Morphology of the stifle in agouti (Dasyprocta prymnolopha,Wagler 1831)

The agouti (Dasyprocta prymnolopha, Wagler 1831) is a wild rodent of great zootechnical potential, a fact that enables anatomical and morphological studies to support management actions with this animal. In this perspective, this study aimed to describe the anatomy and histology of the agouti stifle joint. Four adult agoutis were used, two females and two males. The animals were submitted to dissection and identification of the structures of the stifle joint. For light microscopy study, samples of the patellar ligament, cranial and caudal cruciate ligaments, medial and lateral collateral ligaments were used. Agouti has a highly congruent patellofemoral joint; elongated patella; medial and lateral fabellae at the proximal insertion of the gastrocnemius muscle; medial and lateral meniscus with lunula; in addition to the presence of the following ligament structures: patellar ligament, cranial and caudal cruciate ligaments, medial and lateral collateral ligaments, meniscofemoral ligament, caudal meniscal ligament of the medial meniscus, and medial and lateral cranial ligaments. The patellar ligament presents bundles of parallel collagen fibers with a straight path and coated fibroblasts; collateral and cruciate ligaments had loose and dense connective tissue, coated fibroblasts and collagen bundle undulations, the latter most expressive in the caudal cruciate ligament. Thus, except for the shape and angulation of the stifle, which allows specific movements, the agouti stifle has structures analogous to that of other rodents and domestic animals.     

The morphology of the pineal gland of the yellow‐toothed cavy (Galea Spixii Wagler, 1831) and red‐rumped agouti (Dasyprocta leporina linnaeus, 1758)

The pineal gland is an endocrine gland found in all mammals. This article describes the morphology of this important gland in two species of Caviideae, namely the yellow‐toothed cavy and the red‐rumped agouti. Ten adult animals of the two species used in current analysis were retrieved from the Center for the Multiplication of Wild Animals (CEMAS/UFERSA) and euthanized. The glands were removed and photographed in situ and ex situ. They were fixed in a paraformaldehyde solution 4% or glutaraldehyde 2.5% solution and submitted to routine histological techniques respectively for light and scanning electron microscopy. Macroscopically, the pineal gland with its elongated structure may be found between the cerebral hemispheres facing the rostral colliculi. Microscopically, pinealocytes and some glia cells were predominant. Contrastingly, to the cavy's pineal gland, a capsule covered the organ in the agouti, with the emission of incomplete septa to the interior, which divided it into two lobules. Light and scanning electron microscopes failed to show calcareous concretions in the pineal gland. Based on the topography of the cavy's and agouti's pineal gland, it may be classified as supra‐callosum and ABC type. Microsc. Res. Tech. 78:660–666, 2015. © 2015 Wiley Periodicals, Inc.     

Morphological and morphometric characterization of agoutis' peripheral blood cells (Dasyprocta prymnolopha, Wagler, 1831) raised in captivity

Thirty adult agoutis (Dasyprocta primnolopha) from the Nucleus of Study and Preservation of Wild Animals at the Federal University of Piauí were used. Blood scrubs of these animals were colored by the Leishman method and analyzed in light microscopy. The cells had been measured using programs that analyze images (Leica QWin - Image Processing and Analysis Software). Mature erythrocytes, basophil reticulocytes, lymphocytes, eosinophils, neutrophils, monocytes, and thrombocytes were identified. Agoutis' erythrocytes presented elliptical form, without nucleus with an average diameter of 5.64 micromeres ± 0.38. The lymphocytes are spherical cells with scarce cytoplasm, dense and with a very centralized rounded nucleus measuring an average diameter of 13.20 micromeres ± 0.35. The monocytes are slightly basophilic, with a spherical nucleus, central constriction, and an average diameter of 20.59 micromeres ± 0.32. The neutrophils are spherical, with a polymorphic lobulated nucleus, with an average diameter of 11.2 micromeres ± 0.20. The eosinophils are spherical with lobulated nucleus and with an average diameter of 14.25 micromeres ± 0.36. Only five basophils were observed, with abundance of cytoplasmic granules with 9.8 micrometers of diameter ± 0.30. Thrombocytopenic pleomorphism was frequent. There were similarities in the cellular constituents in peripheral blood of agoutis and of other rodents and humans. The cellular types from the peripheral blood, the morphology, and morphometry of the blood's cells did not vary according to sex.     

Immunophenotypic,immunocytochemistry, ultrastructural,and cytogenetic characterization of mesenchymal stem cells from equine bone marrow

The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium‐rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n = 64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies. Microsc. Res. Tech. 76:618–624, 2013. © 2013 Wiley Periodicals, Inc.     

Effects of rotational culture on morphology,nitric oxide production and cell cycle of endothelial cells

Devices for the rotational culture of cells and the study of biological reactions have been widely applied in tissue engineering. However, there are few reports exploring the effects of rotational culture on cell morphology, nitric oxide (NO) production, and cell cycle of the endothelial cells from human umbilical vein on the stent surface. This study focuses on these parameters after the cells are seeded on the stents. Results showed that covering of stents by endothelial cells was improved by rotational culture. NO production decreased within 24 h in both rotational and static culture groups. In addition, rotational culture significantly increased NO production by 37.9% at 36 h and 28.9% at 48 h compared with static culture. Flow cytometry showed that the cell cycle was not obviously influenced by rotational culture. Results indicate that rotational culture may be helpful for preparation of cell-seeded vascular grafts and intravascular stents, which are expected to be the most frequently implanted materials in the future.     

Bones Morphogenic Protein‐4 and retinoic acid combined treatment comparative analysis for in vitro differentiation potential of murine mesenchymal stem cells derived from bone marrow and adipose tissue into germ cells

Nowadays, infertility is no longer considered as an unsolvable disorder due to progresses in germ cells derived from stem lineage with diverse origins. Technical and ethical challenges push researchers to investigate various tissue sources to approach more efficient gametes. The purpose of the current study is to investigate the efficacy of a combined medium, retinoic acid (RA) together with Bone Morphogenic Protein‐4 (BMP4), on differentiation of Bone Marrow Mesenchymal Stem Cells (BMMSCs) and adipose‐derived mesenchymal stem cells (ADMSCs) into germ cells. Murine MSCs were obtained from both Bone Marrow (BM) and Adipose Tissue (AT) samples and were analyzed for surface markers to get further verification of their nature. BMMSCs and ADMSCs were induced into osteogenic and adipogenic lineage cells respectively, to examine their multipotency. They were finally differentiated into germ cells using media enriched with BMP4 for 4 days followed by addition of RA for 7 days (11 days in total). Analyzing of differentiation potential of BMMSCs‐ and ADMSCs were performed via Immunofluorescence, Flowcytometry and Real time‐PCR techniques for germ cell‐specific markers (Mvh, Dazl, Stra8 and Scp3). Mesenchymal surface markers (CD90 and CD44) were expressed on both BMMSCs and ADMSCs, while endothelial and hematopoietic cell markers (CD31 and CD45) had no expression. Finally, all germ‐specific markers were expressed in both BM and AT. Although germ cells differentiated from ADMSCs showed faster growth and proliferation as well as easy collection, they significantly expressed germ‐specific markers lower than BMMSCs. This suggests stronger differentiation potential of murine BMMSCs than ADMSCs.     

Effects of tricyclazole (5-methyl-1,2,4-triazol[3,4] benzothiazole), a specific DHN-melanin inhibitor, on the morphology of Fonsecaea pedrosoi conidia and sclerotic cells

The influence of tricyclazole (5-methyl-1,2,4-triazol[3,4]benzothiazole), a specific DHN-melanin inhibitor, on the cell walls and intracellular structures of Fonsecaea pedrosoi conidia and sclerotic cells was analyzed by transmission electron microscopy (TEM), deep-etching, and field emission scanning electron microscopy. The treatment of the fungus with 16 microg mL(-1) of tricyclazole (TC) did not significantly affect fungal viability, but electron microscopy observations showed several important morphological differences between TC-treated and non-TC treated cells. Control sclerotic cells presented patched granules, with an average diameter of 47 nm, on the cell surface, which were absent in TC-treated cells. Also, TC-treated sclerotic cells showed an undulated relief. TC treatment leads to an accumulation of electron lucent vacuoles in the fungal cytoplasm of both conidia and sclerotic cells, and treated conidia observed by deep etching showed a relevant thickening of the fungal cell wall. Together, these observations support the previous data of our group that F. pedrosoi synthesizes melanin in intracellular organelles. In addition, we suggest that melanin is not only an extracellular constituent but could also be dispersing all over the cell walls and could have an effective role in cross-linking different cell wall compounds that help maintain the regular shape of the cell wall.     

Lateral magnocellular nucleus of the anterior neostriatum (LMAN) in the zebra finch: neuronal connectivity and the emergence of sex differences in cell morphology.

The song system of birds provides a model system to study basic mechanisms of neuronal plasticity and development underlying learned behavior. Song learning and production involve discrete sets of interconnected nuclei in the avian brain. One of these nuclei, the lateral magnocellular nucleus of the anterior neostriatum (LMAN), is the output of the so-called anterior forebrain pathway known to be essential for learning and maintenance of song, both processes depending on auditory feedback. In zebra finches, only males sing and this sexually dimorphic behavior is mirrored by sexual dimorphism in neuronal structure that develops during ontogeny. Female zebra finches are not able to sing and nuclei of the song system are strongly reduced in size or even lacking, when compared to male brains. Only LMAN can be delineated as easily in females as in males. Since female zebra finches, despite being unable to sing, recognize song just as males do and form a memory for song (model acquisition) early in life, LMAN is a putative candidate for song acquisition in both sexes. Therefore, development of LMAN was studied at the cellular and ultrastructural level in both male and female zebra finches. Regressive development of dendritic spines, enlargement of neuronal cell body and nuclei size, as well as changes at the nucleolar level are events all occurring exclusively in males, when song learning progresses. The decline in synapse number and the augmentation in synaptic contact length at synapses in LMAN in males are indicative for synaptic plasticity, whereas in females synapse number and synaptic contact length remain unchanged.     

Video-rate confocal reflection microscopy of neoplastic cells: Rate of intracellular movement and peripheral motility characteristic of neoplastic cell line (RSK4) with high degree of growth independence in vitro

Video-rate laser confocal interference reflection microscopy was used to demonstrate rapid motion of intracellular organelles and features at the cell periphery in a fully transformed neoplastic cell line, RSK4, and in four other neoplastic cell populations. In the RSK4 cells, vibrational and trafficking movements of intracellular particles at a rate greater than 25 Hz and ranging down to 5 Hz were recorded. Rapidly moving processes changed to ruffles, then microspikes, and previously undetectable ephemeral intercellular contacts were seen. Dynamic cyclical changes were revealed in the sizes of the podosomal close contacts of the transformed cells. The visibility of such features and the temporal and spatial resolution are improved over earlier methods. The fact that fast cellular and intracellular movements can be detected with this microscopic technique offers new possibilities in attempting to recognise differences between unimpaired living cells, and it may prove useful in the identification of malignant cells.     

A new method for cell culture on an electron-transparent melamine foil suitable for successive LM,TEM and SEM studies of whole cells

A new cell culture technique is described which is based on the observation that foils cast from the melamine resin hexamethylol-melamine-ether are suitable for the cultivation of beating heart muscle cells and fibroblasts of the rat. This foil can be flamed for sterilization, is about 80 nm in thickness, homogeneous and smooth, withstands dehydration and critical point-drying, can be removed from glass and permits the imaging of whole cells successively by light microscopy, transmission and scanning electron microscopy. The method is capable of narrowing the gap between light and electron microscopy, yielding excellent whole cell preparations in various kinds of microscopic studies to be performed on one and the same cell.     

Determining the effect of calcium on cell death rate and perforation formation during leaf development in the novel model system,the lace plant (Aponogeton madagascariensis)

Programmed cell death (PCD) is the destruction of unwanted cells through an intracellularly mediated process. Perforation formation in the lace plant (Aponogeton madagascariensis) provides an excellent model for studying developmentally regulated PCD. Ca²⁺ fluxes have previously been identified as important signals for PCD in plants and mammals. The fundamental goal of this project was to determine the influence of Ca²⁺ on the rate of cell death and perforation formation during leaf development in the lace plant. This was investigated using the application of various known calcium modulators including lanthanum III chloride (LaCl₃), ruthenium red and calcium ionophore A23187. Detached lace plant leaves at an early stage of development were treated with these modulators in both short- and long-term exposure assays and analysed using live cell imaging. Results from this study indicate that calcium plays a vital role in developmentally regulated PCD in the lace plant as application of the modulators significantly altered the rate of cell death and perforation formation during leaf development. In conclusion, this study exemplifies the suitability of the lace plant for live cell imaging and detached leaf experiments to study cell death and provides insight into the importance of Ca²⁺ in developmentally regulated PCD in planta.