Membranes Of Organs Research Articles

Laminin Fibrils in Newt Gastrulae Visualized by the Immunofluorescent Staining: (gastrulation/laminin/immunofluorescent staining/newt/amphibian).

An anastomosing network of extracellular fibrils on the inner surface of the ectoderm layer of amphibian gastrulae has been shown to provide an adequate substratum for attachment and migration by the mesodermal cells. These fibrils contain fibronectin as shown by immunostaining at the light and electron microscope levels. Now we report the presence of laminin, another cell adhesion glycoprotein, as a fibrillar network on the inner surface of the ectoderm layer in gastrulae of the Japanese newt (Cynops pyrrhogaster), but its absence on the blastula ectoderm layer, by the immunofluorescent staining using an antiserum specific for mouse laminin. The same antiserum was shown to stain basement membranes of adult newt organs as expected.

The basement membrane of the enamel organ in human odontogenesis

This study was designed to record findings of a basement membrane in human fetal jaws relative to the oral epithelium, dental lamina, and developing teeth. With the use of Lillie's allochrome stain, this could be followed from the oral epithelium and dental lamina to the tooth jaws. The basement membrane was easily traced along the inner enamel epithelium, but only after elimination of the counterstaining phases was it visible adjacent to the outer enamel epithelium.

Organization of acetylcholine receptors in quick-frozen, deep-etched, and rotary-replicated Torpedo postsynaptic membrane.

The receptor-rich postsynaptic membrane of the elasmobranch electric organ was fixed by quick-freezing and then viewed by freeze-fracture, deep-etching and rotary-replication. Traditional freeze-fracture revealed a distinct, geometrical pattern of shallow 8.5-nm bumps on the E fracture-face, similar to the lattice which has been seen before in chemically fixed material, but seen less clearly than after quick-freezing. Fracture plus deep-etching brought into view on the true outside of this membrane a similar geometrical pattern of 8.5-nm projections rising out of the membrane surface. The individual projections looked like structures that have been seen in negatively stained or deep-etched membrane fragments and have been identified as individual acetylcholine receptor molecules. The surface protrusions were twice as abundant as the large intramembrane particles that characterize the fracture faces of this membrane, which have also been considered to be receptor molecules. Particle counts have always been too low to match the estimates of postsynaptic receptor density derived from physiological and biochemical studies; counts of surface projections, however, more closely matched these estimates. Rotary-replication of quick-frozen, etched postsynaptic membranes enhanced the visibility of these surface protuberances and illustrated that they often occur in dimers, tetramers, and ordered rows. The variations in these surface patterns suggested that in vivo, receptors in the postsynaptic membrane may tend to pack into "liquid crystals" which constantly appear, flow, and disappear in the fluid environment of the membrane. Additionally, deep-etching revealed a distinct web of cytoplasmic filaments beneath the postsynaptic membrane, and revealed the basal lamina above it; and delineated possible points of contact between these structures and the membrane proper.

The mechanism of serum alkaline phosphatase elevation by chloroform

The oral ingestion of low levels of chloroform (45 mg/kg/day) by female beagles caused an elevation of serum alkaline phosphatase. No histopathologic lesions were noted, and all other routine clinical parameters were normal. The isoenzyme appearing in the serum was the hepatic type. Alkaline phosphatase was shown to be lost from the brush border of the small intestine while the serum and hepatic enzyme activities increased. Bile volumes and alkaline phosphatase activities were normal. Other brush-border enzymes were lost from the small intestine, but not all of these losses were accompanied by elevations in serum activities. Alkaline phosphatase did not appear to be lost into the lumen of the intestine. These results suggest that the intestine may be the source of the elevated serum alkaline phosphatase activity caused by chloroform. Action of chloroform on the membranes of other organs and organelles is supported by the elevation of other serum enzymes.

Anti-basement membrane antibodies and antigen-antibody complexes in rabbits injected with mercuric chloride

New Zealand rabbits were injected three times a week with 2 mg of mercuric chloride (HgCl 2) (MC) to test the hypothesis that toxic agents release autologous antigens, generating antibody response and tissue injury. After 2 weeks of injections the rabbits developed anti-basement membrane antibodies binding to renal and extrarenal basement membranes and to the peri- and endo-mysium of skeletal muscles. Glomerular histology appeared normal. Membranous glomerulonephritis developed 1–2 months later with granular subepithelial deposits containing rabbit IgG and C3. Similar deposits were present in the basement membranes of other organs. Raji cell tests for immune complexes in the sera were negative in the initial stages and positive in the late stages of the disease. Radioactive MC was found in the cytoplasm of renal tubular, intestinal, and hepatic epithelial cells but not in glomerular deposits. Renal eluates and selected sera reacted with the basement membranes and the collagen matrix in normal or collagenase-digested sections of renal or extrarenal normal rabbit tissues. This reactivity was reduced or abolished by washing tissue sections in PBS or by absorption of renal eluates with whole kidney homogenates. The findings are consistent with the hypothesis that MC induces a biphasic disease characterized first by the production of antibodies to basement membranes and the extracellular collagen matrix and subsequently by antigen-antibody complexes formed in situ and in the circulation and presumably containing soluble polysaccharide components of the collagen matrix.

A morphological study of the cholinergic receptor protein from Torpedo marmorata in its membrane environment and in its detergent-extracted purified form.

ABSTRACT The highly specialized structural features of the postsynaptic membrane of Torpedo mar-morata electric organ have been studied by freeze-fracturing, freeze-etching and negative staining applied both to intact electroplaques and to isolated purified excitable membrane fragments. The external surface of the postsynaptic membrane, revealed by deep-etching and negative staining, is characterized by the presence of geometrically packed arrays of repeating particles having a subunit structure and a central pit. The isolated receptor protein molecules (molecular weight : 300 000) show structural features identical to those characteristic of the repeating particles visualized on the external surface of the postsynaptic membrane. Freeze-fracture cleaves the postsynaptic membrane into 2 asymmetric halves, showing that numerous identical particulate entities are more strongly anchored to the cytoplasmic side of the bilayer. On the other hand, geometrically packed arrays of pits or depressions found on the external fracture face suggest that the receptor protein extends across the outer leaflet and is exposed to the external surface facing the synaptic cleft. The homogeneous size-distribution of the particulate entities associated with the postsynaptic membrane element is consistent with the finding that the receptor protein accounts for at least 40% of the total protein of purified excitable membrane fractions. These results strongly favour the assumption that the particulate entities represent the receptor protein, which forms oligomeric complexes either of identical polypeptides or quasi-equivalently related proteins. Thus, the cholinergic receptor oligomers forming a bidimensional lattice of repeating particles are exposed both to the synaptic cleft and to the cytoplasmic environment. It is proposed that the regulating properties of the excitable membrane depend upon oligomeric or polymeric associations of receptor proteins spanning the bilayer.

A quantitative structure-activity approach to chemoreception: Importance of lipophilic properties

A multiparameter regression analysis utilizing physico-chemical parameters, and commonly known as the Hansch approach has been extended to a set of existing data on olfactive threshold and olfactive stimulatory effectiveness of organic molecules. The compatibility of various parameters has been studied with adequate statistical methods, and we conclude that the lipophilic character (expressed by the octanol/water partition coefficient) of the molecules is an essential parameter related to preliminary non-specific action and transport across the muceous/lipid interphase and the membrane of the olfactive organ. Such an approach allows the refocusing of the problem of the relationship between structure and olfactive properties.

The pathogenesis of experimental membranous glomerulonephritis induced with homologous nephritogenic tubular antigen.

The renal tubular epithelial antigen (Tub-Ag) of rats was solublized by Pronase and purified by gel filtration and acrylamide gel electrophoresis. Purified Tub-Ag was a glycoprotein with S20,W value of 8.4. Utilizing radiolabeled Tug-Ag, a sensitive radioimmunoassay for Tub-Ag and homologous antibody (anti-Tub-Ag) was developed. Tub-Ag activity associated with a protein of the same molecular size was demonstrated in the serum, as well as in Pronase extracts of all the organs tested, including kidney, liver, lung, spleen, intestine, stomach, and heart. The physiochemical properties of the Tub-Ag of rats and its distribution were essentially the same as the Tub-Ag of humans, which had been found in immune deposits in the kidney of some patients with idiopathic membranous glomerulonephritis. Rats were immunized with the purified Tub-Ag emulsified in Freund's complete adjuvant and followed for Tub-Ag and anti-Tub-Ag in the serum, as well as for proteinuria and immunohistological changes in the kidney. Serum Tub-Ag dropped sharply after 20 days, when anti-Tub-Ag appeared in the circulation. Persistent, massive proteinuria appeared still later, more than 30 days after injection, when anti-Tub-Ag disappeared and Tub-Ag reappeared in the serum of some of those rats. In others, anti-Tub-Ag in the serum persisted throughout the observation period of 90 days. The pathology of the kidney of the rats with proteinuria was that of a typical membranous glomerulonephritis; thickening of glomerular capillary walls with granular deposits of gamma-globulin and Tub-Ag was observed. On the basis of these results, Tub-Ag in the serum, probably released from cellular membranes of various organs as a physiological metabolite, is considered to maintain the pathological process in the kidney by providing the antigen continuously to form immune complexes.

Effect of denervation on the organization of the postsynaptic membrane of the electric organ of Torpedo marmorata

Effect of denervation on the organization of the postsynaptic membrane of the electric organ of Torpedo marmorata

Electron microscopic localization of alkaline phosphatase in the enamel organ of the young rat

The localization of alkaline phosphatase in the enamel organs of 1-day-old rats was investigated using the lead-citrate method introduced by Mayahara et al. (1967). The majority of enzyme activity was associated with the cell membranes of the stratum intermedium and the stellate reticulum, and with part of the cell membranes covering the proximal ends of the cells of the inner dental epithelium or the ameloblasts. The enzyme activity of those cells seemed to have occurred in a stage immediately prior to the development of the stratum intermedium. Weak enzyme activity was also observed in association with the basal cell membrane of the enamel organ in the growing end of the tooth, though no enzyme activity was observed in these zones in light-microscopic sections. Some cytoplasmic bodies seen in the cells of the stratum intermedium occasionally showed enzyme activity.

Proliferative lesions of serous membranes in ovariectomised female and entire male dogs after stilboestrol administration.

It was shown previously that ovarian tumours, which frequently metastasise to serous surfaces, can be produced in dogs by prolonged administration of stilboestrol. The ability of non-ovarian serous surfaces to proliferate in response to stilboestrol, independently of ovarian metastases, was studied in 5 ovariectomised females and 4 entire males. Papillary proliferations occurred on the uterine serosae in 3 females and on the testicular and cpididymal serosae in 2 males. In no case was there papillary proliferation on serous membranes of non-genital organs. One of 2 entire females given stillboestrol had proliferations regarded as metastases of ovarian origin on non-genital serous membranes. It is concluded that, in addition to ovarian changes, stilboestrol can cause primary proliferative lesions in the serous membranes of certain genital organs in both sexes. There is no evidence that serous membranes of non-genital organs be capable of a primary proliferative response to stilboestrol.

Distribution of C 14-labeled dimethyl sulfoxide in tissues of intact animals

Summary Adult guinea pigs were loaded with 2 ml of 50% C14-DMSO in saline and sacrificed at predetermined intervals. Liquid scintillation assay of tissues and body fluids indicated presence of C14-DMSO in all assayed tissues and fluids. In blood serum DMSO was not bound to serum proteins. Autoradiographic results indicated over-whelming tendency of C14-DMSO to be confined to interstitial spaces. Localization of C14-DMSO activity sites along cell membranes of various tissues and organs is interpreted in terms of relative impermeability of cell membranes to pharmacological doses of DMSO solvated in body fluids.

Studies on sodium-potassium-activated adenosinetriphosphatase. V. Correlation of enzyme activity with cation flux in six tissues

The activity of the enzyme Na-K-activated ATPase, which is closely related to the active transport system of sodium and potassium in human erythrocytes, was determined in human erythrocytes, frog toe muscle, squid giant axone, frog skin, toad bladder, and in the noninnervated membrane of the Sachs organ of the electric eel. The temperature coefficient of the enzyme was determined in each tissue except the erythrocytes. The results were expressed in moles/sq. cm./sec. and compared with the active cation fluxes, expressed in the same units, for these tissues reported by other investigators. The enzyme activities were corrected for the temperature, at which the flux values had been obtained. A significant correlation was found between Na-K-activated ATPase activity and active cation flux over a 25,000-fold range. The average ratio of equivalents cation transported per mole ATP hydrolyzed (cation/ATP ratio) was 2.56 ± 0.19, which is significantly higher than 2, but not significantly different from 3. The total ATPase activity was not significantly correlated with the flux values. The value of 2.56 for the cation/ATP ratio agrees with many other determinations of this ratio for various tissues reported in the literature. These findings further strengthen the assumption that the Na-K-activated ATPase is closely related to the active cation transport involved in maintenance of ion gradients in single cells, transport of salt and water across epithelial membranes, and repolarization processes in muscle, nerve, and electric organs.

Studies on diabetic beta-1 lipoprotein

We prepared β1-lipoprotein from human diabetic serum and studied its diabetogenic action. The Following results were obtained.1) Intravenously injected insulin I131 was retained in the blood of diabetic patients with and without insulin treatment.Therefore, it was suggested that diabetics had common unknown extracellular factors in their serum which impeded the distribution of insulin I131 to the tissue. At the same time, the blood sugar reduction by insulin was lower in diabetics than in normals.2) Diabetic β1-lipoprotein inhibited the glucose output of rat liver-slices and the glucose uptake of rat diaphragms, but in normal ones it accelerated both.3) Diabetic β1-lipoprotein inhibited the binding of insulin I131 to liver, kidney and muscle slices of rats. Consequently it was suggested that the retardative factors of insulin I131 in blood might be diabetic β1-lipoprotein.4) Diabetic β1-lipoprotein intravenously injected, was more distributed than normal to target organs, and the blood concentration of the former was lower than that of the latter. These findings were in contrast with the concentration curve of insulin I131 (mentioned in 1).At this experiment, after an injection of diabetic β1-lipoprotein the blood sugar increased but decreased after normal.5) The greatest amount of injected diabetic β1-lipoprotein I131 distributed to cell membrane and was not washed out by diffusion.Therefore, chemical binding of lipoprotein with cell membrane was thought to be the reason.6) According to the above mentioned findings, it was thought that, Diabetic β1-lipoprotein intravenously injected, rapidly bound with cell membranes of target organs of insulin and thus inhibited the active transport of glucose of insulin and elevated the blood sugar level.7) By an intravenous injection of diabetic β1-lipoprotein blood sugar was slowly increased and reached its maximum after 4 hours of injection and after 6 hours hyperglycemia still continued. By daily injection, continuous hyperglycemia was obtained.The mechanism of hyperglycemia was thought due to the inhibition of peripheral glucose utilization.8) An intravenous injection of diabetic β1-lipoprotein inhibited the action of insulin and elevated the blood sugar level.9) By the use of infrared-spectrophotometry, it was observed that diabetic β1-lipoprotein did not contain any unknown substance and the proportion of chemical composition of its fraction was not abnormal.10) As the result of chemical analysis of diabetic β1-lipoprotein, cholesterol, phospholipid, neutral fat and protein were slightly larger in quantity than normal.11) By the use of metachromasy of methylorange it was made clear, that the protein fraction of diabetic β1-lipoprotein was different from normal.Therefore the structure of protein of diabetic β1-lipoprotein seemed to be abnormal.

Antigenicity of connective tissue extracts. II. Stimulation of auto-and iso-antibodies by heterologous antigen.

SummaryTissue fixed globulin has been demonstrated in interstitial tissue and basement membranes of various guinea pig organs following inoculation of guinea pigs with saline extracts of rabbit tendon. These animals had a significantly diminished growth. It is likely that the fixed globulin represents “auto-antibody” induced by heterologous antigen.

ABCDEF

At the present time, ascariasis is to be considered as one of the serious medical problems in the oriental countries including Japan. In many human experiments in our country, it is found that both santonin and hexylresorcinol are reliable in their anthelminthic action (1). From the result of comparable studies on the pharmacological as well as clinical benefits of both drugs, it is confirmed that santonin is markedly superior to the other in view of its negligible side action. Effective doses of santonin never induce ill effects except a slight and temporary disturbance in vision, while oral administration of hexylresorcinol induced some local irritations on the mucous membrane of the digestive organ, even when it was administered under a perfect condition of coating on it. After taking santonin, ascaris are expelled alive and active. Some authorities, therefore, assume that santonin is not directly toxic to the parasites, but that rather they are irritated by the drug and migrate from the small intestine to the colon to be expelled (2-6). According to others, the drug is excreted in the intestine as an unknown compound, possibly an oxidation product, on which the ascaricidal properties may depend (7-12). But there has been no positive evidence to support both of the views until recently. We have now been able to find out a characteristic locomotion of ascaris in a glass tube which is made to imitate the shape of the bowel, and have good reasons to connect this movement of the worm closely with the anthelminthic activity of santonin (13).

LOCOMOTION OF ASCARIS SUILLA ET LUMBRICOIDES AND THE INFLUENCE OF ANTHELMINTICS UPON THEM

At the present time, ascariasis is to be considered as one of the serious medical problems in the oriental countries including Japan. In many human experiments in our country, it is found that both santonin and hexylresorcinol are reliable in their anthelminthic action (1). From the result of comparable studies on the pharmacological as well as clinical benefits of both drugs, it is confirmed that santonin is markedly superior to the other in view of its negligible side action. Effective doses of santonin never induce ill effects except a slight and temporary disturbance in vision, while oral administration of hexylresorcinol induced some local irritations on the mucous membrane of the digestive organ, even when it was administered under a perfect condition of coating on it. After taking santonin, ascaris are expelled alive and active. Some authorities, therefore, assume that santonin is not directly toxic to the parasites, but that rather they are irritated by the drug and migrate from the small intestine to the colon to be expelled (2-6). According to others, the drug is excreted in the intestine as an unknown compound, possibly an oxidation product, on which the ascaricidal properties may depend (7-12). But there has been no positive evidence to support both of the views until recently. We have now been able to find out a characteristic locomotion of ascaris in a glass tube which is made to imitate the shape of the bowel, and have good reasons to connect this movement of the worm closely with the anthelminthic activity of santonin (13).

Does the Rabbit Chew the Cud?

THE curious food pellets described by Morot and mentioned in Dr. Madsen's letter must be familiar to all laboratory workers who have occasion to open the rabbit's stomach. They have frequently been seen in the parasitological department of this Laboratory during the course of work carried out on the stomach worm Graphidium strigosum, and we have wondered how it could be possible for the mucous membrane of a large hollow organ to form such perfectly rounded pellets.

Studies in Experimental Hypersensitiveness in the Rhesus Monkey

Abstract In recent studies by Walzer, Sherman, Feldman, and Kaplan (1, 2) it was demonstrated that the mucous membranes of the eyes and of the nose in humans could be passively sensitized at will with sera containing atopic reagins. Gray and Walzer (3) have shown that the mucous membranes of the rectum in man are also subject to passive local sensitization with such sera. Similar studies in human beings on the mucous membranes of organs in the thorax and abdomen were naturally limited because of the inaccessibility of such tissues. Recently, Straus (4) demonstrated conclusively that the skin of the Macacus rhesus monkey could be passively sensitized with human atopic antibodies. This finding suggested the use of the rhesus monkey for studying passive sensitization of those organs which could not be experimentally explored in human subjects. The present communication is a brief preliminary report of the results of such experiments in passive sensitization performed on some of the abdominal organs in the rhesus monkey.

BERLIN

<h3>Predisposition to Colds</h3> P. Schmidt, hygienist of Halle, undertook recently a series of experiments on the origin of ordinary colds in man. His main object was to discover what persons have a special predisposition for colds. He found that colds occur most commonly in persons whose heat regulatory system easily breaks down. When such a person is chilled for some time, there is brought about, through the action of the cold, exactly as in a normal person up to this point, a contraction of the blood vessels of the skin and also of the mucous membranes of the respiratory organs. This vascular contraction of the mucosae soon retrogresses, however, in a healthy person, whereas in the person who is predisposed to colds it continues much longer. Owing to this contraction of the mucous membranes, the defense forces can no longer be adequately mobilized, and as a result of the disturbance