Electron Microscopic Observations on the Morphological Changes of Rat Mesentric Mast Cells Induced by Morphine HCl

The effects of morphine HCI on the rat mesenteric mast cells were studied with the electron microscopy. The materials were prepared for electron microscopy by osmium tetroxide fixation and embedding in Epon. The rat mesenteric mast cells showed no distinct morphological changes due to morphine HCl, but the mast cell granlues were changed in various ways. For instance, they formed dusters, showed granular lysis, and an appearance of electron transparency. Frequently, some granules appeared in the extracellular space and the boundary of the granules was not evident. From the results mentioned above, it was suggested that rat mesenteric mast cell granules were affected by morphine HCl in the shape, the granular matrix, and the granular boundaries.

Effect of sodium selenite on the hepatotoxicity induced with carbon tetrachloride

The authors have demonstrated the effect of sodium selenite on the hepatotoxicity due to carbon tetrachloride, by observing the distribution and disaggregation of the pyroninophilic granules in the hepatic cell of the mature male albino mice. Each experimental mouse of the selenite and the selenite plus carbon tetrachloride groups was given a single dose of 4 ug. of sodium selenite per kilogram of body weight and that of the control and the carbon tetrachloride groups was given 0.1 ml. of distilled water alone. Six hours after the first administration of distilled water or sodium selenite, the experimental mice of the carbon tetrachloride and the selenite plus carbon tetrachloride groups were given a single dose of l.0 ml. of carbon tetrachloride per kilogram of body weight and those of the selenite groups were given 0.l ml. of paraffin oil alone. Following the 1ast administration of carbon tetrachloride or paraffin oil, the mice were sacrificed by bleeding (cutting the common carotid artery) at the intervals of 2,3,4,6,8, and 12 hours respectively. Histochemical preparations were stained by the methyl-green and pyronin method and oil red 0 method. The hepatotoxicity due to the administration of carbon tetrachoride was evident in the hepatic cells; the pyroninophilic granlues were partly reduced in volume in the hepatic cells of the centrilobular and the intermediate zones as early as the 3 hour-period, and markedly reduced or disappeared in the centrilobular and some part of the intermediate zones associated with hydropic degeneration as well as in the 6 hour-period. Thereafter marked reduction or dissolution of the pyroninophilic granules was found and extended as the periportal zone at the 12 hour-period. However, the pyroninophilic granules in the hepatic cells of selenite plus carbon tetrachbride group showed no significant changes in the hepatic cells of these zones, compared to the histochemical feature of the granules in the hepatic cells of the control and the selenite groups. Consequently it is suggested that the lipid peroxidative decomposition of the microsomal membranes, which is induced with carbon tetrachloride, would be prevented by a previous administration of sodium selenite.

Pyroninophilic Granules in Liver Cells of the Mice Treated with Alpha-Tocopherol and Thioacetamide

In an attempt to clarify the protective action of an antioxidant agent against acute toxicity of thioacetamide (TAA) and in order to throw some light on an satisfying concept of the mechanism of its action, a single dose of alphatocopherol (200 mg per kg) was given orally by stomch tube to male mice prior to the administration of thioacetamide in a dose of 200 mg per kg of body weight. Sections of liver samples, obtained from the mice which were sacrificed at intervals of 3, 6, 9, or 12 hours after TAA administration, were stained using the methyl green-pyronin technique. At 3 hours following TAA administration, the pretreatment with alpha-tocopherol inhibited almost completely such alterations of the hepatocytes in the animals given TAA alone, as revealed by loss and clumping of cytoplasmic pyroninophilic granules in the periportal zone of the lobule. At 6, 9, and l2 hours, the prevention of alpha-tocopherol was incomplete in degree and extent. The changes of the hepatocytes were more intense and extensive in the TAA-treated 6 to 12 hour-groups than in the 3 hour-group of TAA-treated ones. Some discussion is given of the mechanism of TAA toxicity, with respect to the microsoma1 lipid peroxidation.

Liver Cells of Cordotomized Rats after Single Doses of Carbon Tetrachloride

In attempting to ellucidate the mechanism of action of CCl4 toxicity on the liver, the histobgical and histochemical studies were carried out, at the cellular or ultrastructural level, rats were given a single oral dose of 1.25 ml/kg of CCl4 one hour after cervical spinal cordotomy. Hepatic lesions induced by CCl4 administration such as the fatty change of hepatic cells and the sinusoidal congestion were abolished by cordotomy. The decreased activities of adenosine triphosphatase and alkaline phosphatase in the hepatic cells and bile canaliculi of the poisoned animals were restored to a large extent by the operation. Cordotomy also prevented some liver cell changes as seen by the electron microscope in the CCl4-intoxicated rats. It is evident that the hepatotoxic effects of carbon tetrachloride can be inhibited or prevented by cervical cordotomy.

On the Effect of Morphine Hydrochloride on the Mesenteric Mast cells of Albino Rats

Histological studies were carried out on the degranulation of mesenteric mast cells of white rats caused by injections of morphine and nalorphine hydrochloride intravenously and the following conclusions were obtained. 1. By the injection of morphine hydrochloride fairly significant degranulation of the mesenteric mast cell was observed. 2. In various experimental doses of morphine hydrochloride the cytological change of the degranulation was not proportional to the doses of it in cases given more than 12mg./kg. of body weight. 3. The degranulating effect of the mesenteric mast cell by the injection of morphine hydrochloride was significantly inhibited by an adrenalectomy.

An Electron Microscopic Study on the Junctional Complex in Frog Epithelia

Electron microscopy on the skin of young frogs, Rana temporaria, has been carried out with particular reference to cellular attachment sites. For the first time now several technical developments allow a more detailed visualization of the fine structure within the cellular attachment sites as well as making it possible to show the ultrastructural morphology of the junctional complexes, and to demonstrate that the desmosomes are regularly distributed around each skin cell, especially in the S. granulosum. The relations of these findings to those of previous investigations concerning the functional organization of the junctional complexes and to the findings in skin cancer from a cellular adhesion view point have been briefly discussed.

Cytomorphic Effects of Chemical and Hormonal Agents, and Electronic Stimulation on the Peritoneal Mast Cells of the Rat

After the intraperitoneal injections of alloxan, carbon tetrachloride, cortisone acetate, adrenocorticotrophic hormone, morphine hydrochloride, toluidin blue, physiological saline solution, distilled water, and direct stimulation with electronic current, the peritoneal mast cells of the rat were observed in order to document and study the cytomorphic changes. Adult Sprague-Dawley strain albion rats were used. The substances tested were dissolved in physiological saline solution and injected into the abdominal cavity. Three to twenty four hours later the rats were sacrificed and the morphological changes of the peritoneal mast cells were observed by means of phase contrast microscopy and ordinary light microscopy. Cytomorphic effects of alloxan on the mast cells were comparatively marked and those effects of CC14, cortisone, ACTH, morphine-HCI, and physiological saline solution were slight and similar to each other. But the distructive effects of toluidin blue, distilled water, and electronic stimulation on the mast cells were severe and noticeable in this study. These results indicate that the intraperitmeal mast cells of rats show more sensitive reactions to a metabolic poisn alloxan, a low osmotic pressured-material distilled water, and a histamine liberator toluidin blue, and a physical stimulus electronic stimulation than the other similar chemical agents.

On the Degranulation of Mesenteric Mast Cells Caused by Antihistamine in Albino Rats: Effects of Various Dosages of Antihistamine

Degranulation of the mast cell has been reported by the injection of histamine liberators and other chemical agents. Chlorpheniramine maleate (1.2mg./kg. and 0.3mg./kg. comprising 1/74and 1/290 of LD50 respectively), which is an antihistamine agent, in physiological saline solution for intravenous injection and in Tyrode solution for intraperitoneal injection were given in single dose. The mesenteric mast cells stained in Pugh solution, as applied by Lee (1968), were counted according to the classification of An (1964) in 4 types; the typical normal mast cell, the Grade I type of mast cell, the Grade II type of mast cell and the Grade III type of mast cell. In the experimental rats given 1.2mg./kg. of chlorpheniramine intravenously, more mesenteric mast cells were s1ightly degranulated than those cells of the rats given 0.3mg./kg. of chlorpheniramine and the control rats. In the experimental rats given 1.2mg./kg. and 0.3 mg./kg. of chlorpheniramine intraperitoneally, more mesenteric mast cells were slightly degranulated than those cells of the control rats. However, in this intraperitoneal study the degree, or severity, of degranulation of the mesenteric mast cell was not in direct proportion to the dosage of this antihistamine. Consequently it is deduced that the experimental dosage of the antihistamine chlorpheniramine maleate, applied 1/74 and l/290 of LD50, caused an evient degranulation of mesenteric mast cells of the albino rats associated with probable histamine liberation.

A Histochemical Study of Cholinesterase Activity in Rabbit's Retinae

In the present study the specific and nonspecific cholinesterase activities of the rabbit's retinae in the fetus, the neonatal, the light-isolated, and the reopened group, which consisted of 65 healthy young rabbits, weighing about 300 to 500 gm, 33 rabbit's fetuses, and neonatal rabbits, were histochemically ovserved by means of the cholinesterase method recommended by Gerebtzoff (1953) and the embedding and sectioning method pesented by Koelle and Friedenwald (1950). Cholinesterase activity of the retinae in the 15 days fetuses was not present but began to develop in the 20 days fetuses. In the 1 week group after suturing the eyelids, the most remarkable activity of specific and nonspecific cholinesterase was observed in the posterior polar area. The nearer to the peripheral area of the retina the weaker the enzymetic activities became. In the 2 weeks group after suturing eyelids, the enzymatic activity was reduced. In the 4, and 8weeks groups after suturing the eyelides, the enzymatic activities were remarkably reduced. In the l4 days after reopening eyelide, which group has previously been kept under the condition of light isolation for 4 weeks, enzymatic activities were fairly recovered and compared with the normal control group. Consequently it is histochemically deduced that the gradual change of specific cholinesterase activities in the rabbit's retinae was closely related to the visual function.

An Autoradiographic Study on Metabolism of Serotonin in Cerebral Nerve Cells Using 5-Hydroxytryptophan-C

In order to demonstrate autoradiographically the sites of serotonin metabolism in the brain, DL 5-HTP and DL 5-HTP-C14 were intraperitoneally administered to healthy adult mice. In order to distinguish histochemically serotonin-like substances which have staining characteristics similar to the enterochromaffin cells of the gastrointestinal tract, serotonin-releaser reserpine was administered intravenously to healthy adult rabbits. A stripping film technique for autoradiography, a ferric ferricyanide reduction test by Schmorl, and a Gomori-Burtner methenamine silver staining method for argentaffin cells were used in this study. In the brain tissues of mice treated with 5-HTP, it was observed that the cytoplasm of the nerve cells, of the cerebral cortex had blue positive staining substances by the ferric ferricyanide technique. In similar tissue sites in mice treated with 5-HTP-C14, a number of blackened-par-ticles reduced by beta rays were easily found. especially in the cytoplasm of nerve cells and neuroglia cells. It is suggested that the serotonin precursor, DL 5-hydrox-ytryptophan is metabolized the cerebral tissue, and serotonin is synthesized also in the nerve cells and the neuroglia cells.

A Phase Contrast Microscopic Study on the Induction of Cellular Deformation in Mast Cells

The influence of colchicine, colcemid, ginseng-alcohol extract, saponin, garlic alcohol-ether extract, and X-irradiation on free floating peritoneal mast cells were studied in rats by means of a phase contrast microscope. After the intraperitoneal injection of colchicine or colcemid the mast cells of the albino rat peritoneal fluid show an altered morphology. Ginseng-alcohol extract, allicin extracted from garlic, and X-irradiation do not elicit this cytological response.Mast cells exposed to saponin exhibit a severe destruction and degranulation. The relation of the mast cell-damaging agents, metabolic poisons, and X1-irradiation to the mast cell response is discussed with reference to the morphological changes. These are compared with the data of Padawer (1960, 1966).

Differentiation of Heterosexually Transplanted Embryonic Ovary Associated with Embryonic Testes of the Albino Rat into Castrated Hosts

The authors studied Witschi's theory of "corticomedullary inductors" with the; heterosexual grafts of the embryonal gonads of the rats in very close proximity or in remote distance each other for the effect of the inductor substance and the possibility of the substance acting as blood-borne agent when multiple embryonal testes and one or two ovaries were separately grafted in distant sites in the mammalian level. The heterosexual grafts of the embryonal gonads aging 16 days old were performed as the methods Macintyre (1956) used. Additionally the author grafted one or two embryonal ovaries of the same age in the subcapsular site and multiple embryonal testes of the same age in the similar site of the opposite kidney of the same host and allowed to develop at the sites for 3 weeks. The explants removed from the host, were fixed in Bouin's fluid, embedded in paraffin, sectioned serially at 6mu, and stained with hematoxylin and eosin. The embryonal transplanted ovary with testis in close contact, was inhibited and depressed to the one side probably due to the more rapid growth and differentiation of the testis as compared with the ovary and contained a tubular structure (seminiferous-like tubule) in which the degenerating oocytes were found. The author presumed the testicular effect upon the mutual ovary as the activity of the diffused inductor substance derived from the testis. In the group, which more than 15 embryonal testes (maximally 25 testes were grafted) were transplanted in the subcapsular site and one or two ovaries in the opposite site of the kidney of the same host, the ovarian grafts, which were at a distant site from the multiple testicular grafts, showed inhibited growth and differentiation by the similar appearance of the transplanted embryonal ovary with testis in mutual contact. By this observation the author considered the inhibited growth and differentiation of the effect of blood-borne inductor substance derived from the multiple testicular grafts of the opposite site of the host kidney.

Effect of Cortisone and X-ray Irradiation on Mast Cells of the Adrenalectomized Albinor Rats

The effects of cortisone administration and whole body irradiation by X-ray upon mesenteric mast cells of intact and adrenalectomized albino rats were studied. In intact rats, the administration of cortisone (50mg./kg) and whole body irradiation by X-ray (800r) caused severe degranulation and disruption of mesenteric mast cells within 24 hours. However their degranulation and disrupting effects upon mesenteric mast cells were markedly inhibited after the removal of the adrenal gland. Although the adrenalectomy alone hardly caused these morphological changes of mesenteric mast cells of the albino rats. According to this experiment it is fairly clear that the effects of cortisone and whole body X-ray irradiation inducing degranulation and disruption of mesenteric mast cells of the albino rats, were not direct phenomena but they indirectly affected the mesenteric mast cells through some special mechanism mediated by the adrenal gland.

The Histochemical Changes of Cholinesterase Activity in Rabbit's Retinae at Early Postnatal Stages

The authors studied histochemically the enzymatic sites of retinal cholinesterase activity in the albino rabbit during different early postnatal periods. The retinae of young rabbits weighing approximately 60gm. whose palpebrae were still fused and unable to function for sight, and of rabbits weighing approximately 300gm. which were able to see, were obtained immediately after killing the animals by the intravenous injection of air or by decapitation and fixed in formalin-sucrose ammonia fixative, as recommended by Pearson (1963), for about 24 hours and incubated in the substrate containing acetylthiocholine, iodide, as presented by Gerebtzoff (1953), at 37degrees C. for 2 to 3 hours. After the treatment of the retinae, as devised by Koelle and Friedenwald (1950), the retinae were sectioned at 5 micra and mounted both without counterstaining and after counterstaining with hematoxylin alone. In young rabbits weighing approximately 60gm., moderate cholinesterase activity was observed only in the ganglion cells of the retinae and slight enzymatic activity was faintly apparent in the layer of the optic nerve fibers. No enzymatic activity was recognized in the remaining layers of the retinae. In rabbits weighing approximately 300gm. the retinae showed different enzymatic distribution compared to the former. The cholinesterase activity was diversely distributed compared to the former and was localized in the inner nuclear layer, inner plexiform layer, and the ganglion cells showing moderate to slight activity.

Mast Cell Degranulation with Special Reference to the Effect of a Saponin Extract of Ginseng upon the Mesenteric Mast Cell of Albino Rats

Water extract of dried ginseng, which is known as a histamine liberator and induces degranulation and disruption of mesenteric mast cells, is thought to contain many different chemical factors. The essential component, a saponin extract of dried ginseng, was obtained and administered to albino rats. Even minute amounts (l mg in 0.0l cc of normal saline solution) when locally injected into the mesentery of albino rats caused degranulation of mesenteric mast cells. Degranulation of mesenteric mast cells followed the intraperitoneal injection of a crude water extract, of an alcohol extract of dried ginseng, and a direct injection of both extracts into the connective tissue of mesentery. This degranulation is believed to be a saponin fraction of ginseng in the ginseng extracts.

Histological Studies on the Degranulation of Mesenteric Mast Cells of the Rat by Water Extracts of Ginseng

Ever since Fujidani made his report in 1905, many workers have studied the chemical components of Panax Ginseng and their effects on depression of blood pressure. Ri1ey (1952) and other workers have demonstrated the degranulation of mast cells in experimental animaIs treated with some histamine liberators, and the existence of a histamine liberator in the water extract of ginseng has been demonstrated by pharmacological assay by Lee et aI (1960). This present study was intended observe the disruption and degranulation of mesenteric mast cells of rats administered the water extract of ginseng, which might contain the histamine liberator. Variab1e doses of the water extract were injected intraperitoneally, and the degranulation of mesenteric mast cells was histologically demonstrated by means of toluidin blue, Giemsa, May-Gr nwa1d and Wright's stains. Degranulation began in the experimental group given 4ml of the extract mixed with 16cc. of Tyrode solution; the severity of degranulation increased probably with the dose of the extract, and extreme degranulation took place in the groups injected with dose of 6 or 8ml of the extract.

A Histological Study of the Blood Vessels in the Rabbit Spleen

No abstract available.