sensory peripheral nerves

  • Published:
Anatomy and Embryology Aims and scope Submit manuscript

Summary

Dorsal root ganglion neurons supplying peroneus longus, soleus and gastrocnemius medius muscles and the sural nerve of the rat were labelled with horseradish peroxidase and analysed for their carbonic anhydrase content. Staining of the sections was done either on the same or on alternate slides. Both methods led to the same results, despite a slight fading of the carbonic anhydrase reaction in double-stained sections. The data indicated that the musles under study were supplied by approximately the same number of horseradish peroxidase-labelled cells, irrespective of their differences in size. 74.9% of these labelled neurons had diameters exceeding 30 μm and 52.4% of them also stained for carbonic anhydrase. The double-labelled cells represented 66.9% of the population of large neurons (>30 μm) and comprised most of those measuring over 47.5 μm. Richness in carbonic anhydrase of the large muscle afferent neurons may be linked to their innervation of the stretch receptors, as components of an active apparatus which includes the gamma motor axons which also stain positively for carbonic anhydrase. in contrast, the ganglion cells supplying the sural nerve were almost totally devoid of carbonic anhydrase, as only 6.4% showed double labelling. This contingent possibly represents the muscle afferents of the small motoneural population which supplies, through this nerve, part of the foot musculature of the rat.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adams JC (1977) Technical considerations on the use of horseradish peroxidase as a neuronal marker. Neurocience 2:141–145

    Google Scholar 

  • Cabelguen JM, Orsal D, Perret C (1984) Discharges of forelimb spindle primary afferents during locomotor activity in the decorticate cat. Brain Res 306:359–364

    Google Scholar 

  • Cammer W, Sacchi R, Sapirstein V (1985) Immunocytochemical localization of carbonic anhydrase in the spinal cords of normal and mutant (Shiverer) adult mice with comparisons among fixation methods. J Histochem Cytochem 33:45–54

    Google Scholar 

  • Carter MJ (1972) Carbonic anhydrase: isoenzymes, properties, distribution, and functional significance. Biochem Soc Trans 47:465–513

    Google Scholar 

  • Carter N, Jeffery S (1985) Carbonic anhydrase: uptake and new horizons. Biochem Soc Trans 13:531–533

    Google Scholar 

  • Carter ND, Jeffery S, Shiels A, Edwards Y, Tipler T, Hopkinson DA (1979) Characterization of human carbonic anhydrase III from skeletal muscle. Biochem Genet 17:837–854

    Google Scholar 

  • Coleman JE (1973) Carbonic anhydrase. In: Eichorn GL (ed) Inorganic Biochemistry. Elsevier Science, Amsterdam, pp 449–498

    Google Scholar 

  • Deutsch HF (1987) Carbonic anhydrases. Int J Biochem 19:101–113

    Google Scholar 

  • Dodd J, Jahr CE, Hamilton PN, Heath MJS, Matthew WD, Jessell TM (1983) Cytochemical and physiological properties of sensory and dorsal horn neurons that transmit cutaneous sensation. Cold Spring Harbor Symp Quant Biol 48:865–695

    Google Scholar 

  • Foehring RC, Sypert GW, Munson JB (1986) Anomolous path taken by branch of medial gastrocnemius nerve. Exp Neurol 92:440–444

    Google Scholar 

  • Gottschall J, Zenker W, Neuhuber W, Mysicka A, Müntener A (1980a) The sternomastoid muscle of the rat and its innervation. Muscle fiber composition, perikarya and axons of efferent and afferent neurons. Anat Embryol 160:285–300

    Google Scholar 

  • Gottschall J, Neuhuber W, Müntener M, Mysicka A (1980b) The Ansa cervicalis and the infrahyoid muscles of the rat. Anat Embryol 159:59–69

    Google Scholar 

  • 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

    Google Scholar 

  • Hagbarth KE, Wohlfart G (1952) The number of muscle-spindles in certain muscles in cat in relation to the composition of the muscle nerves. Acta Anat (Basel) 15:85–104

    Google Scholar 

  • Hansson HPJ (1967) Histochemical demonstration of carbonic anhydrase activity. Histochemie 11:112–128

    Google Scholar 

  • Harper AA, Lawson SN (1985a) Conduction velocity is related to morphological cell type in rat dorsal root ganglion neurons. J Physiol 359:31–46

    Google Scholar 

  • Harper AA, Lawson SN (1985b) Electrical properties of rat dorsal root ganglion neurons with different peripheral nerve conduction velocities. J Physiol 359:47–63

    Google Scholar 

  • Holmes RS (1976) Mammalian carbonic anhydrase isozymes: evidence for a third locus. J Exp Zool 197:289–295

    Google Scholar 

  • Ibi T, Haimoto H, Nagura H, Sahaski K, Kato K, Mokuno K, Sugimura K, Matsuoka Y (1985) Immunohistochemical demonstration of carbonic anhydrase III and muscle-specific enolase in paraffin-embedded human skeletal muscle sections. Acta Neuropathol (Berl) 68:74–76

    Google Scholar 

  • Ikejima T, Ito S (1984) Carbonic anhydrase in mouse salivary glands and saliva: a histochemical, immunohistochemical, and enzyme activity study. J Histochem Cytochem 32:625–635

    Google Scholar 

  • Jänig W, McLachlan EM (1986a) Identification of distinct topographical distributions of lumbar sympathetic and sensory neurons projecting to end organs with different functions in the cat. J Comp Neurol 246:104–112

    Google Scholar 

  • Jänig W, McLachlan EM (1986b) The sympathetic and sensory components of the caudal lumbar sympathetic trunk in the cat. J Comp Neurol 245:62–73

    Google Scholar 

  • Jeffery S, Carter ND, Smith A (1986) Immunocytochemical localization of carbonic anhydrase isozymes I, II, and III in rat skeletal muscle. J Histochem Cytochem 34:513–516

    Google Scholar 

  • Jessell TM, Dodd J (1986) Neurotransmitters and differentiation antigens in subsets of sensory neurons projecting to the spinal dorsal horn. In: Martin JB, Barchas D (eds) Neuropeptides in neurologic and psychiatric disease. Raven Press, New York, pp 111–133

    Google Scholar 

  • Johansson H (1985) Reflex integration in the γ-motor system. In: Boyd IA, Gladden MH (eds) The muscle spindle. Stockton Press, New York, pp 297–302

    Google Scholar 

  • Kai-Kai MA, Anderton BH, Keen P (1986) A quantitative analysis of the interrelationships between subpopulations of rat sensory neurons containing arginine vasopressin or oxytocin and those containing substance P, fluoride-resistant acid phosphatase or neurofilament protein. Neuroscience 18:475–486

    Google Scholar 

  • Kazimierczak J, Sommer EW, Philippe E, Droz B (1986) Carbonic anhydrase activity in primary sensory neurons. I. Requirements for the cytochemical localization in the dorsal root ganglion of chicken and mouse by light and electron microscopy. Cell Tissue Res 245:487–495

    Google Scholar 

  • Knyihar-Csillik E, Bezzegh A, Böti S, Csillik B (1986) Thiamine monophosphatase: a genuine marker for transganglionic regulation of primary sensory neurons. J Histochem Cytochem 34:363–371

    Google Scholar 

  • Knyihar-Csillik E, Csillik B (1981) FRAP: Histochemistry of the primary nociceptive neuron. Prog Histochem Cytochem 14:1–137

    Google Scholar 

  • Kumpulainen T, Korhonen LK (1982) Immunohistochemical localization of carbonic anhydrase isoenzyme C in the central and peripheral nervous system of the mouse. J Histochem Cytochem 30:283–292

    Google Scholar 

  • Laurent G, Venot-Giraud N, Marrig C, Limozin N, Garcon D, Filippi D (1977) Selective tissue distribution of carbonic anhydrase isozymes in the ox. Biochim Biophys Acta 481:222–226

    Google Scholar 

  • Loeb GE, Hoffer JA (1985) Activity of spindle afferents from cat anterior thigh muscles. II. Effects of fusimotor blockade. J Neurophysiol 54:565–577

    Google Scholar 

  • Lönnerholm G (1974) Carbonic anhydrase histochemistry, a critical study of Hansson's cobalt-phosphate method. Acta Physiol Scand [Suppl] 418:1–43

    Google Scholar 

  • Lönnerholm G (1984) Histochemical localization of carbonic anhydrase in mammalian tissue. Ann NY Acad Sci USA 429:369–381

    Google Scholar 

  • Maren TH (1967) Carbonic anhydrase chemistry, phisiology, and inhibition. Physiol Rev 47:595–781

    Google Scholar 

  • Martin TL, Mufson EJ, Mesulam MM (1984) The light side of horseradish peroxidase histochemistry. J Histochem Cytochem 32:793

    Google Scholar 

  • Matthews PBC (1972) The structure of the receptors. In: Davson H, Greenfield ADM, Whittam R, Brindley GS (eds) Mammalian muscle receptors and their central actions. Edward Arnold Ltd, London, pp 1–66

    Google Scholar 

  • McLachlan EM, Jänig W (1983) The cell bodies of origin of sympathetic and sensory axons in some skin and muscle nerves of the cat hindlimb. J Comp Neurol 214:115–130

    Google Scholar 

  • Mesulam MM (1978) Tetramethylbenzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction-product with superior sensitivity for visualizing neural afferents and efferents. J Histochem Cytochem 26:106–117

    Google Scholar 

  • Mesulam MM, Hegarty E, Barbas H, Carson KA, Gower EC, Knapp AG, Moss MB, Mufson EJ (1980) Additional factors influencing sensitivity in the tetramethylbenzidine method for horseradish peroxidase neurohistochemistry. J Histochem Cytochem 28:1255–1259

    Google Scholar 

  • Mesulam MM, Mufson EJ, Wainer BH (1986) Three-dimensional representation and cortical projection topography of the nucleus basalis (Ch4) in the macaque: concurrent demonstration of choline acetyltransferase and retrograde transport with a stabilized tetramethylbenzidine method for horseradish peroxidase. Brain Res 367:301–308

    Google Scholar 

  • Morrell JI, Greenberger LM, Pfaff DW (1981) Comparison of horseradish peroxidase visualization methods: quantitative results and further technical specifics. J Histochem Cytochem 29:903–916

    Google Scholar 

  • Moyle S, Jeffrey S, Carter ND (1984) Localization of human muscle carbonic anhydrase isozymes using immunofluorescence. J Histochem Cytochem 32:1262–1264

    Google Scholar 

  • Murphy PR, Stein RB, Taylor J (1985) Phasic and tonic modulation of impulse rates in motoneurones during locomotion in the premammillary cat and their implications for control of locomotion. In: Boyd IA, Gladden MH (eds) The muscle spindle, Stockton Press, New York, pp 309–310

    Google Scholar 

  • Nagy JI, Daddona PE (1985) Anatomical and cytochemical relationships of adenosine deaminase-containing primary afferent neurons in the rat. Neuroscience 15:799–813

    Google Scholar 

  • Neuhuber W, Mysicka A (1980) Afferent neurons of the hypoglossal nerve of the rat as demonstrated by horseradish peroxidase tracing. Anat Embryol 158:349–360

    Google Scholar 

  • Neuhuber W, Niederle B (1979) Spinal ganglion cells innervating the stomach of the rat as demonstrated by somatopetal transport of horseradish peroxidase (HRP). Anat Embryol 155:355–362

    Google Scholar 

  • Ovalle WK, Dow PR (1985) Morphological aspects of the muscle spindle capsule and its functional significance. In: Boyd IA, Gladden MH (eds) The muscle spindle. Stockton Press, New York, pp 3–22

    Google Scholar 

  • Peyronnard JM, Charron L (1982) Motor and sensory neurons of the rat sural nerve: a horseradish peroxidase study. Muscle Nerve 5:654–660

    Google Scholar 

  • Peyronnard JM, Charron L (1983) Motoneuronal and motor axonal innervation in the rat hindlimb: a comparative study using horseradish peroxidase. Exp Brain Res 50:125–132

    Google Scholar 

  • Peyronnard JM, Charron LF, Lavoie J, Messier JP (1986a) Motor, sympathetic and sensory innervation of rat skeletal muscles. Brain Res 373:288–302

    Google Scholar 

  • Peyronnard JM, Charron L, Lavoie J, Messier JP (1986b) Differences in horseradish peroxidase labeling of sensory, motor and sympathetic neurons following chronic axotomy of the rat sural nerve. Brain Res 364:137–150

    Google Scholar 

  • Peyronnard JM, Messier JP, Charron L, Lavoie J, Bergouignan FX, Dubreuil M (1986c) Carbonic anhydrase activity in the normal and injured peripheral nervous system of the rat. Exp Neurol 93:481–499

    Google Scholar 

  • Peyronnard JM, Charron L, Messier JP, Lavoie J, Faraco-Cantin F (1987) Histochemical localization of CA in normal and diseased human muscle. Muscle Nerve (in press)

  • Register AM, Koester MK, Noltmann EA (1978) Discovery of carbonic anhydrase in rabbit skeletal muscle and evidence for its identity with basic muscle protein. J Biol Chem 253:4143–4152

    Google Scholar 

  • Ridderstrale Y (1976) Intracellular localization of carbonic anhydrase in the frog nephron. Acta Physiol Scand 98:465–469

    Google Scholar 

  • Riley DA, Ellis S, Bain J (1982) Carbonic anhydrase activity in skeletal muscle fiber types, axons, spindles, and capillaries of rat soleus and extensor digitorum longus muscles. J Histochem Cytochem 30:1275–1288

    Google Scholar 

  • Riley DA, Ellis S, Bain JLW (1984) Ultrastructural cytochemical localization of carbonic anhydrase activity in rat peripheral sensory and motor nerves, dorsal root ganglia and dorsal column nuclei. Neuroscience 13:189–206

    Google Scholar 

  • Rye DB, Saper CB, Wainer BH (1984) Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry. J Histochem Cytochem 32:1145–1153

    Google Scholar 

  • Sugai N, Ito S (1980) Carbonic anhydrase, ultrastructural localization in the mouse gastric mucosa and improvements in the technique. J Histochem Cytochem 28:511–525

    Google Scholar 

  • Swett JE, Wikholm RP, Blanks RHI, Swett AI, Conley LC (1986) Motoneurons of the rat sciatic nerve. Exp Neurol 93:227–252

    Google Scholar 

  • Tashian RE, Hewett-Emmett D, Goodman M (1983) On the evolution and genetics of carbonic anhydrases I, II, and III. In: Rattazzi MC, Scandalios JG, WHitt GS (eds) Isozymes: current topics in biological and medical research, vol 7. A R Liss, New York, pp 79–100

    Google Scholar 

  • Taylor DCM, Korf HW, Pierau FrK (1982) Distribution of sensory neurones of the pudendal nerve in the dorsal root ganglia and their projection to the spinal cord. Cell Tissue Res 226:555–564

    Google Scholar 

  • Tuchscherer MM, Seybold VS (1985) Immunohistochemical studies of substance P, cholecystokinin-octapeptide and somatostatin in dorsal root ganglion of the rat. Neuroscience 2:593–605

    Google Scholar 

  • Väänänen HK, Paloniemi M, Vuori J (1985) Purification and localization of human carbonic anhydrase III. Typing of skeletal muscle fibers in paraffin embedded sections. Histochemistry 83:231–235

    Google Scholar 

  • Walro JM, Kucera J (1985) Fusimotor innervation in the rat: evidence for taxonomic differences in the organization of spindles. In: Boyd IA, Gladden MH (eds) The muscle spindle. Stockton Press, New York, pp 39–43

    Google Scholar 

  • Wong V, Barrett CP, Donati EJ, Eng LF, Guth L (1983) Carbonic anhydrase activity in first-order sensory neurons of the rat. J Histochem Cytochem 31:293–300

    Google Scholar 

  • Wong V, Barrett CP, Donati EJ, Guth L (1987) Distribution of carbonic anhydrase activity in neurons of the rat. J Comp Neurol 257:122–129

    Google Scholar 

  • Wong-Riley MTT, Kageyama GH (1986) Localization of cytochrome oxidase in the mammalian spinal cord and dorsal root ganglia, with quantitative analysis of ventral horn cells in monkeys. J Comp Neurol 245:41–61

    Google Scholar 

  • Yoshida S, MAtsuda Y (1979) Studies on sensory neurons of the mouse with intracellular recording and horseradish peroxidase-injection techniques. J Neurophysiol 42:1134–1145

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre de recherche en sciences neurologiques, Université de Montréal, Quebec, Canada

    J. M. Peyronnard, L. Charron, J. Lavoie, J. P. Messier & M. Dubreuil

  2. Hôpital Hôtel-Dieu, Université de Montréal, Quebec, Canada

    J. M. Peyronnard, L. Charron, J. Lavoie, J. P. Messier & M. Dubreuil

Authors
  1. J. M. Peyronnard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L. Charron
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Lavoie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. P. Messier
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Dubreuil
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Peyronnard, J.M., Charron, L., Lavoie, J. et al. Carbonic anhydrase and horseradish peroxidase: double labelling of rat dorsal root ganglion neurons innervating motor and sensory peripheral nerves. Anat Embryol 177, 353–359 (1988). https://doi.org/10.1007/BF00315844

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00315844

Key words

Search

Navigation