Der Geschlechtsdimorphismus der Nebennierenrinde der Ratte

Zusammenfassung An 260 männlichen und weiblichen Sprague-Daweley-Ratten vom 0. Lebenstag bis zur 12. Lebenswoche wird die geschlechtsdifferente Entwicklung der Nebennierenrinde histologisch und histometrisch verfolgt.Von der 4. Lebenswoche an ist die weibliche Fasciculatazelle durch ein helles, fein-granuläres Cytoplasma ohne distinkt abgrenzbare Fettvakuolen ausgezeichnet, während beim Männchen in einem kompakten Cytoplasma zunehmend größere, deutlich begrenzte Vakuolen auftreten. Im Blindversuch gelingt aufgrund dieser Kriterien die Geschlechtsdiagnose mit großer Treffsicherheit.Nach histometrischen Untersuchungen sind die weiblichen Fasciculatakerne größer als die männlichen. Bei größerem Zellvolumen ist die Kernzahl/Flächeneinheit beim Weibchen von der Pubertät an kleiner. Die Glomerulosa und die inneren Rindenschichten zeigen keine Geschlechtsdifferenzen. Fermenthistochemisch ist eine Geschlechtsdifferenz nicht nachweisbar.Die relative Zellhypertrophie in der Fasciculata der weiblichen Nebennierenrinde ist das morphologische Äquivalent eines geschlechtsdifferenten Steroidmetabolismus bei der Ratte.

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Sex differences in the adrenal cortex of the ratLight microscopic and histometric studies

Summary In 260 male and female Sprague-Dawley CD rats the sex dependent development of the adrenal cortex was studied histologically and by histometric procedures.Beginning in the 4th week of life the parenchymal cells in the fasciculata of the female animals are characterized by a light, finely granular cytoplasm without distinct lipid vacuoles. In the males, however, the cells show a dense, compact cytoplasm with clearly visible vacuoles increasing in size. In a blind test the sex diagnosis can be established with great accuracy.Histometrically the nuclei in the female fasciculata cells are larger than in the males. Furthermore, caused by a larger cell volume the number of nuclei per square unit is smaller in postpuberal females.The glomerulosa and the inner parts of the cortex do not show any sex differences, nor does enzyme histochemistry.The relative cellular hypertrophy in the fasciculata of the female adrenal cortex represents the morphologic equivalent of the sexual differences in the rats steroid metabolism.

Die Untersuchungen wurden mit Hilfe der Deutschen Forschungsgemeinschaft durchgeführt. — Für die sorgfältige und unermüdliche technische Mithilfe danken wir Frau B. Kubatsch.     

Autoradiographische Untersuchungen zum Wachstum der Nebennierenrinde der Ratte

Zusammenfassung Anhand des Verlaufs des ³H-Index und der Mitoserate in Anhängigkeit vom Lebensalter wurde die Proliferationsaktivität in den Zonen der Nebennierenrinde untersucht. 84 SPF-Ratten erhielten 2 Ci/g ³H-Thymidin i.p.; der Untersuchungszeitraum erstreckte sich vom 18. Trächtigkeitstag bis zur 12. Lebenswoche. Alle Zonen zeigten in der Rangfolge Glomerulosa — Fasciculata — Reticularis eine Abnahme DNS-synthetisierender Zellkerne. Geschlechtsspezifische Unterschiede im Ausmaß der Proliferationsaktivität konnten zu keiner Zeit nachgewiesen werden. Aus der Dissoziation der Kurven des ³H-Index und der Mitoserate bei vergleichbaren DNS-Syntheseraten wird auf eine Änderung der G₂-Phase des Generationszyklus der NNR-Zellen in der teilexponentiellen Wachstumsphase geschlossen.

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Radioautographic studies on the growth of the adrenal cortex in rats

Summary In 84 SPF-rats the poliferative activity of the adrenal cortical cells was studied from the 18th day of pregnancy up to 12 weeks post partum. Rats were given 2 C/g tritiated thymidine and killed 1 hour thereafter. It was shown that there was no sex-related difference in the degree of proliferation that could explain the sexual dimorphism in adrenal weights. In all cortical zones a decrease in the number of labeled cells was seen during the obvservation period. The highest percentage of labeled cells was found in the glomerulosa. There exists no parallelism of the mitotic and labeling index, which gives evidence for a change in length of the G₂-phase of the cell generation cycle.

Herrn Fleissig und Herrn Kürsamer danken wir für Ihre Mithilfe bei der technischen Durchführung der Experimente.     

Investigations on the turnover of adrenocortical mitochondria

Summary The effects of chronic administration of ACTH (up to 36 consecutive days) on the mitochondria of the zona reticularis of the rat adrenal cortex were investigated by stereologic techniques. It was found that ACTH induces two phases of hypertrophy of mitochondria alternating with two proliferative stages, which are associated with a significant decrease in the average volume of the organelles. It is suggested that, as in the zona fasciculata, ACTH controls the processes of growth and division of mitochondria in the zona reticularis. The mechanism underlying this action of ACTH as well as the differences between the responses to ACTH of the mitochondrial population of the two adrenal zones are discussed in the light of evidence indicating that mitochondria contain a complete genetic apparatus largely independent of nuclear control.The authors wish to thank Miss A. Coi and Mr. G. Gottardo for their excellent technical assistance. This work was partly supported by a contract with the CNR (C.T. 73.00663.04)     

Primary tissue culture of human adult adrenocortical cells. Methodology and electron microscopic observations on ACTH-deprived and ACTH-treated cortical cells

Summary A method of primary tissue culture involving both disaggregation of cells by repeated exposure of small tissue fragments to a solution of trypsin, collagenase and hyaluronidase and explantation of the residual tissue fragments intermingled with isolated cells onto polyethylene discs, has been shown to be adequate for the prolonged maintenance (up to 30 days) in vitro of cells arising from decapsulated adult human adrenocortical tissue. The technique and its critical points are discussed.Adrenocortical cells were organized both as outgrowing columns from microexplants or as variously sized islets of monolayered cells. The ultrastructural features of ACTH-deprived adrenocortical cells (i.e., mitochondria with laminar cristae, endoplasmic reticulum mainly consisting of rough profiles, abundance of lipid droplets and -glycogen particles) suggest that the cells dedifferentiate and retain practically no steroidogenic activity.After 2 days of ACTH-treatment, cultured parenchymal cells were found to be quite similar to the zona fasciculata elements of the normal human adrenal cortex. They were grouped in islets of about 50–100 cells. Rough endoplasmic reticulum had decreased, but smooth endoplasmic reticulum showed focal proliferation. The pleomorphic mitochondria with laminar cristae, transformed into a homogeneous population of round or ovoid mitochondria containing tubulo-vesicular cristae. Lipid droplets and glycogen particles were decreased in number.After 7 days of daily treatment with ACTH, the cortical elements, whose nucleus and cytoplasm seemed to be enlarged, were arranged in clusters formed by up to 300 monolayered elements, in which dividing cells were consistently observed. Their cytoplasm was filled with a meshwork of smooth reticulum tubules, in which scantly ribosome-studded profiles and occasional small stacks of granular cisternae were embedded. Mitochondria were similar to those of the 2 days ACTH-treated cultures. Lipid droplets and glycogen particles were absent. The functional significance of these structural changes as well as the possible mechanism underlying the differentiative effect of ACTH are discussed.Primary cultures of human adult adrenals are proposed as a new tool for studies into the physiopathology of the adrenocortical cells under carefully controlled experimental conditions.mis|It is a pleasure to acknowledge our thanks to Drs. F. Mantero and C. Eccher for kindly supplying the normal human adrenocortical tissue. Thanks are also due to Mr. G. Gottardo for his excellent technical assistance.mis|This work was partly supported by a contract with the CNR-Italy (C.T. 73.00663.04).A preliminary report on part of this work was given at the Annual Meeting of the European Tissue Culture Society, Genua, May 1974, and at the Annual Meeting of the Société Française de Microscopie Electronique, Rennes, May 1974, and published as short communication in The Journal of Endocrinology 63, 247, 1974.     

Die Feinstruktur der interstitiellen Grundsubstanz im inneren Nierenmark der Ratte bei verschiedenen Fixierungsarten

Zusammenfassung Das Interstitium des inneren Nierenmarkes von Ratten wurde nach Anwendung verschiedener Fixierungsarten und Versuchsanordnungen elektronenmikroskopisch untersucht. Neben Bindegewebszellen und kollagenen Fibrillen enthält der interstitielle Raum bandförmige, basalmembranartige Strukturen, in deren weiträumigen Maschen zwei weitere Komponenten, nämlich ein feingranuläres und feinfilamentäres Material sowie kontrastreiche, globuläre Partikel liegen. Diese beiden Formelemente stellen sich bei den verschiedenen Fixierungsarten und Versuchsanordnungen unterschiedlich dar. Bei der feingranulären und feinfilamentären Komponente handelt es sich wahrscheinlich um interstitielles Blutplasma. Diese Deutung wird gestützt durch die morphologische Ähnlichkeit mit dem Blutplasma der Blutgefäße, die unterschiedliche Darstellbarkeit nach Glutardialdehyd-und Formalinfixierung sowie durch die Befunde nach Gefäßperfusion und Nierenvenenstauung. Das Verhalten der globulären Partikel nach OsO₄-Einwirkung steht in Einklang mit der schon früher gegebenen Interpretation als präzipitierte saure Mukopolysaccharide. Ihr elektronenoptisches Bild wird im Vergleich zur bekannten makromolekularen Struktur der Mukopolysaccharid-Protein-Moleküle diskutiert.

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The ultrastructure of the interstitial ground substance in the inner medulla of the rat kidney investigated after various fixations

Summary The interstitial ground substance in the inner medulla of the rat kidney is examined electron microscopically after fixation with various fixation agents and under different experimental conditions. The papillary interstitium contains besides interstitial cells and few collagen fibrils basement membrane like material and two other components, fine granular-fibrillar elements and globular particles with heavy contrast. The appearance of the two latter interstitial components varies according to the different fixation agents used and to the experimental conditions. The fine granular and fine fibrillar material surrounds the globular particles numerously after glutardialdehyde-fixation but scarcely after formaldehyde-fixation. It increases after renal vein occlusion, but is diminished or absent, if the kidney is rinsed bloodfree. These results allow to conclude that this material represents interstitial bloodplasma. The behaviour of the globular particles after OsO₄-fixation agrees well with the earlier interpretation according to which they represent precipitated acid mucopolysaccharides. Their electron microscopic appearance is discussed in comparison with the known macromolecular structure of the acid mucopolysaccharides.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Über die Glia im Hypoglossuskern der Ratte nach Axotomie

Zusammenfassung Es wird unter besonderer Berücksichtigung der Glia über den Feinbau des Hypoglossuskerns normaler Albinoratten und nach Axotomie berichtet. Untersucht wurden Tiere vom 12.–90. Lebenstag.Folgende Ergebnisse sind neu: Weder bei jungen noch bei erwachsenen Ratten kommt im Hypoglossuskern Mikroglia vor. Etwa ab. 3. Tag nach Axotomie kann bei erwachsenen Tieren Mikroglia nachgewiesen werden. Sie entsteht aus aktivierten und proliferierten Oligodendrocyten und pericytenähnlichen Bindegewebszellen. Die Mikroglia umhüllt die Nervenzellen. Eine Aktivierung und Hypertrophie von Astrocyten erfolgt etwa ab 8. Tag nach Axotomie. Von diesem Tage an drängen sich Astrocytenfortsätze zwischen Mikroglia und Nervenzelloberfläche. Die Mikroglia beginnt zu degenerieren. Bei Jungtieren bewirkt die Axotomie nur geringgradige Veränderungen. Schon beim Normaltier findet man aktive Oligodendroglia und Astrocyten sowie häufig pericytenähnliche Bindegewebszellen.

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Glia in the hypoglossal nucleus of the rat following axotomy

Summary The ultrastructure of the Nucleus hypoglossi in normal albino rats and following axotomy is described with special reference to the glia. Animals 12 to 90 days old were investigated. The following results are new: Normally in young and adult rats microglia is absent in the nucleus. Following axotomy from the 3rd day no microglia can be demonstrated in adult rats. It originates from activated and proliferated oligodendrocytes and pericyte-like cells of the vascular connective tissue. These cells surround the neurons. An activation and hypertrophy of astrocytes is visible from the 8th postoperative day on. Astrocyte processes make contacts with the perikarya and separate the microglia from the neurons. Microglia cells then show signs of degeneration. In young animals alterations following axotomy are only sparse. Already normally activated oligodendrocytes and astrocytes as well as pericyte-like connective tissue cells can be seen.

Herrn Prof. Schiebler zum 50. Geburtstag gewidmet.     

Tissue culture of rat adult decapsulated adrenal glands

Summary A method of cultivation involving both repeated trypsinisations (at room temperature) and explantation of the tissue fragments on polythene discs has been shown to be apt to the growth in vitro of rat adult decapsulated adreno-cortical tissue. This is the first time that the successful cultivation of such a tissue is reported. The technique and its applications are discussed.The effects of _1–24 corticotrophin (ACTH_1–24) on the rat adult adrenal cultures have been examined by both electron microscopy and autoradiography. Zona fasciculata and reticularis cells grown in the absence of ACTH for long terms (15–16 days) survive and proliferate as dedifferentiated elements. If ACTH_1–24 is added to the cultures, adrenocortical cells will, within 2 days, simultaneously increase their proliferation rate and differentiate. After 7 days of treatment, cortical cells exhibit not only fully differentiated but even hypertrophic morphologic features. Significant stimulations of adrenal DNA, RNA and gross protein synthesis have been found to take place at different times after the starting of the ACTH_1–24 treatment. These data are discussed in relation to the findings previously reported in literature.Rat adult adrenal gland tissue cultures are proposed as a non-previously available tool for investigations into the physiopathology of the adrenal cells to be carried out in a carefully controlled environment.A Preliminary report on part of this material was given at the annual meeting of the Société Française de Microscopie Elécronique, Nantes, May 1972.Authors wish to thank Drs. P. G. Andreis and A. S. Belloni for their skilful assistance in the autoradiographic experiments. Thanks are also due to Mr. G. Gottardo for his excellent technical help in electron microscopic work. This work was partly supported by a contract with the CNR-Italy (No 69.0172/115.3439).     

Evidence that an extrahypothalamic pituitary corticotropin-releasing hormone (CRH)/adrenocorticotropin (ACTH) system controls adrenal growth and secretion in rats

Within two weeks, hypophysectomy induced in rats a striking decrease in the level of circulating ACTH (the concentration of which was at the limit of sensitivity of our assay system), coupled with a net reduction in the plasma corticosterone concentration and an evident adrenal atrophy. Zona fasciculata, the main producer of glucocorticoids, was decreased in volume, due to a lowering in both the number and average volume of its parenchymal cells. Subcutaneous ACTH infusion (0.1 pmol·min^-1), administered during the last week following hypophysectomy, restored the normal blood level of ACTH and completely reversed all effects of hypophysectomy on the adrenals. Subcutaneous infusion for one week with -helical-CRH or corticotropin-inhibiting peptide (1 nmol·min^-1), which are competitive inhibitors of CRH and ACTH, evoked a further significant lowering of plasma corticosterone concentration and markedly enhanced adrenal atrophy in hypophysectomized rats. These findings strongly suggest that an extrahypothalamic pituitary CRH/ACTH system may be involved in the maintenance of the growth and steroidogenic secretory activity of the rat adrenal cortex.     

Investigations on the turnover of adrenocortical mitochondria

The effects of chronic administration of ACTH and dexamethasone on the morphology of mitochondria in zona glomerulosa cells of the rat adrenal cortex were investigated by stereological techniques. It was found that the volume of the mitochondrial compartment as well as the surface of the outer and inner mitochondrial membranes were significantly increased or decreased in relation to the number of days of ACTH- or dexamethasone-treatment. In ACTH-administered rats, the average volume of individual mitochondria decreased significantly up to the 6th day of treatment and then showed a conspicuous increase from the 6th to the 15th day, whereas in dexamethasone administered animals this parameter, after a small increase during the first 6 days of treatment, displayed a significant decrease. The number of mitochondria per cell showed a dramatic increase during the first 6 days of treatment with ACTH and continued to increase, but only slightly, with the subsequent treatment. In contrast, this parameter showed a parabolic decrease as a function of the duration of treatment in animals receiving dexamethasone. In the light of evidence showing that dexamethasone blocks ACTH-release, these findings are discussed and interpreted to indicate that ACTH is involved in the maintenance and stimulation of the growth and proliferative activity of mitochondria in rat adrenal zona glomerulosa.     

Morphometrisch-ultrastrukturelle Untersuchungen am Hypophysenhinterlappen der Ratte nach Dehydratation

Zusammenfassung Der Hypophysenhinterlappen von Ratten, die einer Dehydratation unterworfen waren, wurde ultrastrukturell-morphometrisch untersucht. Als wichtigster Befund wurde bereits nach 36stündiger Dehydratation eine massive Abnahme der Neurosekretgranula festgestellt. Gleichzeitig ist ein signifikanter Anstieg der Fettkörper in den Pituicyten zu verzeichnen. Die Fettkörper können nach Ansicht der Autoren als Endprodukt der Membranreste oder der Trägersubstanz des Neurohormons interpretiert werden. Eine Mitbeteiligung der Lysosomen bei ihrer Verarbeitung wird diskutiert.

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Morphometric-ultrastructural investigations on the neural lobe of dehydrated rats

Summary The neurohypophysis of the rat after dehydration has been studied by electron microscopic and morphometric methods. As the main result we found a massive decrease of the neurosecretory granules already at a dehydration of 36 hours. At the same time one can note a significant increase of fat vacuoles in the pituicytes. These fat vacuoles could be the final product of residual membranes or of the carrier substance of the neurohormones. The possibility of a functional role of lysosomes in the catabolism is discussed.

     

Morphometrisch-ultrastrukturelle Untersuchungen am Nucleus supraopticus der Ratte nach Dehydratation

Zusammenfassung Der Nucleus supraopticus der Ratte, die einer Dehydratation ausgesetzt war, wurde ultrastrukturell-morphometrisch analysiert. Dabei zeigte sich, daß die relativen Volumenanteile der einzelnen Zellkompartimente während der fünftägigen Durstperiode eine auffallende Konstanz aufweisen. Hingegen läßt sich eine absolute Zunahme der Einzelzellvolumina und somit auch der an der Synthese und Sekretion der Neurohormone beteiligten Zellkompartimente feststellen. Die vorliegenden Befunde sprechen für einen beschleunigten Abtransport des neurosekretorischen Materials bei gesteigerter Synthese. Auf eine optimale Standardisierung der Perfusionsmethode bei Untersuchungen am neurosekretorischen Zwischenhirnsystem wird hingewiesen.

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Morphometric-ultrastructural investigations on the supraoptic nucleus of dehydrated rats

Summary The supraoptic nucleus of the dehydrated rat has been analysed by electron microscopy and morphometry. With that it appears, that the relative volumes of the different cell compartments are striking constant. Otherwise one can see an absolute increase of the cell volume together with the cell compartments which take part at the synthesis and secretion of the neurohormones. These results are expression of an accelerated move of the neurosecretory material during increased synthesis. The importance of an optimal standardization of the perfusion-method in investigations of the neurosecretory system is demonstrated.

Wesentliche Teile der vorliegenden Arbeit werden von H. F. Reinhardt der Medizinischen Fakultät der Universität Basel als Dissertation vorgelegt.     

Elektronenmikroskopische Untersuchungen am Organon vasculosum laminae terminalis der Ratte

Zusammenfassung Das OVLT stellt jenen Abschnitt der Lamina terminalis dar, der durch eine eigentümliche, reiche Vaskularisation auffällt. Die von der Basalmembran bedeckte äußere Hirnoberfläche dringt an einer oder mehreren Stellen tief und spaltenartig in das OVLT ein. Dieser Spalt, der Bindegewebselemente und Gefäße enthält, verzweigt sich immer mehr und bildet ein aus 0,1–0,2 breiten Spalten bestehendes, labyrinthartiges System. Zum großen Teil füllen Gefäße vom Kapillartyp die größeren bindegewebigen Räume aus. Das Endothel der Kapillaren ist allgemein dünn und z. T. fenestriert. Die Bindegewebsräume und mit ihnen die Gefäße können sich dem Ventrikel derart nähern, daß sie von ihm durch nur eine einzige kubische Ependymzelle getrennt werden.Der Stützapparat des Organs wird in erster Linie von den Ependymzellen gebildet. Ihre langen basalen Fortsätze durchschneiden die Gehirnwand im Gebiet des OVLT und nehmen mit ihren Endigungen am Aufbau der Wand der Bindegewebsspalten und der äußeren Hirnoberfläche teil. In einem Teil der Ependymfüße findet man zahlreiche längliche, lysosomenartige Körper. Häufig kommen in die Tiefe der Substanz des OVLT eingedrungene Ependymzellen vor, welche nicht selten Zilien enthalten. Unter den Gliazellen konnten in erster Linie Astrozyten identifiziert werden.Die lichtmikroskopisch im OVLT beschriebenen sog. Parenchymzellen erweisen sich im Elektronenmikroskop als kleine, primitive Neurone. Ein großer Teil der Nervenfasern des Neuropils enthält granulierte Vesikel (Durchmesser zwischen 650 und 950 Å), die im allgemeinen eine runde oder ovoide Gestalt besitzen, obwohl auch tubulös ausgezogene Formen vorkommen. Die Nervenfasern welche die granulierten Vesikel enthalten, verlaufen nahe zur Kammeroberfläche, allgemein in der Längsachse des OVLT, wobei sie die länglichen Ependymzellen überkreuzen; in der Nähe der Endigung der basalen Ependymfortsätze wenden sie sich parallel zu letzteren und endigen zusammen mit ihnen frei am Rand der Bindegewebsspalten. Je ein solches, aus basalen Ependymfasern und Axonen bestehendes Bündel wird mehr oder weniger vom bindegewebigen Spalt umfaßt. Die Axonendigungen enthalten außer den granulierten Vesikeln und Mitochondrien auch zahlreiche synaptische Vesikel. Einige freie Axonendigungen wurden auch auf der freien Oberfläche des OVLT gefunden.Die Frage nach der Funktion des Organs wird an Hand der elektronenmikroskopischen Befunde diskutiert. Es wird für möglich gehalten, daß humorale Faktoren — ähnlich wie in der Eminentia mediana — aus den Axonendigungen in die Blutbahn gelangen; darauf scheinen die freien Endigungen am Rande des bindegewebigen Spaltensystems, die granulierte und synaptische Vesikel enthalten und die teilweise fenestrierten Kapillaren hinzuweisen, welche den aufgezweigten Bindegewebsraum drainieren.

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Electron microscopical investigation of the organon vasculosum laminae terminalis (OVLT) (Supraoptic crest) in the rat

Summary OVLT is that part of the terminal plate which is characterized by its rich vascular supply. The brain surface covered by a basement membrane forms deep, cleft-like invaginations containing vessels and connective tissue elements. These connective tissue spaces dividing into 0.1 to 0.2 end branches are parts of a labyrinthic system in the interior of the organ. The vessels, mostly of the capillary type, are situated in the main clefts; their endothelium often shows fenestration. Some of the capillaries may approach the ventricle to such an extent that they are separated from it by a single ependymal cell.The supporting apparatus of the OVLT is mainly represented by elongated ependymal cells. Their long basal processes traverse the terminal plate to take part with their foot-like endings in the formation of the brain surface and that of the connective tissue spaces. Groups of special ependymal cells often exhibiting cilia may occur in the interior of the organ. Glial cells are mainly represented by astrocytes.The so-called parenchymal cells described in the light microscopy can be identified as small, primitive neurons. A great part of the nerve fibres in the OVLT contains granulated vesicles the diameter of which varies between 650 and 950 Å. The nerve fibres are mainly running vertically between the ependymal processes while at their terminal portion they assume a parallel course to the ependymal processes and end with them at the margin of the connective tissue spaces. Besides granulated vesicles, these free axon terminals contain numerous synaptic-like vesicles and several mitochondria. Some of the free terminals may occur also on the outer surface of the OVLT.The possible functions of the organ are discussed on the basis of the present findings. The hypothesis is raised that — similarly to the median eminence — humoral controlling factors may be released into the vessels. This hypothesis seems to be supported by the presence of free axon terminals containing granulated and synaptic vesicles and the existence of numerous, partly fenestrated capillaries draining the connective tissue spaces.

     

Fine structure of trypsin-dissociated rat adrenal cells

Summary Electron micrographs of trypsin-dissociated rat adrenal showed predominantly intact rounded cells without internal damage. The population contained cells from the glomerular, intermediary and fascicular zones with cells from the zona fasciculata predominant. The presence or absence of cells from the reticular zone could not be determined. Cells from the medullary zone were absent. The addition of adrenocorticotropic hormone (ACTH) to the cellular suspension for 2 hours produced corticosterone. However, these stimulated cells did not display any significant ultrastructural change.Supported by research grants from the U.S.P.H.S. (No. GM15872), and the Research Council of Rutgers University, and the National Science Foundation (No. GB7427 to Dr. George Sayers). We are indebted to Mrs. Jean A. Gibney, Miss Rose-Marie Ma, and Mrs. Mary Vegh for their excellent technical assistance.Postdoctoral Trainee, U.S.P.H.S. Training Grant No. 5T01-GM00899-11.     

The effects of ageing on the morphology and function of the zonae fasciculata and reticularis of the rat adrenal cortex

Summary The morphological counterpart of the well-known age-dependent marked impairment of glucocorticoid secretion of rat adrenals was investigated by use of morphometric techniques. For this purpose 4-, 8-, 16- and 24-month-old rats were studied. Despite the notable lowering of both basal and ACTH-stimulated production of corticosterone by collagenase-dispersed inner adrenocortical cells, ACTH and corticosterone plasma concentrations displayed significant increases with ageing. Zona fasciculata (ZF) and zona reticularis (ZR) showed a notable hypertrophy in aged rats, which was due to rises in both the average volume and number of their parenchymal cells. The hypertrophy of ZF and ZR cells was in turn associated with increase in the volume of the mitochondrial compartment and proliferation of smooth endoplasmic reticulum, i.e., the two organelles involved in steroid-hormone synthesis. All these morphologic changes, conceivably due to the chronic exposure to high levels of circulating ACTH, are interpreted as a response enabling ZF and ZR to compensate for their age-dependent lowering in glucocorticoid secretion. Stereology also demonstrated that ZF and ZR cells underwent a striking age-related lipid-droplet repletion. Lipid droplets are the intracellular stores of cholesterol esters, the obligate precursors of steroid hormones in rats. This finding is in keeping with the contention that the mechanism underlying the age-dependent decline in rat-adrenal glucocorticoid secretion mainly involves impairments of the utilization of intracellular cholesterol previous to its intramitochondrial transformation to pregnenolone.     

Sex differences in adrenocortical structure and function

Summary Adrenal glands of adult female rats are heavier than the glands of corresponding male rats. Postpubertal orchiectomy increases the adrenal weight, an effect restored by testosterone replacement. Under the same conditions ovariectomy of 8 weeks duration does not change the adrenal weight while estradiol replacement enhances the relative adrenal weight.Karyometric studies have shown that nuclei in the female zona fasciculata cells are larger (app. 18%) than those of the male. Similar but only slight differences (2%) were observed in the zona reticularis. Orchiectomy results in enlargement of cell nuclei within all zones of the adrenal cortex; testosterone replacement has the opposite effect. Ovariectomy of 8 weeks duration slightly enhances the volume of nuclei of the zona glomerulosa cells, has no effect on the nuclei of the zona fasciculata and reduces the volume of nuclei in the zona reticularis. Estradiol replacement reduces the volume of nuclei of the zona glomerulosa cells but increases nuclear volume in the zona fasciculata and in the zona reticularis.Thus testosterone has an inhibitory effect on the adrenal cortex of the rat while the physiologic effect of estradiol on the morphology of this gland, particularly on the zona fasciculata cells is rather dubious.The author wishes to thank Mrs. B. Westerska and Miss K. Siejak for excellent technical assistance.This research was supported in part by a grant from the Zoological Committee, 2nd Department, Polish Academy of Sciences.This paper is dedicated to the memory of late Kazimierz Mietkiewski, M.D., Ph.D. whose encouragements and suggestions were most stimulating for my work.     

Über den Peroxydasetransport im Darmepithel der Ratte

Zusammenfassung Bei Ratten tritt 3 min nach intravenöser Injektion von Peroxydase elektronenmikroskopisch ein entsprechendes Reaktionsprodukt im Kapillarlumen der Lamina propria des Dünndarms und an der Basalmembrangrenze der Saumepithelzellen auf. 5 min nach der Injektion finden sich im basalen Abschnitt des Darmepithels pinozytotische Bläschen mit dem Peroxydase-Reaktionsprodukt. — 10–30 min nach der Injektion erreichen die Partikel die apikalen Teile der Zelle. Sie dringen in den interzellulären Spalten bis zu den Haftplatten vor, erreichen jedoch nie das Darmlumen. Im Dünndarm existiert vermutlich auch ein der Resorption entgegengesetzter Saftstrom, der durch Peroxydase markiert werden kann.

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The transport of horseradish peroxidase in the epithelium of the small intestine

Summary In rats, 3 minutes after intravenous injection of peroxidase the reaction product can be observed electronmicroscopically in the lumina of the capillaries of the small intestine as well as at the border of the basement membrane of the epithelial border cells. Pinocytotic vesicles containing peroxidase particles occur in the basal portion of the epithelium of the small intestine 5 minutes after injection. 10–30 minutes later, the peroxidase reaches the apical region of the cell. The particles infiltrate into the intercellular spaces as far as the tight junctions but never reach the intestinal lumen. In the small intestine there probably exists a flow of fluid in opposite direction to the resorption, which can be marked by peroxidase.

     

The fine structure of the newborn rat adrenal cortex

Summary The structural changes of the zona juxtamedullaris of the rat adrenal cortex at birth, have been examined by the light and the electron microscope. In this zone clusters of medullary cells lying among the strands of cortical tissue were observed. In the inner portion of the zona juxtamedullaris two types of adrenocortical cells were found: light and very-dark cells. The latter are smaller than the light cells and are always in close connection with the medullary tissue. The ultrastructural features of the very-dark cells suggest that these elements are in degeneration. This finding supports the hypothesis that at birth there is a partial degeneration of the rat zona juxtamedullaris, i.e. the zone corresponding to the fetal zone of some mammalian species.It is proposed that in all mammalian species at birth there is a partial regression of the zona juxtamedullaris and that the regression of the fetal zone is only the quantitative increase of this phenomenon. This hypothesis is discussed in relation to numerous data demonstrating that there are enzymatic conditions in the rat during fetal life, which permit a discrete hypertrophy of the adrenal cortex.The author wishes to express his appreciation to Dr. A. Gambino and to Mr. G. Gottardo for technical assistance.     

Intramitochondrial bodies in bovine adrenocortical cells

Summary Electron dense, homogeneous, mostly round intramitochondrial bodies were found in bovine adrenal glands, predominantly in the zona glomerulosa. The histochemical results obtained suggest that these bodies might contain a considerable amount of protein, although the possibility that substances other than proteins are contained in these bodies cannot be completely ruled out. The physiological significance of the intramitochondrial bodies remains unclear.This work was supported by a Scientific Research Grant, No. 144017, from the Ministry of Education of Japan to Prof. M. YasudaThe authors are deeply indebted to Dr. Kazuyori Yamada, Department of Anatomy, Nagoya University for his generosity in allowing us to perform a part of the histochemical tests in his laboratory and for the use of the facilities     

Gestational changes in hamster adrenal cortex: Morphometric and ultrastructural stereologic studies

Summary In the hamster, the weight of the adrenal glands increases during the course of gestation, with the highest value at day 5. In comparison to non-pregnant control animals, there were no changes in the volume of the zona glomerulosa (ZG) and zona fasciculata (ZF), while the volume of the zona reticularis (ZR) increased notably. The average volume of ZG-cells rose at day 5 of pregnancy and thereafter gradually decreased to that of control hamsters. A marked drop in the volume of ZF-cells was seen at days 5 and 10 of pregnancy, whereas at day 15 the cells were larger than in controls. At day 5 of pregnancy, a conspicuous increase in the cell volume was found in ZR, followed by lower values at day 10 and again higher than in control hamsters at day 15. The total number of parenchymal cells in hamster adrenal cortex increased at day 10 of gestation, then underwent a marked decrease, reaching the control value at the final day of pregnancy; this drop was mainly due to a reduction in the number of ZF-cells. The changes in the cell volume were paralleled by rather proportional changes in the volume of the mitochondrial compartment and in the quantity of smooth endoplasmic reticulum. The volume of the lipid-droplet compartment significantly rose in the course of gestation in both ZF- and ZR-cells. The cortisol output by adrenal homogenates gradually decreased during pregnancy.Study performed in partial fulfillment of the Ph.D. Thesis of Magdalena Nowak     

Short- and long-term effects of ACTH on the adrenal zona glomerulosa of the rat

Summary Short-term ACTH treatment provoked a decrease in volume of the lipid-droplet compartment in rat zona glomerulosa cells, and a rise in plasma and intracellular concentrations of corticosterone and aldosterone. It enhanced activities of 3-hydroxysteroid dehydrogenase (3HSD), 11-hydroxylase (11OH) and 18-hydroxylase (18OH). Long-term ACTH administration produced a hypertrophy of the zona glomerulosa and its parenchymal cells, a result of the increase in volume of the smooth endoplasmic reticulum and the mitochondrial compartment. The surface area per cell of mitochondrial inner membranes increased; the tubular cristae were transformed into a homogeneous population of vesicles. The plasma and intracellular concentrations of corticosterone further increased, whereas those of aldosterone fell below basal levels (the aldosterone-escape phenomenon). The activities of 3HSD and 11OH were enhanced, that of 180H decreased. Therefore, ACTH stimulates zona glomerulosa growth and transforms parenchymal elements into zona fasciculata celltypes. Cyanoketone nullified acute ACTH effects on plasma and intracellular concentrations of corticosterone and aldosterone, but did not affect the activities of 11OH and 18OH. Chronic ACTH treatment produced similar results, although 18OH activity was not suppressed. The mechanism underlying the aldosterone-escape phenomenon may thus involve a rise in the intracellular concentration of corticosterone, caused by the enhanced synthesis and activation of 3HSD and 11OH.