Liver ultrastructure and a new cell type in the Japanese newt, Cynops pyrrhogaster.

The liver of the Japanese newt, Cynops pyrrhogaster, has been investigated using light, scanning, and transmission electron microscopy. Hepatic parenchyma was composed of clusters and cords or tubules of polyhedral cells separated by a sinusoidal net. Hepatocytes had spherical, euchromatic nuclei with one or more nucleoli and stacked mitochondria with sparse cristae and dense bodies. Rough endoplasmic reticula formed peribiliary stacks and diffusely scattered vesicles and tubules. Smooth endoplasmic reticula were more pronounced in glycogen-rich hepatocytes. Most hepatocytes contained peroxisomes, Golgi complexes and large numbers of fat droplets within the cytoplasm along with glycogen. Some cells were mainly glycogen-storing and contained few or no fat droplets. A special feature of the newt liver was biliary atresia. Bile canaliculi had short, stout microvilli which were entirely atretic in some canaliculi. Canaliculi were sealed off by junctional complexes including zonulae occludentes and maculae adherentes. The latter showed extraordinary wider desmosomal gaps in the vicinity of the atretic bile canaliculi. The sinusoid wall was non-distinctive and contained fenestrated endothelial cells connected to Kupffer cells by zonulae occludentes. A distinctive new cell type (OG cell) was observed in the newt liver. These cells were found individually or in small clusters in proximity with the sinusoidal surfaces. They had small nuclei, a paucity of cytoplasmic organelles, but numerous, unique, osmiophilic granules of two distinct types. Less numerous Type I granules contained homogeneous electron-dense material, and a predominant Type II granule contained circumferentially arranged subparticulation. Granules of both types were detected within the cytoplasm of endothelial cells and within sinusoids together with blood elements. The function of this secretory type cell remains obscure, though it may represent a stage of melanophore.     

Changes in the structure of Nippostrongylus brasiliensis intestinal cells during development from the free-living to the parasitic stages

The ultrastructure of Nippostrongylus brasiliensis intestinal cells was examined in free-living, feeding second-stage larvae, infective, nonfeeding third-stage larvae, and parasitic, feeding third-stage larvae. The intestinal cells of second-stage larvae were characterized by a well-developed microvillar border, large numbers of ribosomes, Golgi complexes, rough endoplasmic reticulum, and nuclei with prominent nucleoli. The intestinal cells of infective, third-stage larvae had very few microvilli and the cells were extremely narrow. Few ribosomes, Golgi complexes, and little rough endoplasmic reticulum were present. Nuclei did not contain nucleoli. When worms were introduced into an in vitro culture system, development of intestinal cells began. By 36 hr, microvilli were well differentiated and the cysoplasm contained numerous ribosomes and Golgi complexes, and rough endoplasmic reticulum, mitochondria, and nucleoli were prominent. These morphological changes were related to changes in the physiology of Nippostrongylus brasiliensis which occur during development from a free-living to parasitic form.     

Ultrastructure and histochemistry of the supportive structures associated with the radula of the slug,Limax maximus

The odontophore and connective tissue-filled portion of the radular sac (called the “collostyle”) of the slug, Limax maximus, were examined by light and electron microscopy. While both of these structures grossly resemble vertebrate cartilage, neither is composed of a type of tissue with the microscopic appearance and histochemical properties of cartilage. The roughly U-shaped odontophore possesses a thin capsule composed of connective tissue. The parenchyma of the odontophore consists of modified muscle cells which are organized into irregular groups by incomplete trabeculae composed of conventional muscle cells. The odontophoral cells are variable in size; they contain glycogen-filled “cores” as well as bundles of peripherally located filaments resembling myofilaments; and they are innervated like muscle cells. The nuclei of the cells are located eccentrically in the glycogen-filled portions of the cells and typically contain prominent nucleoli. The nuclei are surrounded by multiple small Golgi complexes and pleomorphic dense bodies resembling lysosomes. The extracellular matrix of the odontophore is very sparse and contains glycogen and fibrillar material but no histochemically demonstrable acidic mucosubstances. The collostyle consists of a gelatinous type of tissue somewhat like vertebrate mucoid connective tissue. The abundant extracellular matrix contains cross banded filaments, a flocculent material disposed in wavy indefinite strands, and small electron-dense particles. The matrix contains histochemically demonstrable neutral and weakly acidic mucosubstances. The cell population of the collostyle includes solitary muscle cells and fibrocytes containing large quantities of glycogen.     

Stellate cells storing retinol in the liver of adult lamprey,Lampetra japonica

Summary Distribution, localization and fine structure of the stellate cells in the liver of lamprey, Lampetra japonica, were studied during the spawning migration by use of Kupffer's gold-chloride method, fluorescence microscopy for vitamin A (retinol) and electron microscopy. The stellate cells in the lamprey liver differ in some of their properties from those in mammalian livers. Stellate cells which store abundant retinol in lipid droplets, occur not only in the hepatic parenchyma, but also in the dense perivascular and capsular connective tissue of the liver and in the interstitium of pancreatic tissue. In the hepatic parenchyma these cells are located perisinusoidally or along thick bundles of collagen fibrils. The stellate cells display a number of large retinol-containing lipid droplets, granular endoplasmic reticulum, tubular structures, dense bodies, Golgi complex, microtubules, and microfilaments. In the space of Disse, the stellate cells and extracellular fibrilar components such as collagen fibrils and microfibrils (11–12 nm in diameter) are intervened between the two layers of basal laminae. Differentiation and possible functions of the stellate cells in the lamprey liver are discussed.     

Immunohistochemical evidence for cell surface and Golgi localization of galactosyltransferase in human stomach, jejunum, liver and pancreas

Immunohistochemical localization of galactosyltransferase (UDP-galactose: 2-acetamido-2-deoxy beta-D-glucopyranose beta (1-4) transferase) in human tissue specimens of gastric and jejunal mucosa, exocrine pancreas, and liver was carried out at the light microscopic level using affinity purified rabbit anti-human milk galactosyltransferase antibodies. Intracellular localization of galactosyltransferase in epithelial cells appeared as a triangular compact structure close to the apical pole of the nucleus. In hepatocytes, the enzyme was found in discrete spots in the cytoplasm between the nuclei and the bile canaliculi. In addition to the intracellular, juxtanuclear location an intense reaction at the luminal part of the cell surface was found in the lining epithelium of the stomach, in enterocytes of the jejunal villus tips, and in ductular cells of the pancreas. Enterocytes located in the middle portion along the cryptvillus gradient exhibited cytoplasmic staining adjacent to the brush borders. Basolateral membranes appeared negative. Little or no enzyme could be demonstrated in cells belonging to the connective tissue. These results show that secretory cells contain a Golgi apparatus which can be visualized at the light microscopic level by virtue of its content in galactosyltransferase. Presence of galactosyltransferase antigen on the surface of certain cells supports the assumption that ectoglycosyltransferases do exist, at sites, however, apparently not involved in cell contact and adhesion.     

四川短尾鼩肝脏和胰腺的组织解剖学研究

对四川短尾鼩肝脏和胰腺的组织解剖结构进行了研究,结果表明:肝脏分为4叶,它们的肝实质完全分开.肝小叶的分界极不明显,1个肝小叶内的肝细胞大小不一致,靠近中央静脉的肝细胞体积较小,而肝小叶外围的肝细胞体积较大.胰腺的形态松散,无法区分出胰头、胰体和胰尾.胰腺小叶非常明显,大小不等.胰腺的组织结构包括外分泌部和胰岛两部分.     

Electron microscopic study on the innervation of the pancreas of the domestic fowl

Summary The innervation of the pancreas of the domestic fowl was studied electron microscopically. The extrapancreatic nerve is composed mostly of unmyelinated nerve fibers with a smaller component of myelinated nerve fibers. The latter are not found in the parenchyma. The pancreas contains ganglion cells in the interlobular connective tissue. The unmyelinated nerve fibers branch off along blood vessels. Their synaptic terminals contact with the exocrine and endocrine tissues. The synaptic terminals can be divided into four types based on a combination of three kinds of synaptic vesicles. Type I synaptic terminals contain only small clear vesicles about 600 Å in diameter. Type II terminals are characterized by small clear and large dense core vesicles 1,000 Å in diameter. Type III terminals contain small clear vesicles and small dense core vesicles 500 Å in diameter. Type IV terminals are characterized by small and large dense core vesicles. The exocrine tissue receives a richer nervous supply than the endocrine tissue. Type II and IV terminals are distributed in the acinus, and they contact A and D cells of the islets. B cells and pancreatic ducts are supplied mainly by Type II terminals, the blood vessels by Type IV terminals.This work was supported by a scientific research grant (No. 144017) and (No. 136031) from the Ministry of Education of Japan to Prof. M. Yasuda     

Study of RNA synthesis in the livers of aging mice by means of electron microscopic radioautography.

The application of 3H-uridine radioautography results in labeling of the liver cells in which RNA is synthesized at various ages of the mouse. Quantitative changes of RNA synthesis in the hepatocytes of aging mice were studied by electron microscopic radioautography. The silver grains were mainly located in the nucleoli and nuclei and a few in the mitochondria and rough surfaced endoplasmic reticulum of almost all of the cell populations at various ages. The number of silver grains in the hepatocyte gradually increased after birth, reached the maximum at 14 days of postnatal age, then decreased to 24 months with aging. The number of silver grains of the euchromatin was more than those of the heterochromatin of the hepatocyte nuclei at various ages. The number of silver grains of the granular components was more than those of the fibrillar components of the hepatocyte nucleoli at various ages. However, the ratio of silver grains among euchromatin, heterochromatin, granular components and fibrillar components remained approximately constant.     

Ultrastructural location of albumin and fibrinogen in primary cultures of new-born rat liver tissue

In primary cultures of new-born rat liver tissue, albumin and frbrinogen, two proteins normally synthesized by the liver and secreted into plasma were demonstrated by specific antibodies labelled with peroxidase in about 50 and 70% of the hepatocytes; these proteins were not demonstrated in the other types of cells, in particular fibroblasts, present in primary cultures. These two proteins were detected on the ribosomes of the rough endoplasmic reticulum and were also present in the lumina of the rough and smooth endoplasmic reticulum and in the Golgi apparatus. It is concluded that

1. 1. In primary cultures of liver tissue, only the hepatocytes synthesize albumin and fibrinogen.

2. 2. Proliferating cultured hepatocytes are able to synthesize albumin and fibrinogen.

3. 3. The presence of detectable albumin and fibrinogen in the lumina of the rough and smooth endoplasmic reticulum and in the Golgi apparatus in hepatocytes of primary cultures and their absence in the lumina of the rough and smooth endoplasmic reticulum and in the Golgi apparatus in the hepatocytes of adult rat liver might indicate an alteration in the translocation of albumin and fibrinogen through these organelles in cultured hepatocytes.

     

The electron-histochemical detection of the activity of thiol proteinases in normal and cirrhotically altered liver]

The distribution co-activity of thiol proteinases (cathepsin B and cathepsin H) was investigated in normal and cirrhotic liver by the electron-cytochemical method. The reaction product was localized on lysosomes of Kupffer's cells, hepatocytes, endotheliocytes and fibroblasts. Extracellular activity of thiol proteinases was revealed in normal as well as in cirrhotic liver. The reaction product was situated on collagen fibrils near hepatocytes and connective tissue cells. The results evidence that besides participation in intracellular degradation of different proteins, thiol proteinases are secreted by hepatocytes and connective tissue cells of the liver to the intercellular space and can take part in extracellular collagen resorption.     

The structure of the chambered nautilus siphuncle: The siphuncular epithelium

The siphuncle of the chambered nautilus (Nautilus macromphalus) is composed of a layer of columnar epithelial cells resting on a vascularized connective tissue base. The siphuncular epithelium taken from chambers that have not yet begun to be emptied of cameral liquid has a dense apical brush border. The great number of apical cell junctions (zonula adherens) compared to the number of nuclei suggests extensive interdigitation of these cells. The perinuclear cytoplasm of these preemptying cells is rich in rough endoplasmic reticulum. The siphuncular epithelium of both emptying and “old” siphuncle (which has already completed emptying its chamber) both show little rough endoplasmic reticulum but do contain extensive systems of mitochondria-lined infoldings of the basolateral plasma membranes. Active transport of NaCl into the extracellular space of this tubular system probably entrains the water transport involved in the chamber-emptying process. Both emptying and old siphuncular epithelium also show large basal infoldings (canaliculi) continuous with the hemocoel, which appear to be filled with hemocyanin. The apical cell junctions of emptying and old siphuncular epithelium contain septate desmosomes that may help to prevent back-flow of cameral liquid into the chambers.     

商城肥鲵肝及胰的解剖学与组织学观察

利用常规H.E和改良甲苯胺蓝染色方法(modified toluidine staining,MTB),对商城肥鲵(Pachyhynobius shangchengensis)肝、胆囊和胰的解剖学、组织学以及上述器官的肥大细胞进行了观察。结果表明,商城肥鲵的肝小叶不明显,肝索互相连接成网状,肝细胞核大,单核,少数2～3核,肝侧缘细胞胞体大,有的达70μm,但核质比小。肝中存在大量的色素细胞。胆囊黏膜上皮为单层扁平或立方上皮,未见形成皱襞。胰中结缔组织不发达,细胞界限不清,胞核大。除肝的门管区、肝门静脉区和胰的胰管周围分布有较多的肥大细胞外,肝和胰中其他区域分布很少,胆囊壁内有较多的肥大细胞。     

An immunohistochemical study of the pancreatic endocrine cells of the nude mouse, Balb/c-nu/nu

The regional distribution and frequency of the pancreatic endocrine cells in the nude mouse, Balb/c-nu/nu were studied by immunohistochemical (peroxidase anti-peroxidase; PAP) methods using specific antisera against insulin, glucagon, somatostatin and human pancreatic polypeptide (hPP). The pancreas of the mouse was divided into two lobes, the splenic and duodenal lobes, and each lobe was subdivided into three regions, the pancreatic islets (central and peripheral regions), the exocrine region and the pancreatic duct region (consisting of duct epithelium and surrounding connective tissue--sub-epithelial connective tissue). In the pancreatic islets, most of insulin-immunoreactive (IR) cells were located in the central region, and glucagon-, somatostatin and hPP-IR cells were located in the peripheral region regardless of the lobe. In the splenic part, glucagon-IR cells were also located in the central regions, and more numerous somatostatin-IR cells were detected in the central regions compared to those of the duodenal part. hPP-IR cells were restricted to the peripheral regions in both lobes but more numerous cells were detected in the duodenal portion as compared to those of the splenic portion. In the exocrine parenchyma of the splenic lobe, only insulin-, glucagon- and somatostatin-IR cells were detected.. Here, the insulin- and glucagon-IR cells formed cell clusters, while somatostatin-IR cells were present as solitary cells. In the exocrine region of the duodenal portion, only insulin-, somatostatin- and hPP-IR cells were observed, with the same distributional pattern as that found in the splenic lobe. However, clusters of cells consisting only of hPP-IR cells were distributed in the pancreas parenchyma as small islets. In the pancreatic duct region, only solitary hPP-IR cells were demonstrated in the sub-epithelial connective tissue regions of the splenic portion. In conclusion, some strain-dependent characteristic distributional patterns of pancreatic endocrine cells, especially of the hPP-IR cells, were found in the nude mouse. In addition, somewhat different distributional patterns were found between the two pancreatic lobes.     

白鲟肝脏和胰脏的组织学与形态学研究

白鲟肝脏较大,可分为左右两大叶及小的中叶,胆囊位于右叶的凹缺内。肝板多由双层细胞构成,肝小叶不明显。肝内毛细胆小管由4个肝实质细胞围成。胰脏有3支,被厚的浆膜。胰岛明显。胰管与胆管汇合后共同开口于小肠最前部背面。对肝、胰实质细胞的显微或亚显微结构进行了描述。     

Studies on satellite nucleoli in rat hepatocytes and Novikoff hepatoma cells

Summary The present study was undertaken to provide information on the presence and frequency of satellite nucleoli in cells with increased nucleolar biosynthetic activity. The number of hepatocytes containing satellite nucleoli was analyzed in rat liver, regenerating liver 18 h after partial hepatectomy and in Novikoff hepatoma ascites cells. In comparison with hepatocytes of normal liver, the number of both stimulated hepatocytes and malignant hepatoma cells containing satellite nucleoli was significantly reduced. The results also indicated that whereas most satellite nucleoli contain protein C23, a smaller percentage contain protein B23.     

Cytological observations on the so-called neoblasts in the serpulid Spirorbis

The purported “neoblasts” of the serpulid Spirorbis have been studied in Spirorbis (Paradexiospira) vitreus and Spirorbis (Laeospira) borealis at both the light and electron microscopic levels. These perivasal cells occur in greatest abundance around the ventral blood vessel of the achaetous region. In light microscope preparations, the perivasal cells are intensely basophilic, containing basally situated nuclei, and relatively large nucleoli. The fine structure of the perivasal cells reveals that they contain an abundance of rough endoplasmic reticulum, well-developed Golgi complex, heterogeneous dense bodies, and cytolysomes. The respiratory pigment chlorocruorin, which has a diameter of about 230 Å and is believed to be composed of two superimposed hexagonal components, has been localized within: cisternae of the rough endoplasmic reticulum, elements of the Golgi complex, and membrane-bounded vesicles at the base of the perivasal cells. Evidence is advanced which strongly suggests that molecules of chlorocruorin are transported from the perivasal cells into the lumen of the vessel by reverse pinocytosis. It is concluded that whatever other functional role(s) the perivasal cells of Spirorbis may have, a major function is the synthesis and secretion of chlorocruorin. Whether the perivasal cells can be considered to be pluripotent is discussed.     

不同生殖期鳜肝脏超微结构变化的观察

应用透射电镜对生殖季节与非生殖季节鳜肝脏超微结构的变化进行了观察。鳜肝细胞含有单个卵圆形的核,核仁清楚;细胞质内含有粗面内质网、线粒体、糖原颗粒和脂滴等细胞器和内含物。胆小管由相邻的数个肝细胞质膜凹陷围成,而肝血窦则由内皮细胞的胞质构成。还发现了贮脂细胞、枯否氏细胞和成纤维细胞。胆小管腔和窦周隙内浸润许多由肝细胞发出的微绒毛结构。鳜肝细胞的超微结构在产卵前后呈现明显变化：产卵前的肝细胞内富含线粒体、糖原颗粒和脂滴,粗面内质网发达;而产卵后的肝细胞内核仁发生迁移,部分细胞核囊泡化,糖原颗粒和脂滴排空,少数肝细胞具双核结构。非生殖期多数肝细胞核含有双核仁结构,胞质内溶酶体数量增多。     

唐鱼肝脏显微和超微结构观察

应用光镜和透射电镜对繁殖期间唐鱼Tanichthys albonubes肝脏组织的显微和超微结构进行了观察。结果显示,唐鱼肝细胞具单核,中央核仁显著;细胞质内分布着粗面内质网、线粒体、糖原颗粒和脂滴等细胞器和内含物。胆小管由2～3个相邻肝细胞质膜凹陷围成,而肝血窦则由内皮细胞的胞质、成纤维细胞等参与构成。肝细胞与周边细胞通过3种不同方式进行联系:肝细胞之间的紧密连接;与血窦的间接连接;与胆小管的邻接。这些联系方式显示了肝脏具有内分泌腺和外分泌腺功能的特点。研究还发现雌性唐鱼肝脏具有"暗"细胞和"淡"细胞两种类型。本文还讨论了唐鱼肝脏与其他硬骨鱼类肝脏一般组织结构和超微结构的异同点。     

Cytochemical contributions to differentiating GERL from the Golgi apparatus

Synopsis Recent studies from our laboratory are described which deal with endocrine cells (insulinoma, -cells of the pancreas, thyroid epithelial cells), pancreatic exocrine cells, and hepatocytes. These emphasize the importance of the hydrolase-rich specialized region of endoplasmic reticulum, known as GERL, in secretory cells. Also reviewed in this paper are the varied molecular transformations which apparently occur in GERL in different cell types, as reported from other laboratories as well as our own. Evidence of the continuity of GERL with rough endoplasmic reticulum is presented. Two hydrolytic enzyme activities in GERL, in addition to acid phosphatase activity, are recorded. Finally, the use of cytochemical staining procedures in the study of microperoxisomes is briefly described. The Histochemical Journal lecture 1976. Delivered to the Histochemistry and Cytochemistry Section of the Royal Microscopical Society on 14 September 1976     

Tissue and sex differences in the expression of nucleoli in mouse somatic cells.

In Mus musculus, the nucleolus organizer regions (NORs), or sites of ribosomal RNA-encoding genes, map at three chromosomal pairs. A silver procedure was modified to stain nucleoli in interphasic somatic cells of mice. The number of nucleoli per cell nucleus was determined in squashed cells of kidney, liver and pancreas obtained from male and female mice. In liver and pancreas cells the average number of nucleoli per cell was 4.84 and 4.66, respectively, and only 2.83 in kidney cells (p < 0.001). Less than 8% of pancreas cells and about 15% of liver cells contained more than 6 nucleoli per cell, which was the maximum expected number. In addition, the number of nucleoli per cell was significatively different (p < 0.01) when male and female liver or pancreas cells (not kidney cells) were compared. In both cases, female cells presented more nucleoli than the respective male cells. Assuming that the available NORs are the same, the variable number of nucleoli in the examined cell types would be the consequence of a tissue specific NOR regulation. The apparent influence of sex on this regulation is noted.