AN ELECTRON MICROSCOPIC STUDY OF THE BLOOD-BRAIN BARRIER IN THE RAT, EMPLOYING SILVER NITRATE AS A VITAL STAIN

The intravital deposition of silver in the chorioid plexuses, area postrema, intercolumnar tubercle, neurohypophysis, and pineal body of rats, given 1.5 gm. of silver nitrate per liter of drinking water for periods of up to one year, has been investigated by electron microscopy. Unlike other parts of the central nervous system, these regions store large amounts of silver. In all of these structures, silver is deposited in the form of dense granules in the basement membrane upon which the capillary endothelium rests, in and upon the connective tissue cells and fibers constituting a loose pericapillary sheath, and in an outer membrane separating this sheath from the parenchymatous cells. Parts of the central nervous system which do not store silver, for example the spinal cord, cerebellar cortex, cerebral cortex, and reticular formation, lack a connective tissue investment of the capillaries. In these locations, the glial processes or end-feet are closely applied to the walls of the capillaries. Only a narrow space, filled by an amorphous, moderately electron-dense substance, separates the plasma membranes of the endothelial cells and glial processes. The significance of these observations is discussed with respect to the questions of the Virchow-Robin perivascular spaces, the interstitial ground-substance of the brain, and the location of the hematoencephalic barrier.     

SILVER DEPOSITION IN THE CENTRAL NERVOUS SYSTEM AND THE HEMATOENCEPHALIC BARRIER STUDIED WITH THE ELECTRON MICROSCOPE

For the purpose of studying the hematoencephalic barrier as it is concerned with silver circulating in the blood stream, silver nitrate was vitally administered to rats in their drinking water over periods of 6 to 8 months. The cerebrum, cerebellum, medulla, area postrema, and choroid plexus were prepared for light and electron microscopy. Silver deposition was found in the perivascular spaces in the choroid plexus, area postrema, in the medulla surrounding the area postrema, and in minute quantities in the cerebrum, cerebellum, and most of the medulla. Two levels of the hematoencephalic barrier were apparently demonstrated in our investigations. The endothelial linings of the vessels in the cerebrum, cerebellum, and medulla constitute the first threshold of the hematoencephalic barrier (specifically here, blood-brain barrier). The cell membranes adjacent to the perivascular spaces form the second threshold, as follows:—the neuroglial cell membranes in the cerebrum, cerebellum, and medulla (blood-brain barrier); the membranes of the neuroglial cells in the area postrema (blood-brain barrier); and the membranes of the epithelial cells of the choroid plexus (blood-cerebrospinal fluid barrier). This study deals with silver deposition and does not infer that the penetration of ionic silver, if present in the blood stream, would necessarily be limited to the regions described. Bleb-like structures were observed to cover the epithelial cell surfaces in the choroid plexus. They may be cellular projections increasing the cell surface area or they may be secretory droplets.     

THE FINE STRUCTURE OF THE PIGMENT EPITHELIUM IN THE ALBINO RAT

In this report, particular attention is paid to the inclusion bodies found in the apical cytoplasm of the pigment epithelial cell. These bodies are of variable size and form. The smallest (0.4 µ diameter) consist of a granular matrix enclosed by a single membrane, and are similar to the lysosomes of hepatic cells. Larger inclusion bodies contain areas of lamellated material in addition to granular matrix. The largest particles seen (2 µ diameter) are almost entirely lamellar. These different forms seem closely related, for it is possible to find all transitional stages between the smallest and largest particles. The relationship between the lamellar inclusion bodies and the rod outer segments is discussed.     

THE EXPERIMENTAL DEMONSTRATION OF PRE-MIGRATION ACTIVITY IN THE ABSENCE OF FAT DEPOSITION IN BIRDS

Under dietary conditions that experimentally prevented the accumulation of "pre-migratory" depot fat, typical nocturnal migratory restlessness developed in the Brambling Fringilla montifringilla. Subsidiary data are presented from Junco hyemalis and Zonotrichia albicollis.
In captive controls, the rapid accumulation and subsequent utilization of body fat closely paralleled the onset and waning of nocturnal unrest.
Although the Brambling accumulates fat reserves before both nuptial and contra-nuptial flights, it would seem that the presence of such is not an essential element of the so-called "total physiological state" necessary for the onset of migration.     

THE DURATION OF MITOSIS IN THE CRYPTS OF LIEBERKÜHN OF THE TERMINAL ILEUM OF THE ALBINO RAT

The duration of mitosis in the epithelium of the terminal ileum of the adult rat has been determined by counting normal mitotic figures, and colchicine-metaphases, in Feulgen-stained squashes of whole, micro-dissected crypts of Lieberkühn. The duration, which depends on the morphological definition of mitosis adopted, was calculated to be 23 min. This value is similar to those determined by other authors using other methods, except in the case of tritiated-thymidine autoradiography, where the reported duration of mitosis is considerably longer. The possibility that this is due to an effect of thymidine on mitosis is discussed.     

A STUDY OF THE STRUCTURE OF SPLENIC SINUSES IN MAN AND IN THE ALBINO RAT WITH THE LIGHT MICROSCOPE AND THE ELECTRON MICROSCOPE

The splenic sinuses in the spleens of 5 human beings and 7 albino rats have been studied in the light microscope and electron microscope after fixation in Dalton's fluid and Palade's fluid and embedding in n-butyl methacrylate. Splenic sinuses are tortuous vascular channels of large but variable diameter which represent the first venous vessels in the spleen and make up almost the entire red pulp in man and in rats. These vessels are composed of reticulo-endothelial cells flattened to endothelial form and sheathed by a netted reticulum. The luminal surface of the endothelium is made highly irregular by delicate and variable cytoplasmic protrusions, slender corridors separating adjacent endothelial cells, anastomotic openings to other sinuses, bulgings of entire cells, and even thrusts of endothelium spanning the sinai lumen. The supporting reticulum presents a well developed latticed appearance in tangential sections of sinuses, but in most cuts is punctate or linear. The reticulum is composed of strands without limiting membranes, which, in substance, are amorphous and resemble basement membrane. Material identical in appearance to the substance of the reticulum may be present in the endothelium, suggesting that the reticulum is formed by endothelial cells. The endothelium also contains deposits of presumed ferritin and hemosiderin. The extreme luminal bulgings of endothelium suggest production of circulating monocytes or lymphocytes by detachment of endothelial cells. Sinuses are patent and collapsed to varying degrees. Patent sinuses are separated by collapsed sinuses and these collapsed sinuses appear to constitute splenic (Billroth) cords.     

BIOCHEMICAL CHANGES IN RAT BRAIN ASSOCIATED WITH THE DEVELOPMENT OF THE BLOOD-BRAIN BARRIER

—Subcellular fractions from brains of 5, 10, 13, 16, 21, 30 day-old and adult rats were prepared. Protein content and various enzyme activities were assayed in all fractions and brain homogenates. γ-Glutamyl transpeptidase activity and 5′-nucleotidase were very low at 5 days of life but steadily increased, reaching adult concentrations at about 30 days after birth. Alkaline phosphatase, instead slowly decreased with maturation, while monoamine oxidase after an initial decrease, increased rapidly to adult levels. The relation between the appearance of enzymatic activity in brain and the blood-brain barrier function is discussed.     

大白鼠视网膜中向同侧中枢投射的神经节细胞

在大白鼠视束和上丘注射HRP作逆行标记表明,向对侧中枢投射的神经节细胞遍布视网膜各处,而同侧投射的细胞大部分位于颞下侧边缘的新月形区域内,少数细胞分布在此区以外。与视束注射比较,上丘注射时同侧视网膜颞新月区内标记细胞数大为减少。统计测量表明,同侧投射细胞胞体直径平均比对侧投射细胞的略长。文中对上述结果的生理学意义进行了讨论。     

STAGES IN THE ORIGIN OF VERTEBRATES: ANALYSIS BY MEANS OF SCENARIOS

Vertebrates lack an epidermal nerve plexus. This feature is common to many invertebrates from which vertebrates differ by an extensive set of shared-derived characters (synapomorphies) derived from the neural crest and epidermal neurogenic placodes. Hence, the hypothesis that the developmental precursor of the epidermal nerve plexus may be homologous to the neural crest and epidermal neurogenic placodes. This account attempts to generate a nested set of scenarios for the prevertebrate-vertebrate transition, associating a presumed sequence of behavioural and environmental changes with the observed phenotypic ones. Toward this end, it integrates morphological, developmental, functional (physiological/behavioural) and some ecological data, as many phenotypic shifts apparently involved associated transitions in several aspects of the animals. The scenarios deal with the origin of embryonic and adult tissues and such major organs as the notochord, the CNS, grills and kidneys and propose a sequence of associated changes. Alternative scenarios are stated as the evidence often remains insufficient for decision. The analysis points to gaps in comprehension of the biology of the animals and therefore suggests further research.     

IDENTIFICATION OF THE ADRENOCORTICOTROPHIN-PRODUCING CELLS IN THE RAT HYPOPHYSIS BY AUTORADIOGRAPHY

The relative rates of protein (hormone) synthesis and secretion by the various cell types in the anterior hypophysis of the rat have been studied by means of autoradiography. Normal and adrenalectomized male rats were injected with tritiated glycine and their hypophyses removed and fixed at 20, 40, and 90 minutes and 15 hours after injection. Autoradiograms of the hypophysial sections were prepared and autoradiographic grains were counted in the film overlying the cytoplasm of individual cells. With the aid of this method, a unique cell type was identified in the hypophyses of adrenalectomized rats. This cell is morphologically distinct from "gonadectomy cells," "thyroidectomy cells," and from previously described normal cell types, and is therefore designated as the "adrenalectomy cell." Among the 7 cell types differentiated in this study, the "adrenalectomy cell" had the highest tritium content and, in addition, at the time intervals studied this cell had the fastest rate of appearance and disappearance of protein tritium. This autoradiographic evidence of rapid protein (or polypeptide) turnover following adrenalectomy indicates that the "adrenalectomy cell" is the site of adrenocorticotrophin production in the adrenalectomized rat. Further autoradiographic and cytological evidence is presented which suggests that the "adrenalectomy cells" may be derived from chromophobes, and that a portion of the "large chromophobes" as defined in this study may be the site of adrenocorticotrophin production in the normal rat.