Characterization and biological role of cysteine-rich venom protein belonging to CRISPs from turkey seminal plasma

2021 ◽  
Author(s):  
Mariola Słowińska ◽  
Laura Pardyak ◽  
Ewa Liszewska ◽  
Sylwia Judycka ◽  
Joanna Bukowska ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Reproductive Tract ◽  
Protein Localization ◽  
Mass Spectrometry Analysis ◽  
Etiologic Factor ◽  
Secretory Proteins ◽  
Ductus Deferens ◽  
Venom Protein

Abstract Turkey semen contains cysteine-rich secretory proteins (CRISPs) that belong to the dominant seminal plasma proteins. We aimed to isolate and characterize CRISP from turkey seminal plasma and evaluate its possible involvement in yellow semen syndrome (YSS). YSS, which is well characterized, causes reduced fertility and hatchability. The protein was purified using hydrophobic interaction, gel filtration, and reverse phase chromatography. It then was subjected to identification by mass spectrometry, analysis of physicochemical properties and specific antibody production. The biological function of the isolated protein was tested and included its effects on sperm motility and migration and sperm-egg interactions. Sperm motility was measured with the CASA system using Hobson Sperm Tracker. The reproductive tract of turkey toms was analyzed for gene expression; immunohistochemistry was used for protein localization in the male reproductive tract, spermatozoa, and inner perivitelline layer. The isolated protein was identified as cysteine-rich venom protein-like isoform X2 (CRVP X2; XP_010706464.1) and contained feature motifs of CRISP family proteins. Turkey CRVP X2 was present in both spermatozoa and seminal plasma. The extensive secretion of CRVP X2 by the epithelial cells of the epididymis and ductus deferens suggests its involvement in post-testicular sperm maturation. The internally localized CRVP X2 in the proximal part of the sperm tail might be responsible for stimulation of sperm motility. CRVP X2 on the sperm head might be involved in several events prior to fusion and may also participate in gamete fusion itself. Although the mechanisms by which CRPV X2 mediates fertilization are still unknown, the involvement of complementary sites cannot be excluded. The disturbance of CRVP X2 expression can serve as an etiologic factor of YSS in the turkey. This study expands the understanding of the detailed mechanism of fertilization in birds by clarifying the specific role of CRVP X2.

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<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).

scholarly journals Fidelity of Cotranslational Protein Targeting to the Endoplasmic Reticulum

2021 ◽  
Vol 23 (1) ◽  
pp. 281
Author(s):  
Hao-Hsuan Hsieh ◽  
Shu-ou Shan
Keyword(s):  
Molecular Mechanisms ◽  
Protein Targeting ◽  
Protein Localization ◽  
Signal Recognition Particle ◽  
Cellular Membrane ◽  
Secretory Proteins ◽  
Signal Recognition ◽  
Substrate Selection ◽  
Selection Of

Fidelity of protein targeting is essential for the proper biogenesis and functioning of organelles. Unlike replication, transcription and translation processes, in which multiple mechanisms to recognize and reject noncognate substrates are established in energetic and molecular detail, the mechanisms by which cells achieve a high fidelity in protein localization remain incompletely understood. Signal recognition particle (SRP), a conserved pathway to mediate the localization of membrane and secretory proteins to the appropriate cellular membrane, provides a paradigm to understand the molecular basis of protein localization in the cell. In this chapter, we review recent progress in deciphering the molecular mechanisms and substrate selection of the mammalian SRP pathway, with an emphasis on the key role of the cotranslational chaperone NAC in preventing protein mistargeting to the ER and in ensuring the organelle specificity of protein localization.

scholarly journals The role of pH values in porcine reproductive tracts of male and female individuals

2008 ◽  
Vol 24 (3-4) ◽  
pp. 101-108 ◽  
Author(s):  
T. Smiljakovic ◽  
S. Josipovic ◽  
O. Kosovac ◽  
N. Delic ◽  
S. Aleksic ◽  
...  
Keyword(s):  
Reproductive Tract ◽  
Ph Value ◽  
Rete Testis ◽  
Female Reproductive Tract ◽  
Ductus Deferens ◽  
Male And Female ◽  
Ph Values ◽  
Small Peak

For a long time, in practice, and science, has been known that pH values of sperm and vagina are important for successful fertilization. In this investigation this fact was confirmed, and the goal was to investigate the role of pH values through whole reproductive tract of male and female individuals: testis, rete testis, epididymis, ductus deferens, Cowper's gland, vesicula seminalis, prostata, corpus cavernosus, corpus spongiosus, epitel tissue of penis tube, sperm, vagina, uterus, horn of uterus, oviduct, fimbrie ovarica, ovarium, follicular fluid. Measurement was performed in reproductive active males as well as before and after ovulation in females. Porcine reproductive tracts (per 15 female and male individuals) were collected from institute's slaughterhouse, immediately post mortem dissected, homogenised and pH values were measured (according to method Rede&Rahelic (1969)). Ovarium and follicular liquid have the highest pH values (7,4) in females, but a small peak in preovulatory oviduct is also present and corresponded to pH of sperm of reproductive fully active male individuals (pH=app.7,2). After fertilization pH in surrounding of zygot (through depolarisation of its membrane) in oviduct, and zygot which then has external decreased pH value moves to less pH values regions by the same principle, that means to uterus, (pH between 7,2 (horn) and 7,07(cervix)) in postovulatory female reproductive tract, where nidation of blastocyst occurs. This investigation could help to elucidate knowledge about reproductive physiology in vivo, giving importance to role of pH values along reproductive tract of male and female individuals.

Induction of prostatic morphology and secretion in urothelium by seminal vesicle mesenchyme

Development ◽  
1995 ◽  
Vol 121 (7) ◽  
pp. 2199-2207 ◽  
Author(s):  
A.A. Donjacour ◽  
G.R. Cunha
Keyword(s):  
Seminal Vesicle ◽  
Reproductive Tract ◽  
Urogenital Sinus ◽  
Secretory Proteins ◽  
Ductus Deferens ◽  
Male Reproductive Tract ◽  
Mesodermal Origin ◽  
Tissue Recombination ◽  
Endodermal Origin ◽  
Recombination Experiments

Mesenchymal-epithelial interactions are essential for the development of the male reproductive tract. Tissue recombination experiments have been used to define the characteristics of these interactions. When mesenchyme, embryonic connective tissue, is recombined with epithelium from another organ an instructive induction may occur in which the developmental fate of the epithelium is altered. Instructive inductions are most common when the epithelium that is removed from the mesenchyme and the epithelium that is recombined with the mesenchyme are from the same germ layer. All of the mesenchyme of the male reproductive tract is of mesodermal origin. The epithelia of these organs are derived from either the mesodermal Wolffian duct epithelium or the endodermal urogenital sinus epithelium. Urogenital sinus mesenchyme can instructively induce bladder and urethral epithelium to form prostate (Donjacour, A. A. and Cunha, G. R. (1993) Endocrinol. 132, 2342–2350) and seminal vesicle mesenchyme can instructively induce epithelium from the ductus deferens and ureter (Cunha, G. R., Young, P., Higgins, S. J. and Cooke, P. S. (1991) Development 111, 145–158) to form seminal vesicle. To see whether inductive interactions could occur across germ layers in this system, seminal vesicle mesenchyme, normally associated with a mesodermal epithelium, was recombined with epithelium from neonatal or adult bladder or urethra, which are of endodermal origin. The resulting tissue recombinants were analyzed histologically and by immunocytochemistry and western blotting with antibodies to prostatic and seminal vesicle secretory proteins. Full prostatic differentiation was observed in tissue recombinants made with seminal vesicle mesenchyme plus either adult or neonatal bladder or urethral epithelium.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Spermatozoa motility in the Persian sturgeon, Acipenser persicus: effects of pH, dilution rate, ions and osmolality

Reproduction ◽  
2004 ◽  
Vol 128 (6) ◽  
pp. 819-828 ◽  
Author(s):  
Sayyed Mohammad Hadi Alavi ◽  
Jacky Cosson ◽  
Mahmoud Karami ◽  
Bagher Mojazi Amiri ◽  
Mohammd Ali Akhoundzadeh
Keyword(s):  
Dilution Rate ◽  
Sperm Motility ◽  
Seminal Plasma ◽  
Seminal Fluid ◽  
Semen Quality ◽  
Total Duration ◽  
Basic Knowledge ◽  
Acipenser Persicus ◽  
High Osmolality

Sperm motility is a prerequisite factor determining semen quality and fertilizing capacity. The effects of environmental factors including pH, cations and osmolality as well as the role of dilution rate on sperm motility parameters in Acipenser persicus were studied. The best pH and dilution rate for activation of spermatozoa were pH 8.0 and dilution ratio 1:50. Ionic factors can stimulate the initiation of sperm activation. The maximum percentage of motile sperm and total duration of sperm motility were observed in solutions containing 25 mM NaCl, 0.2 mM KCl, 3 mM CaSO4, 10 mM MgSO4 and sucrose with an osmolality of 50 mosmol kg−1. The present study provides us with some basic knowledge about sturgeon spermatozoa biosensitivity to ionic and osmolality effects. A sensitivity of A. persicus sperm was observed after induction of activation of sperm motility in solution containing cations or sucrose with high osmolality. Concentrations more than 50 mM Na+, more than 1 mM K+, more than 3 mM Ca2+ and more than 10 mM Mg2+ had negative effects on sperm motility. Also, osmolality more than 100 mosmol kg−1 had an inhibitory effect. It is clear that ions and osmolality stimulate the motility of spermatozoa by changes in the properties of the plasma membrane including its potential and its ionic conductance. The inhibitory role of high osmolality of the swimming medium (more than 100 mosmol kg−1) and insufficient osmolality of the seminal plasma to inhibit semen motility suggested that osmolality is not the principal factor preventing sperm motility in seminal fluid but that K+ is a major inhibitory factor of sperm motility in seminal plasma.

scholarly journals Role of antimicrobial peptide DEFB126 mutation in pathogenesis of male idiopathic infertility

2021 ◽  
Vol 23 (5) ◽  
pp. 1115-1124
Author(s):  
E. M. Khasanova ◽  
L. V. Gankovskaya ◽  
V. V. Burmakina
Keyword(s):  
Gene Expression ◽  
Male Infertility ◽  
Sperm Motility ◽  
Mutant Allele ◽  
Reproductive Tract ◽  
Control Group ◽  
Idiopathic Infertility ◽  
Fold Reduction ◽  
Distribution Frequency

Male infertility is a multifactorial disease, and elucidation of etiopathogenetic mechanisms of its progression is a topical issue. High percentage of the “idiopathic infertility” diagnosis is largely cased by inability to establish etiology of decrease in reproductive spermatic function. Mutation of в-defensin DEFB126 gene is supposed to affect the fertilizing ability of spermatozoa at different levels: it may decrease their ability to migrate through the cervical mucus and reduce binding capacity to epithelial layer of upper female reproductive tract, and it may also increase susceptibility for infections of reproductive tract, due to impairment of local protective function of defensins. Thus, the aim of the present study was to examine possible role of rs11468374 gene polymorphism of the DEFB126 gene in pathogenesis of male idiopathic infertility. Patients and methods: The group of patient with decreased fertility included 54 male subjects, ages 34 to 42, with a control group of 19 ejaculate donors without acute or chronic disease aged 28 to 36. The indicators of sperm motility in the Moscow population were compared with individual levels of DEFB126 gene expression, as well as with estimated distribution frequency of rs11468374 alleles and genotypes among the subjects.As compared with the control group, the infertile patients exhibited a more than seven-fold reduction of DEFB126 gene expression. Analysis of distribution frequency for alleles and genotypes rs11468374 polymorphic marker of the DEFB126 gene revealed that the mutant allele is detected almost twice as often in males with infertility, as compared with control group. No cases with the DEFB126 del/del genotype were found among the control group, in contrary to 16.1% in the group of patients. The patients with DEFB126 del/del genotype exhibited 5.2-fold reduction of sperm motility. Thus, the data obtained may be used to extend our knowledge on the pathogenetic mechanisms of male idiopathic infertility and to improve techniques for its diagnostics, as well as to provide personalized approach to the treatment of male reproductive disorders. The association between carriage of del mutant allele and decreased level of sperm motility suggests a role of this polymorphism in pathogenesis of male infertility. A general decrease in the level of DEFB126 gene expression in the patients affected by infertility also presumes a contribution of defensin 126 to pathogenesis of the disorder.

scholarly journals Functional insights into the role of seminal plasma proteins on sperm motility of buffalo

2018 ◽  
Vol 195 ◽  
pp. 251-258 ◽  
Author(s):  
Viviane Maria Codognoto ◽  
Paulo Henrique Yamada ◽  
Rúbia Alves Schmith ◽  
Felipe Rydygier de Ruediger ◽  
Caroline Scott ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Seminal Plasma Proteins

The Effect of PDE5 Inhibitors on the Male Reproductive Tract

2020 ◽  
Vol 26 ◽  
Author(s):  
Nikolaos Sofikitis ◽  
Aris Kaltsas ◽  
Fotios Dimitriadis ◽  
Jens Rassweiler ◽  
Nikolaos Grivas ◽  
...  
Keyword(s):  
Male Infertility ◽  
Reproductive Tract ◽  
Tunica Albuginea ◽  
Scientific Data ◽  
Secretory Function ◽  
Pde5 Inhibitors ◽  
Male Reproductive Tract ◽  
Review Study ◽  
Phosphodiesterase 5

The therapeutic range of cyclic nucleotide phosphodiesterase 5 inhibitors (PDE5) inhibitors is getting wider in the last years. This review study focuses on the potential employment of PDE5 inhibitors as an adjunct tool for the therapeutic management of male infertility. The literature tends to suggest a beneficial effect of PDE5 inhibitors on Leydig and Sertoli cells secretory function. It also appears that PDE5 inhibitors play a role in the regulation of the contractility of the testicular tunica albuginea and the epididymis. Moreover scientific data suggest that PDE5 inhibitors enhance the prostatic secretory function leading to an improvement in sperm motility. Other studies additionally demonstrate a role of PDE5 inhibitors in the regulation of sperm capacitation process. Placebo-controlled, randomized, blind studies are necessary to unambiguously incorporate PDE5 inhibitors as an adjunct tool for the pharmaceutical treatment of semen disorders and male infertility.

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