Shotgun proteomics of rainbow trout ovarian fluid

2015 ◽  
Vol 27 (3) ◽  
pp. 504 ◽  
Author(s):  
Joanna Nynca ◽  
Georg J. Arnold ◽  
Thomas Fröhlich ◽  
Andrzej Ciereszko
Keyword(s):  
Rainbow Trout ◽  
Cell Structure ◽  
Cell Lineage ◽  
Myeloid Cell ◽  
Shotgun Proteomics ◽  
Oocyte Quality ◽  
Lipid Binding ◽  
Ovarian Fluid ◽  
Proteomic Approach ◽  
Vitelline Envelope

In the present study we used a shotgun proteomic approach to identify 54 proteins of rainbow trout ovarian fluid. The study has unravelled the identity of several proteins not previously reported in fish ovarian fluid. The proteome of trout ovarian fluid consists of diverse proteins participating in lipid binding and metabolism, carbohydrate and ion transport, innate immunity, maturation and ovulation processes. Most trout ovarian fluid proteins correspond to follicular fluid proteins of higher vertebrates, but 15% of the proteins were found to be different, such as those related to the immune system (precerebellin-like protein), proteolysis (myeloid cell lineage chitinase), carbohydrate and lipid binding and metabolism (vitellogenins), cell structure and shape (vitelline envelope protein gamma) and a protein with unknown functions (UPF0762 protein C6orf58 homologue). The present study could help in the decoding of the biological function of these proteins and in the discovery of potential biomarkers of oocyte quality.

scholarly journals Physiological And Proteomic Analyses Reveals That Brassinosteroids Application Improves The Chilling Stress Tolerance of Pepper Seedlings

2021 ◽  
Author(s):  
Jie Li ◽  
Hamza Sohail ◽  
Muhammad Azher Nawaz ◽  
Chaowei Liu ◽  
Ping Yang
Keyword(s):  
Foliar Application ◽  
Protein Biosynthesis ◽  
Cell Structure ◽  
Chilling Stress ◽  
Calvin Cycle ◽  
Mesophyll Cell ◽  
Limited Information ◽  
Proteomic Approach ◽  
Gsh Metabolism ◽  
Transfer Chain

Abstract Brassinosteroids (BRs) are important in plant resistance to chilling stress. However, limited information is available regarding the specific mechanisms involved at proteomic level. We utilized iTRAQ proteomic approach, physiological assays and information obtained from cellular ultrastructure to clarify the underlying molecular mechanism of BRs to alleviate chilling stress in pepper (Capsicum annuum L.). Foliar application of 24-epibrassinolide (EBR) improved photosynthesis and improved cell structure by presenting a distinct mesophyll cell and chloroplast with well-developed thylakoid membranes in the leaves of pepper seedlings. We identified 346 differentially expressed proteins (DEPs), including 217 up-regulated proteins and 129 down-regulated proteins in plants under chilling (Chill) and Chill + EBR treated plants. Most of the DEPs were related to multiple pathways, including photosynthesis, carbohydrates metabolism, energy metabolism, protein biosynthesis, amino acids synthesis, redox and stress defence (ascorbate peroxidase, glutathione peroxidase, and superoxide dismutase). Up-regulated DEPs were associated with photosynthetic electron transfer chain, oxidative phosphorylation, GSH metabolism pathway, Calvin cycle and signaling pathway. The physiochemical analysis showed that EBR treatment improved the tolerance of pepper seedlings to chilling stress.

scholarly journals Post-ovulatory secretion of pituitary gonadotropins GtH I and GtH II in the rainbow trout (Oncorhynchus mykiss): regulation by steroids and possible role of non-steroidal gonadal factors

1999 ◽  
Vol 163 (1) ◽  
pp. 87-97 ◽  
Author(s):  
J Chyb ◽  
T Mikolajczyk ◽  
B Breton
Keyword(s):  
Rainbow Trout ◽  
Oncorhynchus Mykiss ◽  
Body Cavity ◽  
The Body ◽  
Ovarian Fluid ◽  
Multiple Injections ◽  
Rainbow Trout Oncorhynchus Mykiss

In order to determine the factors of ovarian origin which can modulate the postovulatory secretion of the FSH-like gonadotropin (GtH I) and the LH-like gonadotropin (GtH II), freshly ovulated female rainbow trout were divided into two groups. In the first group the fish were stripped in order to eliminate the eggs and ovarian fluid from the body cavity, while in the second group the eggs were kept in the body cavity. Subsequently, fish from both groups were implanted with testosterone (10 mg/kg), 17beta-estradiol (10 mg/kg) or 17,20beta-ddihydroxy-4-regnen-3-one (17,20betaP) (1 mg/kg) or injected every 2 days with desteroidized ovarian fluid (1.5 ml/kg). The secretion of GtH I dramatically increased in stripped fish, reaching its maximum levels 2 weeks after ovulation. The preservation of eggs in the body cavity led to the suppression of this increase. The profiles of GtH II secretion were opposite to those encountered for GtH I because the increase of GtH II was observed only in unstripped fish. The administration of steroids showed that testosterone is able to inhibit GtH I release and stimulate that of GtH II in stripped fish, having no effect on the release of these gonadotropins in non-stripped animals. 17beta-Estradiol failed to modify GtH I secretion, however it decreased the release of GtH II in fish containing retained eggs in the body cavity. 17,20betaP had a delayed stimulating influence on GtH I release in unstripped fish. Finally, multiple injections of desteroidized ovarian fluid into stripped fish led to a significant decrease of GtH I release and to an increase of GtH II secretion. This study demonstrates that factors, which are present in ovarian fluid, modulate the post-ovulatory secretion of both gonadotropins--their net action is negative on GtH I and positive on GtH II. Among the steroids, testosterone is of major importance, being able to inhibit GtH I release and to stimulate that of GtH II. We also show that non-steroidal factors present in the ovarian fluid can influence the release of both gonadotropins, which indirectly supports the previous findings about the existence of inhibin/activin-like factors in fish.

24 GLOBAL TRANSCRIPT PROFILING OF CLONED BOVINE BLASTOCYSTS USING AFFYMETRIX GENECHIP TECHNOLOGY

2006 ◽  
Vol 18 (2) ◽  
pp. 120
Author(s):  
Z. Beyhan ◽  
P. Ross ◽  
A. Iager ◽  
A. Kocabas ◽  
K. Cunniff ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Expression Profiles ◽  
Cell Structure ◽  
Donor Cell ◽  
Lipid Binding ◽  
Differentially Expressed ◽  
Gtp Binding ◽  
Cloned Bovine

Identification of genes implicated in the biological processes of somatic cell nuclear transfer will improve our understanding of reprogramming events, i.e. the transformation of a lineage-committed cell into a pluripotent one. In addition, the gene expression profile of cloned embryos can help explain the widely reported developmental failures in cloned animals. In this study, we investigated global gene expression profiles of bovine in vitro-fertilized and cloned embryos using Gene Chip Bovine Genome Arrays (Affymetrix, Inc., Santa Clara, CA, USA). For the generation of cloned bovine blastocysts from two adult fibroblast lines (C and D), we employed methods previously proven to generate live offspring and compared these offspring to in vitro-produced blastocysts. Total RNA isolated from groups of 10 blastocysts was amplified by a template-switching PCR. Amplified cDNAs were used to synthesize biotin-labeled antisense RNAs (aRNAs) during and in vitro transcription reaction. Labeled aRNAs were hybridized to microarrays as described by the manufacturer. Experiments were performed in four replicates. Expression data were analyzed using the Significance Analysis of Microarrays (SAM; Tusher et al. 2001 Proc. Natl. Acad. Sci. 98, 5116-5121) procedure and software. Overall, 48.4% and 46% of 23 000 bovine transcripts spotted on the arrays were present in cloned and in in vitro-produced control blastocysts, respectively. The SAM procedure identified 43 genes that changed at least 1.5-fold, with an estimated false discovery rate (FDR) of 20%. Comparison of gene expression between NT embryos produced from two different cell lines and IVF controls with the same criteria revealed 6 (clones from cell line C vs. IVF) and 46 (clones from cell line D vs. IVF) differentially expressed genes. The number of transcripts expressed differentially between the cloned embryos with different donor cell origin was 437. Of the 43 differentially expressed transcripts in cloned blastocysts, 13 have unknown functions and the rest of the genes related to cell structure (tuftelin, desmoplakin), cell cycle/mitosis (Kinesin like 4, katanin, stathmin, PCNA), energy metabolism (lactate dehydrogenase, ATPsynthase, lipid-binding protein, keto acid dehydrogenase E1, metallothionein), and cell signaling (GTP-binding protein1, GTP binding stimulatory protein). Our results indicate that expression profiles of cloned blastocysts could be affected by somatic donor cell.

scholarly journals Proteomic analysis of palmitoylated platelet proteins

Blood ◽  
2011 ◽  
Vol 118 (13) ◽  
pp. e62-e73 ◽  
Author(s):  
Louisa Dowal ◽  
Wei Yang ◽  
Michael R. Freeman ◽  
Hanno Steen ◽  
Robert Flaumenhaft
Keyword(s):  
Protein Interactions ◽  
Protein Identification ◽  
Global Analysis ◽  
Critical Role ◽  
Myeloid Cell ◽  
Metabolic Labeling ◽  
Protein Protein Interactions ◽  
Proteomic Approach ◽  
Protein Palmitoylation ◽  
Liquid Chromatography Tandem Mass

Abstract Protein palmitoylation is a dynamic process that regulates membrane targeting of proteins and protein-protein interactions. We have previously demonstrated a critical role for protein palmitoylation in platelet activation and have identified palmitoylation machinery in platelets. Using a novel proteomic approach, Palmitoyl Protein Identification and Site Characterization, we have begun to characterize the human platelet palmitoylome. Palmitoylated proteins were enriched from membranes isolated from resting platelets using acyl-biotinyl exchange chemistry, followed by identification using liquid chromatography-tandem mass spectrometry. This global analysis identified > 1300 proteins, of which 215 met criteria for significance and represent the platelet palmitoylome. This collection includes 51 known palmitoylated proteins, 61 putative palmitoylated proteins identified in other palmitoylation-specific proteomic studies, and 103 new putative palmitoylated proteins. Of these candidates, we chose to validate the palmitoylation of triggering receptors expressed on myeloid cell (TREM)–like transcript-1 (TLT-1) as its expression is restricted to platelets and megakaryocytes. We determined that TLT-1 is a palmitoylated protein using metabolic labeling with [3H]palmitate and identified the site of TLT-1 palmitoylation as cysteine 196. The discovery of new platelet palmitoyl protein candidates will provide a resource for subsequent investigations to validate the palmitoylation of these proteins and to determine the role palmitoylation plays in their function.

Effects of ovarian fluid on motility characteristics of rainbow trout (Oncorhynchus mykissWalbaum) spermatozoa

2008 ◽  
Vol 24 (4) ◽  
pp. 503-507 ◽  
Author(s):  
G. J. Dietrich ◽  
M. Wojtczak ◽  
M. Słowińska ◽  
S. Dobosz ◽  
H. Kuźmiński ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Ovarian Fluid

scholarly journals IFN Regulatory Factor 8 Represses GM-CSF Expression in T Cells To Affect Myeloid Cell Lineage Differentiation

2015 ◽  
Vol 194 (5) ◽  
pp. 2369-2379 ◽  
Author(s):  
Amy V. Paschall ◽  
Ruihua Zhang ◽  
Chen-Feng Qi ◽  
Kankana Bardhan ◽  
Liang Peng ◽  
...  
Keyword(s):  
T Cells ◽  
Cell Lineage ◽  
Myeloid Cell ◽  
Regulatory Factor ◽  
Gm Csf ◽  
Lineage Differentiation

scholarly journals Immune System in the Brain: A Modulatory Role on Dendritic Spine Morphophysiology?

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Oscar Kurt Bitzer-Quintero ◽  
Ignacio González-Burgos
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Immune System ◽  
Dendritic Spines ◽  
Cell Lineage ◽  
Myeloid Cell ◽  
Brain Parenchyma ◽  
Immune Mediators ◽  
The Central Nervous System ◽  
The Brain

The central nervous system is closely linked to the immune system at several levels. The brain parenchyma is separated from the periphery by the blood brain barrier, which under normal conditions prevents the entry of mediators such as activated leukocytes, antibodies, complement factors, and cytokines. The myeloid cell lineage plays a crucial role in the development of immune responses at the central level, and it comprises two main subtypes: (1) resident microglia, distributed throughout the brain parenchyma; (2) perivascular macrophages located in the brain capillaries of the basal lamina and the choroid plexus. In addition, astrocytes, oligodendrocytes, endothelial cells, and, to a lesser extent, neurons are implicated in the immune response in the central nervous system. By modulating synaptogenesis, microglia are most specifically involved in restoring neuronal connectivity following injury. These cells release immune mediators, such as cytokines, that modulate synaptic transmission and that alter the morphology of dendritic spines during the inflammatory process following injury. Thus, the expression and release of immune mediators in the brain parenchyma are closely linked to plastic morphophysiological changes in neuronal dendritic spines. Based on these observations, it has been proposed that these immune mediators are also implicated in learning and memory processes.

scholarly journals Inter-clutch egg differences and androgenesis in rainbow trout (Oncorhynchus mykiss, Walbaum 1792)

2021 ◽  
Vol 50 (2) ◽  
pp. 160-168
Author(s):  
Marcin Polonis ◽  
Agata Błaszczyk ◽  
Krzysztof Jagiełło ◽  
Ligia Panasiak ◽  
Stefan Dobosz ◽  
...  
Keyword(s):  
Antioxidant Enzymes ◽  
Rainbow Trout ◽  
Ph Value ◽  
Survival Rates ◽  
Nuclear Genome ◽  
Developmental Competence ◽  
X Rays ◽  
Ovarian Fluid ◽  
Developmental Potential ◽  
Rainbow Trout Oncorhynchus Mykiss

Abstract Ionizing radiation (IR) is applied to inactivate the nuclear genome in rainbow trout eggs during induced androgenetic development. However, IR-generated reactive oxygen species (ROS) may affect developmental potential of eggs and reduce the effectiveness of androgenesis. To verify this assumption, androgenetic development of rainbow trout was induced in eggs irradiated with 350 Gy of X-rays. Survival rates, pH of the ovarian fluid and activity of antioxidant enzymes, including SOD, CAT and GPx, were examined in non-irradiated and irradiated eggs originating from four females. Survival rates of androgenetic embryos developing in eggs produced by different females varied from 1% to 57% and these inter-clutch differences were significant. Eggs from female F4, which showed the highest developmental competence for androgenesis, also showed increased activities of SOD, CAT and GPx enzymes. The pH value of the ovarian fluid of each female was over 8 before and after irradiation, therefore it seems that radiation did not affect the ovarian fluid pH. Considering the above-mentioned inter-clutch differences, a strong maternal effect on the effectiveness of androgenesis can be assumed. Eggs with increased activity of antioxidant enzymes before irradiation should be expected to show increased developmental competence for androgenesis.

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