Non-homogeneous stereological properties of the rat hippocampus from high-resolution 3D serial reconstruction of thin histological sections

doi:10.1016/j.neuroscience.2011.12.055

Neuroscience

Volume 205, 15 March 2012, Pages 91-111

https://doi.org/10.1016/j.neuroscience.2011.12.055 Get rights and content

Abstract

Integrating hippocampal anatomy from neuronal dendrites to whole system may help elucidate its relation to function. Toward this aim, we digitally traced the cytoarchitectonic boundaries of the dentate gyrus (DG) and areas CA3/CA1 throughout their entire longitudinal extent from high-resolution images of thin cryostatic sections of adult rat brain. The 3D computational reconstruction identified all isotropic 16 μm voxels with appropriate subregions and layers (http://krasnow1.gmu.edu/cn3/hippocampus3d). Overall, DG, CA3, and CA1 occupied comparable volumes (15.3, 12.2, and 18.8 mm³, respectively), but displayed substantial rostrocaudal volumetric gradients: CA1 made up more than half of the posterior hippocampus, whereas CA3 and DG were more prominent in the anterior regions. The CA3/CA1 ratio increased from ∼0.4 to ∼1 septo-temporally because of a specific change in stratum radiatum volume. Next we virtually embedded 1.8 million neuronal morphologies stochastically resampled from 244 digital reconstructions, emulating the dense packing of granular and pyramidal layers, and appropriately orienting the principal dendritic axes relative to local curvature. The resulting neuropil occupancy reproduced recent electron microscopy data measured in a restricted location. Extension of this analysis across each layer and subregion over the whole hippocampus revealed highly non-homogeneous dendritic density. In CA1, dendritic occupancy was >60% higher temporally than septally (0.46 vs. 0.28, s.e.m. ∼0.05). CA3 values varied both across subfields (from 0.35 in CA3b/CA3c to 0.50 in CA3a) and layers (0.48, 0.34, and 0.27 in oriens, radiatum, and lacunosum-moleculare, respectively). Dendritic occupancy was substantially lower in DG, especially in the supra-pyramidal blade (0.18). The computed probability of dendrodendritic collision significantly correlated with expression of the membrane repulsion signal Down syndrome cell adhesion molecule (DSCAM). These heterogeneous stereological properties reflect and complement the non-uniform molecular composition, circuit connectivity, and computational function of the hippocampus across its transverse, longitudinal, and laminar organization.

Highlights

▶Novel integration of hippocampal anatomy from dendritic arbors to whole system. ▶Non-homogeneous volume and dendritic occupancy in DG, CA3/CA1 subregions and layers. ▶CA3/CA1 volumetric ratio increased septo-temporally because of a specific change in s.r. ▶Probability of dendrodendritic collision significantly correlated with DSCAM. ▶Complements non-uniform molecular and functional properties of the hippocampus.

Section snippets

Experimental procedure and raw image acquisition

Four male Long–Evans hooded rats (22 day old, 226–237 g; Harlan, Indianapolis, IN, USA) were housed in metal wire-hanging cages at 22–24 °C on a 12 h light/dark cycle (lights on at 0700 h.) with access to ad libitum food and water. Animal care was in accordance with George Mason University and National Institute of Health Guide for the Care and Use of Laboratory Animals guidelines. Animals were sacrificed by decapitation with a guillotine when 45 days old. The brains were quickly removed,

Results

The reconstructed 3D hippocampus model (Fig. 3) spanned approximately 5 mm in the rostrocaudal axis and 7 mm both mediolaterally and dorsoventrally. Along its internal coordinates, the maximum length was approximately 6 mm longitudinally, 6 mm transversally from CA3c to the CA1/subiculum border, and 4 mm transversally from the infra-pyramidal to the supra-pyramidal DG blade tips. As commonly observed in histological preparations, virtual sectioning of the model in any of the three canonical

Discussion

Because of the tight packing of the principal cell layers and the laminar organization of interneurons, fibers, and neuropil, volumetric distributions can place considerable constraints on hippocampal function. Many pathological conditions and animal models are characterized by changes in hippocampus volumes (Wolf et al., 2002a, Lee et al., 2009). Numerous studies have been carried out to estimate hippocampal layer volumes by unbiased stereology (Schmitz and Hof, 2005, Boss et al., 1987,

Author contributions

D.R. carried out the crystatic sectioning, histological experiments, imaging, segmentation, computational framework development and database development, and data analysis. S.E.B. carried out, helped, and advised D.R. in the cryostatic sectioning, cover slipping, and Nissl staining. G.A.A. conceived, designed, and guided the data analysis and 3D hippocampus model development. D.R. and G.A.A. wrote the paper.

Acknowledgments

We are indebted to Dr. Ruggero Scorcioni for valuable help throughout this project. We are also thankful to Todd Gillette, Michele Ferrante, and Sridevi Polavaram for critical feedback on an earlier version of this manuscript. This research was supported by National Institute of Health grants NS39600 and NS058816 and Office of Naval Research MURI grant N00014-10-1-0198 to G.A.A.

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Cognitive, Behavioral, and Systems NeuroscienceResearch PaperNon-homogeneous stereological properties of the rat hippocampus from high-resolution 3D serial reconstruction of thin histological sections

Abstract

Highlights

Section snippets

Experimental procedure and raw image acquisition

Results

Discussion

Author contributions

Acknowledgments

Brain Res Mol Brain Res

Neuron

Brain Res Bull

Prog Brain Res

Biochem Biophys Res Commun

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Curr Biol

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Neuron

Neurobiol Aging

J Neurosci Methods

Neurobiol Learn Mem

Cell

Brain Res

Neuroscience

Cell

Neuroscience

Curr Opin Neurobiol

Neuron

J Neurosci Methods

Neuron

Brain Res

Neurobiol Aging

Prog Brain Res

Curr Biol

Brain Res Dev Brain Res

Neuroscience

A decrease in cell size accompanies a loss of docosahexaenoate in the rat hippocampus

Nutr Neurosci

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Neuroanatomical algorithms for dendritic modelling

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NeuroMorphoOrg: A central resource for neuronal morphologies

J Neurosci

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Philos Trans R Soc Lond B Biol Sci

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Behav Neurosci

Cortex: statistics and geometry of neuronal connectivity

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J Comp Neurol

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Neuroinformatics

Dendrites of classes of hippocampal neurons differ in structural complexity and branching patterns

J Comp Neurol

Comparative electrotonic analysis of three classes of rat hippocampal neurons

J Neurophysiol

Quantitative, three-dimensional analysis of granule cell dendrites in the rat dentate gyrus

J Comp Neurol

Automatic matching of homologous histological sections

IEEE Trans Biomed Eng

Growth of neurites toward neurite–neurite contact sites increases synaptic clustering and secretion and is regulated by synaptic activity

Cereb Cortex

Quantitative analysis of delayed neuronal death in the hippocampal subfields of SHRSP and SHR

Cell Mol Neurobiol

A molecular blueprint of gene expression in hippocampal subregions CA1, CA3, and DG is conserved in the brain of the common marmoset

Hippocampus

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Genomic-anatomic evidence for distinct functional domains in hippocampal field CA1

Proc Natl Acad Sci U S A

From conditioning to conscious recollection: Memory systems of the brain

Cognitive, Behavioral, and Systems Neuroscience
Research Paper
Non-homogeneous stereological properties of the rat hippocampus from high-resolution 3D serial reconstruction of thin histological sections