Effect of estradiol-17α and diethylstilbestrol on adrenal cortex and anterior pituitary gland of the mongolian gerbil

Injection of diethylstilbestrol (DES) for 7 days in female gerbils significantly increased the number of vacuolar and membranous intranuclear inclusions within mammotrophs of the anterior pituitary gland. The presence of multiple inclusions adjacent to the nuclear membrane suggests that vacuolar inclusions arise from the inner nuclear membrane. DES also increased significantly the number of lipid droplets in adrenocortical parenchymal cells and in macrophages of the zona reticularis. In contrast, estradiol-17α induced only a few intranuclear inclusions in mammotrophs of castrated male gerbils after 14 days of injection. Estradiol-17α injection was totally ineffective in causing accumulation of lipid droplets in macrophages or parenchymal cells of the zona reticularis, emphasizing the importance of stereo-specificity for inducing ultrastructural changes in the anterior pituitary and adrenal cortex of the Mongolian gerbil.     

Intranuclear filamentous inclusions in the gastro-entero-pancreatic (GEP) endocrine cells of birds

Summary Intranuclear filamentous inclusions were found in the normal endocrine cells of the avian stomach and pancreas. These inclusions were composed of a bundle of closely packed filaments (6–8 nm in diameter), being ultrastructurally similar to those found in the nucleus of various neurons. Most of them appeared as single rod- or spindle-shaped bodies; aggregations of two or more inclusions were rarely seen within a single nucleus. Cells with an intranuclear inclusion often contained a cytoplasmic fibrillar bundle similar to the intranuclear inclusion.     

斜带髭鲷外周血嗜中性粒细胞核内包涵体的超微结构

目的在对斜带髭鲷（Hapalogenys nitens）外周血细胞结构进行透射电镜观察时发现嗜中性粒细胞存在大量的核内包涵体,系统研究了这些核内包涵体的超微结构,以探讨其来源和形成过程。方法应用电镜技术对这些核内包涵体的超微结构进行研究。结果斜带髭鲷外周血嗜中性粒细胞的核内包涵体可分为假包涵体和真包涵体两种类型,包涵体中的内含物来自胞质。胞核首先是以核膜内陷的方式将胞质及其各种有形成分包绕进核内而在核质外层形成具有双层膜包裹的典型假包涵体,随后假包涵体双层膜降解消失而转化成无被膜包裹的真包涵体,即核内糖原包涵体。结论假包涵体是形成真包涵体的开始阶段。随着假包涵体向真包涵体的转变,包涵体内含物的组成及其超微结构也出现了显著变化。     

Effect of dose of estradiol and age of animals on intranuclear inclusions in mammotrophs of the Mongolian gerbil.

Intranuclear inclusions have been examined in mammotrophs of the Mongolian gerbil. No inclusions were identified in nuclei of newborn females, although inclusions were seen in the pituitary gland of young mature females. The number of inclusions in retired breeders was similar to that in young females. Estradiol benzoate increased the number of inclusions, although 10 mug/day for 5 or 14 days induced more inclusions than 5 or 100 mug for 14 days; significantly fewer inclusions were seen with the higher dose (2.81 +/- 0.10 inclusions per field for 10 mug and 2.19 +/- 0.01 inclusions for 100 mug). Significantly fewer inclusions were present at 3 and 4 weeks. The reduced number of inclusions may be attributable at least in part to cellular hypertrophy of mammotrophs which was especially prominent. Vacuolar inclusions predominated at 3 and 4 weeks and there were fewer membranous types than at previous times.     

Effect of partial thyroidectomy, propylthiouracil or thyroxine on estrogen-induced intranuclear inclusions in mammotrophs of the Mongolian gerbil

Partial thyroidectomy caused a significant increase in the number of intranuclear inclusions per field (2-83 +/- 0-10 inclusions as compared with 0-15 +/- 0-03 inclusions for sham-controls). The inclusions occurred exclusively within mammotrophs. Thyrotrophs were stimulated: an increased cell size, numerous secretory granules, an enlarged Golgi apparatus and hypertrophic rough endoplasmic reticulum. Propylthiouracil caused similar ultrastructural changes although sighificantly fewer inclusions were observed (0-64 +/- 0-06 inclusions). Suppression of thyroid function with thyroxine produced no change in the number of inclusions (0-17 +/- 0-03 inclusions) although when combined with estrogen there were significantly fewer inclusions (1-62 +/- 0-07) when compared to estrogen alone (2-69 +/- 0-09). Intranuclear inclusions appear to be a unique reaction of mammotrophs to cellular hyperfunction in the Mongolian gerbil.     

Intranuclear inclusions in the developing neurons of the rat cuneate nuclei

Summary The ultrastructure of intranuclear rodlets, microtubules, fibrillar lattices and membranous inclusions found in the developing cuneate nuclei of rats is described. Rodlets, ranging in diameter from 96–312 nm and in length from 1–2 m, are made up of tightly packed straight filaments measuring 5–8 nm in diameter. Microtubules with a diameter of 26 nm are clustered together. Fibrillar lattices are made up of fibrils with a diameter of 9 nm arranged in layers or sets. Two to nine sets make up a lattice, with a maximum width of 68 nm, in which the adjacent sets are arranged at an angle to each other. Rodlets and fibrillar lattices occur in 6.8% of the neurons. Membranous inclusions, reported here for the first time in normal neurons, are of 2 types: small vesicles of 0.1–0.6 m and large vacuoles measuring 1–2 m. Both types are bounded by either a single or a double membrane and generally have an electron lucent content. Membranous inclusions occur in 25.3 % of the neurons. Changes in the frequency of occurrence of the various intranuclear inclusions in the course of postnatal development are also reported.     

Subcellular localization of Aleutian mink disease parvovirus proteins and DNA during permissive infection of Crandell feline kidney cells.

Confocal microscopy allowed us to localize viral nonstructural (NS) and capsid (VP) proteins and DNA simultaneously in cells permissively infected with Aleutian mink disease parvovirus (ADV). Early after infection, NS proteins colocalized with viral DNA to form intranuclear inclusions, whereas VP proteins formed hollow intranuclear shells around the inclusions. Later, nuclei had irregular outlines and were virtually free of ADV products. In these cells, inclusions of viral DNA with or without associated NS protein were embedded in cytoplasmic VP protein. These findings implied that ADV replication within an infected cell is regulated spatially as well as temporally.     

Intranuclear inclusions in mammotrophs of the Mongolian gerbil: Effect of low doses of estradiol benzoate and a study of females before weaning

The effect of dose of estrogen on induction of intranuclear inclusions was studied in mammotrophs of the Mongolian gerbil. The number of inclusions per high power field increased significantly with 0-5 and 1-0 mug/day of estradiol benzoate but not with the 0-05 mug dose. A form of inclusion was seen in young female gerbils before weaning which suggests that inclusions arise from the cytoplasm. Intranuclear inclusions develop soon after weaning of gerbils (0-06 +/- 0-01 vs. 0-19 +/- 0-03 for young mature females).     

Intranuclear cytoplasmic inclusions in aspirates of follicular neoplasms of the thyroid. A report of two cases

Two cases are reviewed in which the presence of intranuclear cytoplasmic inclusions in fine needle aspiration biopsy specimens of the thyroid suggested a diagnosis of papillary carcinoma. Subsequent surgical specimens were consistent with diagnoses of microfollicular adenoma and follicular carcinoma. These cases suggest that the presence of intranuclear inclusions in epithelial thyroid cells is not limited to papillary and medullary carcinoma, as some authors have suggested.     

Intranuclear vacuoles in nonpapillary carcinoma of the thyroid. A report of three cases

Three cases of nonpapillary carcinoma of the thyroid are reported in which intranuclear vacuoles or (pseudo)inclusions were observed in fine needle aspiration smears. Two of the cases had medullary carcinoma; one had a poorly differentiated follicular carcinoma. The cytologic, histologic and ultrastructural appearances of intranuclear vacuoles and of the so-called ground-glass nuclei of papillary carcinoma are described, and the diagnostic significance of these findings is discussed briefly. Intranuclear vacuoles are distinct and different from ground-glass nuclei.     

Intranuclear crystalline inclusions in spermatozoa ofLumbricus terrestris

Summary In the nuclei of atypical spermatids ofLumbricus terrestris granular or filamentous inclusions are surrounded by dense chromatin. Aggregation and condensation of chromatin in nuclei during spermatid differentiation coincide with increase in density, granularity, and the subsequent crystallization of the intranuclear inclusion. In mature spermatozoa, the crystalline inclusion displaying an irregular shape is composed of parallel repeating units measuring 50–80 Å. The subunits sometimes possess a clear central cavity.Atypical spermatozoa, possessing inclusions that distort their normally cylindrical shape, possess typical acrosomes, middle pieces, and flagella. Spermatozoa bearing intranuclear crystals are rarely observed in the seminal receptacles ofLumbricus.These intranuclear inclusions probably represent proteinaceous material that is not eliminated during nuclear differentiation. Their sole existence in the nuclei of spermatozoa, their transformation into crystalline structures during spermiogenesis, and their similarity to crystals in virus infected plant and animal cells suggest a viral origin.Supported by a training grant (GM-00582-07) from the Public Health Service.     

Membrane-bound intranuclear inclusions in the Leydig cell of the Chinese hamster (Cricetulus griseus)

Membrane-bound intranuclear inclusions have been described, for the first time, in the Leydig cell of the Chinese hamster (Cricetulus griseus). The inclusions were not found in the 1-day-old animal, rarely found prior to sexual maturity, and commonly found in the sexually mature animals. The incidence of inclusions increases with aging. Their size and content varies greatly. They are surrounded by a single membrane and completely enclosed by nucleoplasm. Their close association with nuclear invaginations of cytoplasmic material, and their content of cytoplasmic structures along with some exhibiting the presence of trimetaphosphatase reaction product, suggest a cytoplasmic origin. This phenomenon involves the migration of cytoplasmic structures into the nucleus followed by detachment on the nucleoplasmic side. The presence of the inclusions is not an indication of an abnormality of the Leydig cell. The Leydig cell of the Chinese hamster may be an excellent model to study factors that initiate inclusion formation, and to determine the functional role of membrane-bound intranuclear inclusions.     

Intranuclear and Cytoplasmic Inclusions in Embryogenic and Embryoidal Cells of Panax quinque folius

The presence of intranuclear and cytoplasmic inclusions in cells of embryogenic callus and developing embryoids from tissue cultures of Panax quinquefolius L. was described. These cellular structures were not found in non-embryogenic cells. The size of intranuclear and cytoplasmic inclusions seemed to be related to the developmental status of the cell.     

Visualization of actin in situ by rhodamine-conjugated phalloin in the fungus Uromyces phaseoli

Rhodamine-conjugated phalloin, a derivative of phalloidin which binds to F-actin, was associated with three types of structures in uredospore germlings of the bean rust fungus, Uromyces phaseoli. The structures were filaments, peripheral plaques, and intranuclear inclusions. The filaments, located throughout the germ tube but especially in the more basipetal regions, were observed as either barely perceivable, fine elements or as easily detectable, coarser structures. The plaques, which we suggest to be equivalent to filasomes, occurred near the periphery of the cell's cytoplasm. They were most numerous in the hyphal tip regions. Nuclear inclusions occurred within the nucleoplasm subjacent to the spindle pole body. Treatments with KI and phalloidin substantiated that the fluorescently labelled sites were F-actin. Treatment of the germlings with cytochalasin E caused the intranuclear inclusions to become extended often with branched, fine filaments. Similar treatments led to a disappearance of cytoplasmic filaments, but had no perceivable effect on the peripheral plaques.     

Immunofluorescent labelling of K-papovavirus antigens in glycol methacrylate embedded material: a method for studying infected cell populations by fluorescence microscopy and histological staining of adjacent sections

Trypsin and protease V (pronase) were studied for their ability to enhance immunofluorescent labelling of papovavirus antigens in glycol methacrylate embedded sections of organs infected with murine K-papovavirus. Treatment of Bouin's fixed sections with 0.4% trypsin for 30 minutes resulted in specific immunofluorescent staining equal to that seen in frozen sections and produced little if any loss of histological detail. Treatment with protease V resulted in less brilliant fluorescence and less satisfactory tissue preservation. Studies were then conducted to determine the fluorescence and less satisfactory tissue preservation. Studies were then conducted to determine the fixative which would produce brightest specific fluorescent antibody staining of papovavirus-infected cells while providing clearest definition of intranuclear inclusions and best morphological detail in histologically stained adjacent sections. Brightest immunofluorescence staining was accomplished on material fixed in 96% ethanol/1% glacial acetic acid or Bouin's solution. These fixatives also gave clear definition of intranuclear inclusions with histological stains and provided excellent morphological detail. Phosphate buffered paraformaldehyde/picric acid and 3.7% formalin gave less satisfactory fluorescence and obscured intranuclear inclusions in histological preparations. Sections fixed in 4% paraformaldehyde, 4% paraformaldehyde/1% glutaraldehyde, and 0.5 M p-toluenesulfonic acid were negative for specific fluorescence. Glycol methacrylate, used with proper fixation and trypsin pretreatment of sections, provides a useful embedding medium for immunofluorescent identification of virus-infected cells, and the 1.0-2.0 micron sections routinely obtainable with GMA permit study of individual infected cells by fluorescent antibody and histological staining of adjacent sections.     

Ultrastructural observations of experimental Naegleria meningoencephalitis in mice: intranuclear inclusions in amebae and host cells.

Primary amebic meningoencephalitis was experimentallly produced in mice through intranasal instillation of pathogenic Naegleria fowleri. Experimental animals had a 64% mortality with average time of onset of symtoms of death occurring on the 7-8th day following inoculation. Ultrastructural studies of the olfactory lobes from brains of dead (or sacrificed) animals revealed major concentrations of amebae in the perivascular regions; amebae were also seen to be under attack by host polymorphonuclear leukocytes, and in the lumina of blood vessels. Amebae in brain tissue contained 30 nm intranuclear particles arranged in clusters. In the brains of some mice, dead presumably as a result of amebic meningoencephalitis, particles and crystalloids were observed in the nuclei of degenerating cells of the central nervous system. Some alternatives are examined to explain a possible relationship between ameba intranuclear particles and mouse brain cell intranuclear inclusions.     

Ligand promotes intranuclear inclusions in a novel cell model of spinal and bulbar muscular atrophy

Spinal and bulbar muscular atrophy (SBMA, Kennedy's disease) is one of a group of progressive neurodegenerative diseases resulting from a polyglutamine repeat expansion. In SBMA the polymorphic trinucleotide CAG repeat in exon 1 of the androgen receptor (AR) gene is increased, resulting in expansion of a polyglutamine tract. Patient autopsy material reveals neuronal intranuclear inclusions (NII) in affected regions that contain only amino-terminal epitopes of the AR. Cell models have previously been unable to produce intranuclear inclusions containing only a portion of the AR. We report here the creation of an inducible cell model of SBMA that reproduces this important characteristic of disease pathology. PC12 cells expressing highly expanded AR form ubiquitinated intranuclear inclusions containing amino-terminal epitopes of the AR as well as heat shock proteins. Inclusions appear as distinct granular electron-dense structures in the nucleus by immunoelectron microscopy. Dihydrotestosterone treatment of mutant AR-expressing cells results in increased inclusion load. This model mimics the formation of ubiquitinated intranuclear inclusions containing the amino-terminal portion of AR observed in patient tissue and reveals a role for ligand in the pathogenesis of SBMA.     

Comparative observations on inclusions in nerve fibres of the mammalian neurohypophysis

Summary A comparative study has been made of the inclusions within neurohypophysial nerve fibres of eight species of mammals.Four classes of small inclusions have been found in each of these species, namely small and large membranous vesicles, and small and large osmiophilic membrane-bound granules. No inclusions resembling the rod-shaped inclusions in the hedgehog's neurohypophysis have been found. It seems probable that more than one type of neurosecretory fibre occurs in the infundibular process of all these species.Large lamellated structures were found in some species, and these possibly give rise to membranous vesicles.     

西洋参胚性细胞和胚状体细胞中的核内含体与细胞质内含体

１９２１年,Ｍｏｌｉｓｈ首先在光镜下发现植物叶片细胞核中有一种晶体状的内含物（见Ｗｅｉｎｔｒａｕｂ等［１］）。后来人们陆续在某些动、植物的细胞核中观察到了这种结构［２—４］,并称之为核内含体（ｉｎｔｒａｎｕｃｌｅａｒｉｎｃｌｕｓｉｏｎｓ）。Ｂｉｇａｚｚｉ［３］曾在４５种桔梗科植物的细胞中看到了核内含体。但在离体培养的植物细胞中发现内含体的报道很少,至今仅在榛子组织培养分生细胞中看到了类似的结构［５］。我们在对西洋参体细胞胚胎发生进行超微结构研究的过程中发现,在胚性愈伤组织和胚状体细胞的核和细胞质…