Electron-lucent Areas Research Articles

Degradation of Mutant Protein Aggregates within the Endoplasmic Reticulum of Vasopressin Neurons.

SummaryMisfolded or unfolded proteins in the ER are said to be degraded only after translocation or isolation from the ER. Here, we describe a mechanism by which mutant proteins are degraded within the ER. Aggregates of mutant arginine vasopressin (AVP) precursor were confined to ER-associated compartments (ERACs) connected to the ER in AVP neurons of a mouse model of familial neurohypophysial diabetes insipidus. The ERACs were enclosed by membranes, an ER chaperone and marker protein of phagophores and autophagosomes were expressed around the aggregates, and lysosomes fused with the ERACs. Moreover, lysosome-related molecules were present within the ERACs, and aggregate degradation within the ERACs was dependent on autophagic-lysosomal activity. Thus, we demonstrate that protein aggregates can be degraded by autophagic-lysosomal machinery within specialized compartments of the ER.

A new in vitro model of polymyositis reveals CD8+ T cell invasion into muscle cells and its cytotoxic role

ObjectivesThe hallmark histopathology of PM is the presence of CD8+ T cells in the non-necrotic muscle cells. The aim of this study was to clarify the pathological significance of CD8+ T cells in muscle cells.MethodsC2C12 cells were transduced retrovirally with the genes encoding MHC class I (H2Kb) and SIINFEKL peptide derived from ovalbumin (OVA), and then differentiated to myotubes (H2KbOVA-myotubes). H2KbOVA-myotubes were co-cultured with OT-I CD8+ T cells derived from OVA-specific class I restricted T cell receptor transgenic mice as an in vitro model of PM to examine whether the CD8+ T cells invade into the myotubes and if the myotubes with the invasion are more prone to die than those without. Muscle biopsy samples from patients with PM were examined for the presence of CD8+ T cells in muscle cells. The clinical profiles were compared between the patients with and without CD8+ T cells in muscle cells.ResultsAnalysis of the in vitro model of PM with confocal microscopy demonstrated the invasion of OT-I CD8+ T cells into H2KbOVA-myotubes. Transmission electron microscopic analysis revealed an electron-lucent area between the invaded CD8+ T cell and the cytoplasm of H2KbOVA-myotubes. The myotubes invaded with OT-I CD8+ T cells died earlier than the uninvaded myotubes. The level of serum creatinine kinase was higher in patients with CD8+ T cells in muscle cells than those without these cells.ConclusionCD8+ T cells invade into muscle cells and contribute to muscle injury in PM. Our in vitro model of PM is useful to examine the mechanisms underlying muscle injury induced by CD8+ T cells.

Dose‐specific morphological changes in the Sertoli cell of the adult male Japanese quails testes exposed to di(n‐butyl)phthalate DBP prepubertally

Di(n‐butyl) phthalates (DBP) are endocrine‐disrupting chemicals (EDCs) implicated in a number of reproductive disorders in humans and rodents. Previously, we have shown that DBP effects is anti‐androgenic and causing alterations in seminiferous tubular histology and impairment of Leydig cell steroidogenesis in the avian model. However, the ultrastructural changes on the Sertoli cell remain unclear. The present study was designed to assess, morphologically, the dose‐dependent toxic effect of pre‐pubertal exposure of the plasticizer‐DBP, on the histology and ultrastructure of the Sertoli cell. DBP was administered to 10‐week old male quail birds by oral gavage, with a control group given corn oil vehicle(1mL/kg) only, while the other five experimental groups were fed (1, 10, 50, 200– and 400mg/kgbwt/d DBP(dissolved in corn oil) , for a period of 30days. Testes were fixed in Bouin's liquid or in a mixture of 2.5% glutaraldehyde and 2% formaldehyde for analysis under light and electron microscopes(TEM), respectively. Histologically, the basal Sertoli cell cytoplasm in high DBP dosage levels (200mg and 400mg/kgbwt/d), increasingly displayed aggregations of lipid droplets, but only a few in the apical Sertoli cell cytoplasm. In addition, there was marked vacuolations in the lining germinal epithelium. Ultrastructurally, there were notable, dose‐dependent changes seen in the Sertoli cell in DBP‐treated birds, with low dosage groups displaying no obvious structural changes, while the DBP high‐dose resulted in a remarkable accumulation of numerous, scattered or usually, closely packed lipid droplets, found predominantly in the basal part of the Sertoli cell cytoplasm. In addition, there were several mis‐shaped or swollen mitochondria with electron lucent areas, indicative of early cellular degeneration. These degenerating mitochondria were characterized by swelling and deformation of the tubular cristae, resulting in a “moth‐eaten” appearance. These findings showed that high dose of DBP (200 and 400mg/kgbwt/d) administered daily for 30days significantly resulted in degenerative characteristics, such as the increase in lipid droplets, intra‐epithelial vacuolations and mitochondrial damage in the Sertoli cell. Thus, this study has provided a clear evidence that alterations in Sertoli cell morphology and function were involved in spermatogenic failure induced by DBP, in this specie.Support or Funding InformationThe research was supported by Ahmadu Bello University Board of Research and grants from the South African Veterinary/Novartis Wildlife Foundation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Dystrophic neurites accumulating autophagic vacuoles show early stages of neuritic destruction.

We re-examined the database of some 20,000 electron micrographs from the Echigo-1, the 263K-strain or the 22C-H of scrapie-infected hamsters to look for the cytoplasmic clearance. We reevaluated the largest database in the world of photographed dystrophic neurites for the presence of cytoplasmic clearance as shown in transgenic fruit flies transfected with A-42. In several neurites, we found electron-lucent areas not bound by any membranes or only partially bound; thus, they were not autophagic vacuoles as the latter are membrane-bound and contain cargo. Those changes were not observed in every examined neurite and no correlation with any other changes were noticed. In some neurites, which could be traced over several sections, the electron-lucent areas were evident to change size, i.e. to expand.

Evaluation of the effect of Gallic acid against carbon tetrachloride-induced liver injury in albino rats: histological, immunohistochemical and biochemical study

Introduction: Carbon tetrachloride (CCL4) is used for induction of liver fibrosis. Liver fibrosis has no standard treatment. Gallic acid (GA) is a naturally occurring phenolic compound having anti-inflammatory and anti-cancer activities.Aim of the work: This work aimed to evaluate the preventive role of GA against CCL4 induced liver fibrosis.Materials and Methods: Forty adult male albino rats were divided into 4 groups; Group I: without treatment .GroupII: received olive oil. Group III: received 1.5 mL/kg of CCL4 twice a week. Group IV: received CCL4 with GA 100mg/kg. Animals were sacrificed after 8 weeks. Liver specimens were processed for light and electron microscopic (EM) study. Area % of collagen and α-SMA expression, levels of AST, ALT, ALP, reduced GSH, MDA, SOD and hydroxyproline were measured and evaluated statistically.Results: Group III revealed foci of altered hepatocytes with dense nuclei and vacuolated cytoplasm. Dilatation and congestion of central and portal veins with mononuclear cell infiltration were observed. There was increased collagen deposition around the central vein. Intense stained α-SMA-positive cells were observed. CCL4 increased collagen and α-SMA expression area %. EM showed electronlucent areas in the cytoplasm of the hepatocytes, vacuolation and margination of nuclear chromatin. CCL4 increased AST, ALT, ALP, hepatic MDA and hydroxyproline levels. Moreover, it decreased the activities of SOD and reduced GSH. The co-administration of GA prevented most of these histological and biochemical changes.Conclusions: The use of natural antioxidants as GA can be promising in ameliorating liver fibrosis better than the drugs and their side effects.

Seed structure in Canavalia brasiliensis Mart. ex Benth. (Leguminosae) and subcellular localization of ConBr lectin: Implications for ConBr biological functions

Lectins are proteins capable of specific and reversible recognition of carbohydrates without modifying them. Many studies have isolated these molecules from Leguminosae seeds but little attention was given to subcellular localization and biological function of these molecules. Therefore, this work aimed to describe Canavalia brasiliensis seed structure and the subcellular localization of ConBr. In addition, we tested the affinity of anti-ConBr antibody for other lectins. To accomplish this, seed fragments were processed for light, scanning and transmission electron microscopy, as well as immunocytochemistry. The anti-ConBr affinity was also tested. Under SEM, the testa showed a regular contour of anticlinal walls without trichomes and inner integument of 10–20 cell layers. The cotyledons presented many parenchymatic cell layers with starch grains and cytoplasmic protein bodies. The immunological identity of anti-ConBr immunoglobulin to ConBr and other lectins was confirmed by immunodiffusion and Western blotting. Ultrastructurally, the cotyledons showed protein bodies characterized by heterogeneous appearance of irregularly formed electron-dense and electron-lucent areas. Immunolocalization showed lectin at both protein bodies and cell wall, however, more studies are required for the elucidation of ConBr functions. Moreover, the affinity of anti-ConBr for different lectins can make this molecule a useful biotechnological tool for lectin studies.

Unveiling the effects of berenil, a DNA-binding drug, on Trypanosoma cruzi: implications for kDNA ultrastructure and replication

Trypanosoma cruzi, the etiological agent of Chagas disease, exhibits a single mitochondrion with an enlarged portion termed kinetoplast. This unique structure harbors the mitochondrial DNA (kDNA), composed of interlocked molecules: minicircles and maxicircles. kDNA is a hallmark of kinetoplastids and for this reason constitutes a valuable target in chemotherapeutic and cell biology studies. In the present work, we analyzed the effects of berenil, a minor-groove-binding agent that acts preferentially at the kDNA, thereby affecting cell proliferation, ultrastructure, and mitochondrial activity of T. cruzi epimastigote form. Our results showed that berenil promoted a reduction on parasite growth when high concentrations were used; however, cell viability was not affected. This compound caused significant changes in kDNA arrangement, including the appearance of membrane profiles in the network and electron-lucent areas in the kinetoplast matrix, but nuclear ultrastructure was not modified. The use of the TdT technique, which specifically labels DNA, conjugated to atomic force microscopy analysis indicates that berenil prevents the minicircle decatenation of the network, thus impairing DNA replication and culminating in the appearance of dyskinetoplastic cells. Alterations in the kinetoplast network may be associated with kDNA lesions, as suggested by the quantitative PCR (qPCR) technique. Furthermore, parasites treated with berenil presented higher levels of reactive oxygen species and a slight decrease in the mitochondrial membrane potential and oxygen consumption. Taken together, our results reveal that this DNA-binding drug mainly affects kDNA topology and replication, reinforcing the idea that the kinetoplast represents a potential target for chemotherapy against trypanosomatids.

WHIRLY1 is a major organizer of chloroplast nucleoids.

WHIRLY1 is an abundant protein of chloroplast nucleoids, which has also been named pTAC-1 with regard to its detection in the proteome of transcriptionally active chromosomes (TAC). In barley primary foliage leaves, expression of the WHIRLY1 gene is highest at the base whereas protein accumulation is highest in the middle of the leaf where young developing chloroplasts are found. In order to elucidate the function of WHIRLY1 in chloroplast nucleoids, transgenic barley plants with an RNAi-mediated knock-down of the HvWHIRLY1 gene (RNAi-W1) were generated. The homozygous RNAi-W1-7 plants, barely containing traces of the WHIRLY1 protein, were chosen for detailed analyses of nucleoids. Nucleic acid specific-staining with YO-PRO®-1 revealed that in comparison to wild type chloroplasts, which have multiple small nucleoids attached to thylakoids, chloroplasts of the transgenic plants contain large irregularly formed patches of DNA besides nucleoids that are similar in size and shape to those of wild type chloroplasts. In large electron lucent areas, filamentous structures were detected by conventional transmission electron microscopy. Analyses of ptDNA levels by both DNA dot-blot hybridization and quantitative PCR showed that leaves of the transgenic plants have a two- to three-fold higher level of ptDNA than the wild type. The higher ptDNA level in RNAi-W1 plants coincided with an enhanced expression of the gene encoding a putative organelle targeted DNA polymerase in the mid part of primary foliage leaves. Furthermore, overexpression of the barley WHIRLY1 gene in E. coli cells revealed a higher compaction of bacterial nucleoids. These results suggest that WHIRLY1 belongs to the group of plastid nucleoid associated proteins (ptNAP) having a function in compacting a subpopulation of chloroplast nucleoids thereby affecting DNA replication.

Pre-spermiogenic initiation of flagellar growth and correlative ultrastructural observations on nuage, nuclear and mitochondrial developmental morphology in the zebrafish Danio rerio

The microstructural and ultrastructural changes of germ cells during spermatogenesis of zebrafish (Danio rerio) were examined using light microscopy (LM) and transmission electron microscopy (TEM). Generally the process of spermatogenesis in zebrafish is similar to that of other teleosts, however, here we describe some peculiar features of zebrafish spermatogenic cells which have a limited report in this species. (1) The basic events of spermiogenesis are asynchronous, location of flagellum finished in initial stage, while chromatin condensation sharply occurred in intermediate stage and elimination of excess cytoplasm mainly taken place in final stages. (2) Surprisingly, the cilia or initial flagellae are created in spermatocytes, approach toward the nucleus of early stage spermatids, and then the centrioles depress into nuclear fossa and change their orientation to each other from right angle to obtuse angle about 125°. (3) During spermatogenesis, the chromatin compaction performs in a distinctive pattern, condensed heterogeneously from granular into chromatin clumps with central electron-lucent areas, round or long, which diminished to small nuclear vacuoles in spermatozoa. This finding demonstrates the origin of nuclear vacuoles in zebrafish spermatozoa for the first time. (4) Nuages are observed in both spermatogonia and spermatocytes. They are connected with the mitochondria and nuclear membrane, and are even located in the perinuclear spaces of spermatogonia nuclei. (5) Mitochondrial morphology and distribution shows diversity in different germ cells. The condensed mitochondria appear in pachytene spermatocytes, and mitochondria including membrane conglomerate exist in both spermatocytes and spermatids. This study was undertaken in order to disclose specific spermatogenic cells features in zebrafish that could be helpful for understanding the correlative function in this model species.

Ontogenic development of spermatids during spermiogenesis in the high altitude bunchgrass lizard (Sceloporus bicanthalis)

The body of ultrastructural data on spermatid characters during spermiogenesis continues to grow in reptiles, but is still relatively limited within the squamates. This study focuses on the ontogenic events of spermiogenesis within a viviparous and continually spermatogenic lizard, from high altitude in Mexico. Between the months of June and August, testicular tissues were collected from eight spermatogenically active bunchgrass lizards (Sceloporus bicanthalis) from Nevado de Toluca, México. The testicular tissues were processed for transmission electron microscopy and analyzed to access the ultrastructural differences between spermatid generations during spermiogenesis. Interestingly, few differences exist between S. bicanthalis spermiogenesis when compared with what has been described for other saurian squamates. Degrading and coiling membrane structures similar to myelin figures were visible within the developing acrosome that are likely remnants from Golgi body vesicles. During spermiogenesis, an electron lucent area between the subacrosomal space and the acrosomal medulla was observed, which has been observed in other squamates but not accurately described. Thus, we elect to term this region the acrosomal lucent ridge. This study furthers the existing knowledge of spermatid development in squamates, which could be useful in future work on the reproductive systems in high altitude viviparous lizard species.

Ultrastructural topochemistry of cell wall polymers in Populus nigra by transmission electron microscopy and Raman imaging

The topochemical distribution of lignin and cellulose in individual cell wall layers of Populus nigra stem was determined by transmission electron microscopy (TEM) and confocal Raman microscopy. TEM images exhibited the fiber wall as being typically differentiated into three layers: middle lamella (ML), primary wall (P), and secondary wall (S1, S2, and S3). Higher magnification views showed the S2 layer to be differentiated into electron lucent and dense areas in the radial direction. In situ Raman images calculated by integrating over the intensity of characteristic spectral bands enabled visualization of the spatial variation in lignin and cellulose. Raman images acquired by integrating over the spectral band at 1605 cm-1 suggested that higher lignin content was visualized in the cell corner (CC), the compound middle lamella (CML), and the secondary wall of ray parenchyma. Cellulose distribution followed by taking the band regions around 2897 cm-1 into account showed the opposite pattern, with the highest content in fiber secondary wall. The SEM-EDXA provided semi-quantitative results, showing that the lignin content ratio in various cell wall layers was 1.4 (CC):1.1(CML):1(S2).

Piscirickettsia-Like Organisms as a Cause of Acute Necrotic Lesions in Colombian Tilapia Larvae

Rickettsial organisms are well-known fish pathogens in both natural and culture environments. This study reports an outbreak of disease in red tilapia larvae caused by piscirickettsia-like organisms (PLOs), which lasted from June until October 2009. Severe mortality was recorded almost exclusively in larvae and postlarvae aged 1-22 days old. Although clinical or gross findings were not evident in diseased fish, histopathology revealed severe necrosis of the epidermis and gill epithelium, with concomitant changes in the underlying skeletal muscle as being the most relevant microscopic lesions. Although PLOs were visible with the routine hematoxylin eosin technique, they were better observed with Giemsa and toluidine blue stains. Transmission electron microscopy revealed that the bacterium was located within the cytoplasm and phagolysosoma-like structures of epithelial cells from the gills and the skin. The bacteria measured 0.9 ± 0.2 µm × 2.1 ± 0.6 µm and had a double cell membrane (the outer one having undulating projections), with variable electron-dense and electron-lucent areas. Ultrastructurally, abundant myelin figures surrounded the microorganisms within host cell cytoplasm. Results indicated that Piscirickettsia-like organisms can cause massive epithelial cell damage associated with concomitant alteration of the electrolyte balance.

Chromatin Alterations in Leukocytes of First-episode Schizophrenic Patients

Studies of peripheral blood leukocytes of schizophrenic patients have shown in electron microscopy (EM) that decondensation of the chromatin constitutes a biological marker indicating increased genomic expression. Since this increase depends on chromatin relaxation by dissociation of lysine-rich histone H1 from nucleosomes, with exposure of arginine residues of core histones, the ratio of arginine to lysine residues in each nucleus represents a reliable measure of activation. Lysine- and arginine-rich proteins are demonstrable in light microscopy (LM), differentially, as yellow and black, respectively, with the ammoniacal silver reaction (ASR). Application of ASR on leukocyte pellets before they are dehydrated and embedded in epoxy resins gives reliable results in semithin sections. In thin sections the ASR method localizes only the amino acid arginine by forming deposits of electron-opaque particles, visualized in the EM. Leukocytes of 12 first-episode schizophrenic patients and 5 controls were used. Light micrographs of the semithin sections were inserted in a personal computer. The percentage of lysine and arginine was measured in 300 nuclei per subject. Morphometry showed that lymphocytes of schizophrenic patients have increased ratios of arginine to lysine, compared to controls, indicating activation; neutrophils of the patients have even a higher ratio, indicating an abnormal condition of the genome. Chromatin conformational changes are also evident by phosphotungstic acid hematoxylin (PTAH) block staining, which reveals condensed chromatin as an electron-lucent area in the nuclei, and decondensed chromatin as an electron-dense area. Because decondensed chromatin is a biological marker of schizophrenia, the efficacy of these methods to demonstrate this particular state offers a tool for early diagnosis, since first-episode schizophrenic patients have a better prognosis when treatment is started promptly, at the beginning of the disease.

Fine structure of a septate gregarine trophozoite in the black tiger shrimp Penaeus monodon

Gregarines are parasitic protozoa that occasionally parasitize the gut lumen of penaeid shrimp and other crustaceans. Here we describe the morphology of gregarine trophozoites found in the black tiger shrimp Penaeus monodon using light microscopy (LM) and scanning (SEM) and transmission (TEM) electron microscopy. Using LM with fresh preparations and with paraffin sectioning followed by hematoxylin-eosin staining, several trophozoites were discovered in the foregut and midgut, and gametocysts were found in the hindgut. Trophozoites existed both as solitary individuals and in association and exhibited either caudofrontal or lateral patterns. They were cylindrical in shape, 60 to 300 microm long by 10 to 60 microm wide, and consisted of 2 parts: a small anterior portion known as a protomerite, which was separated by a septum from a larger posterior portion known as a deutomerite. Under SEM, both the protomerite and deutomerite were found to be entirely covered with longitudinal, parallel pellicular folds 0.1 microm thick. Under TEM, the deutomerite was seen to contain a nucleus with a single eccentric nucleolus and several areas of peripherally located chromatin. The cytoplasm of both the protomerite and deutomerite contained abundant vesicles and granules of different sizes. The pellicle consisted of a double layer of electron-dense membranes separated by an electron-lucent area 30 nm wide and containing a microfilament. A few microfilaments were also observed in the cytoplasm underneath the pellicle, possibly serving as locomotive apparatus for the parasite. Based on its morphology, this gregarine appears similar to those of the genus Nematopsis.

Ultrastructural observation on genesis and morphology of cortical granules in Macrobrachium nipponense (Crustacea, Caridea)

Cortical granules are secretory vesicles in oocytes that develop from the Golgi complex. In the freshwater shrimp, Macrobrachium nipponense, mitochondria participates in the formation of cortical granules. We investigated the structural changes of mitochondria and the distribution cortical granules in different stages of oocyte development. Transmission electron microscopy provided evidence for the involvement of mitochondria and a particular spiral lamellar organization and an electron-lucent area in internal cortical granules. The ooplasm provided material for the cortical granules in early oocyte development. We demonstrated that mitochondria play a role in coalescence and maturation of cortical granules in this species. Additionally, a concept of cortical granules regarded as a functional integration is put forward. The genesis of shrimp cortical granules exhibited a particular pathway of maturation. The outer shape and inner organization considering different taxa suggested general as well as specific features of the development of cortical granules.

Late Postoperative Opacification of Hydrogel Intraocular Lenses: Analysis of 13 Explanted Lenses

We report the clinical, morphological, and ultrastructural findings of 13 consecutively explanted opacified Hydroview(R) (hydrogel) intraocular lenses (IOLs). Our purpose was to provide a comprehensive account on the possible factors involved in late postoperative opacification of these IOLs. Thirteen consecutive opacified hydrogel IOLs (Hydroview H 60 M, Bausch & Lomb) were explanted due to the significant visual impairment they caused. The IOLs underwent macroscopical examination, transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and electrophoresis for protein detection. Three unused control Hydroview IOLs served for comparison. Macroscopical examination showed a diffuse or localized grey-whitish opacification within the IOL optic. TEM confirmed the presence of lesions inside the optic in all the explanted IOLs and revealed 3 patterns of deep deposits: a) diffuse, thick, granular, electron-dense ones; b) small, thin, lattice-like ones, with prominent electron-lucent areas; and c) elongated electron-dense formations surrounded by electron-lucent halos. SEM showed surface deposits on four IOLs. EDS revealed oxygen and carbon in all IOLs and documented calcium, phosphorus, silicon and/or iron in the deposits. Two of the patients with iron in their IOLs had eye surgery prior to their phacoemulsification. Iron correlated well with the second TEM pattern of deep lesions, whereas calcium with the third TEM pattern. No protein bands were detected on electrophoresis. Control lenses did not show any ultrastructural or chemical abnormality. The present study supports the presence of chemical alterations inside the polymer of the optic in late postoperative opacification of Hydroview IOLs. This opacification does not follow a unique pathway but may present under different ultrastructular patterns depending on the responsible factors. Mechanical stress during surgery may initiate a sequence of events where ions such as calcium, phosphorus, silicon, and/or iron, participate in a biochemical cascade that leads to gradual alteration of the polymer network. Intraocular inflammation due to previous operation may be a factor inducing opacification through increase of iron-binding capacity in the aqueous humour. Calcification accounts only partially for the opacification noted in this type of IOL.

Ultrastructural Examination of Spermiogenesis and Spermatozoon Ultrastructure in Congo tetra Phenacogrammus interruptus Boulenger, 1899 (Ostariophysi: Characiformes: Alestidae)

Ultrastructural studies of spermiogenesis in Phenacogrammus interruptus using transmission electron microscopy revealed that the process is characterized by flagellum development, formation of a cytoplasmic canal, nuclear rotation, and nuclear fossa formation. Chromatin compaction proceeds during spermatid transformation within the spermatocysts as well as after spermiation within the lumen of the efferent ducts. The spermatozoon is of primitive type and exhibits characters typical for Type I aquasperm. The head consists of a spherical nucleus with highly condensed chromatin and a centrally located electron lucent area connected to a moderate-sized nuclear fossa. The nuclear fossa contain centrioles in perpendicular arrangement, surrounded by osmiophilic fibrous material. In the short midpiece, several mitochondria and vesicles are unevenly distributed in the cytoplasm forming the cytoplasmic collar at the base of the nucleus. The cytoplasmic collar surrounds the initial part of the flagellum, running in the cytoplasmic canal. The flagellar axoneme has a typical pattern (9 x 2 + 2) and the flagellum contains membranous compartments in the portion immediately posterior to the termination of the cytoplasmic canal.

A case report of glomerulopathy-associated podocytic infolding in a patient with tumor lysis syndrome

A 59-year-old man underwent total gastrectomy and splenectomy for gastric cancer 14 months before admission. The pathological diagnosis was neuroendocrine carcinoma of the stomach. Eight months after the operation, systemic chemotherapy with irinotecan and cisplatin was started because of multiple metastases to lymph nodes. After two courses of chemotherapy, renal function continued to decline. Renal biopsy showed acute tubular necrosis with cast formation, where needle crystallization was found. These clinicopathological findings suggested that tumor lysis syndrome was the cause of acute renal insufficiency. Moreover, diffuse, global bubbling and focal segmental spike formation were revealed by periodic acid-silver methenamine stain in the glomerular basement membrane. Electron microscopy showed an infolding of the cytoplasm of podocytes into the basal basement membrane and spotty electron-lucent areas. These ultrastructural findings, but not epimembranous deposits, corresponded with the bubbling on PAM staining. The present case was a rare case of glomerulopathy associated with podocytic infolding, which was not associated with collagen disease but with tumor lysis syndrome.

How are podocytes affected in nail–patella syndrome?

Nail–patella syndrome is an autosomal-dominant hereditary disease named for dysplastic fingernails and toenails and hypoplastic or absent kneecaps evident in patients with the syndrome. Prognosis is determined by the nephropathy that develops in many such patients. Besides podocyte foot-process effacement, pathognomonic changes in the kidney comprise electron-lucent areas and fibrillar inclusions in the glomerular basement membrane. These characteristic symptoms are caused by mutations in the gene encoding the transcription factor LMX1B, a member of the LIM-homeodomain gene family. Comparable with the human syndrome, homozygous Lmx1b knockout mice lack patellae and suffer from severe podocyte damage. In contrast, however, podocin and the α3 and α4 chains of collagen IV are absent in the glomeruli of Lmx1b knockout mice. Further studies with podocyte-specific Lmx1b knockout mice have confirmed the importance of LMX1B in podocytes, as these mice apparently develop foot processes initially but lose them later on. We therefore conclude that LMX1B is essential for the development of metanephric precursor cells into podocytes and possibly also for maintaining the differentiation status of podocytes. LMX1B can serve as a model system to elucidate a genetic program in podocytes.

Ultrastructural comparison of the spermatozoa of the Pacific oyster Crassostrea gigas inhabiting polluted and relatively clean areas in Taiwan

The ultrastructure of spermatozoa of the Pacific oysters Crassostrea gigas from an industrially polluted area on the Hsinchu City coast and a relatively clean aquaculture area on Penghu Island, Taiwan, was studied. Oyster gonads were sectioned and examined with light and transmission electron microscopes. The number of spermatozoa in the acinus lumen was significantly lower in oysters from the polluted area than that from the relatively clean area (160 ± 33 cells per 0.01 mm2 against 280 ± 42 cells per 0.01 mm2, respectively, P < 0.01). Oysters from the polluted area on the Hsinchu City coast had 26.6% spermatozoa with damaged acrosome, coarsely granular chromatin and electron-lucent areas in the nucleus; cell membrane, mitochondria and flagellum were frequently absent in these spermatozoa. By contrast, oysters from the clean area on Penghu Island had 0.4% spermatozoa with the same impairments. Sea water temperature and salinity were similar in the two areas, whereas concentrations of nitrogenous nutrients and heavy metals (e.g., cadmium, copper, and zinc) were higher in the industrially polluted area. It is suggested that high percentage of impairments in the spermatozoa of oysters from the industrial area is attributable to environmental pollution.