scholarly journals Proteomic analysis of uterine fluid of fertile and subfertile hens before and after insemination

Reproduction ◽  
2019 ◽  
Vol 158 (4) ◽  
pp. 335-356
Author(s):  
Cindy Riou ◽  
Aurélien Brionne ◽  
Luiz Cordeiro ◽  
Grégoire Harichaux ◽  
Audrey Gargaros ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Rapid Change ◽  
Apical Part ◽  
Sperm Storage ◽  
Proteomic Data ◽  
Egg Formation ◽  
Before And After ◽  
Uterine Fluid ◽  
Lipid Transporters ◽  
Sperm Storage Tubules

Avian uterine fluid (UF) and uterovaginal sperm storage tubules (SST) are key components in accepting sperm in SST, maintaining sperm function for several weeks, releasing sperm from SST and their ascent through the uterus. To improve the understanding of sperm storage processes requires investigating UF and SST. This study aimed to identify proteins modulated by sperm in the hen’s genital tract and to highlight their role during sperm storage. Two genetic lines of hens exhibiting long (F+) or short (F−) sperm storage ability were used. GeLC MS/MS analysis was used to establish a quantitative inventory of proteins regulated after insemination in both lines. The proteomic data are available via ProteomeXchange with identifier PXD013514. Immunohistochemistry was used to identify high (ANXA4/ANXA5/OCX32) and low (HSPA8/PIGR) fertility markers in the uterovaginal junction. Our results demonstrated that sperm induced a significant and rapid change in the UF proteomic content and also in the SST epithelium. In F+ hens, mobilization of the ANXA4 protein in the apical part of SST cells after insemination was associated with increased levels of some proteoglycans and binding proteins, and also antimicrobial eggshell matrix protein (OCX32) in the UF. We also observed increased levels of lipid transporters involved in egg formation (VTG1-2, APOA1-4-H). In F− hens, insemination induced increased levels of PIGR in both UF and SST, of ANXA5 in SST, of UF enzymes exhibiting metallopeptidase activity and mucins. In conclusion, sperm induced significant changes in the UF proteomic content. This study also provides evidence that the SST immune system plays a major role in regulating sperm storage.

Localization of calcium and zinc in the sperm storage tubules of chicken, quail and turkey using X-ray microanalysis

Reproduction ◽  
2000 ◽  
pp. 331-336 ◽  
Author(s):  
L Holm ◽  
H Ekwall ◽  
GJ Wishart ◽  
Y Ridderstrale
Keyword(s):  
Calcium Concentration ◽  
Sperm Storage ◽  
X Ray ◽  
Elemental Concentrations ◽  
Low Concentrations ◽  
Intracellular Content ◽  
Wet Weight ◽  
The Mean ◽  
Sperm Storage Tubules ◽  
Scanning Electron

Sperm storage tubules from the utero-vaginal junction of chickens, quails and turkeys were analysed for calcium and zinc using X-ray microanalysis of ultra-rapidly frozen tissue in a scanning electron microscope. This technique enabled the tubular fluid surrounding the stored spermatozoa and the intracellular content of the cells of the sperm storage tubules to be analysed separately and, by using standards with known concentrations, their elemental concentrations were estimated. The mean (+/- SEM) concentration of calcium in the tubular fluid from chickens, quails and turkeys was 17 +/- 3, 19 +/- 3 and 17 +/- 4 mmol kg(-1) wet weight, respectively. The intracellular calcium concentration of the cells of the tubules did not differ significantly from these values and was also similar in the mucosal epithelial cells of the utero-vaginal junction. Zinc was localized in the cells of turkey sperm storage tubules and tubular fluid, but at low concentrations. No zinc could be detected in corresponding structures from chickens and quails. The concentration of calcium in the tubular fluid is within the range known to inhibit the motility of spermatozoa, supporting this function for calcium during storage. Zinc is known to depress turkey sperm metabolism and it may also be involved in inducing quiescence of spermatozoa during storage in this species.

scholarly journals Identification of food-derived peptides in human blood after ingestion of corn and wheat gluten hydrolysates

2018 ◽  
Vol 2 ◽  
Author(s):  
Akika Ejima ◽  
Megumi Nakamura ◽  
Yasushi A. Suzuki ◽  
Kenji Sato
Keyword(s):  
Matrix Protein ◽  
Leucine Aminopeptidase ◽  
Wheat Gluten ◽  
Protein Hydrolysates ◽  
The Body ◽  
Size Exclusion ◽  
Extracellular Matrix Protein ◽  
Reverse Phase Hplc ◽  
Before And After ◽  
Exclusion Chromatography

Bioactive peptides in the body after ingestion of plant protein hydrolysates have been speculated but not identified. We aimed to establish an approach to identify small amounts of food-derived peptides in humans after ingestion of non-extracellular matrix protein hydrolysates. Corn and wheat gluten hydrolysates were digested using pancreatin and leucine aminopeptidase; the resultant peptides were identified via size-exclusion chromatography and reverse-phase HPLC-tandem mass spectrometry (MS/MS). Structures of indigestible peptides were confirmed via LC-MS/MS in multi-reaction monitoring mode. All indigestible peptides in the exopeptidase digest were diprolyl and di- and tripyroglutamyl peptides. Blood collected from healthy volunteers (n = 4) before and after ingestion of 9 g of the hydrolysates was assessed for the indigestible peptides via LC-MS/MS. Six peptides (Pro-Ala, Pro-Gly, Pro-Gln, pyroGlu-Pro, pyroGlu-Leu-Pro, and pyroGlu-Gln-Pro) significantly increased in human plasma up to 10–100 nM compared to the baseline. This may hence be a powerful tool for identifying foodderived peptides in blood.

Localization of calcium and zinc in the sperm storage tubules of chicken, quail and turkey using X-ray microanalysis

Reproduction ◽  
2000 ◽  
Vol 118 (2) ◽  
pp. 331-336 ◽  
Author(s):  
L Holm ◽  
H Ekwall ◽  
G. Wishart ◽  
Y Ridderstrale
Keyword(s):  
Sperm Storage ◽  
X Ray ◽  
Sperm Storage Tubules

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.

Isolation and molecular characterisation of sperm storage tubules from the oviducal mucosa of the turkey hen

2000 ◽  
Vol 41 (sup001) ◽  
pp. 14-14
Author(s):  
L. M. King ◽  
T.S. Sonstegard ◽  
T.J. Caperna ◽  
J-P. Brillard ◽  
M.R. Bakst ◽  
...  
Keyword(s):  
Sperm Storage ◽  
Molecular Characterisation ◽  
Sperm Storage Tubules

scholarly journals Comparisons of sperm storage tubule distribution and number in 4 strains of mature broiler breeders and in turkey hens before and after the onset of photostimulation1

2010 ◽  
Vol 89 (5) ◽  
pp. 986-992 ◽  
Author(s):  
M.R. Bakst ◽  
A.M. Donoghue ◽  
D.E. Yoho ◽  
J.R. Moyle ◽  
S.M. Whipple ◽  
...  
Keyword(s):  
Sperm Storage ◽  
Broiler Breeders ◽  
Before And After
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