Bente Pakkenberg

Neocortical neuron number in humans: Effect of sex and age

Modern stereological methods provide precise and reliable estimates of the number of neurons in specific regions of the brain. We decided to estimate the total number of neocortical neurons in the normal human brain and to analyze it with respect to the major macro‐ and microscopical structural components, to study the internal relationships of these components, and to quantitate the influence of important physiological variables on brain structure. The 94 brains reported represent a consecutive collection of brains from the general Danish population. The average numbers of neocortical neurons were 19 billion in female brains and 23 billion in male brains, a 16% difference. In our study, which covered the age range from 20 years to 90 years, approximately 10% of all neocortical neurons are lost over the life span in both sexes. Sex and age were the main determinants of the total number of neurons in the human neocortex, whereas body size, per se, had no influence on neuron number. Some of the data presented have been analyzed by using new mathematical designs. An equation predicting the total neocortical neuron number in any individual in which sex and age are known is provided. J. Comp. Neurol. 384:312‐320, 1997.

Marked loss of myelinated nerve fibers in the human brain with age

The white matter is the structure of the brain that declines most with age—almost 30%, but little is known about the age‐effect on the fibers that constitute the white matter. In the present study, the total length of myelinated fibers was measured with a newly developed stereologic method. Specimens came from 36 normal Danes (18 males and 18 females) with an age ranging between 18 and 93 years. Samples were taken systematically and randomly from the white matter, and the biopsy specimens were randomly rotated before sectioning to avoid bias due to the anisotropic nature of nerve fibers. The fibers were counted at light microscopic level at approximately 10,000× magnification, and the diameter of each counted fiber was measured to get the diameter distribution. Males were found to have a total myelinated fiber length of 176,000 km at the age of 20 and 97,200 km at the age of 80, whereas the total length in females was 149,000 km at the age of 20 and 82,000 km at the age of 80. This finding corresponds to a 10% decrease per decade or a total decrease of 45% from the age of 20 to 80 years, and a sex difference of 16%. The fiber diameter distribution showed that primarily the thinner fibers were lost with a relative preservation of the thicker ones. The marked loss of myelinated nerve fibers with age could explain some of the cognitive decline seen in the elderly. J. Comp. Neurol. 462:144–152, 2003.

Aging and the human neocortex.

Neurostereology has been applied to quantitative anatomical study of the human brain. Such studies have included the total neocortical number of neurons and glial cells, the estimated size distribution of neocortical neurons, the total myelinated fiber length in the brain white matter, the total number of synapses in the neocortex, and the effect of normal aging on these structural elements. The difference in total number of neurons was found to be less than 10% over the age range from 20 to 90 years, while the glial cell number in six elderly individuals, mean age 89.2 years, showed an average number of 36 billion glial cells, which was not statistically significantly different from the 39 billion glial cells in the neocortex of six young individuals with a mean age of 26.2 years. The total myelinated fiber length varied from 150,000 to 180,000 km in young individuals and showed a large reduction as a function of age. The total number of synapses in the human neocortex is approximately 0.15 x 10(15) (0.15 quadrillion). Although the effect of aging is seen in all estimated structural elements, the effect of sex is actually higher. The functional relevance of these differences in neuron numbers in both age and gender is not known.

Age-Induced White Matter Changes in the Human Brain: A Stereological Investigation

In the present pilot study, age-related white matter changes were investigated by the use of design-based stereological methods. In the brains of elderly subjects, the total volume of the white matter and the total volume of the myelinated fibers therein were lower than in those of young subjects (15% and 17%, respectively), but the differences were not statistically significant. The total length of the myelinated fibers of the white matter in the elderly group of 86,000 km was, statistically, significantly decreased by 27% compared with 118,000 km in the young group. This loss of the total nerve fiber length was accompanied in particular by a decline of the myelinated fibers with a small diameter. The mean diameter of the myelinated fibers in the young group was significantly smaller than in the old group, but the relative size distributions of the myelinated fiber diameters between the young and old groups were similar. Our findings show that the atrophy of the human white matter during ageing is probably caused by a loss of myelinated fibers with a small diameter.

Neocortical glial cell numbers in human brains.

Stereological cell counting was applied to post-mortem neocortices of human brains from 31 normal individuals, age 18-93 years, 18 females (average age 65 years, range 18-93) and 13 males (average age 57 years, range 19-87). The cells were differentiated in astrocytes, oligodendrocytes, microglia and neurons and counting were done in each of the four lobes. The study showed that the different subpopulations of glial cells behave differently as a function of age; the number of oligodendrocytes showed a significant 27% decrease over adult life and a strong correlation to the total number of neurons while the total astrocyte number is constant through life; finally males have a 28% higher number of neocortical glial cells and a 19% higher neocortical neuron number than females. The overall total number of neocortical neurons and glial cells was 49.3 billion in females and 65.2 billion in males, a difference of 24% with a high biological variance. These numbers can serve as reference values in quantitative studies of the human neocortex.

Aging of the human cerebellum: A stereological study

Cerebella from 19 normal Caucasian males, ages 19–84 years, were studied using stereological methods. Cerebellum was divided into four different regions: the anterior and posterior lobe, the vermis, and the flocculonodular lobe. Total volume of the cerebellar cortex and white matter, cerebellar surface area, total Purkinje and granule cell number, and the distribution of the volumes of the Purkinje cells and their nuclei were estimated in all four regions. The global white matter was reduced by 26% with age; the mean volume of the Purkinje cell body was decreased by 33% with no decrease in the volume of the Purkinje cell nuclei. A tendency towards a 16% total cerebellar volume loss was seen without a concomitant neuron loss. No global Purkinje or granule cell loss was detected with age, total Purkinje cell number being 28 × 106 (coefficient of variation, CV = 0.16) and total granule cell number 109 × 109 (CV = 0.17). However, a significant change was observed with age in the anterior lobe, where a selective 40% loss of both Purkinje and granule cells was found. Furthermore, a 30% loss of volume, mostly due to a cortical volume loss, was recorded in the anterior lobe, which is predominantly involved in motor control. J. Comp. Neurol. 466:356–365, 2003.

Validation of in vitro probabilistic tractography

Diffusion weighted imaging (DWI) and tractography allow the non-invasive study of anatomical brain connectivity. However, a gold standard for validating tractography of complex connections is lacking. Using the porcine brain as a highly gyrated brain model, we quantitatively and qualitatively assessed the anatomical validity and reproducibility of in vitro multi-fiber probabilistic tractography against two invasive tracers: the histochemically detectable biotinylated dextran amine and manganese enhanced magnetic resonance imaging. Post mortem DWI was used to ensure that most of the sources known to degrade the anatomical accuracy of in vivo DWI did not influence the tracking results. We demonstrate that probabilistic tractography reliably detected specific pathways. Moreover, the applied model allowed identification of the limitations that are likely to appear in many of the current tractography methods. Nevertheless, we conclude that DWI tractography can be a precise tool in studying anatomical brain connectivity.

Increased Islet Volume but Unchanged Islet Number in ob/ob Mice

It is important for our understanding of the pancreatic islets to study whether new islets are able to form in the intact pancreas. We developed a new method to determine the total number and the mean volume of the pancreatic islets, and we used this method to study the expansion of the islet mass in ob/ob mice (n = 8), using ob/+ mice (n = 8) as controls. The total islet volume was increased by a factor of 3.6 in ob/ob mice compared with ob/+ mice, whereas, importantly, the total number of islets did not differ among ob/ob mice and ob/+ mice (3,193 +/- 160 islets in ob/ob mice vs. 3,184 +/- 142 islets in ob/+ mice, P = 0.97). The coefficient of variation in the volume distribution of islets was equal in the two groups, showing that in ob/ob mice, the existing islets expand their volume by the same proportion, without a net formation of new islets. We suggest that the pancreatic islets should be considered as anatomically such complex structures that islet neogenesis does not spontaneously occur in an intact pancreas. Cells within the existing islets are presumably the most important sources for islet cell hyperplasia during expansion of the total islet mass.

No global neocortical nerve cell loss in brains from patients with senile dementia of Alzheimer's type

Precise estimates of total neuron numbers in neocortices of 11 women, mean age 82.6 years (range 79-88) with severe senile dementia of the Alzheimers type (SDAT) were compared with similar estimates in 10 cognitively normal women of comparable mean age (84.1 years; range 74-92). The total mean nerve cell number in the SDAT group was 16.9 x 10(9) with a coefficient of variation (CV = SD/mean) = 0.14, whereas mean total neuron number in the control group was 18.1 x 10(9), CV = 0.18. In a material of this size the reduction of 6% in neocortical cell number in the SDATs is neither statistically nor biologically significant. Nevertheless, all patients with SDAT were severely demented, having a mean score of 5.6 on a 1-7-scale of dementia. This contrasts with the nondemented individuals who had lived an independent life at home until shortly before death. The SDAT patients showed a rather consistent reduction in cortical volume by 14%, an atrophy that was solely due to a reduced cortical thickness. In addition, all had multiple neocortical plaques (Bielschowsky silver stain).

Do alcoholics drink their neurons away

Although it is commonly believed that chronic alcohol abuse results in loss of neocortical neurons, this assumption has not been properly tested. We used new stereological techniques to make a precise and unbiased estimate of the total number of neurons in the neocortex of brains obtained at necropsy from 11 chronic alcoholic men and 11 control men. The groups were matched with respect to age and height. Total mean neocortical neuron numbers in the two groups did not differ (alcoholics 23.4 x 10(9), controls 23.2 x 10(9)). Estimation of macroscopic brain volumes showed significant reductions in alcoholics compared with controls of the volume/weight ratios of white matter (11%, p = 0.013) and of archicortex (30%, p = 0.028). The volume of the ventricles in the alcoholic group was enlarged by 26%, but this was not statistically significant. There was no difference in the volumes of the neocortices. Our study confirms that chronic alcoholics lose white matter, and this could provide the basis for their functional impairment. However, the results also suggest that the observed brain damage in the alcoholic group is potentially reversible since preserved nerve-cell bodies might allow lost or malfunctioning axons to re-established and restored to function after prolonged abstinence and/or treatment. By contrast, lost neocortical neurons cannot be replaced.