244. Effect of batimastat, a specific inhibitor of matrix metalloproteinases, on endometrial breakdown and repair in a mouse model

2005 ◽  
Vol 17 (9) ◽  
pp. 97
Author(s):  
T. J. Kaitu'u ◽  
N. B. Morison ◽  
L. A. Salamonsen
Keyword(s):  
Matrix Metalloproteinases ◽  
Mouse Model ◽  
Expression Patterns ◽  
Specific Inhibitor ◽  
Apparent Difference ◽  
Cross Sectional ◽  
Gelatinase Activity ◽  
Expected Time ◽  
Mmp Inhibitor

Strong correlative evidence supports a role for matrix metalloproteinases (MMPs) in the tissue breakdown at menstruation. As menstruation occurs in very few species besides women, there is a lack of suitable and easily accessible animal models available to examine the functional significance of potential key mediators of this process. A mouse model of endometrial breakdown and repair has been developed,1 which morphologically resembles that of human endometrium at menstruation. Previous studies in our laboratory showed that the expression patterns of various MMPs in the mouse model closely resembled those seen in the human.2 Administration of doxycycline, a broad spectrum MMP inhibitor, decreased gelatinase activity, but had no effect on tissue breakdown in this model. The aim of the present study was to further examine the importance of MMPs in endometrial breakdown and repair via administration of batimistat, a highly potent and specific MMP inhibitor. Batimistat was administered I.P to mice 24 h prior to the expected time of endometrial breakdown. The efficacy of batimistat within the uterus was proven using in situ zymography, which identifies MMP activity (rather than latent forms). This demonstrated that batimistat was reaching its target organ and effectively inhibiting MMP activities (both gelatinase and collagenase). Examination of gross uterine morphology revealed no apparent difference between groups, with batimistat treated uteri displaying a similar extent of tissue breakdown and repair to their control counterparts. Measurement of the breaking down area compared to total endometrial area revealed no difference between control and batimistat treatment, with the breaking down areas being 69 ±13% and 72 ± 9.8% of total endometrial cross-sectional area respectively. There was likewise no effect on endometrial repair. The results of this study together with our previous study using doxycycline, indicate that MMPs are not the key mediators of endometrial breakdown in this model. (1)Shen et al. (2004) Reprod. Fert. Dev. 16(Suppl), A265, p. 97.(2)Brasted et al. (2003) Biol. Reprod. 69, 1273.

scholarly journals 265.Matrix metalloproteinases in the mouse model of menstruation: effect of doxycycline inhibition

2004 ◽  
Vol 16 (9) ◽  
pp. 265
Author(s):  
J. Shen ◽  
J. Zhang ◽  
T. J. Kaitu'u ◽  
M. Brasted ◽  
L. A. Salamonsen
Keyword(s):  
Matrix Metalloproteinases ◽  
Mouse Model ◽  
Functional Significance ◽  
Human Endometrium ◽  
Tissue Destruction ◽  
Apparent Effect ◽  
Gelatinase Activity ◽  
Old World ◽  
Uterine Lumen ◽  
Mmp Inhibitor

Strong correlative evidence supports a role for matrix metalloproteinases (MMP) in the tissue breakdown at menstruation. Because menstruation occurs only in women and a few old-world primates, it has not been possible to examine the functional significance of potentially key mediators of this process. To this end, we developed a mouse model for menstruation (1), in which ovariectomised mice are subjected to a decidualising stimulus: injection of oil into the uterine lumen following appropriate hormone-priming. During the 24 h following withdrawal of progesterone (P), the decidualised tissue progressively breaks down, in a manner that morphologically resembles that of human endometrium at menstruation. The aims of the present study were to examine the pattern of MMP expression during the time from progesterone withdrawal until complete tissue breakdown, and to determine whether administration of doxycycline, (a known MMP modulator), 3 h prior to P withdrawal, affected the expression or activity of the MMPs or restrained the tissue destruction. MMP-3 was present at foci in the decidual zone: these were initially associated with the restructuring at decidualisation and subsequently with the tissue destruction. MMP-7 was detected both in epithelium and in leukocytes, predominantly neutrophils. These were first apparent in the basal zone during the earliest stages of tissue instability, and dramatically increased in numbers as breakdown progressed. MMP-9 was found only in leukocytes, predominantly neutrophils and some macrophages, with greatly increased numbers with time. Zymography revealed a dramatic increase in both latent and active MMP-9 as tissue breakdown proceeded. Doxycycline reduced immunoreactive MMP-3 but not MMP-7 or MMP-9 in the tissue, and also decreased gelatinase activity. However, no apparent effect on tissue breakdown was observed. Further studies with a more potent MMP inhibitor are required to fully establish the importance of MMPs in these processes. (1) Brasted et al. (2003) Biol. Reprod. 69, 1273.

Abstract 212: Fractalkine Neutralization Improves Cardiac Function After Myocardial Infarction

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Xiaosong Gu ◽  
Jiang Xu ◽  
Xiao-Ping Yang ◽  
Edward Peterson ◽  
Pamela Harding
Keyword(s):  
Myocardial Infarction ◽  
Cardiac Function ◽  
Macrophage Infiltration ◽  
Receptor Subtype ◽  
Cross Sectional ◽  
Gelatinase Activity ◽  
Ep4 Receptor ◽  
In Situ Zymography ◽  
Circulating Levels

Background: Circulating levels of the chemokine fractalkine (FKN) are increased in patients with chronic heart failure (HF) and our studies show that aged mice lacking the prostaglandin E2 EP4 receptor subtype (EP4 KO) have increased cardiac FKN coupled with a phenotype of dilated cardiomyopathy. However, whether FKN is a causal factor for HF is not well established. Hypothesis: FKN contributes to the pathogenesis of HF post myocardial infarction (MI) and EP4 KO mice have a better response to anti-FKN treatment due to elevated FKN levels. Methods: Male EP4 KO mice and wild type (WT) littermates underwent surgery to induce MI. Mice were treated with an anti-FKN antibody (40μg/kg/day, ip) or IgG immediately after MI and echocardiography was performed 2 wks post MI. Hearts were excised for infarct size determination, myocyte cross-sectional area (MCSA) and interstitial collagen fraction (ICF) determined by morphometric analysis and macrophage infiltration using immunohistochemistry. Results: Anti-FKN treatment improved cardiac function and prevented remodeling (Table). In situ zymography revealed that gelatinase activity was markedly reduced by anti-FKN treatment in both strains. Moreover, anti-FKN treatment tended to improve survival in EP4 KO mice (p = 0.06, n=20). Conclusions: (1) FKN contributes to the pathogenesis of HF and anti-FKN treatment improves cardiac function and reduces cardiac remodeling. This may be related to reduced macrophage infiltration and decreased gelatinase activity.(2) Contrary to our hypothesis, EP4 KO mice do not have an enhanced response to anti-FKN treatment.

scholarly journals Inhibition of Hippocampal Matrix Metalloproteinase-3 and -9 Disrupts Spatial Memory

2007 ◽  
Vol 2007 ◽  
pp. 1-8 ◽  
Author(s):  
John W. Wright ◽  
Travis E. Brown ◽  
Joseph W. Harding
Keyword(s):  
Matrix Metalloproteinase ◽  
Spatial Memory ◽  
Dorsal Hippocampus ◽  
Gelatinase Activity ◽  
Matrix Metalloproteinase 3 ◽  
Mmp Inhibitor ◽  
Memory Acquisition ◽  
Dependent Learning

Memory consolidation requires synaptic reconfiguration dependent upon extracellular matrix (ECM) molecules interacting with cell adhesion molecules. Matrix metalloproteinase (MMP) activity is responsible for transient alterations in the ECM that may be prerequisite to hippocampal-dependent learning. In support of this hypothesis we have measured increases in MMP-3 and MMP-9 levels within the hippocampus and prefrontal cortex during Morris water maze training. The present investigation extends these findings by determining that infusion of an MMP inhibitor (FN-439) into the dorsal hippocampus disrupted acquisition of this task. In vitro fluorescence enzyme assays to determine the specificity of FN-439 against the catalytic domains of MMP-3 and MMP-9 indicated mean±SEMIC50sof 16.2±7.8 and 210.5±37.8μM, respectively, while in situ zymography using hippocampal sections treated with FN-439 indicated significant reductions in MMP gelatinase activity. These results suggest that compromising the ability of the dorsal hippocampus to reconfigure ECM molecules by inhibiting MMP activity interferes with appropriate spatial memory acquisition, and support a role for hippocampal MMPs in the phenomena of spatial memory acquisition and storage.

scholarly journals Inhibition of matrix metalloproteinases in Siberian hamsters impedes photostimulated recrudescence of ovaries

Reproduction ◽  
2010 ◽  
Vol 140 (6) ◽  
pp. 875-883 ◽  
Author(s):  
Julie Whited ◽  
Asha Shahed ◽  
Carling F McMichael ◽  
Kelly A Young
Keyword(s):  
Matrix Metalloproteinases ◽  
Ovarian Function ◽  
Corpora Lutea ◽  
Gelatinase Activity ◽  
Antral Follicles ◽  
Mmp Inhibitor ◽  
Siberian Hamsters ◽  
First Time ◽  
Long Photoperiod

Exposure of Siberian hamsters to short photoperiod for 14 weeks induces ovarian regression. Subsequent transfer to long photoperiod restores ovarian function, and 2 weeks of photostimulation increases plasma estradiol (E2), antral follicles, and corpora lutea (CL). Because tissue remodeling involved with photostimulated ovarian recrudescence is associated with differential expression of matrix metalloproteinases (MMPs), we hypothesized that inhibiting MMP activity using a broad-spectrumin vivoMMP inhibitor, GM6001, would curtail recrudescence. One group of hamsters was placed in long days (LD; 16 h light:8 h darkness) for 16 weeks. Another group was placed in inhibitory short days (SD; 8 h light:16 h darkness) for 14 weeks. A third group was placed in SD for 14 weeks and transferred to LD for 2 weeks to stimulate recrudescence. During weeks 14–16, animals were either not treated or treated daily with i.p. injections of GM6001 (20 mg/kg) or vehicle (DMSO). GM6001 reduced gelatinase activity and decreased immunohistochemical staining for MMP1, MMP2, and MMP3 compared with vehicle. No differences between controls, vehicle, or GM6001 treatment were observed among LD animals, despite a trend toward reduction in CL and E2with GM6001. Although SD reduced ovarian function, photostimulation of transferred controls increased uterine mass, plasma E2, appearance of antral follicles, and CL. With GM6001 treatment, photostimulation failed to increase uterine mass, plasma E2, antral follicles, or CL. These data show, for the first time, thatin vivoGM6001 administration inhibits MMP activity in hamster ovaries during photostimulation, and indicate that this inhibition may impede photostimulated recrudescence of ovaries. This study suggests an intriguing link between MMP activity and return to ovarian function during photostimulated recrudescence.

The reaction of amorphous Ni-Nb films with Si in the presence of a native SiO2 layer

1990 ◽  
Vol 48 (4) ◽  
pp. 664-665
Author(s):  
N. Rozhanski ◽  
A. Barg
Keyword(s):  
High Temperature ◽  
Crystallization Temperature ◽  
Diffusion Barriers ◽  
Sio2 Layer ◽  
Si Substrate ◽  
Cross Sectional ◽  
Plan View ◽  
Temperature Range ◽  
Sputter Deposited

Amorphous Ni-Nb alloys are of potential interest as diffusion barriers for high temperature metallization for VLSI. In the present work amorphous Ni-Nb films were sputter deposited on Si(100) and their interaction with a substrate was studied in the temperature range (200-700)°C. The crystallization of films was observed on the plan-view specimens heated in-situ in Philips-400ST microscope. Cross-sectional objects were prepared to study the structure of interfaces.The crystallization temperature of Ni5 0 Ni5 0 and Ni8 0 Nb2 0 films was found to be equal to 675°C and 525°C correspondingly. The crystallization of Ni5 0 Ni5 0 films is followed by the formation of Ni6Nb7 and Ni3Nb nucleus. Ni8 0Nb2 0 films crystallise with the formation of Ni and Ni3Nb crystals. No interaction of both films with Si substrate was observed on plan-view specimens up to 700°C, that is due to the barrier action of the native SiO2 layer.

A New Method of Wafer Level Plan View TEM Sample Preparation by DualBeam

2008 ◽  
Author(s):  
Hyoung H. Kang ◽  
Michael A. Gribelyuk ◽  
Oliver D. Patterson ◽  
Steven B. Herschbein ◽  
Corey Senowitz
Keyword(s):  
Sample Preparation ◽  
Ex Situ ◽  
Wafer Level ◽  
Cross Sectional ◽  
Plan View ◽  
Manufacturing Line ◽  
Transmission Electron ◽  
Thin Lamella ◽  
Preparation Techniques

Abstract Cross-sectional style transmission electron microscopy (TEM) sample preparation techniques by DualBeam (SEM/FIB) systems are widely used in both laboratory and manufacturing lines with either in-situ or ex-situ lift out methods. By contrast, however, the plan view TEM sample has only been prepared in the laboratory environment, and only after breaking the wafer. This paper introduces a novel methodology for in-line, plan view TEM sample preparation at the 300mm wafer level that does not require breaking the wafer. It also presents the benefit of the technique on electrically short defects. The methodology of thin lamella TEM sample preparation for plan view work in two different tool configurations is also presented. The detailed procedure of thin lamella sample preparation is also described. In-line, full wafer plan view (S)TEM provides a quick turn around solution for defect analysis in the manufacturing line.

scholarly journals RNA In Situ Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of Drosophila Species

2021 ◽  
Vol 4 (1) ◽  
pp. 20
Author(s):  
Mujeeb Shittu ◽  
Tessa Steenwinkel ◽  
William Dion ◽  
Nathan Ostlund ◽  
Komal Raja ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Drosophila Species ◽  
Gene Expression Patterns ◽  
Probe Design ◽  
Tissue Preparation ◽  
Design And Synthesis ◽  
Rna In Situ Hybridization

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

scholarly journals Spatial expression pattern of serine proteases in the blood fluke Schistosoma mansoni determined by fluorescence RNA in situ hybridization

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Lenka Ulrychová ◽  
Pavel Ostašov ◽  
Marta Chanová ◽  
Michael Mareš ◽  
Martin Horn ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Schistosoma Mansoni ◽  
Rna Sequencing ◽  
Serine Proteases ◽  
Expression Patterns ◽  
High Sensitivity ◽  
Host Parasite Interactions ◽  
Highly Sensitive ◽  
Host Parasite

Abstract Background The blood flukes of genus Schistosoma are the causative agent of schistosomiasis, a parasitic disease that infects more than 200 million people worldwide. Proteases of schistosomes are involved in critical steps of host–parasite interactions and are promising therapeutic targets. We recently identified and characterized a group of S1 family Schistosoma mansoni serine proteases, including SmSP1 to SmSP5. Expression levels of some SmSPs in S. mansoni are low, and by standard genome sequencing technologies they are marginally detectable at the method threshold levels. Here, we report their spatial gene expression patterns in adult S. mansoni by the high-sensitivity localization assay. Methodology Highly sensitive fluorescence in situ RNA hybridization (FISH) was modified and used for the localization of mRNAs encoding individual SmSP proteases (including low-expressed SmSPs) in tissues of adult worms. High sensitivity was obtained due to specifically prepared tissue and probes in combination with the employment of a signal amplification approach. The assay method was validated by detecting the expression patterns of a set of relevant reference genes including SmCB1, SmPOP, SmTSP-2, and Sm29 with localization formerly determined by other techniques. Results FISH analysis revealed interesting expression patterns of SmSPs distributed in multiple tissues of S. mansoni adults. The expression patterns of individual SmSPs were distinct but in part overlapping and were consistent with existing transcriptome sequencing data. The exception were genes with significantly low expression, which were also localized in tissues where they had not previously been detected by RNA sequencing methods. In general, SmSPs were found in various tissues including reproductive organs, parenchymal cells, esophagus, and the tegumental surface. Conclusions The FISH-based assay provided spatial information about the expression of five SmSPs in adult S. mansoni females and males. This highly sensitive method allowed visualization of low-abundantly expressed genes that are below the detection limits of standard in situ hybridization or by RNA sequencing. Thus, this technical approach turned out to be suitable for sensitive localization studies and may also be applicable for other trematodes. The results suggest that SmSPs may play roles in diverse processes of the parasite. Certain SmSPs expressed at the surface may be involved in host–parasite interactions. Graphic abstract

Sign in / Sign up

Export Citation Format

Share Document