TEX101 is shed from the surface of sperm located in the caput epididymidis of the mouse

Zygote ◽  
2005 ◽  
Vol 13 (4) ◽  
pp. 325-333 ◽  
Author(s):  
Takeshi Takayama ◽  
Takuya Mishima ◽  
Miki Mori ◽  
Tomoko Ishikawa ◽  
Takami Takizawa ◽  
...  
Keyword(s):  
Adult Mouse ◽  
Epididymal Sperm ◽  
Cell Surfaces ◽  
Testicular Sperm ◽  
Additional Information ◽  
Reactive Protein ◽  
Surface Molecules ◽  
Specific Manner ◽  
Caput Epididymidis

It is generally believed that cell-to-cell cross-talk and signal transduction are mediated by cell surface molecules that play diverse and important regulatory roles in spermatogenesis and fertilization. Recently, we identified a novel plasma membrane-associated protein, TES101-reactive protein (TES101RP, or TEX101), on mouse testicular germ cells. In this study, we investigate Tex101 mRNA expression in the adult mouse testis using in situ hybridization, and we examine the fate of TEX101 during sperm transport by immunohistochemical and Western blot analyses. Tex101 mRNA was expressed in a stage-specific manner in spermatocytes and in step 1–9 spermatids of the testis, but not in spermatogonia. Although the TEX101 protein remained on the cell surfaces of step 10–16 spermatids and testicular sperm, it was shed from epididymal sperm located in the caput epididymidis. The results of this study provide additional information on germ cell-specific TEX101 expression during spermatogenesis and post-testicular sperm maturation.

scholarly journals Compartmentalization of membrane trafficking, glucose transport, glycolysis, actin, tubulin and the proteasome in the cytoplasmic droplet/Hermes body of epididymal sperm

Open Biology ◽  
2015 ◽  
Vol 5 (8) ◽  
pp. 150080 ◽  
Author(s):  
Catherine E. Au ◽  
Louis Hermo ◽  
Elliot Byrne ◽  
Jeffrey Smirle ◽  
Ali Fazel ◽  
...  
Keyword(s):  
Membrane Trafficking ◽  
Glucose Transporter ◽  
Molecular Level ◽  
Mammalian Species ◽  
Cytoskeletal Proteins ◽  
Epididymal Sperm ◽  
Quantitative Electron Microscopy ◽  
Cytoplasmic Droplet ◽  
Germ Cell Differentiation

Discovered in 1909 by Retzius and described mainly by morphology, the cytoplasmic droplet of sperm (renamed here the Hermes body) is conserved among all mammalian species but largely undefined at the molecular level. Tandem mass spectrometry of the isolated Hermes body from rat epididymal sperm characterized 1511 proteins, 43 of which were localized to the structure in situ by light microscopy and two by quantitative electron microscopy localization. Glucose transporter 3 (GLUT-3) glycolytic enzymes, selected membrane traffic and cytoskeletal proteins were highly abundant and concentrated in the Hermes body. By electron microscope gold antibody labelling, the Golgi trafficking protein TMED7/p27 localized to unstacked flattened cisternae of the Hermes body, as did GLUT-3, the most abundant protein. Its biogenesis was deduced through the mapping of protein expression for all 43 proteins during male germ cell differentiation in the testis. It is at the terminal step 19 of spermiogenesis that the 43 characteristic proteins accumulated in the nascent Hermes body.

scholarly journals AnIn VivoZebrafish Model for Interrogating ROS-Mediated Pancreaticβ-Cell Injury, Response, and Prevention

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Abhishek A. Kulkarni ◽  
Abass M. Conteh ◽  
Cody A. Sorrell ◽  
Anjali Mirmira ◽  
Sarah A. Tersey ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Cell Injury ◽  
Zebrafish Model ◽  
Chemical Genetic ◽  
Regulated Cell Death ◽  
Specific Manner ◽  
High Doses

It is well known that a chronic state of elevated reactive oxygen species (ROS) in pancreaticβ-cells impairs their ability to release insulin in response to elevated plasma glucose. Moreover, at its extreme, unmitigated ROS drives regulated cell death. This dysfunctional state of ROS buildup can result both from genetic predisposition and environmental factors such as obesity and overnutrition. Importantly, excessive ROS buildup may underlie metabolic pathologies such as type 2 diabetes mellitus. The ability to monitor ROS dynamics inβ-cells in situ and to manipulate it via genetic, pharmacological, and environmental means would accelerate the development of novel therapeutics that could abate this pathology. Currently, there is a lack of models with these attributes that are available to the field. In this study, we use a zebrafish model to demonstrate that ROS can be generated in aβ-cell-specific manner using a hybrid chemical genetic approach. Using a transgenic nitroreductase-expressing zebrafish line,Tg(ins:Flag-NTR)s950, treated with the prodrug metronidazole (MTZ), we found that ROS is rapidly and explicitly generated inβ-cells. Furthermore, the level of ROS generated was proportional to the dosage of prodrug added to the system. At high doses of MTZ, caspase 3 was rapidly cleaved,β-cells underwent regulated cell death, and macrophages were recruited to the islet to phagocytose the debris. Based on our findings, we propose a model for the mechanism of NTR/MTZ action in transgenic eukaryotic cells and demonstrate the robust utility of this system to model ROS-related disease pathology.

The testis-specific transcript (ferT) of the tyrosine kinase FER is expressed during spermatogenesis in a stage-specific manner

1990 ◽  
Vol 10 (9) ◽  
pp. 5021-5025
Author(s):  
E Keshet ◽  
A Itin ◽  
K Fischman ◽  
U Nir
Keyword(s):  
In Situ Hybridization ◽  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Meiotic Prophase ◽  
Hybridization Analysis ◽  
Pachytene Stage ◽  
Specific Transcript ◽  
Specific Manner

ferT is a testis-specific transcript of FER encoding a truncated version of the potential tyrosine kinase. Using in situ hybridization analysis, we found that ferT was transiently expressed during spermatogenesis and that expression was restricted to spermatocytes at the pachytene stage of meiotic prophase. This pattern of expression is unprecedented by other tyrosine kinases and suggests a role for ferT in a particular stage of spermatogenesis.

scholarly journals Exo-enzymatic addition of diazirine-modified sialic acid to cell surfaces enables photocrosslinking of glycoproteins

2021 ◽  
Author(s):  
Nageswari Yarravarapu ◽  
Rohit Sai Reddy Konada ◽  
Narek Darabedian ◽  
Nichole J. Pedowtiz ◽  
Soumya N. Krishnamurthy ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Cell Surface ◽  
Cell Types ◽  
Cell Surfaces ◽  
Binding Interactions ◽  
Multiple Cell ◽  
Labeling Method ◽  
Cell Surface Glycoconjugates

Glycan binding often mediates extracellular macromolecular recognition events. Accurate characterization of these binding interactions can be difficult because of dissociation and scrambling that occur during purification and analysis steps. Use of photocrosslinking methods has been pursued to covalently capture glycan-dependent interactions in situ however use of metabolic glycan engineering methods to incorporate photocrosslinking sugar analogs is limited to certain cell types. Here we report an exo-enzymatic labeling method to add a diazirine-modified sialic acid (SiaDAz) to cell surface glycoconjugates. The method involves chemoenzymatic synthesis of diazirine-modified CMP-sialic acid (CMP-SiaDAz), followed by sialyltransferase-catalyzed addition of SiaDAz to desialylated cell surfaces. Cell surface SiaDAz-ylation is compatible with multiple cell types and is facilitated by endogenous extracellular sialyltransferase activity present in Daudi B cells. This method for extracellular addition of α2-6-linked SiaDAz enables UV-induced crosslinking of CD22, demonstrating the utility for covalent capture of glycan-mediated binding interactions.

scholarly journals Double-stranded DNA-scaffolded fluorescent probes for fluorescence imaging of cell-surface molecules

RSC Advances ◽  
2017 ◽  
Vol 7 (83) ◽  
pp. 52581-52587
Author(s):  
Zhanghua Liu ◽  
Yang Liu ◽  
Yanan Sun ◽  
Guo Chen ◽  
Yong Chen
Keyword(s):  
Cell Surface ◽  
Fluorescence Imaging ◽  
Fluorescent Probes ◽  
Cell Surface Molecules ◽  
Cell Surfaces ◽  
Surface Molecules ◽  
Double Stranded Dna

Double-stranded DNA-scaffolded fluorescent probes were developed for fluorescence imaging of molecules on cell surfaces.

scholarly journals Prognostic Metrics Associated with Inflammation and Atherosclerosis Signaling Evaluate the Burden of Adverse Clinical Outcomes in Ischemic Stroke Patients

2020 ◽  
Vol 66 (11) ◽  
pp. 1434-1443
Author(s):  
Daoxia Guo ◽  
Zhengbao Zhu ◽  
Chongke Zhong ◽  
Aili Wang ◽  
Xuewei Xie ◽  
...  
Keyword(s):  
Ischemic Stroke ◽  
Clinical Outcomes ◽  
Pathway Analysis ◽  
Primary Outcome ◽  
Extreme Case ◽  
High Sensitivity ◽  
Additional Information ◽  
Reactive Protein ◽  
Significant Enrichment ◽  
High Level

Abstract Background Conventional prognostic risk factors can only partly explain the adverse clinical outcomes after ischemic stroke. We aimed to establish a set of prognostic metrics and evaluate its public health significance on the burden of adverse clinical outcomes of ischemic stroke. Methods All patients were from the China Antihypertensive Trial in Acute Ischemic Stroke (CATIS). We established prognostic metrics of ischemic stroke from 20 potential biomarkers in a propensity-score-matched extreme case sample (n = 146). Pathway analysis was conducted using Ingenuity Pathway Analysis. In the whole CATIS population (n = 3575), we evaluated effectiveness of these prognostic metrics and estimated their population-attributable fractions (PAFs) related to the risk of clinical outcomes. The primary outcome was a composite outcome of death or major disability (modified Rankin Scale score ≥3) at 3 months after stroke. Results Matrix metalloproteinase-9 (MMP-9), S100A8/A9, high-sensitivity C-reactive protein (hsCRP), and growth differentiation factor-15 (GDF-15) were selected as prognostic metrics for ischemic stroke. Pathway analysis showed significant enrichment in inflammation and atherosclerosis signaling. All 4 prognostic metrics were independently associated with poor prognosis of ischemic stroke. Compared with patients having 1 or 0 high-level prognostic metrics, those with 4 had higher risk of primary outcome (OR: 3.84, 95%CI: 2.67–5.51; PAF: 37.4%, 95%CI: 19.5%–52.9%). Conclusion The set of prognostic metrics, enriching in inflammation and atherosclerosis signaling, could effectively predict the prognosis at 3 months after ischemic stroke and would provide additional information for the burden of adverse clinical outcomes among patients with ischemic stroke.

scholarly journals Intake of 3 Eggs/Day or Equivalent Amount of Choline as Supplement for 4 Weeks Increases Plasma Choline Without Changing Plasma TMAO in Participants with Metabolic Syndrome

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 480-480
Author(s):  
Minu Thomas ◽  
Marissa Dibella ◽  
Olga V Malysheva ◽  
Marie A Caudill ◽  
Christopher Blesso ◽  
...  
Keyword(s):  
Metabolic Syndrome ◽  
Inflammatory Markers ◽  
Liver Enzymes ◽  
Initial Period ◽  
Healthy Population ◽  
Random Allocation ◽  
Additional Information ◽  
Reactive Protein ◽  
Funding Sources ◽  
Egg Period

Abstract Objectives Our previous study in a young healthy population indicated that choline from eggs (phosphatidylcholine) was more bioavailable than choline from a supplement (choline bitartrate) as determined by plasma choline concentrations. The purpose of this study was to compare equivalent amounts of two choline sources on plasma choline and its derivatives including trimethylamine N-oxide (TMAO) and inflammatory markers in men and women aged 32 to 70 years old with metabolic syndrome (MetS). Methods Twenty-three subjects with MetS were included in this randomized, crossover clinical trial. Participants underwent an initial period of 2 weeks without consuming any eggs, which was followed by a random allocation to either 3 eggs/day or a choline-supplement for 4 weeks (both diets had a choline equivalent of 400 mg per day). Following a 3-week washout period, participants were allocated to the alternate diet. We measured plasma choline and plasma TMAO as well as C-reactive protein (CRP), inflammatory markers and liver enzymes. Results Although there was an overall significant increase in plasma choline after egg intake, compared to baseline (P < 0.01), there were no significant differences between egg and supplement at the end of the respective interventions (P > 0.05). Baseline values were 7.9 ± 2.1 nmol/ml compared to 9.9 ± 2.2 and 9.5 ± 2.1 nmol/ml for the egg and supplement, respectively (P < 0.01). In addition, plasma TMAO was not different between baseline, or at the end of the egg and supplement periods (P > 0.1). When we measured inflammatory markers, compared to baseline CRP was lower after the egg period (0.49 ± 0.50 vs. 0.36 ± 0.37 mg/dL, P < 0.01) while no differences in this parameter were observed at the end of the egg or the supplement period. Liver enzymes were not affected by treatment. Conclusions These studies indicate that in contrast to healthy individuals, the plasma choline response appears to be similar in MetS participants, independent of its source or chemical composition. Analyzing the microbiota of these subjects will provide additional information regarding how choline is metabolized in individuals with MetS. Funding Sources The Egg Nutrition Center.

Combined synchrotron x-ray diffraction and micro-Raman for following in situ the growth of solution-deposited YBa2Cu3O7 thin films

2005 ◽  
Vol 20 (12) ◽  
pp. 3270-3273 ◽  
Author(s):  
F. Berberich ◽  
H. Graafsma ◽  
B. Rousseau ◽  
A. Canizares ◽  
R. Ramy Ratiarison ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
X Ray Diffraction ◽  
X Ray ◽  
Additional Information ◽  
Micro Raman Spectroscopy ◽  
High Temperature Process ◽  
Structural Growth ◽  
Metal Organic ◽  
Insight Into

A unique combination of in situ synchrotron x-ray diffraction and in situ micro-Raman spectroscopy was used to study the growth process of YBa2Cu3O6+x films obtained by metal organic decomposition using trifluoroacetate precursor on LaAlO3 substrates. The techniques give complementary information: x-ray diffraction gives insight into the structural growth, whereas micro-Raman spectroscopy gives information of the chemical composition with additional information on the texture. To perform both experiments in situ, a special high-temperature process chamber was designed.

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