In order to try to determine whether differentiated olfactory receptors turn over (die and are replaced by newly differentiated cells) during adult life, mice were injected with a single dose of 3H-thymidine at either 2 or 4 months of age and allowed to survive for up to 12 months; they were caged in a laminar flow unit to prevent rhinitis. Counts of labeled receptor cells detected autoradiographically after injection at 2 months of age revealed that, following an initial decrease from 1 to 3 months of survival, numbers of labeled cells remained approximately constant, at least up to 12 months of survival. Cells still labeled at 12 months of survival were confirmed as receptor cells by electron microscopic examination of reembedded sections. The hypothesis is suggested that in the absence of disease-related destruction of the olfactory epithelium, most or all receptor cell turnover represents newly formed cells that fail to establish synapses with the olfactory bulb; fully differentiated receptor cells may be quite long-lived.

Two regions closely linked synaptically (olfactory epithelium and olfactory bulb) have been compared in an age-graded series of rats. Previous findings of growth and atrophy of constituent elements in Sprague-Dawley Wisconsin (SD) rats have now been confirmed in Charles River (Crl) rats. In the olfactory bulbs of Charles River (Crl) rats, the volume of layers, the number of olfactory axodendritic synapses in the glomeruli, the total volume of glomerular dendrites, and the size of mitral cell bodies all approximately double between 3 and 24--27 months, and then all decrease by 36 months. Unlike SD rats, however, no loss in the number of mitral cells occurs in Crl rats, and the increase in volume of the olfactory bulbs from 3 to 24 months is approximately double that of SD rats. In the olfactory epithelium the total number of septal olfactory receptors more than doubles between 3 and 18--24 months and then declines markedly, as does the volume of olfactory axons in olfactory bulb glomeruli. Comparison of the regression lines for change in number of septal receptors with that of the size of mitral cell bodies discloses that the decline in number of receptors begins several months earlier than the decline in mitral cell size. This suggests that the atrophic changes in the olfactory bulb may in part be secondary to changes in the receptors of the olfactory epithelium. Numbers of synapses in the glomeruli appear to decline less markedly with age than the number of receptors, and a significant increase in number of synapses per receptor occurs in the oldest group studied (33 months), suggesting a compensatory increase in the relative number of synapses per receptor in the surviving receptors.

Earliest origins of macrophage populations in the central nervous system, the liver, and the lungs were studied in rat embryos aged between 10.5-11 days and 14 days of gestation, based on light and electron microscopic identification of macrophages using peroxidase-coupled isolectin B4 of Griffonia simplicifolia (GSA I-B4), which recognizes alpha-D-galactose groups on the cell membrane. During embryonic life macrophages and their precursors are GSA I-B4-positive and generally bereft of peroxidase-positive granules. At 10.5 days the yolk sac and embryonic circulations have just become joined, the brain has five vesicles but nerve cells are little differentiated, the liver exists as a diverticulum of the gut with fingerlike extensions of hepatocytes, and the lungs as a laryngotracheal groove. Macrophages and/or their precursors occurred in small numbers in embryonic mesenchyme and blood vessels but showed no special affinity for either liver or lung rudiments. The developing brain was the first organ to be colonized, beginning on prenatal day 12. The liver followed between days 12 and 13 and was succeeded by the lungs, beginning between days 13 and 14. Dividing macrophages were present in these organs at the outset of colonization and throughout the duration of the embryo series, indicating that from the beginning, replication of resident cells contributes to growth of the local population. Granulocyte precursors were first apparent in the liver around day 13; they are also GSA-positive but are distinguished from macrophages by their content of peroxidase-positive granules. Organ cultures of 13-day liver and lungs, and 14-day brain tissue, indicate that whereas isolated liver fragments support the formation of both granulocytes and macrophages, only the latter develop in brain or lung cultures. A resident population of macrophages evidently is set up very early in these organs, well before white cells colonize the spleen, bone marrow, and other future blood forming regions. The events outlined are seen as stages in an embryo-wide process that leads to establishment of macrophage populations in various organs.

The population dynamics of internal granule cells in the rat olfactory bulb during adult life were analyzed in histological sections and in autoradiograms with (1) counts of granule cells, (2) counts of labeled granule cells 1 month after injection of 3H-thymidine at various ages, and (3) counts of labeled granule cells at varying survival times (up to 18 months) after injection at 3 months and 24 months. The total number of granule cells increases linearly throughout life, approximately doubling between 3 and 31 months. Autoradiographic studies show that the rate of production of new granule cells decreases from 3 to 12 months and then is approximately constant during the rest of the life span. The number of labeled cells found 6 months after injection at 3 and 24 months is about one-fourth and one-half, respectively, that of the number at a 1-month survival, suggesting that many of the cells produced to do not survive. However, at least some granule cells labeled at 3 months survive for 18 months. A model is suggested in which granule cells are produced continuously throughout life and control of the total number of granule cells is effected chiefly through the rate of cell death.

A quantitative study of the rat olfactory bulb during aging was carried out by directly measuring or calculating the following parameters at 3, 12, 24, 27, and 30 months: The volume of the glomerular, external plexiform, and internal granular layers, a relative measure of the size of the olfactory nerve layer, the mean volume of mitral cell nuclei and perikarya, a relative measure of the mean volume of the mitral cell dendritic tree as well as the total length and mean cross-sectional area of its constituent dendrites, and number of mitral cells. In addition, measurements of the size and number of mitral cells in the accessory olfactory bulb were performed. Data were analyzed with analysis of variance, multiple range tests for differences means at the various ages, and simple, partial, and multiple product-moment correlations. From 3 to 24 months a linear increase of approximately 50% occurs in all layers of the olfactory bulb. During this time the mean perikaryal volume and dendritic volume of mitral cells increases, also in a linear fashion, approximately 100%. No significant change occurs in the number of mitral cells. From 24 to 30 months a significant decrease occurs in the volume of the layers. Although the total volume of mitral cell dendritic trees decreases slightly from 24 to 27 months, the volume of individual mitral cell dendritic treess, as well as perikaryal and nuclear size, increases sharply during this period, apparently in compensation for a sharp decrease in the number of mitral cells which occurs at this time. From 27 to 30 months no further decrease in mitral cell number occurs, but the size of mitral cell perikarya, and especially dendritic trees, decreases sharply. The coordinated increase in olfactory bulb size from 3 to 24 months appears to be a comtinuation into adult life of earlier postnatal increases. The atrophy from 24 to 30 months appears not to be associated with peripheral rhinitis, since the glomerular and olfactory nerve layers do not show greater atrophy than the other layers, and atrophy also occurs in the accessory olfactory bulb, which is supplied by nerves from the vomeronasal organ, a structure not normally subject to rhinitis.