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Atrophy of the adrenal gland induced by thyroparathyroidectomy (TPX) is attributable primarily to narrowing of the zona fasciculata. But absolute volumes of the adrenal gland and zona fasciculata, not the zona glomerulosa and zona reticularis, were decreased significantly 7 weeks after TPX. Changes consistent with atrophy were also seen in the zona fasciculata by electron microscopy. A slight decrease in volume of cytoplasm in cells of the zona fasciculata was evident 2 weeks after TPX as shown by quantitative stereological techniques. Seven weeks after TPX, there was a significant decrease in average cytoplasmic volume as well as mitochondrial volume and surface density of mitochondrial membranes. A decrease in total volume of the zona fasciculata, as well as volume per cell of mitochondria, correlates directly with decreased production of corticosterone in response to stress in TPX rats. Parathyroidectomy (PX) had no effect on the volume of the adrenal gland, on the volume of the individual cortical zones, or on the volume per cell of cytoplasm or mitochondria; these observations indicate that changes in TPX rats are due primarily to ablation of the thyroid gland.

The average cell volume of rat adrenocortical zona fasciculata cells, determined using three different stains on semifine sections, was compared to that of dissociated, unfixed zona fasciculata cells. The maximum number of nuclei per area (NA) was obtained by counting nuclear profiles in semifine sections from nonosmicated adrenocortical tissue stained with toluidine blue. In osmicated adrenocortical tissue on the other hand, fewer nuclear profiles were seen and more of the small size classes were missing from the distribution of profiles. The skewness of the nuclear-profile distribution was much greater in osmicated than in nonosmicated tissue, reflecting the missing small profiles. Although the average diameter of nuclei (D) in nonosmicated tissue was smaller than that for osmicated tissue, profiles were in all likelihood more readily identified because they stained quite intensely. Three different methods for correcting nuclear-size profile distribution and determining nuclei per volume (NV) (Giger-Riedwyl, Cruz-Orive, and Weibel-Gomez) were generally in agreement for nonosmicated tissue. Considerable shrinkage occurred during preparation for electron microscopy, with the greater change occurring in nonosmicated tissue. Cell volume in osmicated tissue corrected for shrinkage varied considerably among the different methods, producing significantly greater values for cell volume than those in nonosmicated tissue. The cell volume of nonosmicated tissue, corrected for shrinkage during processing and for missing small nuclear profiles, did not differ significantly from that of freshly dissociated zona fasciculata cells. The one exception was nonosmicated tissue stained by Feulgen-methylene blue and corrected for missing small nuclear profiles by the method of Cruz-Orive. Use of nonosmicated adrenocortical tissue in stereological studies with an appropriate correction for shrinkage for each experiment is recommended for determining cell volume, although osmicated tissue should also be included to facilitate identification of intracellular membranes when electron microscopy is required.