The integument of the Queensland fruit fly,Dacus tryoni (Frogg.)

Summary Testes of the Japanese freshwater turtle Clemmys japonica Temmnick et Schlegel were fixed in 3% potassium permanganate buffered to pH 7.2 with Veronal-acetate buffer, and thin sections of the tissue, embedded in epoxy Epon resin, were studied under the electron or light microscope.At the early stage of differentiation of the spermatid, the cytoplasm contains a few mitochondria provided with cristae which are oriented transversely or longitudinally. As the differentiation of spermatids proceeds, the mitochondrion has been modified into a cupshaped body with a wall consisting of several concentric layers. Such body has been referred to the mitochondrial lamellar body. The formation of such a body is mainly attributed to the mitochondrial cristae, and subsequently to the membrane system of the endoplasmic reticulum. In a more advanced stage of differentiation, the mitochondrial lamellar bodies appear wrapped around a bundle of tail filaments, and seem to present a very wide surface available for the localization of organized enzyme systems to facilitate the motion of spermatozoa.Prior to the formation of the mitochondrial lamellar bodies, the Golgi apparatus has been reorganized into a peculiar body with a floral appearance, consisting of numerous tubular elements, and revealing to be positive in PAS-reaction. The body has been designated as the tubular body which has never been demonstrated in any spermatogenic cells through animal kingdom.One to three tubules oval in cross section, approximately 430 × 700 Å in diameter, have been found in the nucleoplasm along the longitudinal axis of a greatly elongated, cone-shaped nucleus of the spermatid. The tubules open on the apex surface of the nucleus, but they are not encountered in the acrosome. A possible physiological significance of the tubules has been discussed in view of the function of the acrosomal tubules in the decapod and other species spermatozoa as well as on the basis of the metabolism of nucleus.This study was supported by Grant GM-8327 from the United States Public Health Service.We wish to express our gratitude to Dr. B. A. Afzelius, Wenner-Gren Institute, University of Stockholm, for his valuable suggestion to the present work.     

The occurrence of intramitochondrial granules in nerve cells

Summary The mitochondria of spinal cord motoneurons of normal rats have been studied and compared with those of animals given 2.5 mg/kg of strychnine sulfate. The mitochondria in the motoneurons of the treated animals exhibited opaque granules in the matrix. Such granules were observed only when calcium chloride was present in the fixing media and were not present in any other component of the nervous tissue of the spinal cord. Granules were not seen in the mitochondria of motoneurons of untreated control animals. The possible function of these granules in nerve cell metabolism is discussed.Research supported by USPHS Grant NB 05665, 00690 and AM 08222.Recipient of Career Research Development Award 1-K3-GM 28064.     

Development of P-protein in sieve elements ofMimosa pudica

Summary The P-protein in sieve elements of leaves ofMimosa pudica L. is first discernible as fine fibrous material which forms homogeneous aggregates. Ribosomes, rough endoplasmic reticulum, and dictyosomes with associated vesicles occur in the cytoplasm surrounding the aggregates. The plastids and mitochondria are in a parietal position in the parts of the cell where the nascent P-protein accumulates. In a later stage, the fibrillar material is organized into a three-dimensional system of five- and six-sided elongated compartments. The corners of the compartments appear solid at first, then they become electron lucent in the center and assume tubular form. Aggregates of mature P-protein tubules usually occur near the compartmentalized system. Tubules in pentagonal or hexagonal arrangements may be present in the aggregates and may be partly interconnected. The conclusion was drawn that the P-protein tubules are assembled at the corners of compartments within a continuous orderly system. The fully formed tubules occur first in aggregates, the P-protein bodies. Later the aggregates become loose and partly dispersed. Many of the dispersed tubules assume a loose, extended, helical form characteristic of P-protein in older sieve elements.This work was supported in part by National Science Foundation grant GB-5506. I am also grateful to MissHatsume Kosakai and Mr.Robert H.Gill for technical assistance.     

Mitochondrial Polymorphism inAmoeba proteus

Summary The fine structure of the mitochondria ofAmoeba proteus was studied utilizing several methods of fixation. Different forms of mitochondria were detected within the same ameba following aldehyde fixation. Morphological differences between them were most evident in cells fixed withKarnovsky's glutaraldehyde-formaldehyde mixture. Mitochondria with a dense matrix appeared to be rod-shaped and had tubular cristae of variable width. In comparison, other mitochondria had a lighter matrix, an irregular spherical shape, and narrower more uniform tubular cristae. A small percentage of mitochondria with intermediate characteristics was noted. The results obtained with the various preparative procedures are compared and the possible significance of the morphological types is discussed.     

OBSERVATIONS ON PRISMATIC-TYPE MITOCHONDRIA WITHIN ASTROCYTES OF THE SYRIAN HAMSTER BRAIN

This paper describes a special form of mitochondria which has been observed using the electron microscope in certain astrocytes of the Syrian hamster brain. The interior structural pattern of these mitochondria is characterized by the presence of numerous equilateral prismatic tubules in a highly regular hexagonal arrangement, instead of the customary foliate cristae. Such mitochondria have been designated as "prismatic type," in contradistinction to the more usually encountered crista and tubular types. The findings are compared with the occasional similar findings reported in the literature, and their possible functional meaning is briefly discussed.     

The fine structure of the mouse adrenal X zone

Summary Electron microscopically the adrenal X zone was examined in the fourteen SMA female mice aged 40 and 70 days. At these ages, the X zone showed no signs of degeneration. The X zone cell was somewhat smaller than the permanent cortical cell.The mitochondria in the X zone cell were quite bizarre in shape, provided with tubules or cristae. Many intramitochondrial bodies very similar to the cytoplasmic lipid droplets were found in the X zone. A few lipid droplets and globules were also noticed in this zone. The lipid droplets may possibly be formed within the mitochondria.The light and dark cells were differentiated. For the light cells, scant mitochondria and tubular granular endoplasmic reticulum were characteristic in contrast to the abundant mitochondria and multi-lamellated agranular endoplasmic reticulum in the dark cells. The cellular variety in density was discussed with regard to steroid synthesis.The author wishes to express his sincere appreciation to Prof. H. Tauchi, The 2nd Department of Pathology, Nagoya University School of Medicine, for kind advice, to Dr. M. Hoshino for helpful suggestion, and to Mr. J. Aoki for excellent technical assistance.     

Ultrastructural studies of the bovine Graafian Follicle and corpus luteum

Summary Ovaries were obtained from normal adult dairy cows at all days of the estrous cycle. The largest Graafian follicle and corpus luteum were excised and prepared for electron microscopic study.In the follicle wall, membrana granulosa cells contained granular endoplasmic reticulum and mitochondria with villous or lamellar cristae. The theca interna cells during proestrus and estrus contained ribosomes separated from endoplasmic reticulum. The latter during these periods assumed tubular and tortuous shapes. Mitochondria during these periods assumed rounded shapes, were occasionally cup-shaped, and developed tubular cristae.In the corpus luteum, the large luteal cells during metestrus and diestrus contained an abundance of agranular, tubular, branching membranes of endoplasmic reticulum and Golgi apparatus. Mitochondria were large, with tubular cristae, but smaller mitochondria, with irregular or villous cristae, were also present. Transitional bodies of the latter mitochondria to another form were observed. Cup-shaped and annular mitochondria were present during diestrus. In the small luteal cells, large vesicular membrane formations were present and often associated with lipid bodies. The cells were lipid-laden. Lysosomes and granular bodies were present during luteal regression.The observed features of the granulosa cells are related to protein synthesis, those of the pre-ovulatory theca interna cells and metestrus-diestrus large luteal cells to steroid synthesis, and those of the small luteal cells to lipid storage.This investigation was supported by a General Research Support Grant to the College of Veterinary Medicine, University of Minnesota, and Research Grant No. GM-07009, of the United States Public Health Service. Approved for publication as Scientific Journal Series Paper No. 6344, Minnesota Agricultural Experiment Station. The work reported is taken from the senior author's Ph. D. thesis.     

Pigment,abnormal mitochondria and laminar bodies in human sympathetic neurons

Summary A study was made of the fine structure of neurons in lumbar sympathetic ganglia which were surgically removed from a 52 year old male patient. Some samples were fixed by direct immersion into osmium tetroxide, others were first prefixed with an aldehyde solution. Large vesiculated pigment granules were described, compared with similar particles, previously seen in human and animal autonomic neurons, and their glycogen and lipid-soluble components were tentatively identified. These granules increase numerically during aging, are probably derivatives of lysosomes, and may incorporate both lipofuscin and melanin in the same particle. Mitochondrial changes (enlargement, disappearance of cristae and deposit of opaque central material) in sympathetic perikarya occurred under all conditions of preservation, and were interpreted as hypoxia-induced structures which developed during life. Abnormal aggregates of mitochondria were seen in nerve processes. Laminar bodies, chiefly associated with mitochondria were present, especially, in aldehyde prefixed material. It remained undecided whether these structures occurred during life or were artifacts. Compared with direct osmium tetroxide fixation, prefixation with aldehyde, followed by osmium tetroxide proved to be a superior, but not entirely adequate method of preserving deep layers of neuronal tissue. This investigation was supported (in whole) by United States Public Health Service Grant NB 1879-09, Institute for Nervous Diseases and Blindness.The author is greatly indebted to Dr. Jere W. Lord, Jr., Department of Surgery, New York University Medical Center, for the surgically removed tissues used in this study; he is grateful to Dr. Sanford L. Palay, Department of Anatomy, Harvard Medical School, for suggesting the use of the then unpublished method of prefixation with the aldehyde solution. It is a pleasure to acknowledge the expert skill with which Miss Carmen DeLemos and Mr. Arnlod Cianella assisted in completing this work.     

Tubular cytoplasmic bodies in pancreatic islet B-cells of mice

Summary Tubular bodies of varying length and thickness are found in the cytoplasm of pancreatic islet B-cells of obese-hyperglycemic mice and a few of their lean litter mates. Most of these bodies are elongated with tapered ends. There are also some rounded or peculiarly formed variants. They are composed of numerous small electron dense tubular units, often in parallel arrangement. The tubules are embedded in a moderately dense matrix and their interior shows also moderate density. Smaller or larger electron opaque rounded particles are seen in some of the cytoplasmic bodies. Tubular bodies sometimes occur in association with mitochondria, indicating that they might be derived from these cellular organelles. Though the chemical composition and significance of the tubular bodies still are unknown, mitochondrial changes, possibly related to altered metabolic activity, are suggested to form the basis of their development.This work was supported by grants from the Swedish Medical Research Council (Project No. B69-12X-718-04A).     

Steroidogenic characteristics of the adrenal cortex of the mare studied by electron microscopy

The three concentric zones of the horse adrenal cortex (zonae glomerulosa, fasciculata and reticularis) showed marked interpenetration and exhibited a different relative development according to their position in the gland. Whereas the three cortical zones each had a specific histological structure, the ultrastructure of their cells showed a certain qualitative homogeneity. The differences corresponded essentially to the relative abundance of the constituents which are generally considered typical of steroidogeneous cells: mitochondria with vesicular cristae, smooth endoplasmic reticulum and lipid droplets. Their importance increased progressively from the zona glomerulosa to the zona reticularis. In this zone, the presence of gap and septate-like cell junctions, and mitochondria with vesicular cristae in close proximity to a smooth endoplasmic reticulum with numerous dilated tubules, suggested that steroidogenesis may be the most active. Ultrastructural findings were indicative of only quantitative differences between the steroidogenic capacities of the three zones of the mare adrenal cortex.     

Structural changes occurring in interrenal tissue of the duck (Anas platyrhynchos) following adenohypophysectomy and treatment in vivo and in vitro with corticotropin

Adrenal glands from ACTH-treated intact ducks and chronically adenohypophysectomized ducks showed clear zonation into a subcapsular zone (SCZ) and an inner zone (IZ). Adenohypophysectomy caused ultrastructural changes in the IZ but not in the SCZ cells. These included increases in lipid droplets, changes in mitochondrial cristae from tubular to shelf-like, and changes in the shape of the nuclei from spherical to crenated. These changes were reversed by treatment with ACTH. Also, cells of the IZ, but not the SCZ, of adrenals from intact birds given ACTH showed more SER, more dense bodies, fewer lipid droplets and more prominent Golgi complexes. IZ cells incubated in buffer containing no ACTH developed mitochondria with shelf-like cristae and numerous opaque granules in the matrix. Exposure to buffer containing ACTH caused the mitochondrial cristae to become tubular and the matrix granules either decreased in number or disappeared. The granules could be extracted by incubating sections with chelating agents. The mitochondria in SCZ cells did not respond structurally to the presence of ACTH in the incubation medium but the matrix granules, like those in IZ cells, responded to the presence of chelating agents.     

Cytological differentiation of the interrenal tissue of the Japanese quail,Coturnix coturnix

Summary As reported for several other avian species there are clearly distinguishable subcapsular (SCZ) and inner (IZ) zones of interrenal tissue in the Japanese quail. The SCZ contains large columnar cells (type I) with rounded nuclei, polymorphic mitochondria with shelf-like cristae, and relatively small numbers of lipid droplets. The IZ contains two and possibly three types of cells. Type II consists of large columnar cells with moderately dense cytoplasm containing large numbers of lipid droplets and many rounded mitochondria with tubular cristae. Smooth endoplasmic reticulum (SER) and Golgi apparatus are well developed; coated vesicles occur in the Golgi area and at the cell surface. Type-III cells occur in IZ and especially in its more peripheral areas. They are columnar cells with strikingly clear cytoplasm (in comparison with type II) containing mitochondria with plate-like cristae and tubular SER. Type-IV cells are sparsely distributed in IZ and occur rarely in SCZ. Type IV may be a degenerating phase of type III.After adenohypophysectomy or section of portal vessels type-I cells atrophy somewhat with a decrease in lipid droplets; type-II cells, also atrophy with conspicuous increase in size and number of lipid droplets, enlargement of mitochondria, and gradual disappearance of SER; type-III cells decrease in number whereas type-IV cells increase.After injection of ACTH, type-I cells enlarge and their mitochondria, SER and Golgi apparatus become more conspicuous; there is a decrease in lipid droplets in type-II cells and a development of SER, polysomes and Golgi apparatus; there is also a decrease in lipid droplets and a development of SER in type-III cells after injection of 2IU ACTH and an almost complete disappearance of lipid droplets after 4IU ACTH; type-IV cells increase in number.The investigation reported herein was supported by Scientific Research Grants from the Ministry of Education of Japan to Professor Mikami; and by grants from the Japan Society for the Promotion of Science, the National Science Foundation (USA), and the Graduate School Fund of the University of Washington to Professor Farner     

Unusual mitochondrial ultrastructure in the pig adrenal cortex

Unusually large mitochondria with few cristae were observed in the cells of the boundary layer between the zonae fasciculata and reticularis of the pig adrenal. These mitochondria occasionally contained parallel arrays of beaded filaments which appeared to be composed of repetitive electron opaque particles, measuring 10 to 11 nm in diameter. The possibility that these filaments are arranged in closely packed arrays of tubular structures with a central filament is discussed.     

Ultrastructural Characteristics of Adult Rat Adrenocortical Cells Maintained in vitro With and Without ACTH

Bhattacharyya, T. K., Butler, D. G., Price, C. S. 1980. Ultrastructural characteristics of adult rat adrenocortical cells maintained in vitro with and without ACTH. (Ramsay Wright Zoological Laboratories, Department of Zoology, University of Toronto, and Division of Endocrinology, Toronto Western Hospital, and Department of Medicine, University of Toronto, Canada.) — Acta zool. (Stockh.) 61(1): 9–21. The ultrastructural morphology of adrenocortical cells of adult rats maintained in culture for four days with and without corticotropin (ACTH) was studied in comparison to freshly dissociated cells. Identification of cells belonging to zona glomerulosa, zona fasciculata, and zona reticularis was made on the characteristics of the mitochondria and smooth endoplasmic reticulum (SER), and was confirmed by comparison with cellular layers from intact adrenals. Without ACTH, glomerulosa and fasciculata cells showed disappearance of SER, atrophied Golgi apparatus (GA), and a striking proliferation of granular reticulum. Fascicular cells had a loss in mitochondrial matrix density and the mitochondrial cristae showed a tendency to convert to lamellar glomerulosa-type cristae. Zona reticularis cells were not strikingly altered. Maintenance with ACTH led to increased cell size and islet formation of cortical cells. Glomerulosa cells had normal appearance and fasciculata cells manifested a pronounced development of GA and SER, and a normal configuration of mitochondria. The effects of ACTH on glomerulosa cells suggest a trophic influence of ACTH on these cells in vitro. The alterations observed in fascicular cell mitochondria and SER can be explained in terms of known concepts of steroid biosynthesis and basically agree with the patterns of steroid synthesis observed in these cells previously (Price et al. 1975).     

Ultrastructural alterations in skeletal muscle fibers of streptozotocin-diabetic rats

Summary The ultrastructure of fast-twitch-oxidative-glycolytic (FOG), fasttwitch-glycolytic (FG) and slow-twitch-oxidative (SO) fibers in plantaris and soleus muscles of normal and streptozotocin-diabetic rats was studied. In the diabetic animals, the mitochondria of FOG and SO fibers showed a loss of cristae and an increase in electron-dense granules. There was also an increased number of lipid droplets in close proximity to the mitochondria and the nuclei, and a separation of individual muscle nuclei to form satellite cells. Higher incidences of surface projections and sarcoplasmic splittings at the nuclear region were noticed in SO fibers. The FG fibers showed some disorientation of the T-tubular system. It is concluded that streptozotocin-diabetes has differential effects on the fine structure of the three fiber types of rat skeletal muscle.Supported by USPHS Grant AM 18280-04, Boston University Grant GRS-405-BI, and a grant-in-aid award from Sigma Xi Society     

On the relationship of ultrastructural and cytochemical features to color in mammalian skeletal muscle

Summary The semitendinosus muscle of the albino rat is divided grossly into two clearly distinguishable parallel longitudinal bands, one red (anterior) and the other white (posterior). By using mitochondrial content as a criterion for distinguishing fiber types, it is demonstrated that the red portion of the muscle is composed predominantly of red (52%) and intermediate (40%) fibers, while the white portion consists primarily of white fibers (82%). Red fibers have the smallest and white fibers have the largest average diameter. Ultrastructural characteristics of the three fiber types resemble closely those previously described for the rat diaphragm. Red fibers are rich in large mitochondria with abundant cristae, and possess the widest Z lines. In red fibers, the H-band region of the sarcoplasmic reticulum consists of an elaborate network of narrow tubules. In white fibers, mitochondria are smaller, less numerous, and have fewer cristae; Z lines are about half as wide as in red fibers. In the H-band region of the sarcoplasmic reticulum there is a more compact arrangement of broad more or less parallel tubules. Intermediate fibers are similar to red fibers except that their diameters are larger; mitochondria are somewhat smaller and cristae are less abundant; the width of the Z lines is close to that of white fibers. The consistent difference in Z line width establishes this dimension as an important criterion for distinguishing fiber types and facilitates ultrastructural identification, especially of the intermediate fiber.The clear relationship between color of the semitendinosus and cytological features of its component fibers supports the use of the terms red, white, and intermediate as simple and valid designations for fiber types in mammalian skeletal muscle. Measurement of the cross-sectional area contributed by each fiber type to the total area indicates that both red and intermediate fibers may contribute to redness in mammalian skeletal muscle.An early portion of this work was carried out with MissSharon Whelan (Mrs.Bernard Weiss). The author acknowledges the important contribution of Mr.Richard Stearns through his skillful work on the photographic illustrations and the technical assistance of MissAnn Campbell and Mrs.Joan Normington. — This study was supported by Grant No. HD 01026-04 from the United States Public Health Service.     

Distribution and structure of the adrenocortical homologue in the garpike

The microanatomy of the yellow corpuscles (adrenocortical homologue, AH) in the holostean fish, Lepisosteus spp. was studied by serial sectioning, steroid histochemistry, and electron microscopy. The modification of this tissue to short-term ACTH treatment was also observed. The distribution of the AH within the renal tissue of the garpike phylogenetically represents a more advanced condition than that seen in its closest holostean relative, the bowfin, and appears to approximate that in teleosts. The homology of this tissue of vertebrate adrenocortical tissue was established by the positive identification of the enzyme, gamma 5-3 beta-hydroxysteroid dehydrogenase, and by the ultrastructural features of the cells before and after ACTH administration. The AH cells possess fine structural features characteristic of steroidogenic cells, namely, polymorphic mitochondria with tubular cristae, abundant tubules of smooth endoplasmic reticulum, a prominent Golgi complex, and lipid droplets. Other interesting features include the presence of annulate lamellae and a variety of dense bodies. Digitonin perfusion results in the deposition of presumed, cholesterol-digitonide crystalline spicules on the surface microplicae of the cells and as dense accumulations in association with smooth endoplasmic reticulum. ACTH administration results in swelling of mitochondria, a loss of their cristae, and a smooth decrease in electron density of their matrices. Alterations also occur in the smooth and rough endoplasmic reticulum, and large osmiophilic inclusions of irregular profile appear. Some of the ACTH-induced modifications are similar to those observed in the adrenocortical cells of other vertebrate groups following comparable stimulation.     

Crystalloid structures within giant mitochondria of the prothoracic glands of Spodoptera littoralis (Bois) (Lepidoptera)

The fine structure of the prothoracic glands of Spodoptera littoralis was investigated during the first half of the last larval instar. The secretory cells have two types of mitochondria, micromitochondria and macromitochondria. The micromitochondria have rounded to elongated profiles and sometimes branch. They contain lamellar, tubular and also tubulo-vesicular cristae. The macromitochondria appear generally rounded or oval and possess tubular cristae. Many regular parallel membranes appear within macromitochondria. Favorable sections show tubular structures packed in honeycomb fashion. The mitochondrial cristae are in connection with the tubular structures. Honeycomb and parallel membranes increase in number as the size of the macromitochondria increases.     

A correlated thin-section and freeze-fracture analysis of guinea pig adrenocortical cells

Comparison of the fine structural features of guinea pig adrenocortical cells as seen in thin sections with those revealed by freeze-fracture confirms the structural appearance of steroid-secreting cells as interpreted from thin sections and reveals significant new features of the membranous organelles. Smooth-surfaced endoplasmic reticulum appears as a network of tubules, interwoven or in parallel, and as cisternae, fenestrated and non-fenestrated. These elements are tightly packed in the deeper cortical cells, excluding other organelles from their domain. Tubules and fenestrated cisternae possess randomly distributed intramembranous particles on their PF faces, while closely packed non-fenestrated cisternae possess aggregates of particles interspersed with aparticulate regions on their PF faces. These differences in particle distribution suggest functional specialization among the various forms of reticulum. Mitochondria appear as elongated structures of varying shape. Freeze-fracture reveals that all their cristae have circular origins from the inner membrane. Sinuous tubules, which appear as tubules in section, and straight tubules, which appear as lamellae in section, arise from single sites. Flattened sac-like cristae may have multiple circular origins. Definite contact points seen between inner and outer membranes may facilitate passage of molecules, including steroids, into the mitochondrial compartments. Lysosomes and peroxisomes, which are easily identified in thin sections with the aid of cytochemistry, are difficult to identify with certainty by freeze-fracture. Single membrane-bound granules of slightly smaller diameter than mitochondria may represent lysosomes. Smaller granules interconnected with the tubular reticulum, as well as dilated regions of this organelle, may represent peroxisomes. Plasma membranes show no indication of tight junctions but do have abundant gap junctions which show a zonal differentiation: small gap junctions throughout the cortex, medium-sized regularly shaped gap junctions in zona fasciculata externa, and large irregular gap junctions in zona fasciculata interna and zona reticularis. The large junctions cover planar areas as well as surfaces of projections of one cell into another. Such junctions may allow passage of ions as well as of low-molecular-weight substances between the cells, facilitating or even amplifying the response to trophic hormone stimulation.     

Functional differences between two structurally distinct types of steroidogenic cell in the avian adrenal gland

Summary There are two regions of steroidogenic cells in the duck adrenal gland. An outer, subcapsular zone (SCZ), consisting of cells with irregularly shaped nuclei, shows relatively little smooth endoplasmic reticulum and mitochondria with shelf-like cristae. This region surrounds the inner zone (IZ) of the gland which is comprised of smaller cells with rounded nuclei, a more abundant smooth endoplasmic reticulum and mitochondria with tubular cristae. When samples of tissue from these distinct regions of the gland are superfused in vitro with media containing concentrations of 1–24 ACTH ranging from 100 to 1000 ng per ml (0.034 to 0.34 M) the steroidogenic cells in both zones release corticosterone in a dose-dependent manner. The dose-responsiveness of both the SCZ and the IZ cells over this range is a complex quadratic function of the 1–24 ACTH concentration in the medium and the semilogarithmic linear portions of the dose-response curves are restricted to a narrow midrange of ACTH concentrations. Throughout the dose-response range, however, the steroidogenic cells of the IZ are more responsive to corticotropic stimulation than are the cells of the SCZ. The cells of the two zones are further distinguished by their responses to a challenge for a second time with medium containing 1–24 ACTH; the responses of the IZ cells to a second challenge were greater than those of the SCZ cells, and at a high concentration of ACTH the SCZ slices showed no significant second response.This work was supported by a grant from the National Science Foundation (PCM 79-15777) to James Cronshaw and W.N. Holmes