Development of the Polar Filament-Polaroplast Complex in a Microsporidian Parasite*

SYNOPSIS. The development of the polar filament in a microsporidian parasite was studied in the electron microscope. The polar filament is a peculiar and complex organelle with intricate anatomical relationships to other structures in the mature spore. The characteristic ultrastructure of the formative and mature stages of the polar filament made it possible to trace its development and study the interactions among various organelles during its formation. In sporoblasts the polar filament develops sequentially from 3 different regions. The base of the filament appears first and is derived from a dense body. The anterior part of the filament is formed from electron dense material located in the perinuclear cisterna and in agranular endoplasmic reticulum. The base and the anterior part of the filament move toward each other and fuse. Subsequently, the posterior part of the filament develops from the posterior part of the Golgi complex. The polar sac and the polaroplast surrounding the anterior segment of the filament are formed from the anterior region of the Golgi complex.     

Glycogen in the Marine Protozoon Parauronema acutum*

SYNOPSIS. The carbohydrate which accumulates in the cytoplasm of the marine protozoon, Parauronema acutum, during normal growth was isolated, purified and characterized chemically. The highly purified material yielded only glucose residues following hydrolysis in 0.6 N HCl for 3 h at 100 C; measurement of total carbohydrate by the phenol-sulfuric acid method and by treatment with amylo-glucosidase and glucose oxidase gave similar values. Aqueous solutions of the purified material reacted with iodine to form a complex which exhibited an absorption peak at 456 nm with a shift to 484 nm in the presence of 50% saturated (NH₄)₂SO₄. Digestion with α-amylase, β-amylase, and isoamylase yielded 71%, 45% and 8.3% hydrolysis, respectively. Treatment sequentially with both isoamylase and β-amylase gave complete hydrolysis of the polymer. The average chain length (CL) determined by the isoamylase procedure was 12. These observations are consistent with the view that the carbohydrate isolated from the protozoan is a polymer consisting of α-D-glucose residues arranged in chains containing α-(1→4) linkages with branch points containing α-(1→6) linkages occurring once on the average of ～ 12 glucose residues and, as such, is indistinguishable from glycogen isolated from mammalian sources.     

The Differentiation of Herpetomonas megaseliae: Ultrastructural Observations*

Ultrastructure of both undifferentiated (promastigote and paramastigote) and differentiated (opisthomastigote) forms of Herpetomonas megaseliae is described. There is a posterior migration of the kinetoplast at the end of the exponential growth phase. The posterior extension of the flagellar pocket precedes migration of the kinetoplast. Opisthomastigotes have an electron-translucent mitochondrial matrix in comparison with undifferentiated forms. The Golgi body changes from a stack of flattened sacs to an aggregation of vesicles. Several structures previously reported from Trypanosomatidae, e.g. subpellicular organelles, pellicular microtubules, membrane whorls, stored metabolic products, surface blebs, and an intraflagellar body are also present in H. megaseliae.     

Ultrastructural Observations on Aging of Stationary Cultures and Feeding in Ochromonas*

SYNOPSIS. Changes accompanying aging of stationary cultures of Ochromonas danica were examined with the electron microscope. The cultures included light- and dark-grown populations ranging in age from 3 days to 5 weeks. Cells from the youngest cultures contained minimal amounts of lipid and a distinct leucosin vacuole. After 1 week, the number of lipid globules in the cytoplasm increased. The amount of lipid increased progressively in cells from older cultures until the leucosin vacuole was obliterated by the coalescing spheres. Cells from cultures older than 3 weeks showed a general breakdown of cytoplasmic integrity. An area of pinocytotic activity was also present; a relationship between this anterior region and blebs arising from the cell membrane is suggested.     

Microsporidian Spore Wall: Ultrastructural Findings on Encephalitozoon hellem Exospore

A study of the spore wall of Encephalitozoon hellem was performed on thin sections, freeze-fracture, and deep-etched samples to obtain information on spore wall organization and composition. Our observations demonstrate that the spore wall is formed by an inner 30–35 nm electron-lucent endospore and an outer 25–30 nm electron-dense exospore. The exospore is a complex of three layers: an outer spiny layer, an electron-lucent intermediate lamina and an inner fibrous layer. Freeze-fracture and deep-etching techniques reveal that the intermediate lamina and the inner fibrous layer result from the different spatial disposition of the same 4-nm thick fibrils. In thin sections the endospore reveals a scattered electron-dense material that appears in the form of trabecular structures when analyzed in deep-etched samples. The presence of chitin in the exospore is discussed.     

A Protozoon in the Central Nervous System of the Lungfish Polypterus enlicheri*

SYNOPSIS. Trophozoites and what appear to be early spore forms of a parasitic protozoan present in the spinal cord of the lungfish Polypterus enlicheri are described. They are probably intra- and extracellular invaders into the anterior horn and are frequently located near capillaries. They provoke a glial response. The organisms have some myxosporidan features (7) and are tentatively classified as cnidosporidans.     

Ultrastructural and Other Observations Which Suggest Suctorian Affinities for the Taxonomically Enigmatic Ciliate Cyathodinium*

SYNOPSIS. Two species of the taxonomically enigmatic genus Cyathodinium, C. piriforme and C. cunhai, were studied in some detail at both light and electron microscopic levels. Data obtained strongly suggest suctorian affinities for the genus, since a number of structures or features are strikingly reminiscent of similar (if not homologous) structures recently discovered in ciliates belonging to the order Suctorida. Endosprits (suctorial tentacles?) of Cyathodinium show an arrangement of microtubules not unlike that known for several suctorians, especially Acineta and Tokophrya. Haptocysts or missile-like bodies, ca. 600 mμ long, have been observed within endosprits and free in the cytoplasm; again this is reminiscent of the complex organelles recently described from several suctorian groups. Mouthlessness, coupled with the presence of a ventral depression (functioning in gathering prey at distal ends of endosprits?) and the presence of food vacuoles in the cytoplasm, further support a suctorian mode of feeding. Finally, stages in the curious life cycle of Cyathodinium suggest neoteny and a basic similarity to endogenous budding processes in certain suctorians.     

阔鳞瘤蕨(水龙骨科)卵细胞发育超微结构观察

水龙骨科是蕨类植物中最进化的类群,该文采用超微技术对水龙骨科的阔鳞瘤蕨(Phymatosorus hainanensis)卵发育过程进行观察,以完善薄囊蕨植物卵发生的资料,为揭示蕨类植物的有性生殖及演化机制奠定基础。结果表明:(1)幼卵、颈沟细胞、腹沟细胞通过胞间连丝紧密连接。(2)发育过程中,卵与腹沟细胞之间细胞壁显著加厚,将卵细胞与腹沟细胞隔离。(3)在壁的下方产生分离腔,内含大量不定形物质,但卵细胞与腹沟细胞在中间处始终相连。(4)分离腔中的不定形物质沉积在卵细胞质膜外形成了一层加厚的卵膜,而在连接区(孔区)处不形成卵膜,该位置最终形成了受精孔。(5)卵细胞核变得高度不规则,近成熟时卵核产生了大量的核外突。     

Ultrastructural and Morphometric Observations of Cortical Endoplasmic Reticulum in Arbacia,Spisula and Mouse Eggs*

Results of morphometric investigations indicate that Arbacia eggs possess a network of cortical endoplasmic reticulum equal in volume and surface area to that within the subcortex. The cortical endoplasmic reticulum surrounds individual cortical granules and forms associations with the plasma membrane reminiscent of junctions shared by the sarcolemma and sarcoplasmic reticulum. Mouse eggs, which also exhibit a cortical granule reaction, possess endoplasmic reticulum that is associated with cortical granules and the plasmalemma. The same relative volume of cortical endoplasmic reticulum is present in mouse eggs as in Arbacia. Significantly less cortical endoplasmic reticulum is present in Spisula eggs which do not undergo cortical granule discharge upon activation. These observations are discussed in light of the hypothesis that the cortical endoplasmic reticulum transduces the interaction of the gametes into an intracellular calcium release which initiates the cortical granule reaction and the activation of development.     

Ultrastructural Observations on the Origin and Differentiation of Somatic Cells during Gonadal Development in the Frog Rana nigromaculata

The process of gonad development in the frog Rana nigromaculata was observed using the electron microscope. The gonadal medulla was formed by the proliferation and displacement of the epithelial cells within the primordial gonad, and a distinct continuity was observed between the cortical and medullary cells. Sex differentiation of the gonad occurred directly from the sexually indifferent primordial gonads. In the rudimentary testes, the continuity between the cortical and medullary regions increased closer, and the intermingling of cortical and medullary cells was evident. The inner region of the cortex developed into a cord-like structure and subsequently differentiated into rudimentary seminiferous tubules. The medulla differentiated into the testicular rete and efferent duct. In the rudimentary ovaries, the cortex and medulla were separated and the ovarian cavity was formed in the medullary region. In the cortex, the cortical cells surrounding oocytes which had reached the diplotene stage, differentiated into follicular cells. The intrusion of mesenchymal or blastemal cells derived from extragonadal regions into the cortex or medulla was never observed. These findings do not support Witschi's cortico-medullary antagonistic theory of sex differentiation.     

Ultrastructural Observations on the Spermatozoa of Two Temnocephalids (Platyhelminthes)

The spermatozoa of two Temnocephalidae collected in Uruguay, Temnocephala iheringi Haswell, 1893 (Host: Pomacea canaliculata) and Temnocephala axenos Monticelli, 1899 (Host: Parastacus varicosus), were studied with a transmission electron microscope. In both species the spermatozoon is made up of a long sperm body which bears at one extremity two free flagella of the 9+‘1’ flatworm pattern. The sperm body contains the nucleus, mitochondria, dense bodies and parallel, cortical, longitudinal singlet microtubules. Along a part of the sperm body the palissade of the microtubules displays a spiral pattern in transverse sections. A part of the perimeter of the cell is thus lined by two overlapping rows of microtubules. This spiral pattern of the singlets is considered as a synapomorphy of the family Temnocephalidae. The singlet microtubules are interconnected by two kinds of links: tangential links between neighbouring singlets in the same row and radial links between singlets belonging to two rows. The presence of these links suggests that this structure could be a motile system of singlets.     

华东安蕨卵发生的超微结构观察

核心薄囊蕨是蕨类植物中的进化类群,但对受精作用具有显著影响的卵发生研究仍较少,该文利用超微技术对其中蹄盖蕨科的华东安蕨卵发生过程进行了研究,以进一步完善薄囊蕨植物卵发生的科学资料,为理解蕨类植物的有性生殖及演化机制奠定基础。超微结构观察显示:华东安蕨的幼卵和沟细胞在颈卵器中紧密联接;随后,在卵细胞上方出现了分离腔和临时细胞壁,但在卵细胞中间孔区处卵细胞和腹沟细胞始终联接在一起;分离腔中的无定形物质沉积在卵细胞的质膜外形成了1层加厚的卵膜,而在孔区处没有形成卵膜,该位置最后形成了受精孔。在进一步的卵发生过程中,卵细胞核变得高度不规则,形成了大量的核外突和核褶皱。     

华中铁角蕨孢子纹饰形成的超微结构观察

利用透射电子显微镜对铁角蕨科(Aspleniaceae)华中铁角蕨(Asplenium sarelii Hook.)孢子及其纹饰的形成过程进行观察。结果表明:①华中铁角蕨孢子囊发育为薄囊蕨型;②孢子外壁表面光滑,远极面的外壁厚约0.8~1.1μm,近极面的外壁厚约1.4~1.8μm;③孢子周壁厚度约4~5μm,染色较外壁深,分为内层和外层;内层紧帖外壁表面,其上具柱状、瘤状或疣状突起;外层向外隆起形成脊状纹饰的轮廓,脊的下方具空腔,脊的顶端具翅;④铁角蕨型与鳞毛蕨型孢子外壁和周壁纹饰的形成过程具有相似性;⑤孢子的成熟度对于孢子形态的研究是至关重要的,只有完全成熟的孢子的表面纹饰才是稳定的。     

Ultrastructural Observations on the Germ Plasm in the Lizard Podarcis sicula

Ultrastructural studies on embryos and adult females of Podarcis sicula revealed fibrogranular electron-dense aggregates in the cytoplasm of primordial germ cells, oogonia, and oocytes. The ultrastructural similarities of these aggregates to fibrogranular aggregates in germ cells of some animal species and their relationship with mitochondria, free ribosomes, as well as cisternae of the rough endoplasmic reticulum strongly suggest that they correspond to the germ plasm.     

Ultrastructural Observations on Life Cycle Stages of Pneumocystis carinii

An ultrastructural study of life cycle stages of Pneumocystis carinii in infected rat lungs in situ was undertaken utilizing 8 different modes of fixation. Three of the fixatives employed gave good fixation of cysts and intracystic bodies, but for the trophic forms fixation was only fair. Both the trophic forms and intracystic bodies have nuclear pores. The mitochondria of the organism have cristac that appear lamellar. One of the fixation modes revealed a thin, electron-dense layer on the outer surface of the cell wall, a "fuzzy coat" that had not been described previously. This material appears to mediate tight adhesion of trophic forms with other trophic forms, cysts, and with pneumocytcs of the lung alveolus.     

Microscopic Observations on the Filopodia of Entamoeba histolytica*

Living Entamoeba histolytica trophozoites were examined by phase-contrast microscopy. Intact critical point dried trophozoites were examined by transmission electron microscopy at an accelerating voltage of 1000 kV (HVEM) and by scanning electron microscopy (SEM). Half and quarter m? thick sections of epoxy-embedded trophozoites were examined by HVEM. Many of the trophozoites of 2 strains examined had surface filopodia, 1 to over 100 pan in length. The cytoplasm of filopodia was continuous with the cytoplasm and bounded by surface plasmalemma bearing a glycocalyx. Structures called “surface-active lysosomes with trigger,”“dendritic plasmalemmal extensions,” and “extra-amebic vesicles” by previous investigators probably represent portions of filopodia demonstrated in the present study. Filopodia appear to be of frequent normal occurrence in E. histolytica and may function in: (a) endocytosis or pinocytosis; (b) exocytosis; (c) attachment to substratum; (d) penetration of tissue; (e) release of cytotoxic substances; or (f) contact cytolysis of host cells.     

Ultrastructural Observations of Experimental Naegleria Meningoencephalitis in Mice: Intranuclear Inclusions in Amebae and Host Cells*

SYNOPSIS. Primary amebic meningoencephalitis was experimentally produced in mice through intranasal instillation of pathogenic Naegleria fowleri. Experimental animals had a 64% mortality, with average time of onset of symptoms or 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.     

Quantitative Ultrastructural Investigations of Mitochondrial Development in Leishmania donovani During Transformation*

SYNOPSIS. The ultrastructure of the mitochondrion during transformation of Leishmania donovani from amastigote to promastigote stages was studied by morphometric analysis. There was a slight but significant decrease in relative mitochondrial volume (Vvli) during transformation. This decrease was linear with time for at least 27 hr. No change in relative lipid inclusion volume (Vvli) was observed during transformation. The ratio of inner to outer mitochondrial surface membrane densities (Svmii:Svmio) also remained unchanged. Although the relative number of cristae remains unchanged after transformation, there is an increase in cristae number in the portion of the kinetoplast opposite the flagellum.     

Ultrastructural and Cytochemical Observations on Sporogenesis of Myxobolus sp. (Myxosporida: Myxobolidae) from the Common Shiner Notropis cornutus*

SYNOPSIS. The structure and cytochemistry of spores of Myxobolus sp. from plasmodia which occur in the gill filaments of the common shiner Notropis cornutus were studied by light microscopy and by scanning and transmission electron microscopy. The thin-walled valves of the pyriform spores are thickened in the lateral sutural and apical regions. Mucous material is associated predominantly with the posterior end of many spores. The plasmodium is surrounded by a syncytial wall bounded by 2 membranes. Pinocytotic channels are formed by the inner membrane and numerous dense vesicles are pinched off at the distal ends of the channels. Sporogenesis is initiated by the envelopment of one vegetative cell by another. The larger, enveloped cell divides to form a disporous pansporoblast, which contains 2 pairs of capsulogenic and valvogenic cells and 2 binucleate sporoplasm cells. Each capsular primordium and connecting external tubule gives rise to a polar capsule which houses a helically coiled polar tubule. The apical end of each polar capsule is plugged by a stopper. The valvogenic cells surround the capsulogenic and posteriorly situated sporoplasm cells to form the spore valves. Iodinophilic (glycogen) inclusions were not seen in spores stained with iodine or Best's carmine. A darkly stained band was observed around the posterior region of most spores stained with Best's carmine. In the electron microscope large aggregates of β glycogen particles were seen in the cytoplasm of sporoplasm cells in mature spores.