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The anatomy of the porcine subthalamic nucleus evaluated with immunohistochemistry and design-based stereology

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Abstract

This study provides a light-microscopic description of the organization, morphology and number of neurons in the subthalamic nucleus (STN) of the Göttingen minipig. It is based on histological material stained with Nissl, Golgi and autometallographic techniques, and employs design-based stereological estimation of the total neuron number. The organization of several neurotransmitters in the STN has been evaluated in histological preparations stained for acetylcholinesterase (AChE) and immunostained for choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), glutamic acid decarboxylase (GAD) and glutamate. In all of the stained preparations the STN appeared as a distinct lens-shaped structure located in the caudal diencephalon, medial to the internal capsule and ventrolateral to the zona incerta. Rostrally, the STN approached the globus pallidus pars interna, whereas caudally the ventromedial part of the STN was adjacent to the rostral part of the substantia nigra pars compacta (SNc), where some of the neurons of the two nuclei merged. The neurons in the STN had medium-sized (25–40 μm) ovoid or fusiform cell bodies, from which three to six large dendrites emanated in a direction predominantly parallel to the long axis of the STN. Immunohistochemistry revealed that most of the subthalamic neurons were glutamatergic and differed significantly in appearance from the large stellate TH-positive cells of the adjacent SNc. Numerous TH-positive bouton-rich fibers traversed the STN. The GAD-staining revealed a large number of terminals within the boundaries of the STN. The STN was highly AChE-positive, reflecting a prominent innervation by ChAT-positive terminals. The total number of subthalamic neurons in one hemisphere was estimated to be approximately 56,000. We conclude that the neuroarchitecture of the porcine STN is similar to primates, including humans, and appears well-suited for further studies examining the role of the STN in movement disorders.

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Abbreviations

AChE:

Acetylcholinesterase

AMG:

Autometallography

asf:

Area sampling fraction

ChAT:

Choline acetyltransferase

CI:

Internal capsule

DBS:

Deep brain stimulation

fl:

Lenticular fasciculus

GAD:

Glutamic acid decarboxylase

GPe:

Globus pallidus pars externa

GPi:

Globus pallidus pars interna

mfb:

Medial forebrain bundle

MPTP:

1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

nsp:

Nigrostriatal pathway

SN:

Substantia nigra

SNc:

Substantia nigra, pars compacta

SNl:

Substantia nigra, pars lateralis

ssf:

Section sampling fraction

STN:

Subthalamic nucleus

TBS:

Tris-buffered saline

TH:

Tyrosine hydroxylase

VTA:

Ventral tegmental area

ZI:

Zona incerta

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Acknowledgments

The authors acknowledge with gratitude the skillful assistance of Ms. D. Jensen, Ms. L. Munkøe, Ms. K. Wiedemann, Mr. A. Meier, Mr. T. Nielsen, and Ms. H. Mikkelsen. We acknowledge the valuable discussions with Dr. M. Stoltenberg, Dr. M.S. Jensen, Dr. G. Danscher, and Dr. F.A. Geneser.

The following supported the study: the Danish Parkinson Association, the Petrus Andersen Foundation, the Foundation of Politimester J.P.N. Colind og Hustru Asmine Colind, the Foundation of Etatsraad C.G. Filtenborg og Hustru Marie Filtenborg, the Helga og Peter Korning Foundation, the Aarhus University Research Foundation, the Alice Brenaa Foundation, the Danish Medical Research Council, the Foundation for the Advancement of Medical Science, the Lily Benthine Lund Foundation, the Medical Society of Aarhus County, the Novo Nordic Foundation, the Aase and Ejnar Danielsen Foundation, the Foundation of 2/7 1984 against Parkinson’s Disease, the Foundation of Direktør Ib Henriksen, and the Foundation of King Christian the Tenth.

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Authors and Affiliations

  1. Department of Neurobiology, Institute of Anatomy, University of Aarhus, 8000, Aarhus C, Denmark

    Mette Larsen, Carsten R. Bjarkam, Mark J. West & Jens C. Sørensen

  2. Department of Neurology, University Hospital of Aarhus, 8000, Aarhus C, Denmark

    Karen Østergaard

  3. Department of Neurosurgery, University Hospital of Aarhus, 8000, Aarhus C, Denmark

    Jens C. Sørensen

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  1. Mette Larsen
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  2. Carsten R. Bjarkam
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Correspondence to Mette Larsen.

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Larsen, M., Bjarkam, C.R., Østergaard, K. et al. The anatomy of the porcine subthalamic nucleus evaluated with immunohistochemistry and design-based stereology. Anat Embryol 208, 239–247 (2004). https://doi.org/10.1007/s00429-004-0395-0

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