Inhibition of mouse fertilization in vivo by intra-oviductal injection of an anti-equatorin monoclonal antibody

The monoclonal antibody mMN9 recognizes an antigenic molecule, equatorin, which is localized at the equatorial segment of the mammalian sperm acrosome. Our previous results using an IVF system indicated that mMN9 blocked sperm-oocyte fusion. Antibody-containing and control solutions were injected directly into the right and left oviductal ampullae, respectively, of anaesthetized female mice to assess the effect of mMN9 on fertilization in vivo. After hCG treatment, the females were mated, and their oviductal eggs and implanted embryos were examined. mMN9 was retained in the oviductal lumen at 20 h after injection. The rates of fertilization and concomitant pregnancy were significantly lower than in the control side (P < 0.05). In addition, histological studies showed no evidence of pathological changes in the female reproductive tract after the injections. These results indicate that mMN9 inhibits mouse fertilization significantly under in vivo conditions and that this injection method should be useful for studying the effects of antibodies and agents on fertilization in vivo.

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Тhe Safety Assessment of the Ultraviolet Scleral Crosslinking with Riboflavin/UVA in Experiment

Ophthalmology in Russia ◽

10.18008/1816-5095-2019-1s-108-111 ◽

2019 ◽

Vol 16 (1S) ◽

pp. 108-111

Author(s):

M. N. Astrelin ◽

V. K. Surkova

Keyword(s):

Aqueous Solution ◽

Side Effects ◽

Functional State ◽

Safety Assessment ◽

Pathological Changes ◽

Ultraviolet A ◽

Total Exposure Time ◽

The Right ◽

And Control

The purpose is to evaluate the safety of the scleral crosslinking with riboflavin/ultraviolet A (UVA) in an experiment in vivo. Materials and methods. The study was carried out on 34 Chinchilla rabbits (68 eyes). The right eyes were crosslinked (34 eyes), the left eyes were served as controls (34 eyes). Scleral crosslinking (SCXL) was performed with sclera saturation with a photosensitizer (0.1 % aqueous solution of riboflavin) for 20 minutes and its subsequent irradiation with ultraviolet A (wavelength of 370 ± 5 nm, irradiance — 3 mW/cm2), total exposure time — 30 minutes (6 cycles of 5 minutes). The effect of the procedure on the anatomical and functional state of the eye layers was assessed with high-resolution optical coherent tomography (OCT) and electroretinography (ERG) before crosslinking, a day, 7 and 30 days after it. Results. OCT did not reveal any pathological changes after scleral crosslinking with riboflavin/UVA. The layers of the retina, choroid and sclera were clearly visualized. The performed morphometric analysis has showed the absence of statistically significant changes in the eye layers thickness after ultraviolet crosslinking. The amplitude-time characteristics and the shape of the electroretinogram of the experimental and control rabbits eyes were identical during all periods of observation, had a classic appearance. All waves of ERG were well expressed. It indicates a satisfactory functional state of the retinal neuroreceptor mechanisms. Conclusion. For ultraviolet crosslinking we used UVA with an irradiance of 3 mW/cm2 for 30 minutes and a 0.1 % aqueous solution of riboflavin without dextran and did not reveal any side effects of the procedure. Thus, scleral crosslinking with these parameters is safe for the eye layers in an experiment in vivo.

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Mammalian sperm hyperactivation regulates navigation via physical boundaries and promotes pseudo-chemotaxis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2107500118 ◽

2021 ◽

Vol 118 (44) ◽

pp. e2107500118

Author(s):

Meisam Zaferani ◽

Susan S. Suarez ◽

Alireza Abbaspourrad

Keyword(s):

Reproductive Tract ◽

Dynamic Environment ◽

High Amplitude ◽

Critical Value ◽

Female Reproductive Tract ◽

Intrinsic Curvature ◽

Mammalian Sperm ◽

Sperm Migration ◽

Microfluidic Chambers

Mammalian sperm migration within the complex and dynamic environment of the female reproductive tract toward the fertilization site requires navigational mechanisms, through which sperm respond to the tract environment and maintain the appropriate swimming behavior. In the oviduct (fallopian tube), sperm undergo a process called “hyperactivation,” which involves switching from a nearly symmetrical, low-amplitude, and flagellar beating pattern to an asymmetrical, high-amplitude beating pattern that is required for fertilization in vivo. Here, exploring bovine sperm motion in high–aspect ratio microfluidic reservoirs as well as theoretical and computational modeling, we demonstrate that sperm hyperactivation, in response to pharmacological agonists, modulates sperm–sidewall interactions and thus navigation via physical boundaries. Prior to hyperactivation, sperm remained swimming along the sidewalls of the reservoirs; however, once hyperactivation caused the intrinsic curvature of sperm to exceed a critical value, swimming along the sidewalls was reduced. We further studied the effect of noise in the intrinsic curvature near the critical value and found that these nonthermal fluctuations yielded an interesting “Run–Stop” motion on the sidewall. Finally, we observed that hyperactivation produced a “pseudo-chemotaxis” behavior, in that sperm stayed longer within microfluidic chambers containing higher concentrations of hyperactivation agonists.

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THE TRANSPORT OF OVA IN RELATION TO THE DOSAGE OF OESTROGEN IN OVARIECTOMIZED RABBITS

Journal of Endocrinology ◽

10.1677/joe.0.0180108 ◽

1959 ◽

Vol 18 (1) ◽

pp. 108-117 ◽

Cited By ~ 27

Author(s):

R. W. NOYES ◽

C. E. ADAMS ◽

A. WALTON

Keyword(s):

Reproductive Tract ◽

Great Variability ◽

Female Reproductive Tract ◽

The Body ◽

Oestrogen Treatment ◽

Uterine Tube ◽

Ovum Transport ◽

Oestradiol Benzoate ◽

The Right ◽

Egg Transport

SUMMARY In order to determine what part the level of oestrogen in the body might play in the passage of ova through the female reproductive tract, 1249 freshly ovulated ova from donor rabbits were transferred into the uterine tubes of seventy-four ovariectomized recipients, fifty-three of which had previously been treated for 5–18 days with small daily injections of oestradiol benzoate. From 10 to 78 hr after transfer, 41% of the ova were recovered from the uterine tubes, 9% from the uterine horns, and 23% from the vaginae of the recipient animals. Twenty-seven% of the ova were lost. The rate of ovum transport varied widely between similarly treated animals, and between the right and left sides of the reproductive tract of the same animal. Larger proportions of ova were retained in the uterine tubes, and smaller proportions in the uterine horns as the oestrogen dose was increased. The great variability in the stage of cleavage and in the thickness of the mucin coat of ova recovered from the uterus and vagina suggested that the ova might be widely dispersed through the uterine tubes and that they probably pass out of the uterine tube at widely different periods of time. Evidence is presented that ovum transport in ovariectomized rabbits with or without oestrogen treatment is very irregular, that ova may be ejected from either end of the uterine tube at almost any time after transfer, and that ova are not normally retained in the uteri of such animals. Approx. 1 μg oestradiol benzoate administered daily for 5–10 days was necessary to maintain the uterine weight of ovariectomized rabbits at about the same level as that in intact oestrous rabbits, and also to reduce the variability in egg transport observed in control and ovariectomized animals.

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In vivo uptake of [14C]leucine by skeletal muscle ribosomes after injury in rats fed two levels of protein

British Journal Of Nutrition ◽

10.1079/bjn19690034 ◽

1969 ◽

Vol 23 (2) ◽

pp. 271-280 ◽

Cited By ~ 6

Author(s):

V. R. Young ◽

P. C. Huang

Keyword(s):

Skeletal Muscle ◽

Specific Activity ◽

Male Rats ◽

Thigh Muscle ◽

Left Femur ◽

The Right ◽

And Control ◽

The Relationship ◽

In Vivo Uptake

1. After 14 days on a diet containing 5 or 25% casein male rats received a fracture of the left femur. Four hours before they were killed the injured and control rats were injected with [1-14C]leucine; the incorporation of radioactivity into an isolated fraction of skeletal muscle ribosomes was studied 6, 12, 24, 48, 72, 96 and 228 h after injury.2. The incorporation of [14C]leucine into the ribosome fraction in right thigh muscles dropped to 40% of control values 72 h after fracture in well-nourished rats and after 96 h with diets containing 5 or 25%, casein.3. The specific activity of the trichloroacetic acid-soluble fraction of muscle from injured rats was equal to or higher than that of the controls during the first 72 h but lower at 96 h.4. These results suggest that a reduced incorporation of amino acids by ribosomes from the right thigh muscle occurred on day 3 after fracture in the group receiving 25% casein but not in the group receiving 5% casein.5. Muscle RNA and DNA concentrations were not affected by the injury.6. The relationship between these findings and the loss of muscle N after injury is discussed.

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Seminal vesicle proteins SVS3 and SVS4 facilitate SVS2 effect on sperm capacitation

Reproduction ◽

10.1530/rep-15-0551 ◽

2016 ◽

Vol 152 (4) ◽

pp. 313-321 ◽

Cited By ~ 8

Author(s):

Naoya Araki ◽

Natsuko Kawano ◽

Woojin Kang ◽

Kenji Miyado ◽

Kaoru Yoshida ◽

...

Keyword(s):

Seminal Vesicle ◽

Reproductive Tract ◽

Female Reproductive Tract ◽

The Other ◽

Sperm Capacitation ◽

Other Hand ◽

Sperm Membrane ◽

Vesicle Secretion ◽

Mammalian Spermatozoa

Mammalian spermatozoa acquire their fertilizing ability in the female reproductive tract (sperm capacitation). On the other hand, seminal vesicle secretion, which is a major component of seminal plasma, inhibits the initiation of sperm capacitation (capacitation inhibition) and reduces the fertility of the capacitated spermatozoa (decapacitation). There are seven major proteins involved in murine seminal vesicle secretion (SVS1-7), and we have previously shown that SVS2 acts as both a capacitation inhibitor and a decapacitation factor, and is indispensable forin vivofertilization. However, the effects of SVSs other than SVS2 on the sperm have not been elucidated. Since mouseSvs2–Svs6genes evolved by gene duplication belong to the same gene family, it is possible that SVSs other than SVS2 also have some effects on sperm capacitation. In this study, we examined the effects of SVS3 and SVS4 on sperm capacitation. Our results showed that both SVS3 and SVS4 are able to bind to spermatozoa, but SVS3 alone showed no effects on sperm capacitation. On the other hand, SVS4 acted as a capacitation inhibitor, although it did not show decapacitation abilities. Interestingly, SVS3 showed an affinity for SVS2 and it facilitated the effects of SVS2. Interaction of SVS2 and spermatozoa is mediated by the ganglioside GM1 in the sperm membrane; however, both SVS3 and SVS4 had weaker affinities for GM1 than SVS2. Therefore, we suggest that separate processes may cause capacitation inhibition and decapacitation, and SVS3 and SVS4 act on sperm capacitation cooperatively with SVS2.

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RON Mediates Tumor-Promoting Effects in Endometrial Adenocarcinoma

BioMed Research International ◽

10.1155/2021/2282916 ◽

2021 ◽

Vol 2021 ◽

pp. 1-10

Author(s):

Qin Yu ◽

Jianzhang Wang ◽

Tiantian Li ◽

Xinxin Xu ◽

Xinyue Guo ◽

...

Keyword(s):

Nude Mice ◽

Reproductive Tract ◽

Epithelial Mesenchymal Transition ◽

Endometrial Adenocarcinoma ◽

Female Reproductive Tract ◽

Migration And Invasion ◽

Smad Pathway ◽

Mesenchymal Transition ◽

Adenocarcinoma Cells

Endometrial adenocarcinoma is one of the most prevalent female reproductive tract cancers in the world, and the development of effective treatment is still the main goal of its current research. Epithelial-mesenchymal transition (EMT) plays a significant part in the occurrence and development of epithelial carcinoma, including endometrial adenocarcinoma. Recepteur d’origine nantais (RON) induces EMT and promotes proliferation, migration, and invasion in various epithelial-derived cancers, but its role in endometrial adenocarcinoma is still poorly studied. The purpose of this study is to verify the overexpression of RON in endometrial adenocarcinoma and to explore its specific roles. RON expression in tumor lesions was verified by immunohistochemical staining, HEC-1B cells were used to construct stable cell lines with RON overexpression or knockdown to investigate the effects of RON on the function of endometrial adenocarcinoma cells, and xenotransplantation experiment was carried out in nude mice to explore the effect of RON on the growth of endometrial adenocarcinoma in vivo. This study revealed that RON could promote the proliferation, migration, and invasion of HEC-1B cells and induce EMT, and these effects were regulated through the Smad pathway. RON overexpression could promote growth of endometrial adenocarcinoma cells in nude mice, while its inhibitor BMS777607 could restrict this role. RON played an important role in endometrial adenocarcinoma and had a potential to become a new therapeutic target for endometrial adenocarcinoma.

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Proteomics and metabolomics in cow fertility: a systematic review

Reproduction ◽

10.1530/rep-20-0047 ◽

2020 ◽

Vol 160 (5) ◽

pp. 639-658

Author(s):

Nicolas Aranciaga ◽

James D Morton ◽

Debra K Berg ◽

Jessica L Gathercole

Keyword(s):

Oxidative Stress ◽

Systematic Review ◽

Large Scale ◽

Reproductive Tract ◽

Early Embryo ◽

Female Reproductive Tract ◽

Full Potential ◽

Technical Standards ◽

The Impact

Cow subfertility is a multi-factorial problem in many countries which is only starting to be unravelled. Molecular biology can provide a substantial source of insight into its causes and potential solutions, particularly through large scale, untargeted omics approaches. In this systematic review, we set out to compile, assess and integrate the latest proteomic and metabolomic research on cow reproduction, specifically that on the female reproductive tract and early embryo. We herein report a general improvement in technical standards throughout the temporal span examined; however, significant methodological limitations are also identified. We propose easily actionable avenues for ameliorating these shortcomings and enhancing the reach of this field. Text mining and pathway analysis corroborate the relevance of proteins and metabolites related to the triad oxidative stress-inflammation-disease on reproductive function. We envisage a breakthrough in cattle reproductive molecular research within the next few years as in vivo sample techniques are improved, omics analysis equipment becomes more affordable and widespread, and software tools for single- and multi-omics data processing are further developed. Additional investigation of the impact of local oxidative stress and inflammation on fertility, both at the local and systemic levels, is key towards realising the full potential of this field.

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CatSperζ regulates the structural continuity of sperm Ca2+ signaling domains and is required for normal fertility

eLife ◽

10.7554/elife.23082 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 54

Author(s):

Jean-Ju Chung ◽

Kiyoshi Miki ◽

Doory Kim ◽

Sang-Hee Shim ◽

Huanan F Shi ◽

...

Keyword(s):

Reproductive Tract ◽

Female Reproductive Tract ◽

Targeted Disruption ◽

Vitro Fertilization ◽

Male Subfertility ◽

Epididymal Spermatozoa ◽

Mammalian Female ◽

Signaling Domains

We report that the Gm7068 (CatSpere) and Tex40 (CatSperz) genes encode novel subunits of a 9-subunit CatSper ion channel complex. Targeted disruption of CatSperz reduces CatSper current and sperm rheotactic efficiency in mice, resulting in severe male subfertility. Normally distributed in linear quadrilateral nanodomains along the flagellum, the complex lacking CatSperζ is disrupted at ~0.8 μm intervals along the flagellum. This disruption renders the proximal flagellum inflexible and alters the 3D flagellar envelope, thus preventing sperm from reorienting against fluid flow in vitro and efficiently migrating in vivo. Ejaculated CatSperz-null sperm cells retrieved from the mated female uterus partially rescue in vitro fertilization (IVF) that failed with epididymal spermatozoa alone. Human CatSperε is quadrilaterally arranged along the flagella, similar to the CatSper complex in mouse sperm. We speculate that the newly identified CatSperζ subunit is a late evolutionary adaptation to maximize fertilization inside the mammalian female reproductive tract.

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301 THE EFFECT OF EQUINE FOLICULLAR FLUID ON IN VITRO INDUCTION OF THE ACROSOME REACTION IN FROZEN - THAWED STALLION SPERMATOZOA

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab301 ◽

2010 ◽

Vol 22 (1) ◽

pp. 307

Author(s):

D. S. Silva ◽

P. Rodriguez ◽

N. S. Arruda ◽

R. Rodrigues ◽

J. L. Rodrigues

Keyword(s):

Contact Area ◽

Follicular Fluid ◽

Acrosome Reaction ◽

Reproductive Tract ◽

Female Reproductive Tract ◽

Heparin Group ◽

Fluorescent Stain ◽

In Vitro Induction

The capacitation process occurs in vivo upon exposure of the spermatozoa through the female reproductive tract, but can be induced in vitro in the presence of several compounds. This study was conducted to assess the effect of heparin or equine follicular fluid on hyperactivated motility and in vitro induction acrosome reaction swim-up method with frozen-thawed stallion semen. Two hundred microliters of frozen-thawed equine semen was placed in a tube (45°C) to increase contact area and incubated at 37°C for 1 h. After incubation 800 μL of the supernatant was collected by centrifugation (500 × g, 10 min) to collect spermatozoa. The resulting pellet was resuspended in capacitation medium Fert-TALP supplemented with 5.0 μg mL-1 heparin or 100% follicular fluid and incubated for different times (1, 2, 3, 4, and 5 h) at 37°C. After incubation the hyperactivated motility and acrosome-reacted spermatozoa were evaluated. Hoechst stain was used to differentiate live and dead spermatozoa, and chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm; data were analyzed by ANOVA. The effect of equine follicular fluid resulted in improved percentage of spermatozoa with acrosome reaction at all times of incubation (60, 63, 57, 52, and 58%) but immediately after 3 h of incubation, the hyperactivated motility decreased in heparin group and follicular fluid (42 and 30%, respectively).

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Effects of GnRH, PGF2α and oxytocin treatments on conception rate at the time of artificial insemination in lactating dairy cows

Czech Journal of Animal Science ◽

10.17221/1283-cjas ◽

2011 ◽

Vol 56 (No. 6) ◽

pp. 279-283 ◽

Cited By ~ 1

Author(s):

A. Gümen ◽

A. Keskin ◽

G. Yilmazbas-Mecitoglu ◽

E. Karakaya ◽

S. Cevik ◽

...

Keyword(s):

Dairy Cows ◽

Artificial Insemination ◽

Reproductive Tract ◽

Female Reproductive Tract ◽

Conception Rate ◽

Beneficial Effects ◽

Lactating Dairy Cows ◽

Conception Rates ◽

Prostaglandin F ◽

And Control

In several studies, hormones such as gonadotropin-releasing hormone (GnRH), prostaglandins and oxytocin were used to increase pregnancy rate by inducing ovulation and improving the sperm transport in the female reproductive tract in lactating dairy cattle. The objective of this study was to compare the effects of GnRH, prostaglandin F2α (PGF2α) and oxytocin treatments at the time of artificial insemination (AI) after spontaneous oestrus on the conception rate (CR) of lactating dairy cows. Oestrus was detected by visual observations by experienced personnel. All cows (n = 430, 308 Holstein-Frisian and 122 Swedish-Red dairy cows) were inseminated based on the am/pm rule by veterinarians of the farm. After AI, cows were alternately assigned to one of the four treatment groups: (1) GnRH (n = 113); (2) PGF2α (n = 106); (3) oxytocin (n = 106) and (4) non-treated control (n = 105). Pregnancy diagnosis was performed 28–34 and 58–64 days post-insemination by transrectal ultrasonography. Conception rates on days 28–34 and 58–64 were not different among GnRH (46.0%; 52/113 and 44.3%; 50/113), PGF2α (37.7%; 40/106 and 35.9%; 38/106) and control (49.5%; 52/105 and 47.6%; 50/105) groups. However, conception rates were lower (P = 0.02) in oxytocin (31.1%; 33/106 and 30.2%; 32/106) than in GnRH and control groups on days 28–34 and 58–64. Other covariant factors, such as milk production, days in milk (DIM), breed, parity, service number did not affect the conception rate. Thus, there were no beneficial effects of treatments with GnRH and PGF2α at the time of AI, and oxytocin had an adverse effect on CR in lactating dairy cows in this study.

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