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Various mutant strains of Escherichia coli have been exposed to a homogeneous static magnetic field of either 0.5 T or 3.0 T and to the time varying magnetic fields found in echo-planar imaging experiments. No evidence of increased DNA damage was detected, even with bacterial strains disabled for DNA repair.

The morphological aspects of chromaffin tissue are reviewed, based mainly on our studies on the mouse adrenal gland. Particular attention was focused on the differential fixation of adrenaline and noradrenaline, and on the uptake and storage of [3H]dopa, [3H]dopamine and related substances in the adrenaline-storing (A) and noradrenaline-storing (NA) cells. Scanning electron microscopy combined with the NaOH-maceration method was useful for demonstrating the 3-dimensional organisation of nerve terminals, chromaffin cells, glial elements and vascular elements. In transmission electron microscopy, 3 types of chromaffin cell were distinguished. They were A, NA and SGC (small granule chromaffin) cells. After glutaraldehyde fixation followed by postosmication, A cell granules showed lower electron density, whereas NA cell granules were solid and dark. This difference in appearance between A and NA cells was first explained by the hypothesis that, after glutaraldehyde fixation, most of the adrenaline dissolved, whereas noradrenaline was precipitated in situ. Later, this hypothesis was supported by a series of autoradiographic and radioisotopic assay studies using [3H]dopa, [3H]dopamine and related substances; when [3H]adrenaline occurred, radioactivity in A cells mostly disappeared in the specimen, whereas that in the NA cells remained. At 15-60 min after an i.p. injection of [3H]dopa or [3H]dopamine, the concentration of radioactivity in A cells was higher than that in NA cells. However, in hypophysectomised mice, the radioactivity was low and evenly distributed in these 2 types of chromaffin cell. It was deduced that the carrier activities for extracellular dopa and dopamine were made greater in the A cells than in the NA cells by the pituitary gland. The hypophysectomy effects were restored by i.p. administration of ACTH.

The pre- and postganglionic sympathetic innervation of the guinea-pig adrenal medulla was investigated using the retrograde neuronal tracers Fast Blue and WGA-HRP. Labelled preganglionic cell bodies were located in the intermediolateral horn of spinal segments T3-L2, the majority (73.9%) were found between T6-T12 representing 70.2% of the total number of labelled cells; the segment T10 contained the largest number of labelled neurons. Labelled postganglionic cell bodies were found in the paravertebral ganglia between vertebral levels T3-T12 (representing 22.6% of the total labelled neurons), the maximum number was found at T10. In addition, labelled neurons were found in the suprarenal ganglion (representing 7.2%). No labelled cells were found in the coeliac ganglia. The labelled neurons were found ipsilateral to the site of injection into the left adrenal gland. It is concluded that the guinea-pig adrenal gland receives both a pre- and a significant postganglionic sympathetic innervation. The destination of these nerve fibres within the adrenal gland has yet to be determined.

The innervation of the adrenal medulla has been investigated in normal Wistar rats from birth to old age and ultrastructural findings compared with biochemical markers of the cholinergic innervation of the adrenal gland and catecholamine storage. Morphological evidence of the immaturity of the innervation during the first postnatal week is provided and using quantitative morphometry the innervation of chromaffin cells is shown to reach a mean total of 5.4 synapses per chromaffin cell during the period 26 days to 12 weeks of age. The variation in contents of synaptic profiles is discussed in the light of recent work that demonstrates a major sensory as well as visceral efferent innervation of the gland. Adrenal medullary neurons usually occur in closely packed groups, intimately associated with Schwann cells. Axodendritic and axosomatic synapses on these neurons are described and the likely origin of axonal processes innervating the neurons discussed. In old age the density of innervation remains the same as in young adult animals even though the medulla shows evidence of hyperplasia and hypertrophy of individual chromaffin cells.

Echo planar imaging (an extremely fast method of magnetic resonance imaging) was used to measure lung volume in a group of nine infants, all of whom had had respiratory problems. The mean echo planar imaging estimate of total lung volume was 44 +/- 9 ml/kg. In each case the right lung was larger than the left (ratio 52.8:47.2%). The mean thoracic gas volume was 36 +/- 8 ml/kg. The entire sequence of images of the thorax (about 400) takes five minutes to complete, infants require no sedation, and there are no side effects.

The fate of tl-[2, 5, 6-3h] dopa, and the intracellular localization of its metabolic products dopamine, noradrenaline and adrenaline, have been determined by the simultaneous use of assay techniques following separation of amines by chromatography and light and electron microscopic autoradiography. During the first 24 h after i.v. or i.p. injection of [3H] DOPA, synthesis of the above catecholamines occurred. Throughout this time the labelled amines were associated with chromaffin granules or immediately adjacent cytosol and not with either the Golgi complex or rough endoplasmic reticulum. Labelling of chromaffin granules occurred simultaneously throughout the cell and there was no evidence of regions containing recently labelled granules and others containing previously charged (older) granules. Adrenaline-storing cells took up [3H] DOPA and its products more rapidly and lost recently synthesized adrenaline more rapidly than noradrenaline-storing cells took up and stored their equivalent amines. This was in keeping with a more rapid turnover of catecholamines in adrenaline-storing elements.

NMR echo-planar imaging (EPI) has been used in a realtime mode to visualise the thorax of a live piglet. Moving pictures are available on an immediate image display system which demonstrates dynamic cardiac function. Frame rates vary from one per cardiac cycle in a prospective stroboscopic mode with immediate visual output to a maximum of 10 frames per second yielding up to six looks in one piglet heart cycle, but using a visual playback mode. A completely new system has been used to obtain these images, features of which include a probe assembly with 22 cm access and an AP400 array processor for real-time data processing.

Adrenal glands of the mouse, fixed either in glutaraldehyde followed by osmium tetroxide or in a mixture of potassium dichromate and glutaraldehyde, and embedded in Epon 812, were investigated by light and electron microscopy. An argentaffin reaction was applied to semi-thin sections for light microscopy and to ultra-thin sections for electron microscopy. Since the mature secretory granules in the Small Granule Chromaffin (SGC) cell were argentaffin and were mainly located along the cell membrane, this cell was clearly distinguishable under the light microscope both from the A (adrenaline) cell whose secretory granules were non-argentaffin and from the NA (noradrenaline) cell whose cytoplasm was rich and was filled with large, strongly argentaffin granules. Chromaffinity of the SGC cell was demonstrated under the light microscope. The SGC cell was intensively stained with toluidine blue without revealing metachromasia. It was demonstrated at the EM level that not only the secretory granules but also the synaptic-like vesicles in the SGC cell contained argentaffin substances. Possible functional relationship between the secretory granules and the synaptic-like vesicles was discussed.

A brief review of recent developments in line scan imaging of large objects is given, together with some representative images showing anatomical detail and a discussion of some spin-lattice relaxation time mapping results. Current progress in high speed imaging by means of the echo planar technique is reported and some preliminary results obtained at both 15.0 and 4.0 MHz are discussed.

New variants of the ultra-high-speed echo-planar imaging technique have been used to obtain snap-shot images of adult patients and volunteers at 0.1 T. Modified pulsed-gradient sequences together with non-linear signal sampling and activity screened gradients have greatly improved the image quality obtainable by single-shot methods. A particular variant, modulus blipped echo-planar single-pulse technique (MBEST), although slightly slower than the blipped echo-planar single-pulse technique (BEST), is experimentally more robust and incorporates intrinsic T2 weighting. An account of these improvements together with some experimental results is presented.

The digital nerves to the thumb and index finger have been studied by dissecting twenty-five embalmed upper limbs. The palmar digital nerves to the thumb were constant in position and course, with a short lateral cutaneous branch from the radial palmar digital nerve in 30 per cent of cases. The palmar digital nerves to the index finger had a variable pattern, the commonest arrangement, well described in Gray's Anatomy, occurring in 74 per cent of cases. The variations and their frequency are described. By examining histological cross-sections of the index finger it was found that of about 5,000 endoneurial tubes entering the finger, 60 per cent passed beyond the distal digital crease to supply the pulp and nail bed. The depth of the palmar digital nerves was about 3 millimetres, but less at the digital creases, and their diameter lay between 1 and 1.5 millimetres as far as the distal digital crease. Clinical applications of the findings are discussed.

Following the injection of 3-5 microliters of horseradish peroxidase or Fast Blue into the left adrenal medulla, labelling of neurons in the vagal sensory ganglia of rats and guinea-pigs has been demonstrated in all cases. Labelled vagal motor neurons with cell bodies in the dorsal motor nucleus of the vagi have been demonstrated in all cases in the guinea-pig and occasionally in the rat. The possible reasons for this variation are discussed. In both rat and guinea-pig the motor and sensory vagal innervation of the rat adrenal is derived from bilaterally situated cell bodies and the proportion of innervation derived from the two sides is approximately equal but with a slight ipsilateral predominance.

The pre- and postnatal development of the adrenal medulla was examined in the rat by immunohistochemistry and by assay of catecholamines. Immunohistochemistry involved the use of antibodies to noradrenaline (NA), adrenaline (A) and the biosynthesizing enzymes dopamine beta-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT). Adrenal glands were obtained from animals from the 16th day of gestation to the 7th postnatal day at daily intervals, and at the 14th postnatal day, and from adult rats. Tissues were fixed in ice-cold, 4% paraformaldehyde, buffered at pH 7.3. Cryostat sections (7 microns) were stained with the indirect immunofluorescence technique. Adrenals from the same developmental stages were assayed for the presence of DA (dopamine), NA and A by ion-pair reversed-phase liquid chromatography with electrochemical detection. In adult adrenals the majority of the medullary cells (approximately 80%) were highly immunoreactive to A and moderately immunoreactive to NA. They also showed immunoreactivity to both DBH and PNMT, i.e., they are synthesizing and storing A. The remaining cell clusters were only stained by antibodies to DBH and NA (NA-synthesizing and -storing cells). These findings correlate well with the relative concentrations of A and NA as determined by assay. Three developmental phases could be distinguished. In the first phase, the 16th and 17th prenatal day, medullary cells were only immunoreactive to DBH and NA, and only very small amounts of A as compared to NA were found. During the second period, from the 18th prenatal day to 2 or 3 days after birth, all medullary cells were immunoreactive to DBH, NA, PNMT and A, and during this phase the adrenaline concentration increased daily and became the predominant amine on the 20th day of gestation. Adrenaline represented 75% of total catecholamine on the 1st to 3rd day after birth. The third phase started at the 2nd or 3rd postnatal day and was characterized by the presence of an increasing number of medullary cells solely immunoreactive to DBH and NA, hence synthesizing and storing NA. The remaining cells were immunoreactive to DBH, NA, PNMT and A. Postnatally, the relative concentration of A continued to rise reaching 79% by the 4th postnatal day. These results indicate that initially the adrenal medullary cells are synthesizing and storing almost exclusively NA.(ABSTRACT TRUNCATED AT 400 WORDS)

Gastrointestinal (GI) tract motility was depicted in four human volunteers with the high-speed echo-planar imaging technique: modulus blipped echo-planar single-pulse technique (MBEST). Data acquisition times of 64 and 128 msec obviate image degradation due to motion without the need for gut paralysis and allow imaging of the GI tract in real time. Peristaltic patterns of the gastric antrum and proximal small intestine were depicted for fasting and fed subjects and subjects in whom peristalsis had been pharmacologically stimulated. The potential for quantitative measurements of GI tract motion with this new technique was demonstrated.

The ultra-high-speed echo-planar imaging technique produces instant snapshot magnetic resonance images in exposure times ranging from approximately 10 to 100 msec. A particular variant, the modulus blipped echo-planar single-pulse technique (MBEST), possesses two advantages: First, it makes the technique experimentally more robust; second, it introduces inherent T2 weighting in the images. The authors have used MBEST with an exposure time of 64 msec to explore the potential range of clinical applications in a series of normal volunteers. Applications include evaluation of the heart, renal function, and gastrointestinal tract. All images were obtained with proton resonances at 22 MHz.

This study has investigated the flow of blood to the adrenal gland of the anaesthetised rat under basal conditions and after the induction of haemorrhagic hypotension. By combining the use of radioactive microspheres with subsequent serial sectioning we have been able not only to assess total adrenal blood flow but also to determine its distribution within the gland. In normal anaesthetised rats approximately 7.4% of the adrenal blood flow passes directly to the medulla, the remainder being distributed to the cortex. This proportional flow is also observed immediately after withdrawal of sufficient venous blood to cause a 32% reduction in systolic blood pressure. However, the latter procedure results in a 52% increase in the total blood flow to the gland despite the associated fall in cardiac output.

An attempt has been made to determine the relative functional activities of adrenal and extra-adrenal chromaffin tissue during the neonatal phase and up to 1 week of age using young rabbits and guinea-pigs whose main extra-adrenal abdominal para-aortic chromaffin bodies are respectively non-innervated and innervated. Amine synthesis and storage were followed by assay and autoradiography after a single intraperitoneal injection of L-[2,5,6-3H]DOPA and the findings correlated with amine content as assessed by high performance liquid chromatography and by volume of tissue. The results indicate that in the guinea-pig, in spite of differences in proportions of adrenaline and noradrenaline in the adrenal gland and para-aortic body (PAB), the loss of labelled catecholamines from the innervated PAB closely follows that from the adrenal medulla, suggesting that both participate in normal sympathoadrenal activity. By comparison, in the rabbit the PAB shows only a minimal decreased labelled amine during the first week of life during which period the amine content and concentration of the PAB doubles: the functional significance of this non-innervated extra-adrenal chromaffin tissue, which persists throughout life, has still to be determined.

The sensory innervation of the guinea-pig adrenal medulla was studied using the fluorescent retrograde tracer Fast Blue. Labelled neurons were found in the dorsal root ganglia at segments T3-L2, the greatest contribution arising from T10 representing 15.2% of the total number of labelled cells. Labelling was ipsilateral to the site of injection of tracer into the adrenal medulla and the labelled neurons ranged in size between 6 and 36 microns. The overall size distribution fell into two groups of between 6 and 15 microns and of between 18 and 36 microns. Presumed sensory nerve endings were found in the adrenal medulla associated with chromaffin cells and vascular elements. The nerve endings were ovoid to spherical and densely packed with mitochondria. It is concluded that events occurring in the guinea-pig adrenal medulla, and possibly cortex, are monitored by sensory nerve terminals whose cell bodies lie within dorsal root ganglia. The exact distribution of the sensory nerve terminals within the adrenal gland is yet to be determined.

Using morphometric analysis and high-pressure liquid chromatography with electrochemical detection of catecholamines the volumes of adrenaline-storing (A) and noradrenaline-storing (NA) cells and the adrenaline and noradrenaline contents of adrenal glands of normal and age-matched hypophysectomized rats were determined and compared. Some hypophysectomized rats were also administered 150 mg metyrapone/kg daily for a 4-week period. Hypophysectomy resulted in an increase in noradrenaline storage in adrenal glands. This was more marked in hypophysectomized animals not treated with metyrapone even though the combination of metyrapone and hypophysectomy resulted in a reduction of blood corticosterone concentration to only approximately 6% normal. There was no evidence of a change in proportion of A and NA cells in the adrenal medulla of hypophysectomized rats or hypophysectomized animals treated with metyrapone and it is concluded that the changes in amine concentrations and proportions observed reflect an increase in the storage of noradrenaline in A cells as a consequence of less efficient methylation of noradrenaline after hypophysectomy. The findings also showed that the left adrenal gland and medulla was usually slightly larger and had a greater catecholamine content and slightly higher proportion of NA cells than the right adrenal gland. During the period of the experiment (animals aged 6-15 weeks) there was an increase in the concentration of both adrenaline and noradrenaline in the normal adrenal glands. In normal Sprague-Dawley rats weighing 140 and 400 g the adrenal medulla accounted for some 5% of adrenal volume; this increased to 13% after hypophysectomy; A and NA cells together accounted for 50-60% of the volume of adrenal medulla.

The high-speed echo-planar imaging (EPI) technique is used to obtain rapid T1 and spin density measurements by a two-point method. It is shown that neglect of edge effects in the slice selection procedure leads to significant systematic errors in T1. T1 maps for two young patients, obtained at 4.0 MHz, are presented. The T1 and spin density values obtained are used to produce computer-generated images in inversion recovery simulations. These results demonstrate marked improvement in image contrast without paying the time penalty incurred in real experiments, thereby greatly increasing patient throughput potential.

The sympathetic innervation of the rat adrenal medulla was studied using the fluorescent tracer Fast Blue. Labelled preganglionic cell bodies were located in the intermediolateral horn of the spinal cord at segments T1 and L1; the greatest number was found in T9. The ILp nucleus contained 95%, the ILf nucleus 4%, and the IC nucleus 1% of the total number of labelled preganglionic cells. Labelled postganglionic cell bodies were found in the sympathetic ganglia at levels T4 and T12; the maximum number were located in ganglia at T9 and T10. In addition, labelled cells were found in the suprarenal ganglion. No labelled cells were found in the ganglia of the coeliac plexus. The number of labelled preganglionic cells in the spinal cord accounted for 88.6%, the labelled cells in the sympathetic chain for 8.7% and those in the suprarenal ganglia for 2.7% of the total number of labelled cells found. The detailed distribution within the gland of postganglionic axons has yet to be determined, but it is thought that some are destined for the adrenal cortex.

A descriptive account and morphometric study of the adrenal medulla and adrenal chromaffin cells of Wistar rats aged from birth to 22 months is presented. Distinct adrenaline and noradrenaline-storing types of chromaffin cells are first identified in 4-day-old animals, prior to that time individual cells contain both adrenaline- and noradrenaline-storing granules. Primitive sympathetic cells and phaeochromoblasts occurs up to 2 days postnatally. Mitotic figures occur in chromaffin cells of a maturity and appearance appropriate to age and most frequently in adrenaline-storing cells. The adrenal medulla continues to increase in volume throughout the period and chromaffin cells increase in number and in some cases volume. There is a corresponding increase in neuronal elements. In 22-month-old rats individual cells or groups of chromaffin cells show evidence of hypertrophy and changes in cytoplasmic organelles, in particular lysosomes, rough endoplasmic reticulum and storage granules and vesicles. Some hypertrophied chromaffin cells contain a heterogeneous population of granules suggesting a degree of functional plasticity in some cells in aged rats.

The development of the noradrenaline (NA)- and adrenaline (A)-storing cells was examined in the adrenal gland of pre- and postnatal rats and pigs. Cryostat sections were immunostained with antibodies to NA and A. Amine levels were estimated in homogenates of adrenals by ion-pair reversed-phase liquid chromatography with electrochemical detection. 1. In adult animals separate NA- and A-storing cells were found. In the rat A-cells comprised about 80% of the parenchyma of the adrenal medulla. NA-cells, the remaining 20%, were randomly arranged in clusters. In the pig, in contrast, the A- and NA-storing cells were equally distributed, with spherical clusters of NA-cells were surrounded by A-cells. 2. In the earliest developmental stages examined (16th day of gestation in the rat, and 42nd day in the pig) the adrenals only contained NA-immunoreactive cells. In the rat separate NA- and A-storing cells were first noticed 2 or 3 days after birth, whereas in the pig separated cells were already present at the 56th prenatal day (full term at 114th day). 3. In both rat and pig adrenals dopamine (DA), NA and A increased in amount rapidly during development. In the rat this change mainly took place after birth (an increase of 13 times from the 17th day of gestation, and a 100-fold increase from birth to adult age). In porcine adrenal increase of the total amount of amines mainly occurred before birth (200 times between the 42nd day of gestation and birth, whereas only 10 times after birth). 4. In both species DA-levels remained low during pre- and postnatal development. On the contrary, the relative concentrations of NA decreased while that of A increased correspondingly. Again, species differences were noticed: in the rat NA decreased from 90% (17th day of gestation) via 35% (just before birth) to 20% at adult age, while the porcine adrenal showed a more gradual decrease, i.e., from 90% (42nd day of gestation) to 70% (birth) ending up with 50% in the adult stage. 5. The immunohistochemical and biochemical data indicate that, in rat adrenal medulla three phases of development can be distinguished. First, up to the 18th day of gestation, medullary cells synthesize and store only NA. Second, from the 18th day to 2 or 3 days after birth, NA and A are synthesized and stored in a single cell type ("mixed cell type"), and third, NA and A are localized in separate cell types.(ABSTRACT TRUNCATED AT 400 WORDS)

The localisation of chromogranins A and B, met-enkephalin-arg6-gly7-leu8 (met-enk 8) and protein gene product 9.5 (PGP 9.5) in the adrenal medulla and extra-adrenal chromaffin tissue has been studied in the developing rat by immunogold-silver staining. In the adult rat adrenal the cytoplasm of all medullary chromaffin cells showed a positive response with chromogranin A and B; in each case occasional groups of cells with a low reactivity that may have been NA cells were seen. Chromogranin A was first detected in adrenal medullary and extra-adrenal chromaffin cells at 18 days of gestation whilst chromogranin B was not detected in animals younger than 7 days. In 15 days old animals the adrenal medullary response to A and B was of the same intensity as that seen in the adult. Less than 1% of adult medullary chromaffin cells were responsive to met-enk 8 staining and medullary cells were unreactive in the fetus, with only extra-adrenal chromaffin tissue responding prenatally. During the first postnatal week immunoreactive cells appeared in the adrenal medulla in considerably greater proportions than in the adult gland. In contrast, positively stained nerve terminals associated with chromaffin cells and abundant in the adult adrenal were not detected during the first week of life. Immunoreactive nerve terminals were first seen early in the second week of life at a time when positive chromaffin cells were becoming less common. PGP 9.5 was located in all chromaffin cells of the adult adrenal and was readily detected in chromaffin cells in the adrenal and in extra-adrenal locations of the earliest stage examined (E16). Our findings suggest that the ontogenesis of the chromogranin-like immunostaining reflects the maturation of chromaffin granules and the PGP 9.5 immunostaining detected a protein common to cells of neuronal origin and expressed at an early stage of differentiation. The reciprocal relationship between the presence of enkephalins in chromaffin cells and in their presynaptic terminals merits further investigation.

Light-microscopic autoradiography has revealed characteristic labelling patterns in adrenal medullary cells following the intravenous administration of different catecholamines. The uptake patterns for [3H] dopa, [3H] dopamine, [3H] noradrenaline and [3H] adrenaline ahve been compared. In all cases A cells were more active than NA cells and cells situated in the zone nearest the cortex demonstrated a markedly higher rate of uptake than central cells. It was concluded that adjacent chromaffin cells with very similar morphology may differ as much as 50 fold in their capacities to incorporate exogenous amines. The adrenergic nature of te innervation of the vessels of the adrenal cortex and capsule in the mouse was confirmed.

Echo planar imaging (EPI) is that form of magnetic resonance imaging which provides very short image acquisition times. EPI also provides very rapid sequential imaging. The EPI method is ideal for imaging the heart and thoracic content because images are devoid of cardio-respiratory motion artefact. Previously an analysis of transectional images has been presented. This paper is concerned with the study of the heart by the use of EPI constructions in the sagittal and coronal planes. Defining connections between ventricle and great artery is of cardinal importance in paediatric cardiology. EPI constructions in the normal heart, transposition, truncus arteriosus and right heart hypoplasia are presented and discussed.

Light-microscopic autoradiographs of the adrenal medulla at various intervals after the intravenous injection of [3H] 5-HTP, [3H] 5-HT, [3H] noradrenaline and [3H] adrenaline have been studied. The distribution of silver grains following [3H] 5-HTP uptake was found to be uniform over each of the two main cell populations, adrenaline-storing (A) cells and noradrenaline-storing (NA) cells in the adrenal medulla, but A cells were twice as active as NA cells in incorporating the isotope, a situation very similar to that found after [3H] dopa uptake. 5-HT administration resulted in a pattern resembling the distribution of [3H] noradrenaline uptake, with A cells being 4 or 5 times more active than NA cells and a gradient of activity from the periphery of the medulla inwards. However, the time-course for the loss of radioactivity was not the same for both amines: levels of 5-HT activity were not significantly reduced after one week whereas the degree of [3H] noradrenaline labelling after one week was less than 10% of that at one hour. Thus 5-HT may be bound to sites in the adrenal medulla normally occupied by noradrenaline but it would appear that the release mechanism is different. There was no evidence of 5-HT uptake by adrenal nerve endings.

Echo-planar imaging (EPI), a distinctive variant of nuclear magnetic resonance, needs only a fraction of a second for an image to be acquired and so is free from movement artifacts caused by respiration or heart beat. Clinical findings in the lungs, heart, and mediastinum of three children with high respiratory and heart rates who were examined by EPI are described.

Adrenal medullary arteries have been followed from their origins in the capsular or subcapsular plexus to the medullary plexus in the mouse, rat, hamster, cat, guinea-pig, rabbit and bovine. Adrenal medullary arteries have a zonal distribution to the medullary capillary plexus and the number of arteries present in species variable and proportional to the size of the medulla. Cortical veins drain through distinct venous channels between groups of chromaffin cells, and may run adjacent to either adrenaline-storing (A) or noradrenaline-storing (NA) cells. In the normal unstimulated gland both A and NA cells lie adjacent to capillaries of the medullary plexus which derive their blood supply from adrenal medullary arteries. In the normal unstimulated gland no evidence of selective supply of arterial or venous blood to either A or NA cells has been obtained.

A morphometric analysis of normal Wistar rat adrenal medulla following perfusion fixation and Araldite embedding, was correlated with catecholamine levels on fresh tissue, measured by high-performance liquid chromatography. The mean volume of whole adrenal is 13.2 mm3 and the mean medullary volume 1.3 mm3. Volume density estimates showed that the medulla is composed of 63% chromaffin tissue with an adrenaline to noradrenaline storing cell ratio of 4.4:1. The vasculature occupies 20%, neuronal tissue 5% and interstitial tissues 12% of the medulla. A comparison was made of cell volumes, cell numbers and volume and surface density estimates of cytoplasmic organelles in adrenaline and noradrenaline storing cells. The mean cell volume of adrenaline storing cells at 1300 micron3 is larger than that of noradrenaline storing cells at 980 micron3. A single adrenal medulla contains 4.4-5.7 X 10(5) adrenaline cells and 1.5-1.9 X 10(5) noradrenaline cells. Chromaffin granules account for approximately 30% of the volume of the cytoplasm; the numerical density of granules at different sites in the cell was calculated for adrenaline cells. The volume density of mitochondria (4%) and the surface density of mitochondrial membranes (the ratio of outer to inner membrane being approximately 1:2.3) were similar in both cell types. Rough endoplasmic reticulum was the only organelle to show a significant difference in volume and surface density between the two cell types. Adrenaline storing cells have stacks of rough endoplasmic reticulum which have two to three times the surface and volume densities of that found diffusely scattered throughout noradrenaline cells. The adrenaline content of an adrenaline storing cell is 0.14 X 10(-6) microM and that of a granule 3.0 X 10(-12) or 3.8 X 10(-12) mumoles depending on the method of calculation. The noradrenaline content of noradrenaline storing cells can only be calculated on the assumption that all noradrenaline is stored in this cell type though it is likely that some is contained within adrenaline cells. Based on this assumption the noradrenaline content is 0.17 X 10(-6) mumoles per cell and 5 X 10(-12) mumoles per granule. The present study provides baseline morphometric data on the rat adrenal medulla at tissue and cellular level correlated with amine levels in adrenaline and noradrenaline storing cells and granules.

Following intravenous or intraperitoneal injections of 3H-l-dopa radioactivity in adrenal medullary chromaffin cells of the mouse has been followed by autoradiographic and radioisotopic assay methods. For autoradiographic purposes tissues were fixed in either phosphate buffered glutaraldehyde or in 1-5% potassium dichromate in glutaraldehyde pH 7-2. A high initial radioactivity is demonstrated by both autoradiography and radioisotopic assay. During the first hour dopamine is the predominant labelled amine. Between 1 and 4 hours after the injection noradrenaline is in highest concentration and from 1 to 24 hours labelled adrenaline appears and becomes the predominant labelled amine (about 70%) after 24 hours. Autoradiographs of tissues fixed in glutaraldehyde alone confirm that adrenaline is lost from sections by this technique. When 1-5% potassium dichromate is added to glutaraldehyde then adrenline is fixed in A cells and these show only a slightly decreased grain count as compared to NA cells at 5 days; this slight decrease is probably due to more rapid turnover of amines by A cells.

The intracellular localization of L-[4,5-3H] leucine in chromaffin cells has been observed using light and electron microscopic autoradiography and the association of the labelled amino acid with particular cell components confirmed by statistical analysis. By making observations at short intervals after a single intravenous pulse of [3H]leucine it has been possible to follow the movement of the isotope from the endoplasmic reticulum through the Golgi complex to the chromaffin granules. No evidence for movement of the label through the Golgi complex was observed in adjacent cortical cells. The time sequence of transport of the amino acid through the various cell organelles was very similar to that observed by previous workers in protein-secreting exocrine cells.

An autoradiographic study was performed on the effects of hydrocortisone, reserpine, propranolol and phentolamine on the uptake of tritiated amines by adrenal medullary cells of the mouse. Oral feeding of hydrocortisone ahd no significant effect on the normal uptake pattern of dopamine, noradrenaline or adrenaline by medullary cells of different type (A cells or NA cells) or location (marginal or central), although the overall amounts taken up were markedly reduced. Handling the animals led to similar reductions in the uptake of all three amines and was thus clearly shown to be the important factor in this effect. Reserpine reduced the uptake of [3H] noradrenaline to 25% of the control value although the relative distribution remained unchanged. Propranolol and phentolamine had no observed effect on [3H] noradrenaline uptake. These results are discussed in the light of the previously reported action of ACTH in reversing the effects of hypophysectomy on medullary amine uptake (Hirano and Kobayashi 1978), and it is concluded that ACTH must exert this effect directly on the adrenal medulla rather than through the secretion of adrenal corticosteroids. It is also suggested that reserpine acts, as in neurons, by blocking amine uptake into intracellular granules rather than by blocking uptake into the cell itself.