How to use the optical fractionator: an example based on the estimation of neurons in the hippocampal CA1 and CA3 regions of tree shrews

Keuker, Jeanine I.H.; Vollmann-Honsdorf, Gesa K; Fuchs, Eberhard

doi:10.1016/s1385-299x(01)00064-2

Cited by 73 publications

(68 citation statements)

References 29 publications

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“…Cell numbers were separately determined for the left and right hemispheres. A modified unbiased stereology protocol (Keuker et al, 2001) was used to determine the total number of BrdU-labeled cells. After randomly selecting a starting point, every sixth section between Bregma 4.70 and 1.60 mm (Paxinos and Watson, 1986) was analyzed, yielding an average of 10 sections per animal for quantification.…”

Section: Quantification Of Brdu-labeled Cellsmentioning

confidence: 99%

“…The modified unbiased stereology protocol (Keuker et al, 2001) we used has been successful in quantifying BrdU labeling (Malberg et al, 2000;Czéh et al, 2001). Every sixth section throughout the dentate gyrus was examined, yielding a mean of 16 sections per animal.…”

Section: Quantification Of Brdu-labeled Cellsmentioning

confidence: 99%

“…Every sixth section throughout the dentate gyrus was examined, yielding a mean of 16 sections per animal. Stereological quantification requires determining the number of all BrdU-positive cells in a predefined fraction throughout the complete region of interest (Keuker et al, 2001). We used a section sampling fraction of 1/6, and an area and a thickness sampling fraction of one.…”

Section: Quantification Of Brdu-labeled Cellsmentioning

confidence: 99%

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Chronic Social Stress Inhibits Cell Proliferation in the Adult Medial Prefrontal Cortex: Hemispheric Asymmetry and Reversal by Fluoxetine Treatment

Czéh

Müller-Keuker

Ryguła

et al. 2006

Neuropsychopharmacol

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318

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Profound neuroplastic changes have been demonstrated in various limbic structures after chronic stress exposure and antidepressant treatment in animal models of mood disorders. Here, we examined in rats the effect of chronic social stress and concomitant antidepressant treatment on cell proliferation in the medial prefrontal cortex (mPFC). We also examined possible hemispheric differences. Animals were subjected to 5 weeks of daily social defeat by an aggressive conspecific and received concomitant, daily, oral fluoxetine (10 mg/kg) during the last 4 weeks. Bromodeoxyuridine (BrdU) labeling and quantitative stereological techniques were used to evaluate the treatment effects on proliferation and survival of newborn cells in limbic structures such as the mPFC and the hippocampal dentate gyrus, in comparison with nonlimbic structures such as the primary motor cortex and the subventricular zone. Phenotypic analysis showed that neurogenesis dominated the dentate gyrus, whereas in the mPFC most newborn cells were glia, with smaller numbers of endothelial cells. Chronic stress significantly suppressed cytogenesis in the mPFC and neurogenesis in the dentate gyrus, but had minor effect in nonlimbic structures. Fluoxetine treatment counteracted the inhibitory effect of stress. Hemispheric comparison revealed that the rate of cytogenesis was significantly higher in the left mPFC of control animals, whereas stress inverted this asymmetry, yielding a significantly higher incidence of newborn cells in the right mPFC. Fluoxetine treatment abolished hemispheric asymmetry in both control and stressed animals. These pronounced changes in gliogenesis after chronic stress exposure may relate to the abnormalities of glial cell numbers reported in the frontolimbic areas of depressed patients.

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Section: Quantification Of Brdu-labeled Cellsmentioning

confidence: 99%

Section: Quantification Of Brdu-labeled Cellsmentioning

confidence: 99%

Section: Quantification Of Brdu-labeled Cellsmentioning

confidence: 99%

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Chronic Social Stress Inhibits Cell Proliferation in the Adult Medial Prefrontal Cortex: Hemispheric Asymmetry and Reversal by Fluoxetine Treatment

Czéh

Müller-Keuker

Ryguła

et al. 2006

Neuropsychopharmacol

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318

221

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“…The position of the knives to the hippocampus along its axis was random. It was decided randomly for each animal whether the even or the odd slabs, containing hippocampal tissue, were embedded in glycolmethacrylate resin (Technovit 7100; Kulzer, Weinheim, Germany; see also [26]). In short, the selected slabs were washed in double distilled water for 24 h, dehydrated through 70 and 96% ethanol (8 and 16 h, respectively), preinfiltrated with 96% ethanol and base liquid Technovit 7100 (1:1, v/v) for 8 h, and infiltrated with pure base liquid Technovit 7100 with hardener I for 48 h. Then, the tissue slabs were polymerized in Technovit 7100 containing hardener I and II in plastic molding cup trays at 4 • C. After 2 days, the polymerized tissue blocks were mounted with Technovit 3400 (Kulzer).…”

Section: Tissue Preparationmentioning

confidence: 99%

“…Because of the mathematical approach, there is a possibility to optimize the counting procedure during a pilot study, and the total group variance can be split into the biological variance and the variance contribution from the method, e.g. when comparing two experimental groups [23,26,62,67,68].…”

Section: Introductionmentioning

confidence: 99%

Preservation of hippocampal neuron numbers in aged rhesus monkeys

Keuker

Luiten

Fuchs

2003

Neurobiology of Aging

123

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Preservation of hippocampal neuron numbers in aged rhesus monkeys Keuker, J.I.H.; Luiten, P.G.M.; Fuchs, E. Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. AbstractTo investigate whether or not aging of nonhuman primates is accompanied by a region-specific neuron loss in the hippocampal formation, we used the optical fractionator technique to obtain stereological estimates of unilateral neuron numbers of the hippocampi of eight young (0-4 years) and five aged (18-31 years) male rhesus monkeys (Macaca mulatta). Our results show a preservation of neurons (mean × 10 3 ± S.D. × 10 3 ) in the subiculum (young = 588 ± 124, aged = 612 ± 207), CA1 (young = 1051 ± 249, aged = 1318 ± 311), CA2 (young = 100 ± 18, aged = 113 ± 12), CA3 (young = 478 ± 125, aged = 509 ± 139), hilus (young = 337 ± 115, aged = 394 ± 90), and dentate gyrus (young = 5550 ± 1725, aged = 7799 ± 2087) of the hippocampal formation. These results confirm a previous stereological study in rhesus monkeys, but are in conflict with data for humans, showing age-dependent region-specific alterations in the hippocampal formation.

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Preservation of hippocampal neuron numbers and hippocampal subfield volumes in behaviorally characterized aged tree shrews

Keuker

Biurrun

Luiten

et al. 2003

J of Comparative Neurology

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Aging is associated with a decreased ability to store and retrieve information. The hippocampal formation plays a critical role in such memory processes, and its integrity is affected during normal aging. We used tree shrews (Tupaia belangeri) as an animal model of aging, because in many characteristics, tree shrews are closer to primates than they are to rodents. Young and aged male tree shrews performed a holeboard spatial memory task, which permits assessment of reference and working memory. Upon completion of the behavioral measurements, we carried out modified stereological analyses of neuronal numbers in various subdivisions of the hippocampus and used the Cavalieri method to calculate the volumes of these subfields. Results showed that the working memory of aged tree shrews was significantly impaired compared with that of young animals, whereas the hippocampus-dependent reference memory remained unchanged by aging. Estimation of the number of neurons revealed preserved neuron numbers in the subiculum, in the subregions CA1, CA2, CA3, and in the hilus of the dentate gyrus. Volume measurements showed no aging-related changes in the volume of any of these hippocampal subregions, or in the molecular and granule cell layers of the dentate gyrus of tree shrews. We conclude that the observed changes in memory performance in aging tree shrews are not accompanied by observable reductions of hippocampal neuron numbers or hippocampal volume, rather, the changes in memory performance are more likely the result of modified subcellular mechanisms that are affected by the aging process.

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How to use the optical fractionator: an example based on the estimation of neurons in the hippocampal CA1 and CA3 regions of tree shrews

Cited by 73 publications

References 29 publications

Chronic Social Stress Inhibits Cell Proliferation in the Adult Medial Prefrontal Cortex: Hemispheric Asymmetry and Reversal by Fluoxetine Treatment

Chronic Social Stress Inhibits Cell Proliferation in the Adult Medial Prefrontal Cortex: Hemispheric Asymmetry and Reversal by Fluoxetine Treatment

Preservation of hippocampal neuron numbers in aged rhesus monkeys

Preservation of hippocampal neuron numbers and hippocampal subfield volumes in behaviorally characterized aged tree shrews

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