scholarly journals Detection of osteopontin on Holstein bull spermatozoa, in cauda epididymal fluid and testis homogenates, and its potential role in bovine fertilization

Reproduction ◽  
2007 ◽  
Vol 133 (5) ◽  
pp. 909-917 ◽  
Author(s):  
David W Erikson ◽  
Amy L Way ◽  
David A Chapman ◽  
Gary J Killian
Keyword(s):  
Reproductive Tract ◽  
Bovine Milk ◽  
High Fertility ◽  
Epididymal Sperm ◽  
Holstein Bull ◽  
Sds Page ◽  
Sperm Membrane ◽  
Epididymal Fluid

Osteopontin (OPN) is a secreted extracellular matrix phosphoprotein identified in various tissues and fluids including those of the male and female reproductive tracts. OPN was previously identified as a 55 kDa high fertility marker in Holstein bull seminal plasma, produced by the ampulla and the vesicular gland. The objectives of this study were to characterize OPN on ejaculated and cauda epididymal sperm using immunofluorescence and western blot analysis, and to assess the role of sperm OPN in fertilization. Solubilized sperm membrane proteins from ejaculated and cauda epididymal sperm were separated by 1D SDS-PAGE, transferred to nitrocellulose, and probed with an antibody to bovine milk OPN. A 35 kDa protein was detected by this antibody in both ejaculated and cauda epididymal sperm membranes. Analyses also recognized OPN at 55 and 25 kDa in cauda epididymal fluid and testicular parenchyma homogenates respectively. Immunofluorescent analysis of ejaculated and cauda epididymal sperm showed OPN localization in a well-defined band in the postacrosomal region of the sperm head and also on the midpiece. Results ofin vitrofertilization experiments showed that sperm treated with an antibody to OPN fertilized fewer oocytes than sperm treated with control medium while increasing incidence of polyspermy, suggesting a role of sperm-associated OPN in fertilization and a block to polyspermy. These studies demonstrate that OPN exists at multiple molecular weight forms in the bull reproductive tract and its presence on ejaculated sperm may signal its importance in fertilization by interacting with integrins or other proteins on the oocyte plasma membrane.

3 ROLE OF β-DEFENSIN 126 IN PROMOTING SPERM MOTILITY IN CATTLE

2016 ◽  
Vol 28 (2) ◽  
pp. 131
Author(s):  
B. Fernandez-Fuertes ◽  
F. Narciandi ◽  
K. G. Meade ◽  
C. O'Farrelly ◽  
S. Fair ◽  
...  
Keyword(s):  
Western Blot ◽  
Reproductive Tract ◽  
Cervical Mucus ◽  
Surface Proteins ◽  
Sperm Function ◽  
Department Of Agriculture ◽  
Sperm Migration ◽  
Epididymal Fluid ◽  
Multiple Species

As immature sperm migrate through the epididymis, they are bathed in region-specific epididymal fluid, which leads to a sequential addition, deletion, and modification of their surface proteins. These changes ultimately result in the acquisition of motility and fertilising abilities. Among the hundreds of proteins secreted by the epididymis, several β-defensins have been identified and correlated with male fertility in multiple species. In cattle, β-defensin 126 (BD126) is exclusively detected in the reproductive tract of pubertal males, with preferential mRNA expression in the epididymis. Both the macaque and human orthologs have been shown to play a role in the ability of sperm to migrate through cervical mucus. The aim of this study was to examine the role of bovine BD126 in sperm function. Western blot revealed that the peptide is uniquely present in both the cauda epididymis sperm and fluid and is absent from sperm recovered from other proximal epididymal regions, or the ejaculate of vasectomised animals. Confocal analysis showed immunofluorescent labelling of BD126 specific to the tail and acrosomal region in cauda sperm only, suggesting a role in motility. We hypothesised that addition of cauda epididymal fluid (CEF) or recombinant BD126 (rBD126) to immature corpus sperm would improve ability to penetrate cervical mucus. Testes from adult bulls were collected at an abattoir, and sperm from the corpus and cauda epididymis, as well as CEF, were recovered. Corpus sperm were incubated for 1 h with CEF in the absence or presence of BD126 antibody, or with different rBD126 concentrations (30 or 60 μg mL–1); untreated corpus and cauda sperm were used as controls. A higher number of cauda than corpus sperm migrated through cervical mucus from oestrus cows (P < 0.001), and addition of CEF increased the number of corpus sperm migrating through this matrix (P < 0.05). The presence of the BD126 antibody in CEF failed to abrogate this effect. Western blot analysis of the sperm samples revealed the antibody was not successful in blocking BD126 from binding onto the sperm surface, which would explain the lack of differences observed. Furthermore, the addition of rBD126 did not increase corpus sperm migration through mucus. In conclusion, we have characterised the expression of bovine BD126 protein in the bovine testis and epididymis. Incubation of sperm from the corpus with CEF from the cauda resulted in enhanced sperm migration through cervical mucus. However, incubation of sperm with rBD126 in the absence of other factors and proteins from the CEF failed to produce the same effect. These results suggest that the role of BD126 in cattle is different from that observed in primates. We are currently investigating other roles of BD126 and related β-defensins in mediating bovine sperm function. This work was supported by a grant from the Irish Department of Agriculture, Food and The Marine under the Research Stimulus Programme (Grant No. 11S 104).

scholarly journals The LH/hCG Axis in Endometrial Cancer: A New Target in the Treatment of Recurrent or Metastatic Disease

2010 ◽  
Vol 2010 ◽  
pp. 1-5 ◽  
Author(s):  
A. Arcangeli ◽  
I. Noci ◽  
A. Fortunato ◽  
G. F. Scarselli
Keyword(s):  
Cell Proliferation ◽  
Endometrial Cancer ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Proliferation And Apoptosis ◽  
Cell Invasiveness ◽  
Activate Protein Kinase ◽  
Kinase A

Endometrial cancer (EC) is a hormone-dependent cancer that currently represents the most frequent malignancy of the female reproductive tract. The involvement of steroid hormones in EC etiology and progression has been reported. More recently, gonadotropins, and, in particular LH/hCG, are emerging as novel regulators of tumor progression. In the present review, we discuss the role of the LH/hCG axis (i.e. LH/hCG and its receptors, LH/hCG-R) in both gonadal and nongonadal tissues, in physiological and neoplastic conditions. In cancer cells, LH/hCG mainly controls cell proliferation and apoptosis. In particular, in EC LH/hCG improves cell invasiveness, through a mechanism which involves the LH/hCG-R, which in turn activate protein kinase A and modulate integrin adhesion receptors. Indeed, the LH/hCG-R mRNA is expressed in primary ECs and this expression correlates with LH/hCG-induced cell invasiveness in vitro. These results lead to hypothesize that recurrent and metastatic ECs, which express LH/hCG-R, could benefit from therapies aimed at decreasing LH levels, through Gn-RH analogues. Hence, the LH/hCG axis could represent a prognostic factor and a new therapeutic target in EC.

scholarly journals In vivo exposure to 17β-estradiol triggers premature sperm capacitation in cauda epididymis

Reproduction ◽  
2013 ◽  
Vol 145 (3) ◽  
pp. 255-263 ◽  
Author(s):  
Lukas Ded ◽  
Natasa Sebkova ◽  
Martina Cerna ◽  
Fatima Elzeinova ◽  
Pavla Dostalova ◽  
...  
Keyword(s):  
Reproductive Tract ◽  
Negative Impact ◽  
Reproductive Potential ◽  
Female Reproductive Tract ◽  
Sperm Capacitation ◽  
Epididymal Sperm ◽  
Knock Out ◽  
17Β Estradiol

Estrogens play a crucial role in spermatogenesis and estrogen receptor α knock-out male mice are infertile. It has been demonstrated that estrogens significantly increase the speed of capacitation in vitro; however this may lead to the reduction of reproductive potential due to the decreased ability of these sperm to undergo the acrosome reaction. To date the in vivo effect of estrogens on the ability of sperm to capacitate has not been investigated. Therefore, in this study, we exposed mice (n=24) to 17β-estradiol (E2) at the concentration of 20 ng/ml either during puberty from the fourth to seventh week of age (n=8), or continuously from birth for a period of 12 weeks (n=8) at which age the animals from both groups were killed. The capacitation status of epididymal and testicular sperm was analysed by tyrosine phosphorylation (TyrP) antibody (immunofluorescence and western blot) and chlortetracycline (CTC) assay. According to our results, in vivo exposure to increased E2 concentrations caused premature sperm capacitation in the epididymis. The effect of E2, however, seems reversible because after the termination of the exposure premature epididymal sperm capacitation is decreased in animals treated during puberty. Furthermore the changes in epididymal sperm capacitation status detected by TyrP and CTC positively correlate with plasma levels of E2 and the expression of the estrogen-dependent trefoil factor 1 (Tff1) gene in testicular tissue. Therefore, our data implicate that in vivo exposure to E2 under specific conditions leads to the premature capacitation of mouse sperm in epididymis with a potential negative impact on the sperm reproductive fitness in the female reproductive tract.

scholarly journals Phage composition of a fermented milk and colostrum product assessed by microbiome array; putative role of open reading frames in reference to cell signaling and neurological development

2019 ◽  
Author(s):  
Stefania Pacini ◽  
Marco Ruggiero
Keyword(s):  
Dna Sequences ◽  
Bovine Milk ◽  
Fermented Milk ◽  
Open Reading Frames ◽  
Neurological Development ◽  
Putative Role ◽  
Biological Entities ◽  
Reading Frames

AbstractBacteriophages (phages), Earth’s most numerous biological entities, are natural constituents of alimentary matrices; in this study we describe the characterization of phage populations in a product obtained by fermentation of bovine milk and colostrum. Such characterizations were achieved using a microarray consisting of a chip covered in short DNA sequences that are specific to certain target organisms for a total of approximately 12,000 species. The only viruses evidenced by the array belonged to Siphoviridae, the largest phage family that targets bacteria and archea. The array yielded 27 iterations corresponding to a unique target. We discuss the putative role of some open reading frames of these phages in conferring health-supporting properties with particular reference to cells signaling and neurological development. We also describe the in vitro interaction of this fermented product with alpha-N-acetylgalactosaminidase, an enzyme whose activity in serum is elevated in neurodevelopmental disorders.

scholarly journals Bovine-associated non-aureus staphylococci suppress Staphylococcus aureus biofilm dispersal in vitro yet not through agr regulation

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Bruno Toledo-Silva ◽  
Fernando N. de Souza ◽  
Kristien Mertens ◽  
Sofie Piepers ◽  
Freddy Haesebrouck ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Communication Systems ◽  
Bovine Milk ◽  
Subclinical Mastitis ◽  
Interspecies Interactions ◽  
Environmental Signals ◽  
Biofilm Dispersion ◽  
Biofilm Development

AbstractBiofilm formation is a significant virulence factor in Staphylococcus (S.) aureus strains causing subclinical mastitis in dairy cows. A role of environmental signals and communication systems in biofilm development, such as the agr system in S. aureus, is suggested. In the context of multispecies biofilm communities, the presence of non-aureus staphylococci (NAS) might influence S. aureus colonization of the bovine mammary gland, yet, such interspecies interactions have been poorly studied. We determined whether 34 S. chromogenes, 11 S. epidermidis, and 14 S. simulans isolates originating from bovine milk samples and teat apices (TA) were able to affect biofilm formation and dispersion of S. aureus, and if so, how isolate traits such as the capacity to regulate the S. aureus agr quorum sensing system are determinants in this process. The capacity of an agr-positive S. aureus strain to form biofilm was increased more in the presence of S. chromogenes than in the presence of S. simulans and S. epidermidis isolates and in the presence of NAS isolates that do not harbor biofilm related genes. On the other hand, biofilm dispersion of this particular S. aureus strain was suppressed by NAS as a group, an effect that was more pronounced by isolates from TA. Furthermore, the observed effects on biofilm formation and dispersion of the agr-positive S. aureus strain as well as of an agr-negative S. aureus strain did not depend on the capacity of NAS to repress the agr system.

scholarly journals Murine binder of sperm protein homolog 1: a new player in HDL-induced capacitation

Reproduction ◽  
2015 ◽  
Vol 149 (4) ◽  
pp. 367-376 ◽  
Author(s):  
Geneviève Plante ◽  
Puttaswamy Manjunath
Keyword(s):  
Specific Interaction ◽  
Main Function ◽  
Sperm Capacitation ◽  
Epididymal Sperm ◽  
Male Genital Tract ◽  
Direct Role ◽  
Bsp Proteins ◽  
Male Genital

Binder of sperm (BSP) proteins are ubiquitous among mammals and are exclusively expressed in male genital tract. The main function associated with BSP proteins is their ability to promote sperm capacitation. In mice, two proteins (BSP protein homolog 1 (BSPH1) and BSPH2) have been studied. Using recombinant strategies, BSPH1 was found to bind to epididymal sperm membranes and promote sperm capacitation in vitro. The goal of this study was to evaluate the role of native murine BSPH1 protein in sperm capacitation induced by BSA and HDLs. The effect of antibodies, antigen-binding fragments (Fabs), and F(ab′)2 specific for murine BSPH1 on BSA- and HDL-induced capacitation was tested. Results indicate that BSPH1 has no direct role in BSA-induced capacitation. However, antibodies, Fabs, and F(ab′)2 could block capacitation induced by HDLs and could inhibit the HDL-induced increase in tyrosine phosphorylation, suggesting a specific interaction between HDLs and BSPH1. Results indicate that murine BSPH1 proteins in mice could be a new important piece of the puzzle in sperm capacitation induced by HDLs. As murine BSPH1 is orthologous to human BSPH1, this study could also lead to new insights into the functions and the importance of the human protein in male fertility.Free French abstractA French translation of this abstract is freely available at http://www.reproduction-online.org/content/149/4/367/suppl/DC1.

150 Effect of bovine seminal plasma and sperm on endometrial gene expression

2019 ◽  
Vol 31 (1) ◽  
pp. 200
Author(s):  
B. Fernandez-Fuertes ◽  
J. M. Sanchez ◽  
S. Bages ◽  
P. Lonergan
Keyword(s):  
Gene Expression ◽  
Seminal Plasma ◽  
Reproductive Tract ◽  
Epididymal Sperm ◽  
Rna Integrity ◽  
Rna Quality ◽  
Rna Integrity Number ◽  
Maternal Immune System ◽  
Negative Effect

In cattle, most pregnancy losses are sustained before implantation. Many factors are involved in implantation failure, but in mice, pigs and humans there is increased evidence of a role for the maternal immune system and its regulation by seminal plasma (SP). However, there is little evidence for a role of SP in bovine fertility, where dilution or removal of SP before AI is routine. Therefore, the aim of this work was to determine the effect of bull SP or sperm on endometrial gene expression. To this end, 6 heifers were oestrous synchronised and slaughtered 12h after the onset of oestrus. Five endometrial explants from the horn ipsilateral to the preovulatory follicle were obtained from each animal. Explants were incubated with (1) RPMI medium (control); (2) epididymal sperm (106 epididymal sperm mL−1); (3) complete ejaculate (106 ejaculated sperm mL−1+25% SP); (4) ejaculated sperm alone (106 ejaculated sperm mL−1); and (5) SP alone (25% SP). Epididymal sperm were collected and pooled from the cauda epididymis of 3 bulls slaughtered in a commercial abattoir. In addition, complete ejaculates were obtained from 3 other bulls using an artificial vagina. After pooling the samples, a small volume was washed through a density gradient to obtain the ejaculated sperm, and the rest of the ejaculate was filtered to obtain sperm-free SP. The RPMI media was used to dilute sperm and SP to the working concentrations. After 6h of incubation, explants were snap frozen. The RNA quality was assessed with an Agilent 2100 bioanalyzer (Agilent, Santa Clara, CA, USA) before RT-qPCR analysis. Interestingly, SP had a dramatic effect on endometrium RNA integrity, as evidenced by a lower RNA integrity number in explants exposed to a complete ejaculate (2.4±0.14) or SP (2.4±0.06), in comparison with the control, epididymal sperm, or ejaculated sperm treatments (6.7±0.43, 6.9±0.32, 6.7±0.30, respectively; P&lt;0.05). Due to the low RNA quality, those treatments were excluded from further analysis. However, this finding is currently being explored, along with the possibility of this effect being inherent to species that ejaculate intravaginally. We then compared the ability of ejaculated sperm (which have been exposed to SP) and epididymal sperm (which have never had contact with SP) to regulate the endometrial expression of IL1A, IL1B, IL8, IL6, PTGES2, TNFA, and LIF. Although IL6, IL1A and LIF increased in all animals when exposed to either ejaculated or epididymal sperm, there was no effect of treatment. In conclusion, these data did not support the notion that exposure of sperm to SP is important for the immune regulation of the bovine uterus. In addition, the negative effect of SP on the endometrium, together with the fact that bulls are intravaginal ejaculators, suggests that any putative immunoregulatory role of this fluid in the cattle uterus is indirect. Further analysis of the effect of SP on the vagina and cervix will help elucidate whether this response is present in this species and whether it can propagate to more distal regions of the reproductive tract. This work was supported by Science Foundation Ireland (13/IA/1983, 16/IA/4474).

Expression of the secretory form of recombinant regucalcin ofBubalus bubalis in methylotropic yeast

2017 ◽  
Author(s):  
S. K. Bhure ◽  
P. Harikrishna ◽  
J. Usharani ◽  
A. M. Shende ◽  
S. Harikumar
Keyword(s):  
Reproductive Tract ◽  
Amino Acid Level ◽  
Initial Step ◽  
Reproductive Organs ◽  
Male Reproduction ◽  
Male Reproductive Tract ◽  
Multifunctional Protein ◽  
Sds Page ◽  
Ascorbic Acid Biosynthesis

Regucalcin is a multifunctional protein having an important role in calcium homeostasis, L-ascorbic acid biosynthesis, anti-prolific, anti-apoptotic functions as well as detoxification of chemical warfare nerve agents. Recently, it has been localized to male reproductive tract of rat and human, and identified as an androgen-target gene. The literature suggests a possible role of regucalcin in male fertility. However, no detailed studies have been conducted on its role in male reproductive organs of domestic animals. As an initial step, we had cloned and expressed regucalcin in Pichia pastoris. The sequence analysis showed 100% homology with regucalcin of Bos tours both at nucleotide and amino acid level. The SDS-PAGE and Western blot studies of recombinant protein probed with anti-regucalcin monoclonal antibody showed a higher molecular weight (56 kDa) than the expected (35.5 kDa) that could be due to hyperglycosylation. The recombinant regucalcin and its antibodies can be used to study the detailed role of the protein in male reproduction.

Role of microRNAs in cancers of the female reproductive tract: insights from recent clinical and experimental discovery studies

2014 ◽  
Vol 128 (3) ◽  
pp. 153-180 ◽  
Author(s):  
Monica Logan ◽  
Shannon M. Hawkins
Keyword(s):  
Reproductive Tract ◽  
Chemotherapy Resistance ◽  
Clinical Information ◽  
Current Data ◽  
Female Reproductive Tract ◽  
Model Systems ◽  
Future Research ◽  
Cancer Type

microRNAs (miRNAs) are small RNA molecules that represent the top of the pyramid of many tumorigenesis cascade pathways as they have the ability to affect multiple, intricate, and still undiscovered downstream targets. Understanding how miRNA molecules serve as master regulators in these important networks involved in cancer initiation and progression open up significant innovative areas for therapy and diagnosis that have been sadly lacking for deadly female reproductive tract cancers. This review will highlight the recent advances in the field of miRNAs in epithelial ovarian cancer, endometrioid endometrial cancer and squamous-cell cervical carcinoma focusing on studies associated with actual clinical information in humans. Importantly, recent miRNA profiling studies have included well-characterized clinical specimens of female reproductive tract cancers, allowing for studies correlating miRNA expression with clinical outcomes. This review will summarize the current thoughts on the role of miRNA processing in unique miRNA species present in these cancers. In addition, this review will focus on current data regarding miRNA molecules as unique biomarkers associated with clinically significant outcomes such as overall survival and chemotherapy resistance. We will also discuss why specific miRNA molecules are not recapitulated across multiple studies of the same cancer type. Although the mechanistic contributions of miRNA molecules to these clinical phenomena have been confirmed using in vitro and pre-clinical mouse model systems, these studies are truly only the beginning of our understanding of the roles miRNAs play in cancers of the female reproductive tract. This review will also highlight useful areas for future research regarding miRNAs as therapeutic targets in cancers of the female reproductive tract.

scholarly journals Leukotrienes modulate secretion of progesterone and prostaglandins during the estrous cycle and early pregnancy in cattle: an in vivo study

Reproduction ◽  
2010 ◽  
Vol 140 (5) ◽  
pp. 767-776 ◽  
Author(s):  
Anna J Korzekwa ◽  
Mamadou M Bah ◽  
Andrzej Kurzynowski ◽  
Karolina Lukasik ◽  
Agnieszka Groblewska ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Estrous Cycle ◽  
Early Pregnancy ◽  
Luteal Phase ◽  
Reproductive Tract ◽  
Cell Function ◽  
Ovarian Cell

Recently, we showed that leukotrienes (LTs) regulate ovarian cell functionin vitro. The aim of this study was to examine the role of LTs in corpus luteum (CL) function during both the estrous cycle and early pregnancyin vivo. mRNA expression of LT receptors (BLTfor LTB4andCYSLTfor LTC4), and 5-lipoxygenase (5-LO) in CL tissue and their localization in the ovary were studied during the estrous cycle and early pregnancy. Moreover, concentrations of LTs (LTB4and C4) in the CL tissue and blood were measured.5-LOandBLTmRNA expression increased on days 16–18 of the cycle, whereasCYSLTmRNA expression increased on days 16–18 of the pregnancy. The level of LTB4was evaluated during pregnancy compared with the level of LTC4, which increased during CL regression. LT antagonists influenced the duration of the estrous cycle: the LTC4antagonist (azelastine) prolonged the luteal phase, whereas the LTB4antagonist (dapsone) caused earlier luteolysisin vivo. Dapsone decreased progesterone (P4) secretion and azelastine increased P4secretion during the estrous cycle. In summary, LT action in the bovine reproductive tract is dependent on LT type: LTB4is luteotropic during the estrous cycle and supports early pregnancy, whereas LTC4is luteolytic, regarded as undesirable in early pregnancy. LTs are produced/secreted in the CL tissue, influence prostaglandin function, and serve as important factors during the estrous cycle and early pregnancy in cattle.

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