15 SPERM STORAGE IN FEMALE REPRODUCTIVE TRACT: STUDY OF MOLECULES INVOLVED

2015 ◽  
Vol 27 (1) ◽  
pp. 100
Author(s):  
C. Riou ◽  
A. Gargaros ◽  
G. Harichaux ◽  
A. Brionne ◽  
J. Gautron ◽  
...  
Keyword(s):  
Protein Content ◽  
Quantitative Methods ◽  
Reproductive Tract ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Protein ◽  
Sperm Storage ◽  
Female Reproductive Tract ◽  
Storage Duration ◽  
Uterine Fluid

Because of prolonged sperm storage in their oviduct, domestic hens can produce fertile eggs for up to 3 weeks following a single AI. The oviduct secretions may have an effect on sperm survival, but its composition during fertilization is unknown. In the present study, we compared the proteomic content of uterine fluid collected from two lines of hens divergent by their duration of fertility period (DFP), which defined sperm-storage duration. The first line displays a shorter period of sperm storage (10 days, line DFP–), whereas the second displays a longer period of sperm storage (21 days, DFP+). The aim was to identify proteins or peptides that may be involved in spermatozoa survival. Uterine fluid was collected 10 h after oviposition either before (n = 5/line) or 24 h after (n = 5/line) AI. Samples were pooled by condition: DFP+ before AI, DFP+ after AI, DFP– before AI, and DFP– after AI. Bottom-up approach using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nano LC-MS/MS was performed (3 replicates). Data were matched against the NCBInr database (2014) using Mascot, and identifications were validated by the peptide and protein Prophet algorithm using Scaffold software. To determine the differences in protein expression, spectral counting and XIC quantitative methods were employed using Scaffold Q+ (P < 0.05, ratio > 2). Two proteins were up-regulated and one was down-regulated in oviductal secretion of both lines in response to AI. However, AI induced a significantly different abundance between protein content of DFP– and DFP+ fluids. A panel of 8 proteins, included one DFP+-specific protein, was more abundant in DFP+ line than in DFP–. Only one protein was less abundant in DFP+ line than in DFP–. In conclusion, the presence of sperm in the genital tract induced quantitative differences of the protein content of the uterine fluid in DFP– and DFP+ hen lines. These differences imply proteins that are known as male proteins (sperm, seminal plasma, testis). Analysis of sperm protein modifications after storage will help us to understand the functional implication of these candidates.

scholarly journals Do spermathecal morphology and inter-mating interval influence paternity in the polyandrous beetle Tribolium castaneum?

Behaviour ◽  
2006 ◽  
Vol 143 (5) ◽  
pp. 643-658 ◽  
Author(s):  
Ludovic Arnaud ◽  
Giorgina Bernasconi ◽  
Yves Brostaux ◽  
Eric P. Meyer
Keyword(s):  
Sperm Competition ◽  
Tribolium Castaneum ◽  
Reproductive Tract ◽  
Sperm Storage ◽  
Female Reproductive Tract ◽  
Phenotypic Marker ◽  
Male And Female ◽  
The Difference ◽  
Different Strains

AbstractIn polyandrous insects, postcopulatory sexual selection is a pervasive evolutionary force favouring male and female traits that allow control of offspring paternity. Males may influence paternity through adaptations for sperm competition, and females through adaptations facilitating cryptic female choice. Yet, the mechanisms are often complex, involving behaviour, physiology or morphology, and they are difficult to identify. In red flour beetles (Tribolium castaneum), paternity varies widely, and evidence suggests that both male and female traits influence the outcome of sperm competition. To test the role of spermathecal morphology and of sperm storage processes on the outcome of sperm competition, we mated each of 26 virgin females with two males, one of which carrying a phenotypic marker to assign offspring paternity. We manipulated the interval between mating with the first and the second male, to create different conditions of sperm storage (overlapping and non-overlapping) in the female reproductive tract. To investigate the role of sperm storage more closely, we examined the relationship between paternity and spermathecal morphology in a subset of 14 experimental females. In addition, we also characterized variation in spermathecal morphology in three different strains, wildtype, Chicago black and Reindeer. No significant influence of the intermating interval was found on the paternity of the focal male, although the direction of the difference was in the expected direction of higher last male paternity for longer intervals. Moreover, paternity was not significantly associated with spermathecal morphology, although spermathecal volume, complexity, and tubule width varied significantly and substantially among individuals in all investigated strains.

119. IDENTIFICATION AND CHARACTERISATION OF SURFACE PROTEIN COMPLEXES IN HUMAN SPERMATOZOA

2010 ◽  
Vol 22 (9) ◽  
pp. 37 ◽  
Author(s):  
K. A. Redgrove ◽  
B. Nixon ◽  
E. A. McLaughlin ◽  
M. K. O'Bryan ◽  
R. J. Aitken
Keyword(s):  
Zona Pellucida ◽  
Reproductive Tract ◽  
Protein Complexes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Human Sperm ◽  
Female Reproductive Tract ◽  
Human Spermatozoa ◽  
Apical Region ◽  
Post Translational Modification ◽  
Cellular Mechanisms

A unique characteristic of mammalian spermatozoa is that upon ejaculation, they are unable to recognise and bind to an ovulated oocyte. These functional attributes are only realised following the sperms ascent of the female reproductive tract whereupon they undergo a myriad of biochemical and biophysical changes collectively referred to as ‘capacitation’. Since spermatozoa are both transcriptionally and translationally quiescent cells, this functional transformation must be engineered by a combination of post-translational modification and spatial reorganisation of existing sperm proteins. Indeed, evidence from our laboratory suggests that a key attribute of capacitation is the remodeling of the sperm surface architecture leading to the assembly and / or presentation of multimeric sperm-oocyte receptor complex(es). Through the novel application of Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE), we have secured the first direct evidence that human spermatozoa express a number of these protein complexes on their surface. Furthermore, we have demonstrated that a subset of these complexes harbour putative zona adhesion proteins and display strong affinity for solubilised zona pellucidae. In this study, we have extended our findings through the characterisation of one such complex containing arylsulfatase A (ASA), a protein with recognised affinity for sulfated ligands present within the zona pellucida. Through the application of immunohistochemistry and flow cytometry we revealed that ASA undergoes a capacitation-associated translocation to become expressed on the apical region of the human sperm head, a location compatible with a role in the mediation of sperm-zona pellucida interactions. This dramatic relocation was completely abolished by incubation of capacitating spermatozoa in exogenous cholesterol, suggesting that it may be driven in part by alteration in the membrane fluidity characteristics. Our current research is focused on confirming the role of ASA in human sperm-zona pellucida adhesion and elucidating the precise cellular mechanisms that underpin the proteins translocation to the cell surface.

scholarly journals Characterization of a Host-Specific Protein Toxin (Ptr ToxB) from Pyrenophora tritici-repentis

1999 ◽  
Vol 12 (8) ◽  
pp. 728-732 ◽  
Author(s):  
Stephen E. Strelkov ◽  
Lakhdar Lamari ◽  
G. Murray Ballance
Keyword(s):  
Amino Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Protein ◽  
Tan Spot ◽  
Partial Purification ◽  
Toxic Activity ◽  
Pyrenophora Tritici Repentis ◽  
Protein Toxin ◽  
Cation Exchange Column

Pyrenophora tritici-repentis, the causal agent of tan spot of wheat, differentially induces tan necrosis and/or chlorosis in wheat. A chlorosis-inducing, host-specific toxin, termed Ptr ToxB (formerly Ptr chlorosis toxin), was purified from the culture filtrates of a race 5 isolate of P. tritici-repentis. Partial purification was performed by 25 to 80% ammonium sulfate precipitation and passage through a carboxy-methyl-Sephadex C-25 cation exchange column. Final purification was performed by fast performance liquid chromatography, with a Mono S HR 5/5 cation exchanger, followed by size fractionation on a Superose 12 HR 10/30 column. The toxin was shown to be proteinaceous in nature, and purity was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular mass of Ptr ToxB was determined to be 6.61 kDa. The amino acid composition and partial N terminus amino acid sequence of the toxin were also obtained. Ptr ToxB was found to be heat stable, maintaining full toxic activity after 1 h at 55°C. Infiltration of toxin concentrations as low as 14 nM produced chlorosis on susceptible cultivars.

scholarly journals B-Cell Lymphoma-2 Localization in the Female Reproductive Tract of the Chinese Soft-Shelled Turtle,Pelodiscus Sinensisand Its Relationship With Sperm Storage

2015 ◽  
Vol 298 (12) ◽  
pp. 2011-2017 ◽  
Author(s):  
Yuan Le ◽  
Shaofan Chen ◽  
Lisi Hu ◽  
Linli Zhang ◽  
Shakeeb Ullah ◽  
...  
Keyword(s):  
B Cell ◽  
Reproductive Tract ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Sperm Storage ◽  
Female Reproductive Tract

scholarly journals Identification of Differentially Expressed Protein from Electrical Stunning of Broiler Chickens Meat Protein

Molekul ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. 71 ◽  
Author(s):  
Sandra Hermanto ◽  
Maya Ina Sholaikah ◽  
Sri Suci Mulyani
Keyword(s):  
Muscle Tissue ◽  
Broiler Chickens ◽  
Broiler Chicken ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Protein ◽  
Differentially Expressed ◽  
Over Expression ◽  
Differentially Expressed Protein ◽  
Sds Page

Identification of differentially expressed protein from the muscle tissue of broiler chicken meat with different conditions of pre-slaughter has been done. Each sample (6 broilers aged 21 days, 1 kg of weight ) was prepared through the process of pre-slaughter with 3 conditions, the first sample slaughtered in a conventional way which untreated electrical stunning, while the second and third sample of the chicken was prepared by using electrical stunning with 1 A and 25 Volts for 5 seconds and 1 A, 125 Volts for 30 seconds. Two biological replicate were done for each of samples. Muscle tissue protein extracted in Tris HCl pH 8.0 and the proteins separation by using SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis). Identification of differentially expressed protein performed by densitometry to identify the profile of the resulting proteins. The results of this study showed that the protein bands constructed in the range of 8.5-140 kDa and 9 dominant protein bands with different relative intensities. Densitogram analysis results showed there are two specific protein bands appear on the results of the electrical stunning which more extensive over expression. This indicates the electrical stunning of slaughter process may triggered the expression levels of certain proteins that do not occur in the nonelectrical stunning.

scholarly journals Specific protein synthesis in isolated rat testis leydig cells. Influence of luteinizing hormone and cycloheximide

1977 ◽  
Vol 162 (2) ◽  
pp. 341-346 ◽  
Author(s):  
F H A Janszen ◽  
B A Cooke ◽  
H J van der Molen
Keyword(s):  
Protein Synthesis ◽  
Luteinizing Hormone ◽  
Leydig Cell ◽  
Half Life ◽  
Leydig Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Specific Protein ◽  
Rat Testis

The effect of luteinizing hormone (luteotropin) and cycloheximide on specific protein synthesis in rat testis Leydig cells has been investigated. Proteins were labelled with either I114C]leucine, [3H]leucine or [35S]methionine during incubation with Leydig-cell suspensions in vitro. Total protein was extracted from the cells and separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. No detectable increase in the synthesis of specific proteins could be observed after incubation of Leydig cells with luteinizing hormone for up to 1 h. However, after a 2h incubation period, an increase in [35S]methionine incorporation was observed in a protein with an apparent mol.wt. of 21000 (referred to as ‘protein 21’). When, after labelling of this protein with [35S]-methionine, Leydig cells were incubated for another 30min with cycloheximide, no decrease in radioactivity of this protein band was observed, indicating that it does not have a short half-life. However, another protein band was detected, which after incubation with cycloheximide disappeared rapidly, the reaction following first-order kinetics, with a half-life of about 11 min. This protein, with an apparent mol.wt. of 33000 (referred to as “protein 33”), was found to be located in the particulate fraction of the Leydig cell, and could not be demonstrated in other rat testis-cell types or blood cells. No effect of luteinizing hormone on molecular weight, subcellular localization or half-life of protein 33 was observed. A possible role for protein 33 and protein 21 in the mechanism of action of luteinizing hormone on testosterone production of Leydig cells is discussed.

Sign in / Sign up

Export Citation Format

Share Document