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Three-dimensional and stereological characterization of the human substantia nigra during aging

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Abstract

The human brain undergoes non-uniform changes during aging. The substantia nigra (SN), the source of major dopaminergic pathways in the brain, is particularly vulnerable to changes in the progression of several age-related neurodegenerative diseases. To establish normative data for high-resolution imaging, and to further clinical and anatomical studies we analyzed SNs from 15 subjects aged 50–91 cognitively normal human subjects without signs of parkinsonism. Complete brains or brainstems with substantia nigra were formalin-fixed, celloidin-mounted, serially cut and Nissl-stained. The shapes of all SNs investigated were reconstructed using fast, high-resolution computer-assisted 3D reconstruction software. We found a negative correlation between age and SN volume (p = 0.04, rho = −0.53), with great variability in neuronal numbers and density across participants. The 3D reconstructions revealed SN inter- and intra-individual variability. Furthermore, we observed that human SN is a neuronal reticulum, rather than a group of isolated neuronal islands. Caution is required when using SN volume as a surrogate for SN status in individual subjects. The use of multimodal sequences including those for fiber tracts may enhance the value of imaging as a diagnostic tool to assess SN in vivo. Further studies with a larger sample size are needed for understanding the structure–function interaction of human SN.

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References

  • Alladi PA, Mahadevan A, Yasha TC, Raju TR, Shankar SK, Muthane U (2009) Absence of age-related changes in nigral dopaminergic neurons of Asian Indians: relevance to lower incidence of Parkinson’s disease. Neuroscience 159:236–245

    Article  CAS  PubMed  Google Scholar 

  • Braak H, Braak E (1986) Nuclear configuration and neuronal types of the nucleus niger in the brain of the human adult. Hum Neurobiol 5:71–82

    CAS  PubMed  Google Scholar 

  • Braak H, Braak E (1991) Neuropathological stageing of Alzheimer-related changes. Acta Neuropathol 82:239–259

    Article  CAS  PubMed  Google Scholar 

  • Braak H, Del Tredici K, Rüb U, de Vos RA, Jansen Steur EN, Braak E (2003a) Staging of brain pathology related to sporadic Parkinson’s disease. Neurobiol Aging 24:197–211

    Article  PubMed  Google Scholar 

  • Braak H, Rüb U, Gai WP, Del Tredici K (2003b) Idiopathic Parkinson’s disease: possible routes by which vulnerable neuronal types may be subject to neuroinvasion by an unknown pathogen. J Neural Transm 110:517–536

    Article  CAS  PubMed  Google Scholar 

  • Cabello CR, Thune JJ, Pakkenberg H, Pakkenberg B (2002) Ageing of substantia nigra in humans: cell loss may be compensated by hypertrophy. Neuropathol Appl Neurobiol 28:283–291

    Article  CAS  PubMed  Google Scholar 

  • Chu Y, Kompoliti K, Cochran EJ, Mufson EJ, Kordower JH (2002) Age-related decreases in Nurr1 immunoreactivity in the human substantia nigra. J Comp Neurol 450:203–214

    Article  CAS  PubMed  Google Scholar 

  • Damier P, Hirsch EC, Agid Y, Graybiel AM (1999) The substantia nigra of the human brain. II. Patterns of loss of dopamine-containing neurons in Parkinson’s disease. Brain 122(Pt 8):1437–1448

    Article  PubMed  Google Scholar 

  • Di Giovanni G, Di Matteo V, Esposito E (2009) Birth, life and death of dopaminergic neurons in the substantia nigra. J Neural Transm Suppl 73:1

    PubMed  Google Scholar 

  • Dickson DW (2012) Parkinson’s disease and parkinsonism: neuropathology. Cold Spring Harb Perspect Med 2(8). doi:10.1101/cshperspect.a009258.1

  • Doraiswamy PM et al (1992) Morphometric changes of the human midbrain with normal aging: MR and stereologic findings. AJNR Am J Neuroradiol 13:383–386

    CAS  PubMed  Google Scholar 

  • Eidelberg D, Galaburda AM (1982) Symmetry and asymmetry in the human posterior thalamus. I. Cytoarchitectonic analysis in normal persons. Arch Neurol 39:325–332

    Article  CAS  PubMed  Google Scholar 

  • Esiri MM (2007) Ageing and the brain. J Pathol 211(2):181–187

    Article  CAS  PubMed  Google Scholar 

  • Fearnley JM, Lees AJ (1991) Ageing and Parkinson’s disease: substantia nigra regional selectivity. Brain 114(Pt 5):2283–2301

    Article  PubMed  Google Scholar 

  • Ferri CP et al (2005) Global prevalence of dementia: a Delphi consensus study. Lancet 366:2112–2117

    Article  PubMed  Google Scholar 

  • Francois C, Percheron G, Yelnik J, Heyner S (1985) A histological atlas of the macaque (Macaca mulatta) substantia nigra in ventricular coordinates. Brain Res Bull 14:349–367

    Article  CAS  PubMed  Google Scholar 

  • German DC, Schlusselberg DS, Woodward DJ (1983) Three-dimensional computer reconstruction of midbrain dopaminergic neuronal populations: from mouse to man. J Neural Transm 57:243–254

    Article  CAS  PubMed  Google Scholar 

  • Glaser EM, Wilson PD (1998) The coefficient of error of optical fractionator population size estimates: a computer simulation comparing three estimators. J Microsc 192(2):163–171

    Article  CAS  PubMed  Google Scholar 

  • Grinberg LT et al (2007) Brain bank of the Brazilian aging brain study group—a milestone reached and more than 1600 collected brains. Cell Tissue Bank 8:151–162

    Article  PubMed  Google Scholar 

  • Grinberg LT et al (2009) The dorsal raphe nucleus shows phospho-tau neurofibrillary changes before the transentorhinal region in Alzheimer’s disease. A precocious onset? Neuropathol Appl Neurobiol 35:406–416

    Article  CAS  PubMed  Google Scholar 

  • Grinberg LT, Rueb U, Heinsen H (2011) Brainstem: neglected locus in neurodegenerative diseases. Front Neurol 2:42

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gundersen HJ, Jensen EB (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147:229–263

    Article  CAS  PubMed  Google Scholar 

  • Haber SN, Behrens TE (2014) The neural network underlying incentive-based learning: implications for interpreting circuit disruptions in psychiatric disorders. Neuron 83:1019–1039

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Haber SN, Fudge JL, McFarland NR (2000) Striatonigrostriatal pathways in primates form an ascending spiral from the shell to the dorsolateral striatum. J Neurosci 20:2369–2382

    CAS  PubMed  Google Scholar 

  • Halliday GM, McRitchie DA, Cartwright H, Pamphlett R, Hely MA, Morris JG (1996) Midbrain neuropathology in idiopathic Parkinson’s disease and diffuse Lewy body disease. J Clin Neurosci 3:52–60

    Article  CAS  PubMed  Google Scholar 

  • Hassler R (1937) Zur Normalanatomie der Substantia nigra. Versuch einer architektonischen Gliederung. J Psychol Neurol 48:1–55

    Google Scholar 

  • Hedreen JC, DeLong MR (1991) Organization of striatopallidal, striatonigral, and nigrostriatal projections in the macaque. J Comp Neurol 304:569–595

    Article  CAS  PubMed  Google Scholar 

  • Heinsen H, Heinsen YL (1991) Serial thick, fronze, gallocyanin stained sections of human central nervous system. J Histotechnol 14:167–173

    Article  Google Scholar 

  • Heinsen H, Henn R, Eisenmenger W, Götz M, Bohl J, Bethke B, Lockemann U, Püschel K (1994) Quantitative investigations on the human entorhinal area: left–right asymmetry and age-related changes. Anat Embryol (Berl) 190(2):181–194

    Article  CAS  Google Scholar 

  • Heinsen H, Arzberger T, Schmitz C (2000) Celloidin mounting (embedding without infiltration)—a new, simple and reliable method for producing serial sections of high thickness through complete human brains and its application to stereological and immunohistochemical investigations. J Chem Neuroanat 20:49–59

    Article  CAS  PubMed  Google Scholar 

  • Heinsen H, Arzberger T, Roggendorf W, Mitrovic T (2004) 3D reconstruction of celloidin-mounted serial sections. Acta Neuropathol 108(4):374

    Google Scholar 

  • Jorm AF (1994) A short form of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE): development and cross-validation. Psychol Med 24:145–153

    Article  CAS  PubMed  Google Scholar 

  • Kouri N, Whitwell JL, Josephs KA, Rademakers R, Dickson DW (2011) Corticobasal degeneration: a pathologically distinct 4R tauopathy. Nat Rev Neurol 7:263–272

    Article  CAS  PubMed  Google Scholar 

  • Kreczmanski P et al (2007) Volume, neuron density and total neuron number in five subcortical regions in schizophrenia. Brain 130:678–692

    Article  PubMed  Google Scholar 

  • Kubis N et al (2000) Preservation of midbrain catecholaminergic neurons in very old human subjects. Brain 123(Pt 2):366–373

    Article  PubMed  Google Scholar 

  • Lynd-Balta E, Haber SN (1994) Primate striatonigral projections: a comparison of the sensorimotor-related striatum and the ventral striatum. J Comp Neurol 345:562–578

    Article  CAS  PubMed  Google Scholar 

  • Lyness SA, Zarow C, Chui HC (2003) Neuron loss in key cholinergic and aminergic nuclei in Alzheimer disease: a meta-analysis. Neurobiol Aging 24:1–23

    Article  CAS  PubMed  Google Scholar 

  • Ma SY, Röyttä M, Rinne JO, Collan Y, Rinne UK (1995) Single section and disector counts in evaluating neuronal loss from the substantia nigra in patients with Parkinson’s disease. Neuropathol Appl Neurobiol 21:341–343

    Article  CAS  PubMed  Google Scholar 

  • Ma SY, Röytt M, Collan Y, Rinne JO (1999) Unbiased morphometrical measurements show loss of pigmented nigral neurones with ageing. Neuropathol Appl Neurobiol 25:394–399

    Article  CAS  PubMed  Google Scholar 

  • McRitchie DA, Halliday GM, Cartwright H (1995) Quantitative analysis of the variability of substantia nigra pigmented cell clusters in the human. Neuroscience 68:539–551

    Article  CAS  PubMed  Google Scholar 

  • Morris JC (1993) The clinical dementia rating (CDR): current version and scoring rules. Neurology 43:2412–2414

    Article  CAS  PubMed  Google Scholar 

  • Mouton PR (2011) Unbiased stereology: a concise guide. The John Hopkins University Press, Baltimore

    Google Scholar 

  • Muthane U, Yasha TC, Shankar SK (1998) Low numbers and no loss of melanized nigral neurons with increasing age in normal human brains from India. Ann Neurol 43(3):283–287

    Article  CAS  PubMed  Google Scholar 

  • Ogisu K et al (2013) 3D neuromelanin-sensitive magnetic resonance imaging with semi-automated volume measurement of the substantia nigra pars compacta for diagnosis of Parkinson’s disease. Neuroradiology 55:719–724

    Article  PubMed  Google Scholar 

  • Pakkenberg B, Møller A, Gundersen HJ, AM Dam, Pakkenberg H (1991) The absolute number of nerve cells in substantia nigra in normal subjects and in patients with Parkinson’s disease estimated with an unbiased stereological method. J Neurol Neurosurg Psychiatry 54:30–33

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Perl DP, Good PF, Bussiere T, Morrison JH, Erwin JM, Hof PR (2000) Practical approaches to stereology in the setting of aging- and disease-related brain banks. J Chem Neuroanat 20:7–19

    Article  CAS  PubMed  Google Scholar 

  • Pujol J, Junqué C, Vendrell P, Grau JM, Capdevila A (1992) Reduction of the substantia nigra width and motor decline in aging and Parkinson’s disease. Arch Neurol 49:1119–1122

    Article  CAS  PubMed  Google Scholar 

  • Rudow G, O’Brien R, Savonenko AV, Resnick SM, Zonderman AB, Pletnikova O, Marsh L, Dawson TM, Crain BJ, West MJ, Troncoso JC (2008) Morphometry of the human substantia nigra in ageing and Parkinson’s disease. Acta Neuropathol 115(4):461–470

    Article  PubMed  PubMed Central  Google Scholar 

  • Schmithorst VJ, Dardzinski BJ, Holland SK (2001) Simultaneous correction of ghost and geometric distortion artifacts in EPI using a multiecho reference scan. IEEE Trans Med Imaging 20:535–539

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schmitz C (1998) Variation of fractionator estimates and its prediction. Anat Embryol 198:371–397

    Article  CAS  PubMed  Google Scholar 

  • Schmitz C, Hof PR (2000) Recommendations for straightforward and rigorous methods of counting neurons based on a computer simulation approach. J Chem Neuroanat 20(1):93–114

    Article  CAS  PubMed  Google Scholar 

  • Schmitz C, Korr H, Heinsen H (1999a) Design-based counting techniques: the real problems (letter). Trends Neurosci 22(8):345

    Article  CAS  PubMed  Google Scholar 

  • Schmitz C, Rüb U, Korr H, Heinsen H (1999b) Nerve cell loss in the thalamic mediodorsal nucleus in Huntington’s disease. II. Optimization of a stereological estimation procedure. Acta Neuropathol 97:623–628

    Article  CAS  PubMed  Google Scholar 

  • Sohmiya M, Tanaka M, Aihara Y, Hirai S, Okamoto K (2001) Age-related structural changes in the human midbrain: an MR image study. Neurobiol Aging 22:595–601

    Article  CAS  PubMed  Google Scholar 

  • Tanner C, Gilley D, Goetz C (1990) A brief screening questionnaire for parkinsonism. Ann Neurol 28:267–268

    Google Scholar 

  • Theofilas P, Polichiso L, Wang X, Lima LC, Alho AT, Leite RE, Suemoto CK, Pasqualucci CA, Jacob Filho W, Heinsen H, Grinberg LT, Group BABS (2014) A novel approach for integrative studies on neurodegenerative diseases in human brains. J Neurosci Methods 226:171–183

    Article  PubMed  PubMed Central  Google Scholar 

  • UN (2013) World population prospects: the 2012 revision, vol Volume I: comprehensive tables ST/ESA/SER.A/336. United Nations

  • van Domburg PH, ten Donkelaar HJ (1991) The human substantia nigra and ventral tegmental area. A neuroanatomical study with notes on aging and aging diseases. Adv Anat Embryol Cell Biol 121:1–132

    Article  PubMed  Google Scholar 

  • West MJ (1993) New stereological methods for counting neurons. Neurobiol Aging 14(4):275–285

    Article  CAS  PubMed  Google Scholar 

  • West MJ, Ostergaard K, Andreassen OA, Finsen B (1996) Estimation of the number of somatostatin neurons in the striatum: an in situ hybridization study using the optical fractionator method. J Comp Neurol 370(1):11–22

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to the families that donated brains for this research. We would also like to thank all the members of the Brain Bank of the Brazilian Aging Brain Study Group. We are grateful to Prof. Dr. Edson Amaro Junior for scientific collaboration and Dr. Eduardo Alho for technical support in 3D reconstruction. We also thank Dr. Janet Johnston (http://www.output.ie) for language editing. Funding sources: Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), Institute for Education and Research of Albert Einstein Hospital, São Paulo Research Foundation (FAPESP), Brazilian National Council for Scientific and Technological Development (CNPq), LIM-22 (HC-FMUSP) for research financial and technical support in Brazil, and the National Institutes of Health, USA (R01AG040311).

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    Authors and Affiliations

    1. Grupo de Estudos em Envelhecimento Cerebral e LIM 22, Department of Pathology, Faculdade de Medicina da Universidade de Sao Paulo, Av. Dr. Arnaldo, 455 sala 1353, São Paulo, CEP 01246-903, Brazil

      Ana Tereza Di Lorenzo Alho, Claudia Kimie Suemoto, Lívia Polichiso, Edilaine Tampellini, Kátia Cristina de Oliveira, Mariana Molina, Glaucia Aparecida Bento Santos, Camila Nascimento, Renata Elaine Paraizo Leite, Renata Eloah de Lucena Ferreti-Rebustini, Alexandre Valotta da Silva, Ricardo Nitrini, Carlos Augusto Pasqualucci, Wilson Jacob-Filho & Lea Tenenholz Grinberg

    2. Labor für Morphologische Hirnforschung der Klinik und Poliklinik für Psychiatrie und Psychotherapie, Institut Rechtsmedizin, Universitätsklinikum Würzburg, Würzburg, Germany

      Ana Tereza Di Lorenzo Alho, Kátia Cristina de Oliveira & Helmut Heinsen

    3. Instituto do Cérebro, Hospital Israelita Albert Einstein, São Paulo, Brazil

      Ana Tereza Di Lorenzo Alho, Glaucia Aparecida Bento Santos & Alexandre Valotta da Silva

    4. Discipline of Geriatrics, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil

      Claudia Kimie Suemoto, Renata Elaine Paraizo Leite, Renata Eloah de Lucena Ferreti-Rebustini & Wilson Jacob-Filho

    5. Department of Neurology, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil

      Ricardo Nitrini

    6. Memory and Aging Center, Department of Neurology, University of California, San Francisco, USA

      Lea Tenenholz Grinberg

    7. Department of Pathology, Faculdade de Medicina da Universidade de Sao Paulo, São Paulo, Brazil

      Carlos Augusto Pasqualucci, Helmut Heinsen & Lea Tenenholz Grinberg

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    1. Ana Tereza Di Lorenzo Alho
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    2. Claudia Kimie Suemoto
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    Correspondence to Lea Tenenholz Grinberg.

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    All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

    This study was approved by the ethical committees _ENREF_23 of the University of Sao Paulo and the University of Würzburg. Written informed consent for brain donation and provision of clinical information was obtained from the next-of-kin.

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    H. Heinsen has contributed equally.

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    Di Lorenzo Alho, A.T., Suemoto, C.K., Polichiso, L. et al. Three-dimensional and stereological characterization of the human substantia nigra during aging. Brain Struct Funct 221, 3393–3403 (2016). https://doi.org/10.1007/s00429-015-1108-6

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