Showing papers in "Cell and Tissue Research in 1977"

Comparison of vascular smooth muscle cells from adult human, monkey and rabbit in primary culture and in subculture.

Abstract: A method is presented for growing large numbers of pure isolated smooth muscle cells from adult human, monkey, and rabbit blood vessels in primary culture. In the first few days in culture these cells closely resembled those in vivo and could be induced to contract with angiotensin II, noradrenaline and mechanical stimulation. They stained intensely with antibodies against smooth muscle actin and myosin. Fibroblasts and endothelial cells did not stain with these antibodies thereby allowing the purity of each batch of cultures to be monitored. This was consistently found to be better than 99%. The smooth muscle cells modified or "dedifferentiated" after about 9 days in culture to morphologically resemble fibroblasts. At this stage cells could no longer be induced to contract and did not stain with the myosin antibodies. Intense proliferation of these cells soon resulted in a confluent monolayer being formed at which stage some differentiated characteristics returned. The modification of "dedifferentiation" process could be inhibited by the presence of a feeder layer of fibroblasts or endothelial cells, or the addition of cAMP to the culture medium. Smooth muscle cells which had migrated from explants in primary culture, and cells in subculture, had morphological and functional properties of "dedifferentiated" cells at all times. The advantages of differentiated rather than "dedifferentiated" smooth muscle cells in culture for the study of mitogenic agents in atherosclerosis is discussed.

The origin of the vasopressinergic and oxytocinergic fibres of the external region of the median eminence of the rat hypophysis.

Abstract: The origin of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence was investigated by means of hypothalamic lesions, adrenalectomy and immunocytochemistry. The results obtained in bilaterally adrenalectomized animals with complete, or incomplete, destruction of the suprachiasmatic nuclei showed that, at least, the great majority of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence do not originate from the suprachiasmatic nuclei. From the observations obtained in bilaterally adrenalectomized animals with total or subtotal destruction of both paraventricular hypothalamic nuclei, it appears that the paraventricular nuclei must be the origin of (nearly) all the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence. The results strongly suggest that both types of fibres originate from all parts of the paraventricular nuclei.

Reformation of the severed septohippocampal cholinergic pathway in the adult rat by transplanted septal neurons.

Abstract: Transplants containing developing cholinergic neurons were obtained from the septum-diagonal band area of rat fetuses and were implanted into a lesion of the septohippocampal cholinergic pathway or into a cavity of the occipital cortex in adult recipient rats. The growth of new cholinergic fibres from the implant into the hippocampal formation was followed with choline acetyltransferase (ChAT) determinations and acetylcholine esterase (AChE) histochemistry. A fimbrial lesion alone, transecting the septohippocampal pathway, caused an almost complete cholinergic denervation of the hippocampal formation that persisted throughout the five month experimental period. A septal transplant implanted into the cavity of the fimbrial lesion restored a new AChE-positive innervation pattern in the hippocampus and the dentate gyrus that closely mimicked the original innervation removed by the lesion. In parallel, there was a progressive recovery in the ChAT levels, starting in the septal end, and progressing in a temporal direction. A new cholinergic fibre supply could be established in the hippocampal formation also along an abnormal route, i.e. from the transplants implanted into a cavity in the occipital cortex (involving also the dorsal part of the entorhinal cortex). Provided the hippocampus previously had been denervated of its normal cholinergic innervation, a partly normal AChE-positive terminal pattern was thus re-established also from this abnormal position. If, on the other hand, the cholinergic afferents were left intact, the ingrowing fibres were restricted mainly to the outer portion of the dentate molecular layer, i.e. the terminal zone of the lesioned entorhinal perforant path fibres. This suggests that the growth of the sprouting AChE-positive fibres into the normal cholinergic terminal fields was blocked by the presence of an intact cholinergic innervation. It is concluded that regrowing cholinergic axons can be guided over large distances within the hippocampal formation, and that their patterning within the terminal fields is very precisely regulated by mechanisms released by deafferentation.

Relationship between structure and function of antennal chemo-, hygro-, and thermoreceptive sensilla in Periplaneta americana

Abstract: On the antennae of Periplaneta americana, 25 chemo-, hygro- or thermosensitive sensilla were investigated electrophysiologically and, after marking, by transmission and scanning electron microscopy. A clear-cut relationship of functional types to structural types was observed. Two different stimulus conducting structures were observed: a) pore tubules which are found only in smooth, single-walled sensory pegs and b) secretion-filled canals which occur only in grooved double-walled sensilla. Temperature- and humidity-sensitive receptors occur only in double-walled sensilla with secretion material as the stimulus conducting system. Olfactory sensory cells were found in both types, however, those with a specific sensitivity for short-chain n-alcohols are restricted to single-walled pegs with pore tubules, while those which are most sensitive to short-chain n-acids and amines are found in double-walled sensilla, sometimes together with thermosensitive units. The stimulus conducting systems may control the access of odorous substances to the dendritic membranes and thus contribute to the discriminatory properties of the sensilla.

Distribution, ontogeny and ultrastructure of somatostatin immunoreactive cells in the pancreas and gut.

Abstract: Somatostatin cells are numerous in the pancreas and digestive tract of mammals as well as birds. In the pancreas of chicken, cat and dog they occur in both the exocrine parenchyma and in the islets. In the rat and rabbit, somatostatin cells have a peripheral location in the islets, whereas in the cat, dog and man the cells are usually more randomly distributed. In the stomach of rabbits and pigs, somatostatin cells are more numerous in the oxyntic gland area than in the pyloric gland area, whereas the reverse is true for the cat, dog and man. In the cat, pig and man, somatostatin cells are fairly numerous in the duodenum, whereas in the rat, rabbit and dog they are few in this location. In the remainder of the intestines somatostatin cells are few but regularly observed. Somatostatin cells are numerous in the human fetal pancreas and gut. In the fetal rat, somatostatin cells first appear in the pancreas and duodenum (at about the 16–17th day of gestation) and subsequently in the remainder of the intestine. Somatostatin cells do not appear in the gastric mucosa until after birth. Three weeks after birth, somatostatin cells show the adult frequency of occurrence and pattern of distribution. In the chicken, somatostatin cells are numerous in the proventriculus, absent from the gizzard, abundant in the gizzard-duodenal junction (antrum), infrequent in the duodenum and virtually absent from the remainder of the intestines. No immunoreactive cells can be observed in the thyroid of any species nor in the ultimobranchial gland of the chicken. In the chick embryo, somatostatin cells are first detected in the pancreas and proventriculus (at about the 12th day of incubation). They appear in the remainder of the gut much later, in the duodenum at the 16th day, in the antrum at about the 19th day and still later in the lower small intestine. The ultrastructure of the somatostatin cells was studied in the chicken, rat, cat and man; the cells were identified by the consecutive semithin/ultrathin section technique. The somatostatin cells display the properties of the D cell. There was no difference in granule ultrastructure between somatostatin cells in the gut and the pancreas. The granules, which are the storage site of the peptide, are round, supplied with a tightly fitting membrane and have a moderately electron-dense, fine-granulated core. The mean diameter of the somatostatin granules is smallest in rat (155–170 nm) and largest in the chicken (270–290 nm).

The effects of aging on satellite cells in skeletal muscles of mice and rats

Abstract: Myosatellite cells were examined and quantified at the fine structural level of resolution during aging of skeletal muscles in mice and rats. Satellite cells in the soleus and gastrocnemius muscles of animals between eight and 30 months of age appeared, according to morphological criteria, metabolically less active than those examined in immature muscles. In the soleus muscle of the mouse, satellite cells decreased in number from 4.6% at eight months of age to 2.4% at 30 months. This decrease appeared to be due to the passage of some satellite cells into the interstitial space as a result of the formation of external lamina material around the entire satellite cell surface.

A neuroanatomical study on the organization of the central antennal pathways in insects

Abstract: Receptor cell axons from the antennal flagellum terminate in the glomeruli of the ipsilateral deutocerebrum in Periplaneta americana and Locusta migratoria. Processes from several groups of deutocerebral neurons also enter the glomeruli and terminate in characteristic branching patterns. There, they contact the antennal axons. Connections are both convergent and divergent. Not only do single central neurons collect the inputs from many receptor cells, but receptor axons were often observed to branch and terminate at more than one deutocerebral neuron. The axons from a portion of the neurons go to form the deutocerebral bundle of the tractus olfactorioglobularis. These axons of the bundle terminate in the ipsilateral calyx of the corpus pedunculatum and in the lateral lobus protocerebri. The processes of the majority of the deutocerebral neurons stay within the deutocerebrum itself and may serve as local interneurons. Part of some antennal fibers terminate in the lobus dorsalis. The lobus glomeratus receives inputs from the maxillary palps and also from processes of deutocerebral neurons.

Ultrastructural changes in rat hypothalamic neurosecretory cells and their associated glia during minimal dehydration and rehydration.

Abstract: A quantitative ultrastructural study was performed to determine the changes in the neurosecretory neurons of the supraoptic (SON) and circularis (NC) nuclei following 4–24 h of water deprivation (WD) and subsequent rehydration (12 and 24 h). In both nuclei, the amount of direct soma-somatic contact increased throughout WD, apparently by retraction of fine glial processes from between the cells. Rehydration reversed these changes. The number of smaller (<1600 A) neurosecretory granules (NSG's) decreased in both nuclei at 4 h of WD but returned to control levels by 24 h of WD and remained so during rehydration. Larger (<1600 A) NSG's decreased in number at 4 h of WD in SON and then returned to control levels by 24 h of WD and remained the same throughout rehydration. In NC, these NSG's did not change in number with WD, but significantly increased between 12 and 24 h of rehydration. No cells with dilated rough endoplasmic reticulum were seen in NC during this study. In SON, however, the percentage of such cells increased at 4 and 12 h of dehydration only to decrease to control levels at 24 h of dehydration and throughout rehydration. Lysosomes decreased at 4 h of dehydration in SON and returned to control levels thereafter. In NC, lysosomes tended to decrease with dehydration and increase with rehydration. These findings indicate that detectable morphological changes take place in the course of alterations in hydration state that are well within the physiological range.

Immunohistochemical localization of neurotensin in endocrine cells of the gut

Abstract: Endocrine cells displaying neurotensin immunoreactivity are found scattered in the jejuno-ileum of all mammals studied, including man. They are rather scarce in rat, guinea pig, rabbit and pig and fairly numerous in cat, dog and man. In most mammals the neurotensin cells predominate on the villi. Only in the dog are they more numerous in the crypts. In the chicken, neurotensin cells occur all along the intestinal tract. They are particularly numerous in the zone that joins the gizzard with the duodenum. The ontogeny of the neurotensin cells in the gut was studied in rats and chickens. In the rat, the cells are first observed in the jejuno-ileum immediately before birth. The adult frequency is reached 4–5 days later. In the chicken, neurotensin cells first appear in the colon in the 18 day old embryo and in the small intestine two days later (i.e. one or two days before hatching). A few days after hatching, the gut has achieved the adult number of neurotensin cells per unit area.

Intrapulmonary neuro-epithelial bodies in newborn rabbits. Influence of hypoxia, hyperoxia, hypercapnia, nicotine, reserpine, L-DOPA and 5-HTP.

Abstract: Neonatal rabbit neuro-epithelial bodies (NEB) were investigated under various experimental conditions with light microscopy, microspectrography, morphometry and electron microscopy. (1) Hypoxia causes a decreased amine fluorescence intensity and an increased secretory exocytosis of dense core vesicles (DCV). Otherwise the NEB appear structurally normal. (2) Hypercapnia also produces a decreased fluorescence and an increased exocytosis; ultrastructurally, however, the dense core of DCV fragmentizes. (3) Hyperoxia does not appear to affect significantly either fluorescence or exocytosis. (4) The uptake of biogenic amines such as 5-HTP and L-DOPA was demonstrated by fluorometry and electron microscopy. (5) Reserpine, on the other hand, provokes an amine depletion with a decrease of the NEB fluorescence and an ultrastructural palor of the DCV. (6) Intratracheally administered nicotine is accompanied by a decreased fluorescence and a distinct exocytosis of fragmented DCV. The reaction of NEB to hypoxia and hypercapnia suggests that these corpuscles could be intrapulmonary chemoreceptors (in addition to the classically known central and peripheral chemoreceptors), inducing a reflex reaction through the liberation of DCV at the corpuscular sensible nerve endings and via the CNS. In addition, they may subserve a local intrapulmonary effect by modulating directly the hypoxic and hypercapnic pulmonary vasoconstriction and thus the V/Q ratio.

Immunocytochemical localization of the vasopressinergic and the oxytocinergic neurons in the human hypothalamus.

Abstract: The human hypothalamic-neurohypophysial hormone-producing nuclei were investigated with the unlabeled antibody peroxidase-antiperoxidase complex (PAP) technique at the light microscopic level. The size, shape and location of the supraoptic, paraventricular, accesssory supraoptic and suprachiasmatic nuclei were determined. It was demonstrated in the human hypothalamus, as well as in the hypothalamus of other mammals, that vasopressin and oxytocin are synthesized in separate neurons. In each of the nuclei of the magnocellular neurosecretory system, the distribution, ratios and structural features of the vasopressinergic and oxytocinergic neurons were determined. It was shown that the human suprachiasmatic nuclei contain numerous neurophysin-vasopressin-producing neurons.

An ultrastructural classification of the neuronal cell bodies of the rat dorsal root ganglion using zinc iodide-osmium impregnation.

Abstract: Zinc iodide-osmium (ZIO) impregnation of rat dorsal root ganglia differentially stained various elements in the neuronal cells, particularly their Golgi bodies. On the basis of this differential ZIO staining dorsal root ganglion neurones have been classified into seven types. The ultrastructure of these is described and the numbers of each type in the L4 dorsal root ganglion have been determined. Prolonged nerve stimulation did not change the relative numbers of the different cell types suggesting that none of the differences between cell types represents differences in their state of activity. The possibility is discussed that differences in morphology may reflect differences in neurotransmitter function.

Structure and function of midgut epithelial cells in culicidae mosquitoes (insecta, diptera).

Abstract: The midgut of female Anophelinae and Culicinae comprises a narrow anterior (A-part) and a wide posterior part (P-part, stomach). Both are lined by a singlelayered epithelium, and in both parts regenerative cells are found between the epithelial cells. “Clear cells”, possibly with an endocrine function, are only found in the P-part.

Neurons and their synaptic organization in the visual cortex of the rat

Abstract: Cells in the visual cortex (area 17) of adult rats were impregnated by the rapid Golgi method and characterized by light microscopy Selected cells were then sectioned for electron microscopy and their cytological characteristics and the pattern of synapses on their cell bodies and dendrites were studied Twelve classical pyramidal cells from layers II–VI, two pyramid-like cells from layer VI, two inverted pyramidal cells from layers V and VI, ten spine-free non-pyramidal cells from layers II–VI and two spinous non-pyramidal cells from layer IV were examined The cytoplasmic features of the identified cells, where these could be discerned, corresponded to those previously reported for the different cell types in conventionally prepared tissue Pyramidal Cells received exclusively type 2 synaptic contacts on their cell bodies, type 1 contacts on their dendritic spines and a mixture of synaptic types (type II predominating) on their shafts, where synaptic density was relatively low This pattern of synaptic contacts was consistent for all portions of the dendritic tree; inverted pyramidal cells and pyramid-like cells showed the same synaptic organization as classical pyramids The axon collaterals of pyramidal cells established type I contacts with dendritic spines (or, rarely, shafts) of unknown origin Non-Pyramidal Cells received both type 1 and type 2 contacts (the former predominating) on their cell bodies and dendrites The spinous variety also received type I contacts on their dendritic spines Axon terminal of spine-free non-pyramidal cells established type II synaptic contacts with dendritic shafts of unknown origin The similarity in synaptic organization between the spine-free and spinous non-pyramidal cells examined in this study suggest that the latter correspond to the sparsely spinous stellate cells rather than to the spinous stellate cells of cat and monkey visual cortex

Extracellular matrix fibrils and cell contacts in the chick embryo. Possible roles in orientation of cell migration and axon extension.

Abstract: The migration of neural crest and sclerotome cells and the extension of ventral root axons in chick embryos at stages 16–20 were studied by light microscopy as well as scanning and transmission electron microscopy at the leg bud level of fixed specimens Extensive cellular movements take place in association with an extracellular matrix consisting of microfibrils The neural crest and sclerotome cells migrate into the large matrix-filled extracellular space surrounding the neural tube and notochord, apparently using microfibril bundles as substratum The cells exhibit pseudopodia which are closely associated with the matrix fibrils The fibrils around the notochord show a spatial arrangement indicating that the sclerotome cells are contact-guided to their subsequent positions Mutual cell contacts, including those established by cell processes, frequently show cytoplasmic electron dense plaques at adjacent membranes These small “plaque contacts” might be correlated to contact inhibition of locomotion between the cells and participate in the guidance of cells The growth cones of extending axons exhibit filopodia contacting both surrounding mesenchyme cells and extracellular fibrils The orientation of the axons might thus be affected by contacts with cell surfaces as well as with extracellular material

Morphological alterations and functional changes of interhepatocellular junctions induced by bile duct ligation

Abstract: The effect of bile duct ligation on the intercellular junctions of hepatocytes was investigated The features and the arrangement of the bile canaliculi and the zonulae occludentes alter concomitant to the increase of the intracanalicular pressure The lumen of the bile canaliculi enlarges and the microvilli disappear The array of the zonulae occludentes becomes irregularly shaped, the number of strands diminishes and interruptions of the strands occur With peroxidase a leakage in the bile-blood barrier is detected Furthermore a disappearance of gap junctions between the hepatocytes after bile duct ligation is observed The present investigation shows that the zonulae occludentes are mobile structures which are changed by increased unilateral pressure Due to their ultrastructural alterations, a leakage of the permeability barrier between physiological compartments is found

Quantitation of fiber growth in transplanted central monoamine neurons

Abstract: Nerve fiber production by central noradrenaline (NA), dopamine (DA), and 5-hydroxytryptamine (5-HT) neurons was studied using immature brain tissue containing locus coeruleus, substantia nigra, or ventro-caudal medulla oblongata respectively, homologously grafted to the anterior chambers of rat eyes. A method was developed for quantitation of the fiber growth that occurs on the sympathetically denervated host irides as observed in whole mounts using Falck-Hillarp fluorescence histochemistry and by the uptake of 3H-metaraminol into the irides. Survival and growth in oculo of the three different areas were characterized by direct observations through the cornea in vivo for a number of pre- and postnatal stages of development of the donors, and the findings correlated to the degree of monoamine nerve fiber production on the host irides. The growth of fetal locus coeruleus transplants on irides was quantified using both fluorescence microscopical measurements of innervated areas and uptake of 3H-metaraminol. The uptake was well correlated to the histochemical measurements on individual irides, thus validating the fluorescence microscopical measurements of fiber production. The fiber growth of fetal locus coeruleus grafts on irides was followed for 20 weeks. The nerves increased in number and uptake capacity approximately linearly for 6 weeks whereafter the increase rapidly levelled off. On average, the final amount of nerve production by fetal locus grafts did not cover more than 1/3 of the host iris surface, and the average uptake of 3H-metaraminol by these nerves did not exceed 60% of that found in sympathetically intact control irides. The locus grafts produced a similar amount of fluorescent fibers in the host iris independent of the crown-rump length stage of the donor fetus and the final size of the transplants in oculo. The survival and growth of NA, DA and 5-HT neurons grafted from various postnatal donor rats was also followed by fluorescence microscopy. Locus coeruleus grafts produced markedly more fibers than the two other types of grafts when the donor was one week old or less, and DA grafts produced the least fibers of the three. Even from one month old donors some MA neurons survived grafting. Also, using prenatal donars, the locus coeruleus grafts produced many more fibers on the irides than did the DA grafts. It was concluded that the intraocular transplantation technique is very suitable for quantitative studies of nerve fiber production by immature monoamine neurons, and that it should be possible to study many other neuron systems in similar ways with this technique.

Isolation of fat cell precursors from adult rat adipose tissue.

Abstract: The possible existence of adipocyte precursors in adult rat adipose tissue was investigated. Cells were isolated from the stromal fraction of adipose tissue and were grown in culture. Skin fibroblasts were used as controls. The stromal fraction cells were initially fusiform and proliferated; in culture, they accumulated lipid inclusions, became rounder and acquired an eccentric nucleus. In contrast, the skin fibroblasts from the same rat and grown under identical culture conditions, did not exhibit any appreciable lipid accumulation. The doubling time for both the stromal fraction cells and skin fibroblasts was 40–60 h. At confluency, the stromal fraction cells contained 5–7 times more glyceride-glycerol than skin fibroblasts.

Fine structure of the olfactory epithelium in the goldfish, Carassius auratus. A study of retrograde degeneration.

Abstract: The fine structure of the goldfish olfactory epithelium was studied by transmission and scanning electron microscopy. Six different cell types were distinguished. Identification of the olfactory receptor cell was accomplished by use of retrograde degeneration studies. Two morphologically distinct types of olfactory receptor cells were identified: one type bears radially oriented cilia (Type I cell); the other type bears microvilli (Type II cell). The other four cell types were not identifiable as olfactory receptor cells: they are ciliated cells (Type III), rod-shaped cells (Type IV), supporting cells (Type V), and basal cells (Type VI).

The migration of osteoblasts.

Abstract: The endocranial matrix surfaces of parietal bones of 2-week old Albino Wistar rats were partly denuded of osteoblasts and then cultured for various periods up to 24 h, in control or PTE-enriched medium. They were examined by scanning electron microscopy and evidence for cell locomotion was found. Osteoblasts traversed the denuded bone surface and cut edges of bone in either medium, and cells also migrated out from vascular channels. Glass spicules were placed on the otherwise undisturbed osteoblast layer in similar organ cultures for 2, 3 or 5 days. Osteoblasts migrated from the bone to populate the glass, negotiating any angle. The cells in PTE-enriched media were always aligned parallel to one another and elongated, tended to align with the edges of the glass and, in time, formed a substrate of aligned fibrils whose axes were parallel to those of the cells. Osteoblasts in control medium on glass showed variable degrees of alignment and elongation and were less influenced by edges of the glass. Non-locomotory, nearly equidiametrical cells on glass in 5d control cultures had formed a substrate of randomly oriented fibrils. Migrating osteoblasts on bone matrix did not have leading edge ruffles; isolated, migrating ones on glass did.

Morphological aspects of the hypothalamic-hypophyseal system. VII. The tanycytes: Their relation to the hypophyseal adrenocorticotrophic function. An ultrastructural study.

Abstract: The ultrastruct of the tanycyte ependyma in male 160-180 g Wistar albino rats was studied under normal conditions and in experiments involving long-term suppression of ACTH secretion and its long-term stimulation. The former was accomplished by daily (for 8 days) intraperitoneal administration of dexamethasone phosphate at low (5 microgram/100 g) and high (100 microgram/100 g) concentrations. The effectiveness of suppression of the hypothalamic-hypophyseal-adrenal system in the experimental animals was judged by their reaction to two-minute ether stress (determination of plasma corticosterone) and by the results of measurement of the adrenal weights. Stimulation of ACTH secretion was achieved by bilateral adrenalectomy; the animals were examined on days 8, 10, 14, and 22 following the operation. The results obtained were in agreement with the previously established fact that there is a negative correlation between tanycyte activity and hypophyseal adrenocorticotropic function (Akmayed and Fidelina, 1974). They also testified to the predominant involvement of the median eminence tanycyte ependyma (beta-tanycytes according to the author's nomenclature) in these relationships. It is supposed that correlations are regulated by a feedback mechanism and attest to the involvement of beta-tanycytes in the inhibiting control of hypophyseal adrenocorticotrophic function. The mechanism of this control may be explained alternatively: either the tanycytes transport ACTH-suppression substances (catecholamines, corticosteroids, ACTH) from the CSF to the hypophyseal portal system or they themselves secrete substances possessing ACTH-suppressive activity. The authors distinguish several types of vesicles in the beta-tanycytes, the number of which changed with experimentally induced shifts in hypophyseal adrenocorticotrophic function. These vesicles are discussed in connection with the transport and secretory activity of the tanycytes and are considered to be a possible substrate of the hypothalamic inhibiting effect on ACTH secretion.

Immunocytochemical investigation of the hypothalamo-neurohypophysial system in birds.

Abstract: The results of an immunohistochemical investigation of the hypothalamo-neurohypophysial system in several species of birds have shown that: (1) mesotocin and vasotocin are synthesized in separate neurons; (2) in all species investigated the distribution of mesotocinergic and vasotocinergic perikarya follows a common pattern; (3) the external zone of the avian anterior median eminence contains exclusively vasotocinergic nerve fibers, originating in supraoptic and ventral paraventricular regions; (4) the distribution of immunoreactive elements in the neural lobe shows a definite species-dependent pattern.

Intestinal enzymes activities in isolated villus and crypt cells during postnatal development of the rat.

Abstract: A modification of Weiser's (1973) cell isolation method was used in order to study the developmental pattern of various intestinal enzyme activities in villus and crypt cells of normal rats from 5 days after birth until 8 weeks. Alkaline phosphatase and enterokinase activities were always located in the upper villus zone during postnatal development. Enterokinase activity was higher in the upper villus cells during the third week of life than after this period. Aminopeptidase activity was located in the crypt cells during the first week, its maximum activity remained in this area until the third week. At this time, sucrase activity appeared in the crypt cells, then aminopeptidase and sucrase activities rose to the villus zone during the fourth week. Amylase activity was detected along the entire crypt-villus axis 5 days after birth, reaching maximum activity in crypt cells at the end of the first week and in the upper villus cells after the fourth week. In contrast with the other enzymes studied almost all amylase activity was soluble in the youngest animals whereas at weaning most of the activity appeared in a particulate form in the villus cells. But in the crypt cells the ratio between particulate and soluble form remained unchanged until the adult stage. Various hypotheses are advanced to explain the patterns of evolution of the different enzymes.

The hypothalamo-choroidal tract. I. Immunohistochemical demonstration of neurophysin pathways to telencephalic choroid plexuses and cerebrospinal fluid.

Abstract: Neurosecretory pathways were examined in normal male rats by the use of the immunoperoxidase technique for the localization of neurophysin in Bouin-fixed, deparaffinized sections. Using this technique two projections of extrahypothalamic neuorosecretory fibers can be traced to the sites of origin of the choid plexuses of both horns of the lateral ventricles. Neurophysin-containing axons originating primarily from the paraventricular field course dorsolaterad to enter the choroid fissure of the dorsal horn. A caudally direced group of fibers course ventrolaterad to enter the ventral horn choroid fissures. The supraoptic nuclear field is the primary contributor to the latter group. Scattered neurosecretory neurons are found in association with both pathways, usually in contact with blood vessels supplying the choroid plexuses, or in the telencephalic subependymal stroma. Neurosecretory fibers and terminals occur within the choroid fissures and juxtaventricular neuropil. The neurosecretory terminals in the choroid fissures appear as Herring-body type neurohemal organs; in the neuropil they appear as punctate peri-neuronal desities suggestive of synaptic contacts. Thes morphologic findings are discussed in relation to reports indicating the presence of antidiuretic, vasopressin-like activity in cerebrospinal fluid and choroid plexus extraxts together with ultrastructural evidence supportive of vasopressin-mediated transchoroidal cerebrospinal fluid absorption. These results and those of others indicated the possible involvement of the neurosecretory system in the regulation of brain interstitial-ventricular cerebrospinal fluid dynamics.

The ventricular system in neuroendocrine mechanisms. III. Supraependymal neuronal networks in the primate brain.

Abstract: This investigation has utilized a correlative scanning-transmission electron microscopic technique in the analysis of the primate cerebral ventricular system. This approach has demonstrated a complex network of supraependymal cellular elements upon the walls of the third cerebral ventricle in direct contact with the ventricular lumen. Type I neuronal-like cells and type II histiocytic-like cells with potential phagocytic capabilities have been observed in large numbers throughout the third ventricle. Type I neuron-like cells are discussed in the context that they may represent a population of receptor-cells which serve to assess ambient changes in the composition of bioactive peptides in the cerebrospinal fluid and may serve as a supraependymal network that integrates the endocrine hypothalamus with other circumventricular organs which may also be sites of neuroendocrine transduction.

An ultrastructural analysis of the gametes and early fertilization in two bivalve molluscs, Chama macerophylla and Spisula solidissima with special reference to gamete binding

Abstract: An ultrastructural investigation of the gametes and their interaction during the early events of fertilization in molluscs has been performed. A gamete binding event involving large numbers of sperm has been identified and examined in detail. The surface of the oocyte is projected into numerous microvilli which extend through the vitelline envelope. Tufts of fibrillar material radiate from the tips of these microvilli, forming a layer external to the vitelline envelope. The acrosomal vesicle of the mature spermatozoon contains two major components, which function differently during fertilization. The vesicle is indented at its adnuclear surface, constituting a preformed acrosomal tubule. This tubule does not elongate during the acrosome reaction. Completion of the reaction results in the formation of an extracellular coat, derived from one component of the acrosomal vesicle, on the anterior surface of the sperm. Sperm-egg binding is accomplished by an association of the extracellular coat on the reacted sperm and the fibrous tufts on the tips of the microvilli of the oocyte. Evidence that gamete membrane fusion occurs by fusion of the acrosomal tubule and a microvillus is presented. These observations provide a generalized pattern of molluscan fertilization.

In vivo and in vitro formation of the junctional complex in choroid epithelium. A freeze-etching study.

Abstract: The Junctional complex of choroid epithelial cells was studied during in vivo formation, disaggregation after trypsin treatment, and in vitro reaggregation. The in vivo formation begins with the occurrence of amorphous patches of particles followed by the formation of small particulate rows and polygonal-ordered particle assemblies. Further arrangement of the zonula occludens continues with the confluence of particles and smooth contoured ridges. At the 9th day stage a fully developed zonula occludens has developed. In a subsequent step nexus become integrated within the tight junction formation. Disaggregation after trypsination results in fragmentation of the zonulae occludentes. Parts of the disassembling aggregates become incorporated in vacuoles indicating an endocytotic mode of “digestion”. The in vitro reconstruction of the zonula occludens proceeds from remnants of the former zonula occludens. On the 3rd to 4th day of cultivation mature tight junctions are visible. In vitro integrations of nexus were observed during a later phase. On the 7th day, cultivated choroid epithelial cells reveal well differentiated Junctional complexes consisting of continuous zonulae occludentes and integrated gap junctions.

Differentiation of chick embryo neuroretina cells in monolayer cultures. An ultrastructural study. I. Seven-day retina.

Abstract: Neuroretinas from 6–7 day-old chick embryos were cultivated after trypsin dissociation as monolayer cultures in Petri dishes, and examined after various intervals of time with the electron microscope. Soon after plating, cells begin to reaggregate in small clumps, and typical rosettes are formed. During the first week in vitro, cells appear to differentiate as neuroblasts and presumed Muller cells; the latter form a continous sheet on the substrate, upon which neuroblasts migrate and grow their neurites. Differentiated ribbon synapses are found after 8 days in vitro, the time at which they normally appear in situ. After 15 and 21 days in vitro, synapses are still found in large numbers, mimicking their “in vivo” counterparts. Photoreceptor cells were identified on the basis of the presence of typical ribbons in their cytoplasm, but no outer segment was found. It appears then that synaptogenesis in the retina is programmed independently of the tissue environment, which is markedly disturbed in the monolayer culture.

Identification of a secretomotor centre in the brain of Locusta migratoria, controlling the secretory activity of the adipokinetic hormone producing cells of the corpus cardiacum.

Abstract: After retrograde filling of axons terminating in the glandular lobe of the corpus cardiacum (CC) of Locusta migratoria with cobalt chloride, a paired group of about 15 cobalt containing cells was demonstrated in the lateral area of the protocerebrum. The axons of these cells run via the NCC II into the glandular lobe of the CC. These small neurons have the characteristics of secretory cells; they contain secretory granules of about 1000 A in diameter. The axon terminals in the glandular lobe, making synaptic contacts with the glandular cells, contain secretory granules of the same size. It is therefore concluded that the cell groups in the protocerebrum control the activity of the glandular cells which produce an adipokinetic hormone. Arborizations of fibers of the lateral secretomotor cells are present in the dorsal neuropile of the protocerebrum, ventral of the mushroom bodies and along the tracts of the NCC I within the brain. It is proposed that these arborizations are sites of synaptic input. It is discussed that the axons of these cells might receive additional synaptic input in the storage lobe of the CC.

Effect of anabolic steroids on rat heart muscle cells. I. Intermediate filaments.

Abstract: Long-term treatment of female rats with the anabolic steroid hormone Methandrostenolone results in a conspicuous increase of intermediate sized, nonmyofibrillar filaments in muscle cells of the left cardiac ventricle, as revealed by electron microscopy. These filaments, measuring 70 - 110 A in diameter, form a characteristic network at the Z-level of the sarcomere, either encircling or penetrating the Z-bands, and appear to insert into the nuclear membrane. The T-system is accompanied by the filaments adjacent to the site of the couplings. Here they are attached to subsarcolemmal electron-dense patches, which may be Z-line precursor material. The filaments may function as a cytoskeleton, to provide passive support in the mechanism of contraction and to mediate nucleo-sarcolemmal and nucleo-myofibrillar exchange.