Epithelio-mesenchymal interface during mouse kidney tubule induction in vivo

Development ◽  
1975 ◽  
Vol 34 (3) ◽  
pp. 695-705
Author(s):  
E. Lehtonen
Keyword(s):  
Mesenchymal Cell ◽  
Ruthenium Red ◽  
Mouse Kidney ◽  
Embryonic Induction ◽  
Metanephric Mesenchyme ◽  
Kidney Tubule ◽  
Extracellular Material ◽  
20 Nm ◽  
Scanning Electron

Transmission and scanning electron microscopy were used to study the epithelio-mesenchymal interface between the interacting mouse ureter-bud and the metanephric mesenchyme. The gap between the epithelial and mesenchymal cells varied in width. At the stalk of the ureter-bud the interspace was often about 1 µm, but in the inductively active areas at the tips of the branching ureter-bud epithelio-mesenchymal contacts were seen through discontinuities in the basal lamina. At these points the gap between the interacting cells was often less than 20 nm, in places less than 10 nm. The amount of electron-dense, ruthenium-red-positive material was greatest at the stalk of the ureter-bud, but only a small amount of extracellular material was found between the interacting cells at the tips. Whether epithelio-mesenchymal cell contacts play a role in kidney tubule induction is not yet known, but their existence in the inductively active areas and their absence in inactive zones suggests that they are morphogenetically significant. The finding also obviates the need to postulate long-range transmission of inductive signals to explain this example of embryonic induction.

LIF, the ES-cell inhibition factor, reversibly blocks nephrogenesis in cultured mouse kidney rudiments

Development ◽  
1991 ◽  
Vol 113 (1) ◽  
pp. 193-198 ◽  
Author(s):  
J.B. Bard ◽  
A.S. Ross
Keyword(s):  
Embryonic Stem ◽  
Mouse Kidney ◽  
Leukaemia Inhibitory Factor ◽  
Es Cell ◽  
Metanephric Mesenchyme ◽  
Ureteric Bud ◽  
Cell Inhibition ◽  
Striking Effect

Mouse kidney induction proceeds in vitro much as it does in vivo: the ureteric bud bifurcates to give collecting ducts while the mesenchyme condenses into aggregates which epithelialise and then elongate into tubules with glomerular and other nephron structures. We report here that the factor known as LIF (leukaemia inhibitory factor), which regulates the differentiation and growth of embryonic-stem (ES) and other cells in culture, has little effect in vitro on growth or on ureteric-bud morphogenesis other than to stimulate the bifurcation process. It does however exert a striking effect on the mesenchyme. At about four times the concentration required to inhibit ES-cell differentiation, LIF strongly but reversibly blocks the effects of metanephric mesenchyme induction: although mesenchyme condenses around growing duct tips, the number of mature nephrons that form over 6 days is reduced by 75% or more. The few nephrons that do develop in the presence of LIF probably come from mesenchyme already induced at the time of culture and are indistinguishable from those that form in controls as assayed by morphology, by X-gal staining of endogenous galactosidase and by antibodies to brush-border and CD15 antigens. There is a further unexpected feature of rudiments cultured in LIF which is absent in controls: they contain an unexpectedly high number of stable epithelialised aggregates that express laminin around their periphery and which do not develop further. These results argue that the process of nephrogenesis involves at least two distinct stages which can be blocked by LIF: the effect of the initial induction and the future development of epithelialised aggregates.(ABSTRACT TRUNCATED AT 250 WORDS)

Osteogenesis in Marrow-Derived Mesenchymal Cell Porous Ceramic Composites Transplanted Subcutaneously: Effect of Fibronectin and Laminin on Cell Retention and Rate of Osteogenic Expression

1992 ◽  
Vol 1 (1) ◽  
pp. 23-32 ◽  
Author(s):  
James E. Dennis ◽  
Stephen E. Haynesworth ◽  
Randell G. Young ◽  
Arnold I. Caplan
Keyword(s):  
Chemical Composition ◽  
Bone Formation ◽  
Pore Structure ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
Cell Binding ◽  
Ceramic Surfaces ◽  
Number Of Cells ◽  
Scanning Electron

Cultured-expanded rat marrow-derived mesenchymal cells differentiate into osteoblasts when combined with a porous calcium phosphate delivery vehicle and subsequently implanted in vivo. In this study, the effects of ceramic pretreatment with the cell-binding proteins fibronectin and laminin on the osteogenic expression of marrow-derived mesenchymal cells were assessed by scanning electron microscopy, [3H]-thymidine-labeled cell quantitation, and histological evaluation of bone formation. Scanning electron microscopic observations showed that marrow-derived mesenchymal cells rapidly spread and attach to both fibronectin- or laminin-adsorbed ceramic surfaces but retain a rounded morphology on untreated ceramic surfaces. Quantitation of [3H]-thymidine labeled cells demonstrated that laminin and fibronectin preadsorbed ceramics retain approximately double the number of marrow-derived mesenchymal cells than do untreated ceramics harvested 1 wk postimplantation. Histological observations indicate that the amount of time required to first detect osteogenesis was shortened significantly by pretreatment of the ceramic with either fibronectin or laminin. Fibronectin- and laminin-coated ceramic composite samples were observed to contain bone within 2 wk postimplantation, while in untreated ceramic the earliest observation of bone was at 4 wk postimplantation. A comparison was made of the initial cell-loading, in vivo cell retention characteristics, and rate of osteogenesis initiation of marrow-derived mesenchymal cells on two types of ceramic with different pore structure and chemical composition, with and without preadsorption with fibronectin or laminin. “Biphasic” ceramics contain randomly distributed pores 200-400 μm in diameter, and “coral-based” ceramics have continuous pores of approximately 200 μm in diameter. Laminin or fibronectin preadsorption significantly increases the number of cells retained in all ceramic test groups by day 7 postimplantation. In addition, by day 7 postimplantation, the biphasic ceramics retain a significantly greater number of cells for all test groups than do coral-based ceramics. The biphasic ceramics consistently have more specimens positive for bone with the identical cell-loading conditions used throughout this study. These results indicate that the retention of cells within the ceramic is an important factor for optimization of marrow mesenchymal cell initiated bone formation. The retention of cells within ceramics is augmented by the adsorption of the cell-binding proteins laminin and fibronectin, but this effect varies depending on ceramic pore structure and/or chemical composition.

A scanning electron microscopic study of the veliger larvae of the scallop argopecten purpuratus

1988 ◽  
Vol 46 ◽  
pp. 284-285
Author(s):  
G.C. Bellolio ◽  
K.S. Lohrmann ◽  
E.M. Dupré
Keyword(s):  
Morphological Description ◽  
Electron Microscopic ◽  
Larval Stages ◽  
Scanning Electron Microscopic Study ◽  
The Pacific ◽  
Argopecten Purpuratus ◽  
Veliger Larvae ◽  
20 Nm ◽  
Ethanol Series ◽  
Scanning Electron

Argopecten purpuratus is a scallop distributed in the Pacific coast of Chile and Peru. Although this species is mass cultured in both countries there is no morphological description available of the development of this bivalve except for few characterizations of some larval stages described for culture purposes. In this work veliger larvae (app. 140 pm length) were examined by the scanning electron microscope (SEM) in order to study some aspects of the organogenesis of this species.Veliger larvae were obtained from hatchery cultures, relaxed with a solution of MgCl2 and killed by slow addition of 21 glutaraldehyde (GA) in seawater (SW). They were fixed in 2% GA in calcium free artificial SW (pH 8.3), rinsed 3 times in calcium free SW, and dehydrated in a graded ethanol series. The larvae were critical point dried and mounted on double scotch tape (DST). To permit internal view, some valves were removed by slightly pressing and lifting the tip of a cactus spine wrapped with DST, The samples were coated with 20 nm gold and examined with a JEOL JSM T-300 operated at 15 KV.

Development of the Foramen Secundum in the Chick as Observed by Scanning Electron Microscopy

1976 ◽  
Vol 34 ◽  
pp. 212-213
Author(s):  
M.J.C. Hendrix ◽  
D.E. Morse
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Procaine Hydrochloride ◽  
Cardiac Anomaly ◽  
Chick Embryos ◽  
Congenital Cardiac Anomaly ◽  
Septal Defects ◽  
Critical Point Drying ◽  
Scanning Electron

Atrial septal defects are considered the most common congenital cardiac anomaly occurring in humans. In studying the normal sequential development of the atrial septum, chick embryos of the White Leghorn strain were prepared for scanning electron microscopy and the results were then extrapolated to the human heart. One-hundred-eighty chick embryos from 2 to 21 days of age were removed from their shells and immersed in cold cacodylate-buffered aldehyde fixative . Twenty-four embryos through the first week post-hatching were perfused in vivo using cold cacodylate-buffered aldehyde fixative with procaine hydrochloride. The hearts were immediately dissected free and remained in the fixative a minimum of 2 hours. In most cases, the lateral atrial walls were removed during this period. The tissues were then dehydrated using a series of ascending grades of ethanol; final dehydration of the tissues was achieved via the critical point drying method followed by sputter-coating with goldpalladium.

scholarly journals Microscopy Methods for Biofilm Imaging: Focus on SEM and VP-SEM Pros and Cons

Biology ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 51
Author(s):  
Michela Relucenti ◽  
Giuseppe Familiari ◽  
Orlando Donfrancesco ◽  
Maurizio Taurino ◽  
Xiaobo Li ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Ion Beam ◽  
Ruthenium Red ◽  
Variable Pressure ◽  
Sem Images ◽  
Advantages And Disadvantages ◽  
Pros And Cons ◽  
Microscopy Techniques ◽  
Scanning Electron

Several imaging methodologies have been used in biofilm studies, contributing to deepening the knowledge on their structure. This review illustrates the most widely used microscopy techniques in biofilm investigations, focusing on traditional and innovative scanning electron microscopy techniques such as scanning electron microscopy (SEM), variable pressure SEM (VP-SEM), environmental SEM (ESEM), and the more recent ambiental SEM (ASEM), ending with the cutting edge Cryo-SEM and focused ion beam SEM (FIB SEM), highlighting the pros and cons of several methods with particular emphasis on conventional SEM and VP-SEM. As each technique has its own advantages and disadvantages, the choice of the most appropriate method must be done carefully, based on the specific aim of the study. The evaluation of the drug effects on biofilm requires imaging methods that show the most detailed ultrastructural features of the biofilm. In this kind of research, the use of scanning electron microscopy with customized protocols such as osmium tetroxide (OsO4), ruthenium red (RR), tannic acid (TA) staining, and ionic liquid (IL) treatment is unrivalled for its image quality, magnification, resolution, minimal sample loss, and actual sample structure preservation. The combined use of innovative SEM protocols and 3-D image analysis software will allow for quantitative data from SEM images to be extracted; in this way, data from images of samples that have undergone different antibiofilm treatments can be compared.

scholarly journals Harnessing PET to track micro- and nanoplastics in vivo

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Outi Keinänen ◽  
Eric J. Dayts ◽  
Cindy Rodriguez ◽  
Samantha M. Sarrett ◽  
James M. Brennan ◽  
...  
Keyword(s):  
Sea Water ◽  
Accurate Determination ◽  
Nuclear Imaging ◽  
Imaging Data ◽  
Biodistribution Studies ◽  
Positron Emission ◽  
20 Nm ◽  
Pharmacokinetic Behavior

AbstractThe proliferation of plastics in the environment continues at an alarming rate. Plastic particles have been found to be persistent and ubiquitous pollutants in a variety of environments, including sea water, fresh water, soil, and air. In light of this phenomenon, the scientific and medical communities have become increasingly wary of the dangers posed to human health by chronic exposure to microplastics (< 5 mm diameter) and nanoplastics (< 100 nm diameter). A critical component of the study of the health effects of these pollutants is the accurate determination of their pharmacokinetic behavior in vivo. Herein, we report the first use of molecular imaging to track polystyrene (PS) micro- and nanoplastic particles in mammals. To this end, we have modified PS particles of several sizes—diameters of 20 nm, 220 nm, 1 µm, and 6 µm—with the chelator desferrioxamine (DFO) and radiolabeled these DFO-bearing particles with the positron-emitting radiometal zirconium-89 (89Zr; t1/2 ~ 3.3 d). Subsequently, positron emission tomography (PET) was used to visualize the biodistribution of these radioplastics in C57BL/6J mice at 6, 12, 24, and 48 h after ingestion. The imaging data reveal that the majority of the radioplastics remain in the gastrointestinal tract and are eliminated through the feces by 48 h post-ingestion, a result reinforced by acute biodistribution studies. Ultimately, this work suggests that nuclear imaging—and PET in particular—can be a sensitive and effective tool in the urgent and rapidly growing effort to study the in vivo behavior and potential toxicity of micro- and nanoplastics.

scholarly journals Apoptosis in metanephric development.

1992 ◽  
Vol 119 (5) ◽  
pp. 1327-1333 ◽  
Author(s):  
C Koseki ◽  
D Herzlinger ◽  
Q al-Awqati
Keyword(s):  
Spinal Cord ◽  
Protein Kinase ◽  
Transcriptional Activation ◽  
Phorbol Esters ◽  
Dna Degradation ◽  
Morphological Evidence ◽  
Metanephric Mesenchyme ◽  
Embryonic Spinal Cord

During metanephric development, non-polarized mesenchymal cells are induced to form the epithelial structures of the nephron following interaction with extracellular matrix proteins and factors produced by the inducing tissue, ureteric bud. This induction can occur in a transfilter organ culture system where it can also be produced by heterologous cells such as the embryonic spinal cord. We found that when embryonic mesenchyme was induced in vitro and in vivo, many of the cells surrounding the new epithelium showed morphological evidence of programmed cell death (apoptosis) such as condensed nuclei, fragmented cytoplasm, and cell shrinking. A biochemical correlate of apoptosis is the transcriptional activation of a calcium-sensitive endonuclease. Indeed, DNA isolated from uninduced mesenchyme showed progressive degradation, a process that was prevented by treatment with actinomycin-D or cycloheximide and by buffering intracellular calcium. These results demonstrate that the metanephric mesenchyme is programmed for apoptosis. Incubation of mesenchyme with a heterologous inducer, embryonic spinal cord prevented this DNA degradation. To investigate the mechanism by which inducers prevented apoptosis we tested the effects of protein kinase C modulators on this process. Phorbol esters mimicked the effects of the inducer and staurosporine, an inhibitor of this protein kinase, prevented the effect of the inducer. EGF also prevented DNA degradation but did not lead to differentiation. These results demonstrate that conversion of mesenchyme to epithelial requires at least two steps, rescue of the mesenchyme from apoptosis and induction of differentiation.

scholarly journals Shift in collagen type as an early response to induction of the metanephric mesenchyme.

1981 ◽  
Vol 89 (2) ◽  
pp. 276-283 ◽  
Author(s):  
P Ekblom ◽  
E Lehtonen ◽  
L Saxén ◽  
R Timpl
Keyword(s):  
Basal Lamina ◽  
Type Iv Collagen ◽  
Early Response ◽  
Type I ◽  
Metanephric Mesenchyme ◽  
Type Iv ◽  
Interstitial Collagens ◽  
Membrane Components

Conversion of the nephrogenic mesenchyme into epithelial tubules requires an inductive stimulus from the ureter bud. Here we show with immunofluorescence techniques that the undifferentiated mesenchyme before induction expresses uniformly type I and type III collagens. Induction both in vivo and in vitro leads to a loss of these proteins and to the appearance of basement membrane components including type IV collagen. This change correlates both spatially and temporally with the determination of the mesenchyme and precedes and morphological events. During morphogenesis, type IV collagen concentrates at the borders of the developing tubular structures where, by electron microscopy, a thin, often discontinuous basal lamina was seen to cover the first pretubular cell aggregates. Subsequently, the differentiating tubules were surrounded by a well-developed basal lamina. No loss of the interstitial collagens was seen in the metanephric mesenchyme when brought into contact with noninducing tissues or when cultured alone. Similar observations were made with nonnephrogenic mesenchyme (salivary, lung) when exposed to various heterotypic tissues known to induce tubules in the nephrogenic mesenchyme. The sequential shift in the composition of the extracellular matrix from an interstitial, mesenchymal type to a differentiated, epithelial type is so far the first detectable response of the nephrogenic mesenchyme to the tubule-inducing signal.

scholarly journals The role of RXFP2 in mediating androgen-induced inguinoscrotal testis descent in LH receptor knockout mice

Reproduction ◽  
2010 ◽  
Vol 139 (4) ◽  
pp. 759-769 ◽  
Author(s):  
F P Yuan ◽  
X Li ◽  
J Lin ◽  
C Schwabe ◽  
E E Büllesbach ◽  
...  
Keyword(s):  
Mesenchymal Cell ◽  
Postnatal Period ◽  
Testosterone Replacement Therapy ◽  
Developmental Defect ◽  
Mrna Levels ◽  
Lh Receptor ◽  
Protein Levels ◽  
Testis Descent

LH receptor knockout (LhrKO) male mice exhibit a bilateral cryptorchidism resulting from a developmental defect in the gubernaculum during the inguinoscrotal phase of testis descent, which is corrected by testosterone replacement therapy (TRT).In vivoandin vitroexperiments were conducted to investigate the roles of the androgen receptor (AR) and RXFP2 signals in regulation of gubernacular development inLhrKO animals. This study demonstrated that AR and RXFP2 proteins were expressed in the gubernaculum during the entire postnatal period. TRT normalized gubernacular RXFP2 protein levels inLhrKO mice. Organ and primary cell cultures of gubernacula showed that 5α-dihydrotestosterone (DHT) upregulated the expression ofRxfp2which was abolished by the addition of an AR antagonist, flutamide. A single s.c. testosterone injection also led to a significant increase inRxfp2mRNA levels in a time-dependent fashion inLhrKO animals. DHT, natural and synthetic insulin-like peptide 3 (INSL3), or relaxin alone did not affect proliferation of gubernacular mesenchymal cells, while co-treatments of DHT with either INSL3 or relaxin resulted in an increase in cell proliferation, and they also enhanced the mesenchymal cell differentiation toward the myogenic pathway, which included a decrease in a mesenchymal cell marker, CD44 and the expression of troponin. These effects were attenuated by the addition of flutamide, siRNA-mediatedRxfp2knockdown, or by an INSL3 antagonist. Co-administration of an INSL3 antagonist curtailed TRT-induced inguinoscrotal testis descent inLhrKO mice. Our findings indicate that the RXFP2 signaling pathway plays an important role in mediating androgen action to stimulate gubernaculum development during inguinoscrotal testis descent.

Differentiation of kidney antigens in the human foetus

Development ◽  
1969 ◽  
Vol 21 (3) ◽  
pp. 517-537
Author(s):  
Ewert Linder
Keyword(s):  
Chick Embryo ◽  
Specific Antigen ◽  
Mouse Kidney ◽  
Human Foetus ◽  
Specific Antigens ◽  
Number Of Authors

The appearance of new antigens in the embryo during differentiation has been investigated by a number of authors. Among the proteins studied were myosin (Holtzer, 1961; Ebert, 1962), Jens crystallin (Ten Cate & Van Doorenmaalen, 1950), chick embryo haemoglobin (Wilt, 1962), and keratin during feather formation in chick embryo (Ben-Or & Bell, 1965). The development of liver proteins in the chick embryo was studied by D'Amelio, Mutolo & Piazza (1963). Okada & Sato (1963) and Okada (1965) studied the appearance of a ‘kidney-specific’ antigen in the developing mesonephros. Lahti & Saxen (1966) demonstrated the appearance of mouse kidney-specific tubule antigens during development both in vivo and in vitro. ‘Kidney-specific’ antigens are found in the metanephric proximal secreting tubules of various mammals (Hill & Cruickshank, 1953; Weiler, 1956; Groupe & Kaplan, 1967; Nairn, Ghose & Maxwell, 1967), including man (Nairn, Ghose, Fothergill & McEntegart, 1962), and in the mesonephric tubules of birds.

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