Ontogeny of the testicular excurrent duct system of male Japanese quail (Coturnix japonica): A histological,ultrastructural, and histometric study

The testicular excurrent duct system undergoes several physiological and morphological changes during the reproductive stage or breeding season in mammals, birds, and reptiles. Studies on normal age-related histomorphological changes in the excurrent duct system of Japanese quails (Coturnix japonica) remain unreported, despite the extensive use of this bird as an avian model in research studies. The current study investigated the histological, ultrastructural, and histometric changes in the testicular excurrent duct system of the Japanese quail during three reproductive stages, namely prepubertal, pubertal, and adult. Simple squamous to low cuboidal cells formed the epithelia of the rete testis in prepubertal and pubertal birds, while in adult birds the lining was low cuboidal to cuboidal. In pubertal and adult birds, the nonciliated Type I epithelial cells of the proximal efferent duct displayed a subapical endocytotic apparatus comprising coated pits, coated apical tubules, and endosomes. There was a significant increase (p ≤ .001) in epithelial heights of all ducts of the excurrent duct system in the mature, sexually active, adult birds when compared to the other age groups. The luminal and tubular diameters, and the cross-sectional areas of efferent ducts and the epididymal duct unit increased significantly (p ≤ .001) with age. It is concluded that the morphology and morphometry of the excurrent ducts of the testis of the Japanese quail change as birds mature.     

Histology and ultrastructure of the testis and vas deferens in Pseudochorthippus parallelus parallelus (Orthoptera,Acrididae)

Pseudochorthippus parallelus parallelus (Zetterstedt, 1821) (Orthoptera, Acrididae) is a widespread species in Europe, and also it is localized in some regions in Turkey such as Bursa, Eski?ehir, Ankara, Bolu, Düzce, and Çank?r?. The features of the reproductive organs such as the numbers and shapes of testes and follicles can be used as taxonomical characters. For this purpose, the ultrastructural and histological features of testis and vas deferens in P. parallelus parallelus were examined with using light microscope, scanning electron microscope, and transmission electron microscope. The mature P. parallelus parallelus has two conjugated testes produce spermatozoa. Each testis is composed of numerous testis follicles in which different stages of spermatogenesis and spermiogenesis develop. First, spermatocytes are formed by the mitosis division of the germ cells at the distal end of the follicles. Then, spermatocytes form spermatids by meiosis division in the middle region of the follicles. Finally, spermatids are differentiated to spermatozoa at the proximal region of the follicles. After maturation of the spermatozoa, sperm tails come together as the sperm bundles called as spermatodesm. Each follicle is connected to vas deferens via vas efferens to discharging spermatozoa. In spite of some differences, the testes and the vas deferens in P. parallelus parallelus are highly similar to the those of other species, especially Orthopteran species.     

Nitric oxide: relation to integrity, injury, and healing of the gastric mucosa

Nitric oxide (NO) plays a multifaceted role in mucosal integrity. The numerous functions of NO and the double-edged role played by NO in most of them provide a great complexity to the NO action. The three enzymatic sources of NO, neuronal NO-synthase (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS), have been characterised in the gastrointestinal tract. The protective properties of the NO derived from constitutive NO-synthases (eNOS and nNOS) have already been well established. Less clear is the role assigned to iNOS. The simplistic initial view of low levels of NO synthesised by constitutive NOS being protective while exaggerated NO levels after iNOS induction leading irremediably to cytotoxicity is being questioned by new evidence. As initially reported for constitutive NOS, iNOS activity may be associated to reduced leukocyte-endothelium interaction and platelet aggregation as well as protection of mucosal microcirculation. Moreover, iNOS activity may be important to resolve inflammation by increasing apoptosis in inflammatory cells. It is entirely possible that a low level of expression of iNOS will reflect a positive host-defense response to challenge, but that exaggerated or uncontrolled expression of iNOS itself becomes detrimental. There is no doubt about the protective role of NO in physiological conditions. However, when the mucosa is threatened, the role of NO becomes multiple and the final effect will probably depend on the nature of the insult, the environment involved, and the interaction with other mediators.     

NO message from muscle.

The synthesis of the free radical gas nitric oxide (NO) is catalyzed by the enzyme NO synthase (NOS). NOS converts arginine and molecular oxygen to NO and citrulline in a reaction that requires NADPH, FAD, FMN, and tetrahydrobiopterin as cofactors. Three types of NOS have been identified by molecular cloning. The activity of the constitutively expressed neuronal NOS (nNOS) and endothelial NOS (eNOS) is Ca(2+)/calmodulin-dependent, whereas that the inducible NOS (iNOS) is Ca(2+)-insensitive. The predominant NOS isoform in skeletal muscle is nNOS. It is present at the sarcolemma of both extra- and intrafusal muscle fibers. An accentuated accumulation of nNOS is found in the endplate area. This strict sarcolemmal localization of nNOS is due its association with the dystrophin-glycoprotein complex, which is mediated by the syntrophins. The activity of nNOS in skeletal muscle is regulated by developmental, myogenic, and neurogenic influences. NO exerts several distinct effects on various aspects of skeletal muscle function, such as excitation-contraction coupling, mitochondrial energy production, glucose metabolism, and autoregulation of blood flow. Inside the striated muscle fibers, NO interacts directly with several classes of proteins, such as soluble guanylate cyclase, ryanodine receptor, sarcoplasmic reticulum Ca(2+)-ATPase, glyceraldehyde-3-phosphate dehydrogenase, and mitochondrial respiratory chain complexes, as well as radical oxygen species. In addition, NO produced and released by contracting muscle fibers diffuses to nearby arterioles where it acts to inhibit reflex sympathetic vasoconstriction.     

Biology of spermatozoa maturation: an overview with an introduction to this issue

Mammalian spermatozoa undergo morphological, biochemical, and physiological modifications initially in the testis (testicular maturation) and later in the epididymis (epididymal maturation). These maturational changes are commensurate with the functional events that occur in developing germ cells and maturing spermatozoa. This special issue reviews the recent, relevant topics dealing with spermatozoa maturation and focuses on the events that occur in internal components such as the nucleus, the acrosome, the perinuclear theca, the fibrous sheath, and the cytoplasmic droplet as well as the plasma membrane. These structures/elements and the constituent proteins of which they are comprised undergo a variety of sequential modifications starting from their origination in developing germ cells up to epididymal maturation. Several steps of the maturation processes on the sperm plasma membrane are mediated by external enzymes and secretions derived from the epithelium lining of the genital tract. Degradation of some of the constituent proteins and the elimination of defective spermatozoa are controlled by the degradation/recycling system, the ubiquitin system. These maturational modifications are necessary for spermatozoa to become fertilization-competent cells and to be stored safely in the male.     

Nitric oxide synthase expression and function in embryonic and adult cardiomyocytes.

Nitric oxide (NO) is an important signalling molecule that plays a relevant role in different cell systems, among them the adult heart. The effects of NO are primarily mediated through modulation of Ca(2+) homeostasis, myofibrillar contractility, and metabolic regulation in cardiomyocytes. Recent evidence also suggests an important role of NO for cardiomyogenesis by modulating proliferation and differentiation and regulating cardiac function. In the embryonic, but also the healthy and diseased, adult mammalian heart, the inducible (iNOS) and the endothelial (eNOS) nitric oxide synthases (NOS) are detected. However, the expression pattern of NO and its function differ during development. Furthermore, under pathophysiological conditions NOS expression can also change and cause impairment of cardiac performance and cytotoxic effects. The present review focuses on the role and function of NO during cardiomyogenesis, the mechanisms responsible for eNOS availability, and the paracrine effects of NO generated by cardiomyocytes.     

1, 2 di‐substituted idopyranose from Vitex negundo l. Protects against streptozotocin‐induced diabetes by inhibiting nuclear factor‐kappa B and inducible nitric oxide synthase expression

The aim of this study is to investigate the mechanism of an action of compound isolated from Vitex negundo in streptozotocin‐induced diabetic mice. Light microscopic examination of liver, kidney and pancreatic sections of streptozotocin‐induced diabetic mice showed changes like coarsening of acinar cells of endoplasmic reticulum, destruction of β‐cells, and alteration in their secretory function were observed in the pancreas. Changes like dilation of vein, unusual concentric arrangement of hepatocytes, and liver fibrosis were observed in the liver. Thickening of tubules and expansion of glomerulus were observed in kidneys. All these altered parameters were reversed close to normal condition upon treatment using idopyranose. The results show the antidiabetic potential of idopyranose. Interestingly, liver, kidney, and pancreatic sections of diabetic mice fed with the isolated 1, 2 di‐substituted idopyranose showed regeneration of hepatocytes, nephrocytes, as well as β‐cells and acinar region appeared normal with increased numbers of β‐cells. To understand the probable mechanism of action of 1, 2 di‐substituted idopyranose, we analyzed proinflammatory inducible nitric oxide synthase (iNOS) and nuclear factor‐kappa B (NF‐κB) expression by immunohistochemistry and the results showed an increased iNOS and NF‐κB levels in streptozotocin‐induced diabetic liver, kidney and pancreas. Such high iNOS and NF‐κB levels were inhibited in 1, 2 di‐substituted idopyranose treated mice. The results suggest that 1, 2 di‐substituted idopyranose helps in the protection of hepatocytes, nephrocytes and pancreatic β‐cells probably by its action against NF‐κB and iNOS mediated inflammation in streptozotocin‐induced diabetes. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

The histomorphological structure of the male reproductive system of maize leaf weevil Tanymecus dilaticollis Gyllenhal, 1834 (Coleoptera: Curculionidae)

The histomorphology of the reproductive system and the germ cells has been useful to establish phylogenetic relationships in many insects. However, these elements remain little known in the Curculionidae. In this study, histomorphological structure of the male reproductive system of Tanymecus dilaticollis, which is economically important, is described, illustrated using stereomicroscopy, light microscopy, and scanning electron microscopy techniques, and discussed in relation to other Coleoptera species. Results showed that distinctive features of the male reproductive system of T. dilaticollis consist of a pair of yellowish testes, a pair of seminal vesicles, a pair of vasa deferentia, an ejaculatory duct, accessory glands, prostate glands, and aedeagus. Each testis is subdivided into two testicular follicles, enclosed by a peritoneal sheath. Each follicle of the mature testes is full sperm cysts with germ cells at various stages development of spermatogenesis. The testes have four types of germ cells (spermatogonia, spermatocytes, spermatids, and spermatozoa). They are occupied by the growth zone containing spermatogonia and spermatocytes, the maturation zone containing spermatids, while differentiation zone containing spermatozoa. There is a seminal vesicle at the center of each testis. Most mature sperms are stored in the seminal vesicle. Each testis is attached to the vas deferens by a stalk‐like seminal vesicle. In the distal part, vasa deferentia fuse with the ejaculatory duct. It is linked to the aedeagus. The provided results will contribute to the understanding of the reproductive cell biology of Curculionidae.     

Expression of the male reproduction‐related gene in spermatic ducts of the blue swimming crab,Portunus pelagicus,and transfer of modified protein to the sperm acrosome

Expression of a sex‐specific gene in Macrobrachium rosenbergii (Mr‐Mrr), encoding a male reproduction‐related (Mrr) protein, has been identified in the spermatic ducts (SDs) and postulated to be involved in sperm maturation processes. M. rosenbergii is the only decapod that the expression and fate of the Mrr protein has been studied. To determine that this protein was conserved in decapods, we firstly used cloning techniques to identify the Mrr gene in two crabs, Portunus pelagicus (Pp‐Mrr) and Scylla serrata (Ss‐Mrr). We then investigated expression of Pp‐Mrr by in situ hybridization, and immunolocalization, as well as phosphorylation and glycosylation modifications, and the fate of the protein in the male reproductive tract. Pp‐Mrr was shown to have 632 nucleotides, and a deduced protein of 110 amino acids, with an unmodified molecular weight of 11.79 kDa and a mature protein with molecular weight of 9.16 kDa. In situ hybridization showed that Pp‐Mrr is expressed in the epithelium of the proximal, middle, distal SDs, and ejaculatory ducts. In Western blotting, proteins of 10.9 and 17.2 kDa from SDs were all positive using anti‐Mrr, antiphosphoserine/threonine, and antiphosphotyrosine. PAS staining showed they were also glycosylated. Immunolocalization studies showed Pp‐Mrr in the SD epithelium, lumen, and on the acrosomes of spermatozoa. Immunofluorescence staining indicated the acrosome of spermatozoa contained the Mrr protein, which is phosphorylated with serine/threonine and tyrosine, and also glycosylated. The Mrr is likely to be involved in acrosomal activation during fertilization of eggs. Microsc. Res. Tech., 2013. © 2012 Wiley Periodicals, Inc.     

Effect of black tea on histological and immunohistochemical changes in pancreatic tissues of normal and streptozotocin‐induced diabetic mice (Mus musculus)

The aim of the present study is to evaluate the effect of hot water extract of black tea in regenerating β cells in streptozotocin‐induced diabetic mice. Light microscopic examination of pancreatic sections of streptozotocin‐induced diabetic mice showed the acinar cells to be small, shrunken, and with deteriorated β cells. The dose of streptozotocin not only altered the function of β cells but also damaged the acinar region. The changes in acinar cells were coarsening of endoplasmic reticulation suggesting alteration in their secretory function. The control pancreatic tissue showed well‐defined granulated islets and dark β cells when stained with chrome hematoxylin and phloxine. Interestingly, pancreatic sections of diabetic mice fed with black‐tea extract showed regeneration of β cells and acinar region appeared normal with increased numbers of β cells. To understand the probable mechanism of action of black‐tea extract, we analyzed inducible nitric oxide synthase (iNOS) expression by immunohistochemistry and the results showed an increased iNOS levels in streptozotocin‐induced diabetic pancreas, and such high iNOS levels were inhibited in black‐tea extract treated mice. According to histological results obtained, it can be concluded that the black‐tea extract helps in regeneration of damaged pancreas and protects pancreatic β cells by its antioxidant action against nitrosative stress in streptozotocin‐induced diabetes. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

Structural and histological observations on the male reproductive system of adult red poplar leaf beetle Chrysomela populi Linnaeus, 1758 (Coleoptera: Chrysomelidae)

This study described the anatomy and histology of the male reproductive system in Chrysomela populi, which is an economically important species belonging to the family Chrysomelidae. Therefore, reproductive biology has been studied to combat this insect. As well as, the characters associated with the reproductive tract have been potential to discuss aspects of the system and to better understand the reproductive dynamics. The male reproductive system of C. populi has a pair of testes, a pair of vas efferentia and deferentia, a pair of seminal vesicles, a pair of accessory glands, an ejaculatory bulb, an ejaculatory duct, and an aedeagus. The testis consists of two flower-shaped lobes. Each testis has 20 sperm tubules (testicular follicles) containing cysts of germ cells at various developmental stages within the light orange peritoneal sheath. Testicular follicles are composed of three different (growth, maturation, and differentiation) zones. In the middle region of each testis joins with the vas efferens. The testis is attached to the seminal vesicle by a small stalk like vas efferens. In the lumen of the vas efferens, seminal vesicle, and vas deferens, sperms form clumps in the form of thin threads. The proximal end of the vas deferens is connected to the common ejaculatory duct. It joins with the ejaculatory bulb. Around the ejaculator bulb, there is a pair of convoluted, flat-surface tubular structure accessory glands. Posterior ejaculatory duct joins with the aedeagus.     

Ethanol exposure during peripubertal period increases the mast cell number and impairs meiotic and spermatic parameters in adult male rats

Puberty is characterized by psychosomatic alterations, whereas chronic ethanol consumption is associated with morphophysiological changes in the male reproductive system. The purpose of this study was to show the toxic effects on testis and epididymal morphophysiology after ethanol administration during peripuberty. To this end, male Wistar rats were divided into two groups: ethanol (E) group: received a 2 g dose of ethanol/kg in 25% (v/v); and control (C) group: received the same volume of filtered water; both were treated by gavage for 54 days. On the 55th day of the experiment, epididymis, and testis were collected for sperm count, histopathology, mast cell count, and morphometry. The vas deferens was collected for sperm motility analysis. The femur and testicle were used for cytogenetic analysis. Ethanol exposure caused reduction in daily sperm production (DSP) and in sperm motility, multinucleated cells or those having no chromosomal content, and late chromosome migrations. No changes were observed in the number of chromosomes in the mitotic analysis. However, some alterations could be seen in meiocytes at different stages of cell division. Stereological analysis of the epididymis indicated reorganization of its component in the 2A and 5A/B regions. The epididymal cauda had greater recruitment, and both degranulated and full mast cells showed an increase in the initial segment, in the ethanol group. In conclusion, ethanol administration during the pubertal phase affects epididymis and testis in adult rats, as indicated mainly by our new findings related to mast cell number and meiotic impact. Microsc. Res. Tech. 79:541–549, 2016. © 2016 Wiley Periodicals, Inc.     

Curcumin protects against hepatic and renal injuries mediated by inducible nitric oxide synthase during selenium‐induced toxicity in Wistar rats

The aim of this study is to evaluate the effect of curcumin in protecting against selenium‐induced toxicity in liver and kidney of Wistar rats. Light microscopy evaluation of selenium alone administered rats showed liver to be infiltrated with mononuclear cells, vacuolation, necrosis, and pronounced degeneration. Control liver sections showed a regular morphology of parenchymal cells with intact hepatocytes and sinusoids. Kidney from selenium alone administered rats showed vacuolar degeneration changes in the epithelial cells, cellular proliferation with fibrosis, thickening of capillary walls, and glomerular tuft atrophy. Such changes were also observed in rats administered with selenium and curcumin simultaneously and rats administered first with selenium and then curcumin 24 h later. Interestingly, such degenerative changes observed in liver and kidney induced by selenium were not seen in rats that were administered with curcumin first and selenium 24 h later. This clearly suggests the protective nature of curcumin against selenium toxicity. To understand the probable mechanism of action of curcumin, we analyzed inducible nitric oxide synthase (iNOS) expression by immunohistochemistry, and the results showed an increased iNOS expression in selenium‐alone induced liver and kidney. Such high iNOS levels were inhibited in liver and kidney of rats pretreated with curcumin and then with selenium 24 h later. Based on the histological results, it can be concluded that curcumin functions as a protective agent against selenium‐induced toxicity in liver as well as kidney, and this action is probably by the regulatory role of curcumin on iNOS expression. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Perinuclear theca during spermatozoa maturation leading to fertilization

Mammalian spermatozoa acquire the capacity to fertilize the ovum and display motility during their passage through the epididymis. At the same time, they undergo changes in metabolic patterns, enzymatic activities, ability to bind to zona pellucida surface, and electrophoretic properties and, furthermore, stabilization of some sperm structures by the establishment of disulphide linkages takes place in several sperm structures. The cytoplasmic perinuclear theca (PT) is a unique extranuclear cytoskeletal element that surrounds the nucleus, which is proposed to be a structural scaffold to the sperm nucleus. The purpose of this review is to describe PT changes related to epididymal sperm maturation. We will focus mainly on the protein components of the PT of eutherian mammalian spermatozoa and on quantitative protein changes during sperm maturation. The protein constituents of the PT have not been completely defined and most of them are different from the cytoskeletal proteins of somatic cells. However, they are proteins with cytoskeletal features. The morphologic changes reported for PT and the proposed functions of PT are discussed.     

Quantitative analysis of surface micro-roughness alterations in human spermatozoa using atomic force microscopy

A new male contraceptive given the name RISUG^® (an acronym for Reversible Inhibition of Sperm Under Guidance) has been developed by our research group. RISUG^® is a bioactive polymer and is injected into the lumen of the vas deferens using a no-scalpel approach. The polyelectrolytic nature of this contraceptive induces a surface charge imbalance on sperm membrane system leading to its destabilization. Complete disintegration of the plasma membrane with subsequent rupture and dispersion of the acrosomal contents is observed on RISUG^® treatment. In the present study, micro-structural properties of human spermatozoa exposed to RISUG^® in vitro have been quantitatively analysed using atomic force microscopy. The parameters used to quantify these morphological changes include amplitude (peak–valley height difference, arithmetic roughness, root mean square roughness) and spatial roughness. Factor loadings (Varimax rotation) have been used to determine the parameters displaying maximum variation. Further, sperm cells have been classified in various principal-component planes using principal-component analysis. The periodic structural features of the atomic force microscopy images have also been obtained using power spectral analysis.     

Can scanning near‐field optical microscopy be compared with confocal laser scanning microscopy? A preliminary study on α‐sarcoglycan and β1D‐integrin in human skeletal muscle

The dystrophin–glycoprotein complex and the vinculin–talin–integrin system constitute, together a protein machinery, called costameres. The dystrophin–glycoprotein complex contains, among other proteins, also dystrophin and the sarcoglycans subcomplex, proteins playing a key role in the pathogenesis of many muscular dystrophies and linking the cytoplasmic myofibrillar contractile elements to the signal transducing molecules of the extracellular matrix, also providing structural support to the sarcolemma. The vinculin–talin–integrin system connects some components of the extracellular matrix with intermediate filaments of desmin, forming transverse bridges between Z and M lines. In our previous reports we always studied these systems by confocal laser scanning microscopy (CLSM). In this paper we report on the first applications of optical near‐field fluorescence microscopy to the spatial localization of α‐sarcoglycan and β1D‐integrin in human skeletal muscle fibres in order to better compare and test the images obtained with conventional CLSM and with scanning near‐field optical microscopy (SNOM). In addition, the analysis of the surface morphology, and the comparison with the fluorescence map is put forward and analyzed for the first time on human muscle fibres. In aperture‐SNOM the sample is excited through the nanometre‐scale aperture produced at the apex of an optical fibre after tapering and subsequent metal coating. The acquisition of the topography map, simultaneously to the optical signal, by SNOM, permits to exactly overlap the fluorescence images obtained from the two consecutive scans needed for the double localization. Besides, the differences between the topography and the optical spatial patterns permit to assess the absence of artefacts in the fluorescence maps. Although the SNOM represented a good method of analysis, this technique remains a complementary method to the CLSM and it can be accepted in order to confirm the hypothesis advanced by CLSM.     

Species-specific localization of actin in mammalian spermatozoa: fact or artifact?

Actin has been characterized and localized in sperm cells of many mammals. Nevertheless, the reported localizations obtained by different methods and/or antibodies varied from species to species and even for the same species. To clarify the question, sperm actin distribution was reinvestigated under uniform technical conditions. Immunogold post-embedding procedures were performed using a polyclonal and two monoclonal antibodies of known specificity to localize actin in spermatids and spermatozoa of rabbit, mouse, rat, monkey, and human. In these species, actin (F-actin) was detected with the three antibodies between the nucleus and the acrosome of round and elongating spermatids. Species-specific changes occurred in maturing spermatids. In the rabbit, actin labeling decreased and disappeared from the tip to the base of the subacrosomal layer. In testicular and epididymal spermatozoa actin was detected only with a monoclonal antibody (Amersham) successively in the neck, postacrosomal area, and subacrosomal bulges. In mouse late spermatids a transitory labeling of the neck was detected only with the polyclonal antiactin. In testicular and epididymal spermatozoa an actin labeling was observed in the principal piece of the tail. In rat, monkey, and human sperm cells actin remained undetected. These results suggest that there is a redistribution of actin in late spermatids and spermatozoa which is a species-specific process but not an artifact of methodological origin. Thus, a function for actin in sperm, if any, remains to be demonstrated.     

Detection of CD133 (prominin‐1) in a human hepatoblastoma cell line (HuH‐6 clone 5)

We examined CD133 distribution in a human hepatoblastoma cell line (HuH‐6 clone 5). We directly observed the cultured cells on a pressure‐resistant thin film (silicon nitride thin film) in a buffer solution by using the newly developed atmospheric scanning electron microscope (ASEM), which features an open sample dish with a silicon nitride thin film window at its base, through which the scanning electron microscope beam scans samples in solution, from below. The ASEM enabled observation of the ventral cell surface, which could not be observed using standard SEM. However, observation of the dorsal cell surface was difficult with the ASEM. Therefore, we developed a new method to observe the dorsal side of cells by using Aclar® plastic film. In this method, cells are cultured on Aclar plastic film and the dorsal side of cells is in contact with the thin silicon nitride film of the ASEM dish. A preliminary study using the ASEM showed that CD133 was mainly localized in membrane ruffles in the peripheral regions of the cell. Standard transmission electron microscopy and scanning electron microscopy revealed that CD133 was preferentially concentrated in a complex structure comprising filopodia and the leading edge of lamellipodia. We also observed co‐localization of CD133 with F‐actin. An antibody against CD133 decreased cell migration. Methyl‐β‐cyclodextrin treatment decreased cell adhesion as well as lamellipodium and filopodium formation. A decrease in the cholesterol level may perturb CD133 membrane localization. The results suggest that CD133 membrane localization plays a role in tumor cell adhesion and migration. Microsc. Res. Tech. 76:844–852, 2013. © 2013 Wiley Periodicals, Inc.