The ultrastructure of the developing neural lobe

Summary The ultrastructure of the developing neural lobe of rats was studied. The results revealed three periods in its development. The first period lasts till the 17th day of fetal life. At its beginning the anlage of the neural lobe is formed as a mass of cells very similar in appearance and in connection with the subependymal cells of the future median eminence. During the first period the cells of the anlage differentiate into pituicytes, and the penetration of the first nerve fibres and blood vessels among them is seen. The second period is from the 18th day of fetal life till one month after birth. At its beginning the first signs of neurosecretory activity were detected. During the period increasing numbers of neurosecretory fibres penetrate into the neural lobe, and the pituicytes show morphological signs characteristic of active cells. An increase in the functional activity of the neural lobe is also detected. The third period is from the end of the 1st till the end of the third month. During this period the development of the neural lobe proceeds and at the end it has the appearance of the adult gland. During this period the pituicytes gradually lose the signs of activity and at the end of the period they look like those observed in adult animals. Considering the results from the study, together with some data from previous investigations it is suggested that the pituicytes exert some stimulating and regulative influences on the process of neurosecretion in the neural lobe.Research fellow of the Alexander von Humboldt Foundation.     

On the fine structure of the perivascular cells in the neural lobe of rats

Summary The purpose of this study is to investigate the ultrastructural features of perivascular cells as found in the neural lobe of the hypophysis. In particular, an inquiry was made into the nature of the relationship of such cells to neurosecretory fibers and endings. The latter, in fact, are often invaginated within the perivascular cells and enveloped by their processes; furthermore, they often reveal a certain number of empty granules as well as characteristics of degenerative nature. In the course of this study the localization of the perivascular cells has been investigated as well as that of their processes within the extensive interlobular network typical of the hypophysial neural lobe of rodents. Based on the data gathered, the hypothesis is put forward that the perivascular cells play an important role in the turnover of neurosecretory endings, both under physiological and experimental conditions, contributing thereby to the release of post-hypophysial hormones.With the technical assistance of Vincenzo Panetta.     

Perivascular regions of the rat neural lobe

Summary Neural lobes and portions of occipital cortex from rats were examined electron microscopically following fixation in 4% tannic acid in 2.5% glutaraldehyde. The procedure allowed a clear demonstration of the perivascular space and intercellular spaces in both tissues. The perivascular spaces in the neurosecretory tissue was far more extensive than in the neural tissue and the role of this region in relation to the process of neurosecretion is discussed.     

Ultrastructure of the rat neural lobe during recovery from hypertonic saline treatment

Summary Neural lobes from rats which had been allowed to drink hypertonic saline for five days were examined electron microscopically and by bioassay of oxytocin levels. The profound changes in the ultrastructure were examined and the reversal of these changes in morphology was followed until the hormone levels returned to normal. The recovery of the gland as measured by the two parameters of structure and hormone content indicated that the morphological recovery apparently preceded the hormone level recovery, a factor which might be explained by continued release of hormone during the period of recovery.     

新生小鼠卵巢移植雄鼠肾囊下卵泡的生长发育

将1日龄小鼠卵巢移植入成年雄鼠肾囊下,分别于移植后18d、36d回收移植卵巢进行形态学、组织学观察,以评价卵巢移植体在成年雄性受体小鼠体内生长及卵泡发育潜能。结果表明:移植体生长增大,有各级生长卵泡发育;18日龄移植体平均直径为1881.1μm±204.7μm,与1日龄卵巢相比差异极显著(P<0.01),卵泡发育到有腔卵泡阶段;36日龄移植体平均直径达2575.3μm±466.4μm,显著大于18日龄移植体(P<0.01),有成熟卵泡出现,未观察到黄体;从移植体分离到GV期卵母细胞和卵丘卵母细胞复合体。研究表明1日龄小鼠卵巢移植体在雄性受体生理环境中具有正常生长发育和形成成熟卵泡的潜能。     

Electron microscopy of neurosecretory nerve fibres in the neural lobe of the embryonic mouse

Summary Nerve fibres of the neurosecretory hypothalamo-hypophyseal tract were studied in embryonic C3H mouse neural lobes; at least four glands at each gestational day 15–19 were examined.Single axons and small bundles of fibres are visible at gestational days 15 and 16. By day 17 large fibre bundles penetrate between glial cells. They increase in number during the next two days.Electron-lucent and electron-dense vesicles are seen in the fibres of the 15th and 16th gestational days. In the 17–19 day-old embryos development is characterized by a successive rise in the number of the two types of vesicles. The mean diameter of the electron-lucent vesicles is approximately unchanged in all the stages examined (50 nm). The electron-dense vesicles increase in size from approximately 80–90 nm at days 15–16 to 140 nm at the 19th gestational day.By day 19 contacts between neurosecretory fibre terminals and the outer basement membrane of internal and peripheral capillaries are occasionally observed. The possibly adrenergic nature of a few terminals contacting peripheral vascular structures in 17 and 18 day-old embryos is suggested.This investigation was supported by grant No. 2180-020 from the Swedish Natural Science Research Council. The skilful technical assistance of Mrs. Ulla Wennerberg is gratefully acknowledged.     

Effects of hormone-releasing stimuli on the area of the perivascular space in the neural lobe of the rat

Summary Neural lobes from rats subjected to neurohypophysial hormone-releasing stimuli were examined electron microscopically following fixation in 4 % tannic acid in 2.5 % glutaraldehyde. This fixation allowed the delineation of the perivascular space in the neural lobe tissue. Measurement of the area of the perivascular space showed that it was significantly increased in the rats subjected to vagal stimulation and intraarterial calcium ions compared to the control rats. The rats which had been subjected to haemorrhage as a hormonereleasing stimulus did not show any significant change in the area of the perivascular space. The significance of these findings in relation to hormone release is discussed.     

Pituitary astrocytes from the neural lobe of rats

Summary Tissue culture preparations of adult and neonatal rat pituitary neural lobes were examined by use of cell-type specific immunohistochemical markers. Cultures obtained from explanted or dissociated adult tissue or explanted neonatal tissue produced cells immunoreactive for endothelial and fibroblast markers. In contrast, dissociated neonatal tissue produced, in addition, two distinct forms of astrocytic glial cells immunoreactive for glial fibrillary acidic protein, one of which was also immunoreactive for the ganglioside GD3.     

Effects of blinding on the ultrastructure of mouse pinealocytes with particular emphasis on the dense-cored vesicles

Summary The mammalian pineal is thought to produce an antigonadotropic principle under conditions of reduced photoperiod, constant darkness or blinding by optic enucleation. A number of previous studies on mammalian pineals have suggested that the dense-cored vesicles present in pinealocytes may represent morphological evidence of secretory activity.In the present study the ultrastructure of pinealocytes was studied in adult Charles River CD-1 mice blinded by optical enucleation. By one month following optic enucleation the mean number of dense-cored vesicles in the cytoplasm of pinealocytes adjacent to pericapillary spaces had significantly decreased by 55% when compared with intact controls, and remained at this low level at two months and six months. A relative increase in the proportion of large agranular vesicles and an increased number of large, irregular vacuoles was observed also in the pinealocytic polar processes of blinded mice. When compared to control mice the pinealocytic Golgi regions appeared to be hypertrophied in blinded mice. The apparent stimulation of pinealocytic organelles coupled with the observed decrease in dense-cored vesicles suggest an increased synthesis and release of secretory product.Supported in part by NIH Grant No. HD 08759     

The cell cycle in the small intestine transplanted beneath the kidney capsule in syngenic mice

Summary Transplantation of a small fragment of the ileum beneath the kidney capsule in syngenic mice results in the formation of a cyst lined with proliferating intestinal epithelium. The duration of the cell cycle in this epithelium was determined (using tritiated thymidine and the FLM method) as 14.5 h, as compared with 11.5 h in the intestinal epithelium in situ. We conclude that the intestinal content has little effect on the cell cycle of epithelial cells of the small intestine.     

Quantitative assessment of GABA-uptake sites in the neural lobe by electron-microscopic autoradiography

Summary Distribution of (³H)GABA in the rat neural lobe was investigated 5 min after intracarotid administration using quantitative electron-microscopic autoradiography. Specificity of (³H)GABA-uptake was tested by pretreatment of control animals with nipecotic acid. It was concluded that, apart from a small fraction in the perivascular spaces, radioactivity was present exclusively in pituicytes. The results confirm and quantify earlier in-vitro observations; they are compared with recent immunocytochemical findings that reveal the presence of glutamate-decarboxylase-containing axons in the neural lobe. It is concluded that there may be GABAergic terminals that lack an uptake mechanism for exogenous transmitter. Nevertheless, (³H)GABA autoradiography is useful in demonstrating other functional components of GABAergic systems, i.e., glial cells.     

骨髓移植小鼠二甲基亚硝胺肝纤维化模型的建立

目的构建绿色荧光蛋白(GFP)标记骨髓细胞的小鼠,并复制其二甲基亚硝胺(DMN)肝纤维化模型。方法 ICR雄性小鼠32只,随机分为正常组6只和移植组26只。移植组接受致死量γ射线照射后,经尾静脉输入GFP转基因小鼠的骨髓细胞;正常组不进行照射和移植,仅尾静脉注射等量生理盐水。两个月后制备血涂片,观察移植组造血重建情况,造血重建动物再分为对照组和造模组,造模组用DMN按每次10mg/kg体重腹腔注射制备肝纤维化模型,隔日一次,正常组和对照组给予等量生理盐水。设染毒后3周和4周两个时间观察点,生化法测定肝功能;Jamall法检测肝组织羟脯氨酸含量;HE染色及天狼猩红染色观察肝组织炎症、坏死及纤维组织增生情况;GFP免疫荧光组织化学观察骨髓源性细胞在肝脏的归巢特点。结果骨髓移植两个月后,移植组外周血中出现满视野GFP+细胞。与正常组比较,两个时间观察点造模组肝功能(ALT、AST、Alb及T.Bil)均明显异常(P<0.05),肝组织羟脯氨酸含量显著增高(P<0.05),造模3周末肝组织出血性坏死,有炎性细胞浸润,但尚未形成完全的纤维间隔;造模4周末肝组织炎症、坏死程度加重,可见完全的纤维间隔,在DMN造模动物肝组...     

The three-dimensional structure of the zona pellucida in growing and atretic ovarian follicles of the mouse

Summary The present study provides further details on the fine-structural three-dimensional architecture of the zona pellucida (ZP) in growing and atretic follicles of mice by use of ruthenium red in combination with the detergents Triton X100 and saponin. These detergents were used for extraction of the soluble fraction of the zonal proteins in an attempt to expose the structural zonal glycoproteins, which in turn can be viewed as minute three-dimensional networks upon transmission- and scanning electron-microscopic examination. By use of these methods, the ZP of growing follicles appeared to be formed by interconnected filaments which also bind to globular structures building up a three-dimensional lattice. In contrast, the ZP of stage I as well as other (II and III) stages of atretic follicles showed a structure characterized by the presence of closely packed granules connected with short filaments to form a close-mesh reticulum. This structural change of the ZP, which in the present study is also associated with the disappearance of gap junctions within the granulosa and cumulus cell population, might represent one of the early events involved in the onset of atresia. These changes, most probably depending on an altered secretory activity of both oocytes and follicle cells, might lead to a degradation of the ZP network structure and to its subsequent increased density (condensation). All these morphodynamic events eventually contribute to a sequestration of the oocyte in the early stage of atresia.     

Rapid refilling of neurosecretory terminals with secretory granules after an acute stimulus to the isolated neural lobe of the rat

Summary Quantitative ultrastructural techniques were used to study changes in the distribution of intracellular organelles in neurosecretory terminals of the rat neurohypophysis during recovery from a depolarising stimulus in vitro. The volumetric density of neurosecretory granules which, in profiles of nerve terminals sectioned through the area of contact with the basal lamina, was decreased as a result of stimulation, returned to control values within 2 h after cessation of stimulation. We conclude that, even in the absence of the cell body and action potentials propagated from it, granules can migrate quickly within the terminal region of the neurosecretory axons to refill a depleted compartment.     

Ultrastructure of the male rat hypophysis chronically grafted under the kidney capsule

Summary Male rats were divided in two experimental groups. In group I two partes distales of the hypophysis were grafted under the kidney capsule and in group II two complete hypophyses were transplanted. Animals were killed 5 to 22 months after the operation. The grafted tissue was excised and processed for light and electron microscopy.The transplanted pars distalis tissue showed a well developed vascularisation in contrast to the pars intermedia which appeared poorly vascularised. Six different cell types were observed in grafted pars distalis. They correspond to the different types of cells found in the rat pars distalis in situ. The predominant cell type in the graft displayed all the morphological characteristics of stimulated prolactotrophs. Pars intermedia cells appeared hypertrophied resembling the MSH cells under stimulation. Two types of syncytial formations were frequently seen. One of them appeared to originate from prolactotrophs and the other from MSH cells. Bodian impregnated fibres and structures resembling either growth cones of axons or typical nerve endings were observed in the pars intermedia of long-term grafted hypophyses. Pituicytes remained as isolated clusters of cells. Canaliculi lined by two or more pituicytes were observed. Saccular formations resembling the hypophyseal cleft appeared in all grafts studied. The present findings suggest that in the male rat the chronically grafted pituitary gland is capable of synthesising most or all the hormones which are known to be produced by the gland in situ. Furthermore, prolactin and MSH seem to be the predominant secretion of the transplanted pituitary.Supported by the National Research Council of Argentina. Grant 7315/74Members of the Research Career of Conicet, Argentina     

The rostral zone of the intermediate lobe of the mouse hypophysis,a zone of particular concentration of corticotrophic cells

Summary The rostral zone of the intermediate lobe of the mouse hypophysis can clearly be distinguished from the other lobes of the adenohypophysis, especially from the pars tuberalis and the remainder of the intermediate lobe. It consists almost exclusively of corticotrophic cells which show reactive changes after adrenalectomy. The hypophysial stalk is entirely surrounded by this zone; laterally it forms large cell aggregations which extend dorsally as thin cell strands. The corticotrophs are also found within the hypophysial stalk which they invade along the blood vessels; frequently they are dispersed among the typical cells of the intermediate lobe, especially along the neural lobe and the hypophysial cleft.On leave from the Department of Veterinary Anatomy, University of Missouri, Columbia, Mo., U.S.A.     

Organization and development of the perivascular space system in the neurohypophysis of the laboratory mouse

Summary The organization of the system of perivascular space around the capillaries in the neurohypophysis was studied in the adult and developing laboratory mouse by the use of histological silver impregnation and electron microscopical techniques.In the median eminence short and long extensions, arising mainly from the shallow space around capillary loops of the primary plexus of the portal system, formed radiations into the adjacent neural tissue of the external zone. The tissue of the neural lobe was separable into non-vascular regions dominated by undilated portions of neurosecretory nerve fibres and pituicytes, and neurovascular regions with perivascular space extensions forming an extensive system of connections between neighbouring capillaries.In the median eminence, the system of extensions of the perivascular space was estimated to increase the neurovascular contact surface area by at least 50%, implying an increased efficiency of the organ without a notable increase of its volume. The possibility that the ramifications of the perivascular space imply an enhanced uptake rate into the bloodstream and a subsequent increased concentration of the neurohormones in the portal blood, was discussed.During development of the median eminence, differentiation of perivascular space extensions of the adult type started in the juvenile of about three weeks of age, when shallow capillary loops had been formed. In the neural lobe, perivascular space ramifications were already present when the internal capillaries were formed and were fairly frequent in ten-day young. At the age of three to four weeks the organization of the system was similar to that of the adult animal.     

The reversal of hyperglycemia after transplantation of mouse embryonic stem cells induced into early hepatocyte-like cells in streptozotocin-induced diabetic mice

Cellular replacement therapy is a potential therapeutic strategy for diabetes. In this study, we investigated the effect of transplantation of induced mouse embryonic stem cells (mESCs) into endoderm and early hepatocyte-like cells in streptozotocin (STZ)-diabetic mice. After embryoid body (EB) formation from mESC, the EBs were cultured in the presence of dexamethasone (DEX) and insulin for 4 days then was added acidic fibroblast growth factor (aFGF), hepatocyte growth factor (HGF) and oncostatin M (OSM) for 10 days, respectively. Blood glucose levels, intraperitoneal glucose tolerance (IGT) test and islet histology were assessed. The result revealed that transplantation of induced mESCs into early hepatocyte-like cells could repair pancreatic islets of control group. Blood glucose levels and intraperitoneal glucose tolerance test were significantly improved in test group compared to control group. Furthermore, there was significant increase in the number of islets in test group compared to control group. The findings declare that induced mESCs into endoderm and early hepatocyte-like cells, are appropriate candidate for regenerative therapy of pancreatic islets in type I diabetes.     

冻融后小鼠休眠胚胎超微结构的变化

目的从亚细胞超微结构的角度揭示其抗冻能力优于正常孵化胚胎的原因。方法利用透射电子显微镜观察小鼠休眠胚胎与正常孵化期胚胎在细胞连接和各细胞器形态与分布上的差异,以及冻融培养后的变化,并进行相关比较分析。结果通过亚细胞结构对比分析发现:冷冻前小鼠休眠胚胎为紧缩状,处于能量代谢较低的"基态",通过冻融后培养,细胞器结构恢复与正常孵化胚胎冷冻前相似;而正常孵化胚胎经过冻融后,线粒体数量减少,细胞核松散,异染色质增多。结论小鼠休眠胚胎与正常孵化胚胎冻融后相比,其细胞状态更有利于物质储存及能量代谢,表明小鼠休眠胚胎从亚细胞超微结构的角度比其正常孵化胚胎更具抗冻性。