Summary
Immunocytochemical detection of glutamate decarboxylase (GAD), the predominant biosynthetic enzyme of gamma-aminobutyric acid (GABA), reveals the presence of a dense GABAergic innervation in all parts of the inferior olive. One brain center that provides a substantial projection to the inferior olive is the cerebellar nuclei, which contain many small GABAergic neurons. These neurons were tested as a source of GABAergic olivary afferents by combining retrograde tract tracing with GAD immunocytochemistry. As expected from previous studies, injections of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) into the inferior olive retrogradely label many small neurons in the interposed and lateral cerebellar nuclei and the dorsal part of the lateral vestibular nucleus, and fewer neurons in the ventro-lateral region of the medial cerebellar nucleus. These projections are predominantly crossed and are topographically arranged. The vast majority, if not all, of these projection neurons are also GAD-positive. The relative contribution of this projection to the GABAergic innervation of the inferior olive was tested by lesion of the cerebellar nuclei, or the superior cerebellar peduncle. Within 10 days the lesion eliminates most GAD-immunoreactive boutons in the principal olive, the rostral lamella of the medial accessory olive, the ventrolateral outgrowth, and the lateral part of the dorsal accessory olive ventral fold. Thus, the effectiveness of this depletion demonstrates that the cerebellar nuclei provide most of the GABAergic innervation to regions of the inferior olive known to receive a cerebellar projection. Moreover, when the lateral vestibular nucleus is damaged, the dorsal fold of the dorsal accessory olive is depleted of GABAergic boutons. The synaptic relations that boutons of the GABAergic cerebello-olivary projection share with olivary neurons were investigated at the electron microscopic level by GAD-immunocytochemistry, anterograde degeneration of the cerebellar axons or anterograde transport of WGA-HRP. All of these methods confirm that GABAergic, cerebello-olivary axon terminals contain pleomorphic vesicles, and synapse on various portions of olivary neurons, and especially on dendritic spines within glomeruli, often in very close proximity to the gap junctions that characteristically couple the dendritic profiles. These results demonstrate four major points: that virtually all of the GABAergic, and presumably inhibitory, neurons of the cerebellar and dorsal lateral vestibular nuclei are projection neurons; that a large portion of the inferior olive receives GABAergic afferents from the cerebellar nuclei; that a portion of the dorsal accessory olive receives GABAergic afferents from the dorsal lateral vestibular nucleus; and that cerebello-olivary fibers often synapse near gap junctions, and therefore could influence electrical coupling of olivary neurons.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- aMAO :
-
subnucleus a of MAO
- beta :
-
beta nucleus
- bMAO :
-
subnucleus b of MAO
- cMAO :
-
subnucleus c of MAO
- dc :
-
dorsal cap
- DC :
-
dorsal cochlear nucleus
- dfDAO :
-
dorsal fold of DAO
- dlh :
-
dorsal lateral hump of cerebellar nuclei
- dIPO :
-
dorsal lamella of PO
- Giαa :
-
gigantocellular reticular nucleus
- dmcc :
-
dorsomedial cell column
- GABA :
-
gamma-aminobutyric acid
- GAD :
-
glutamate decarboxylase
- HRP :
-
horseradish peroxidase
- icp :
-
inferior cerebellar peduncle
- IC :
-
inferior colliculus
- Inf :
-
infracerebellar nucleus
- IntA :
-
anterior interposed cerebellar nucleus
- IntP :
-
posterior interposed cerebellar nucleus
- Lat :
-
lateral cerebellar nucleus
- LRt :
-
lateral reticular nucleus
- LSO :
-
lateral superior olive
- LVe :
-
lateral vestibular nucleus
- MAO :
-
medial accessory olive
- Med :
-
medial cerebellar nucleus
- Me5 :
-
mesencephalic trigeminal nucleus
- MVe :
-
medial vestibular nucleus
- PFl :
-
paraflocculus of the cerebellar cortex
- PO :
-
principle olive
- RMg :
-
raphe magnus
- rMAO :
-
rostral lamella of MAO
- rs :
-
rubrospinal tract
- scp :
-
superior cerebellar peduncle
- SuVe :
-
spinal vestibular nucleus
- SuVe :
-
superior vestibular nucleus
- vfDAO :
-
ventral fold of DAO
- vlo :
-
ventrolateral outgrowth
- vlPO :
-
ventral lamella of PO
- Y :
-
Y, y vestibular nucleus
- WGA :
-
wheatgerm agglutinin
References
Achenbach KE, Goodman DC (1968) Cerebellar projections to pons, medulla and spinal cord in the albino rat. Brain Behav Evol 1: 43–57
Adams JC (1981) Heavy metal intensification of DAB-based HRP reaction product. J Histochem Cytochem 29:775
Adams JC, Mugnaini E (1987) Patterns of glutamate decarboxylase immunostaining in the feline cochlear nuclear complex studied with silver enhancement and electron microscopy. J Comp Neurol 262:375–401
Andersson G, Hesslow G (1986) Evidence for an inhibitory action by cerebellar nuclei cells on the inferior olive. Neurosci Lett [Suppl 26]:S231
Andersson G, Hesslow G (1987a) Inferior olive excitability after high frequency climbin fibre activation in the cat. Exp Brain Res 67:523–532
Andersson G, Hesslow G (1987b) Activity of Purkinje cells and interpositus neurones during and after periods of high frequency climbing fiber activation in the cat. Exp Brain Res 67:533–542
Angaut P, Cicirata F (1982) Cerebello-olivary projections in the rat. Brain Behav Evol 21:24–33
Angaut P, Sotelo C (1987) The dentato-olivary projection in the rat as a presumptive GABAergic link in the olivo-cerebelloolivary loop. An ultrastructural study. Neurosci Lett 83:227–231
Beitz AJ (1976) The topographical organization of the olivo-dentate and dentato-olivary pathways in the cat. Brain Res 115:311–317
Billard JM, Batini C, Buisseret-Demas C, Daniel H (1989) The inferior olive innervation from the cerebellar and lateral vestibular nuclei: evidence for a longitudinal zonal segregation of the cortico-nucleo-olivary connection in the rat. In: Strata P (ed) The olivocerebellar system in motor control. Springer,Berlin. Exp Brain Res 17:117–120
Brown JT, Chan-Palay V, Palay SL (1977) A study of afferent input to the inferior olivary complex in the rat by retrograde axonal transport of horseradish peroxidase. J Comp Neurol 176:1–22
Buisseret-Delmas C, Batini C (1978) Topology of the pathways to the inferior olive: an HRP study in cat. Neurosci Lett 10:207–214
Carleton SC, Carpenter MB (1983) Afferent and efferent connections of the medial, inferior and lateral vestibular nuclei in the cat and monkey. Brain Res 278:29–51
Carpenter MB, Nova HR (1960) Descending division of the brachium conjunctivum in the cat: a cerebello-reticular system. J Comp Neurol 114:295–305
Carrea RME, Mettler FA (1955) Function of the primate brachium conjunctivum and related structures. J Comp Neurol 102:151–322
Chan-Palay V (1977) Cerebellar dentate nucleus, organization, cytology, transmitters. Springer, Berlin
Chan-Palay V (1982) Neurotransmitters and receptors in the cerebellum: immunocytochemical localization of glutamic acid decarboxylase, GABA-transaminase, and cyclic GMP and autoradiography with [3H]-muscimol. In: Palay SL, Chan-Palay V (eds) The cerebellum, new vistas. Springer, Berlin, pp 552–584
Chan-Palay V, Palay SL, Wu J-Y (1979) Gamma-aminobutyric acid pathways in the cerebellum studied by retrograde and anterograde transport of glutamic acid decarboxylase antibody after in vivo injections. Anat Embryol 157:1–14
Chen DH (1978) Qualitative and quantitative study of synaptic displacement in chromatolyzed spinal motoneurons of the cat. J Comp Neurol 177:635–664
Cohen D, Chambers WW, Sprague JM (1958) Experimental study of the efferent projections from the cerebellar nuclei to the brainstem of the cat. J Comp Neurol 109:233–259
Cummings SL, Young WS III, Bishop GA, DeSousa EB, King JS (1989) Distribution of corticotropin-releasing factor in the cerebellum and precerebellar nuclei of the opossum: a study utilizing immunocytochemistry, in situ hybridization histochemistry, and receptor autoradiography. J Comp Neurol 280:501–521
Desclin JC (1974) Histological evidence supporting the inferior olive as the major source of cerebellar climbing fibers in the rat. Brain Res 77:365–384
deZeeuw CI, Holstege JC, Calkoen F, Ruigrok TJH, Voogd J (1988) A new combination of WGA-HRP anterograde tracing and GABA immunocytochemistry applied to afferents of the cat inferior olive at the ultrastructural level. Brain Res 447:369–375
deZeeuw CI, Holstege JC, Ruigrok TJH, Voogd J (1989) Ultrastructural study of the GABAergic, cerebellar, and mesodiencephalic innervation of the cat medial accessory olive: anterograde tracing combined with immunocytochemistry. J Comp Neurol 284:12–35
deZeeuw CI, Holstege JC, Ruigrok TJH, Voogd J (1990a) Mesodiencephalic and cerebellar terminals end upon the same dendritic spines within the glomeruli of the cat and rat inferior olive: an ultrastructural study using a combination of [3H]leu-cine and WGA-HRP anterograde tracing. Neuroscience 34:645–655
deZeeuw CI, Ruigrok TJH, Holstege JC, Jansen HG, Voogd J (1990b) Intracellular labeling of neurons in the medial accessory olive of the cat. II. Ultrastructure of dendritic spines and their GABAergic innervation. J Comp Neurol 300:478–494
deZeeuw CI, Ruigrok TJH, Holstege JC, Schalekanp MPA, Voogd J (1990c) Intracellular labeling of neurons in the medial accessory olive of the cat. III. Ultrastructure of axon hillock and initial segment and their GABAergic innervation. J Comp Neurol 300:495–510
Dietrichs E, Walberg F (1981) The cerebellar nucleo-olivary projection in the cat. Anat Embryol 162:51–67
Dietrichs E, Walberg F (1985) The cerebellar nucleo-olivary and olivo-cerebellar nuclear projection in the cat as studied with anterograde and retrograde transport in the same animal after implantation of crystalline WGA-HRP II. The fastigial nucleus. Anat Embryol 173:253–261
Dietrichs E, Walberg F (1989) Direct bidirectional connections between the inferior olive and the cerebellar nuclei. In: Strata P (ed) The olivocerebellar system in motor control. Springer, Berlin. Exp Brain Res Ser 17:61–81
Flood S, Jansen J (1966) The efferent fibers of the cerebellar nuclei and their distribution on the cerebellar peduncles in the cat. Acta Anat 63:137–166
Fredette BJ, Adams JC, Mugnaini E (1991) GABAergic neurons in the mammalian inferior olive and ventral medulla detected by glutamate decarboxylase immunocytochemistry. J Comp Neurol (in press)
Gerrits NM, Voogd J, Magras IN (1985) Vestibular afferents of the inferior olive and the vestibulo-olivo-cerebellar climbing fiber pathway to the flocculus in the cat. Brain Res 332:325–336
Graybiel AM, Nauta HJW, Lasek RJ, Nauta WJH (1973) A cerebello-olivary pathway in the cat: an experimental study using autoradiographic tracing techniques. Brain Res 58:205–211
Haroian AJ (1982) Cerebello-olivary projections in the rat: an autoradiographic study. Brain Res 235:125–130
Hesslow G (1986) Inhibition of inferior olivary transmission by mesencephalic stimulation in the cat. Neurosci Lett 63:76–80
Houser CR, Barber RP, Vaughn JE (1984) Immunocytochemical localization of glutamic acid decarboxylase in the dorsal lateral vestibular nucleus: evidence for an intrinsic and extrinsic GABAergic innervation. Neurosci Lett 47:213–220
Ito M (1984) The Cerebellum and Neural Control. Raven Press, New York
Kalil K (1979) Projections of the cerebellar and dorsal column nuclei upon the inferior olive in the Rhesus monkey: an autoradiographic study. J Comp Neurol 188:43–62
King JS, Andrezik JA, Falls WM, Martin GF (1976) The synaptic organization of the inferior olivary circuit. Exp Brain Res 26:159–179
Kitai ST, Preston JR, McCrea R (1975) Interposito-olivary and olivo-interpositus relationship: An electrophysiological study. Anat Rec 181:396
Korneliussen HK (1968) On the morphology and subdivision of the cerebellar nuclei of the rat. J Hirnforsch 10:109–122
Krnjevic K (1976) Inhibitory action of GABA and GABA-mimetics on vertebrate neurons. In: Roberts E, Chase TN, Tower DB (eds) GABA in nervous system function. Raven Press, New York, pp 269–281
Lapresle J, Ben Hamida M (1970) The dentato-olivary pathway. Somatotopic relationship between the dentate nucleus and the contralateral inferior olive. Arch Neurol 22:135–143
Legendre A, Courville J (1987) Origin and trajectory of the cerebello-olivary projection: an experimental study with radioactive and fluorescent tracers in the cat. Neurosci 21:877–891
Llinás R (1974) Motor aspects of cerebellar control. Physiologist 17:19–46
Llinás R (1989) Electrophysiological properties of the olivocerebellar system. In: Strata P (ed) The olivocerebellar system in motor control. Springer, Berlin, pp 201–208
Llinds R, Yarom Y (1986) Oscillatory properties of guinea-pig inferior olivary neurones and their pharmacological modulation: an in vitro study. J Physiol (Lond) 376:163–182
Martin GF, Henkel CK, King JS (1976) Cerebello-olivary fibers: their origin, course and distribution in the North American opossum. Exp Brain Res 24:219–236
Matsushita M, Iwahori N (1971) Structural organization of the interpositus and the dentate nuclei. Brain Res 35:17–36
McCrea RA, Bishop GA, Kitai ST (1978) Morphological and electrophysiological characteristics of projection neurons in the nucleus interpositus of the cat cerebellum. J Comp Neurol 181:397–420
Mehler WER (1967) Double descending pathways originating from the superior cerebellar peduncle. An example of neural species differences. Anat Rec 157:374
Mehler WR, Rubertone JA (1985) Anatomy of the vestibular nucleus complex. In: Paxinos G (ed) The rat nervous system. Vol 2 Hindbrain and spinal cord. Academic Press, Sydney, pp 185–219
Mehler WR, Vernier VG, Nauta WJH (1958) Efferent projections from dentate and interpositus nuclei in primates. Anat Rec 130:430–431
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 26:106–116
Monaghan PL, Clements JR, Madl JE, Maley BE, Larson AA, Beitz AJ (1986a) Ultrastructural analysis of GABAergic and glutaminergic cells and processes in the deep cerebellar nuclei of the rat. Soc Neurosci Abstr 12:462
Monaghan PL, Bietz AJ, Larson AA, Altschuler RA, Madl JE, Mullett MA (1986b) Immunocytochemical localization of glutamate-, glutaminase- and aspartate aminotransferase-like immunoreactivity in the rat deep cerebellar nuclei. Brain Res 363:364–370
Mugnaini E, Nelson BJ (1989) Corticotropin-releasing factor (CRF) in the olivocerebellar system and the feline olivary hypertrophy. Exp Brain Res Ser 17:187–190
Mugnaini E, Oertel W (1981) Distribution of glutamate decarboxylase positive neurons in the rat cerebellar nuclei. Soc Neurosci Abstr 7:122
Mugnaini E, Oertel WH (1985) An atlas of the distribution of GABAergic neurons and terminals in the rat CNS as revealed by GAD immunocytochemistry. In: Björklund A, Hökfelt T (eds) Handbook of chemical neuroanatomy, Vol 4: GABA and Neuropeptides in the CNS, Part I. Elsevier, British Vancouver, pp 436–608
Nelson B, Mugnaini E (1985) Loss of GABAergic nerve terminals in the inferior olive of cerebellectomized rats. Soc Neurosci Abstr 11:182
Nelson BJ, Mugnaini E (1988) The rat inferior olive as seen with immunostaining for glutamate decarboxylase. Anat Embryol 179:109–127
Nelson BJ, Mugnaini E (1989) Origins of GABAergic inputs to the inferior olive. In: Strata P (ed) The oivocerebellar system in motor control. Springer, Berlin. Exp Brain Res Ser 17:86–107
Nelson B, Barmack NH, Mugnaini E (1984) A GABAergic cerebello-olivary projection in the rat. Soc Neurosci Abstr 10:539
Nelson B, Barmack NH, Mugnaini E (1986) GABAergic projection from vestibular nuclei to rat inferior olive. Soc Neurosci Abstr 12:225
Nelson BJ, Adams JC, Barmack NH, Mugnaini E (1989) A comparative study of glutamate decarboxylase immunoreactive boutons in the mammalian inferior olive. J Comp Neurol 286:514–539
Oertel WH, Schmechel DE, Tappaz ML, Kopin IJ (1981a) Production of a specific antiserum to rat brain glutamic acid decarboxylase by injection of an antigen-antibody complex. Neuroscience 6:2689–2700
Oertel WH, Schmechel DE, Mugnaini E, Tappaz ML, Kopin IJ (1981b) Immunocytochemical localization of glutamate decarboxylase in rat cerebellum with a new antiserum. Neuroscience 6:2715–2735
Oertel WH, Schmechel DE, Brownstein MJ, Tappaz ML, Ransom DH, Kopin IJ (1981c) Decrease of glutamate decarboxylase (GAD)-immunoreactive nerve terminals in the substantia nigra after kainic acid lesions of the striatum. J Histochem Cytochem 29:977–980
Ohkawa K (1957) Comparative anatomical studies of cerebellar nuclei in mammals. Arch Hist Jpn 13:21–58
Ottersen OP, Storm-Mathisen J (1984) Glutamate- and GABAcontaining neurons in the mouse and rat brain as demonstrated with a new immunocytochemical technique. J Comp Neurol 229:374–392
Peters A, Palay SL, Webster H deF (1991) The fine structure of the nervous system: neurons and their supporting cells, 3rd edn. Oxford University Press, New York Oxford
Ramon Y, Cajal S (1911) Histologie du système nerveux de l'homme et des Vertébrés, Vol I. Maloine, Paris
Ribak CE, Vaughn JE, Roberts E (1980) GABAergic nerve terminals decrease in the substantia nigra following hemitransection of the striatonigral and pallidonigral pathways. Brain Res 192:413–420
Saint-Cyr JA, Courville J (1979) Projection from vestibular nuclei to the inferior olive in the cat: an autoradiographic and horseradish peroxidase study. Brain Res 165:189–200
Sasaki K, Llinás R (1985) Dynamic electrotonic coupling in mammalian inferior olive as determined by simultaneous multiple Purkinje cell recording. Biophys J 47:53a
Sotelo C, Gotow T, Wassef M (1986) Localization of glutamic-aciddecarboxylase-immunoreactive axon terminals in the inferior olive of the rat, with special emphasis on anatomical relations between GABAergic synapses and dendro-dendritic gap junctions. J Comp Neurol 252:32–50
Sugimoto T, Mizuno N, Nomura S, Nakamura Y (1980) Fastigio-olivary fibers in the cat as revealed by the autoradiographic tracing method. Brain Res 199:443–446
Swenson RS, Castro AJ (1983a) The afferent connections of the inferior olivary complex in rats: a study using the retrograde transport of horseradish peroxidase. Am J Anat 166:329–341
Swenson RS, Castro AJ (1983b) The afferent connections of the inferior olivary complex in rats. An anterograde study using autoradiographic and axonal degeneration techniques. Neuroscience 259:275
Tolbert DL (1982) The cerebellar nucleocortical pathway. In: Palay SL, Chan-Palay V (eds) The cerebellum: new vistas. Springer, Berlin, pp 296–317
Tolbert DL, Bantli H (1980) Uptake and transport of3H-GABA (γ-aminobutyric acid) injected into the cat dentate nucleus. Exp Neurol 70:525–538
Tolbert DL, Massopust LC, Murphy MG, Young PA (1976) The anatomical organization of the cerebello-olivary projection in the cat. J Comp Neurol 170:525–544
Tolbert DL, Bantli H, Bloedel JR (1978) Multiple branching of cerebellar efferent projections in cats. Exp Brain Res 31:305–316
Tolbert DL, Bantli H, Hames EG, Ebner TJ, McMullen TA, Bloedel JR (1980) A demonstration of the dentato-reticulospinal projection in the cat. Neuroscience 5:1479–1488
Wiklund L, Toggenburger G, Cuenod M (1982) Aspartate: possible neurotransmitter in cerebellar climbing fibers. Science 216:78–79
Author information
Authors and Affiliations
Additional information
This paper is dedicated to Professor Fred Walberg on the occasion of his 70th hirthdav
Rights and permissions
About this article
Cite this article
Fredette, B.J., Mugnaini, E. The GABAergic cerebello-olivary projection in the rat. Anat Embryol 184, 225–243 (1991). https://doi.org/10.1007/BF01673258
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01673258