Protein Identification of Spermatozoa and Seminal Plasma in Bottlenose Dolphin (Tursiops truncatus)

Proteins play an important role in many reproductive functions such as sperm maturation, sperm transit in the female genital tract or sperm-oocyte interaction. However, in general, little information concerning reproductive features is available in the case of aquatic animals. The present study aims to characterize the proteome of both spermatozoa and seminal plasma of bottlenose dolphins (Tursiops truncatus) as a model organism for cetaceans. Ejaculate samples were obtained from two trained dolphins housed in an aquarium. Spermatozoa and seminal plasma were analyzed by means of proteomic analyses using an LC-MS/MS, and a list with the gene symbols corresponding to each protein was submitted to the DAVID database. Of the 419 proteins identified in spermatozoa and 303 in seminal plasma, 111 proteins were shared by both. Furthermore, 70 proteins were identified as involved in reproductive processes, 39 in spermatozoa, and 31 in seminal plasma. The five most abundant proteins were also identified in these samples: AKAP3, ODF2, TUBB, GSTM3, ROPN1 for spermatozoa and CST11, LTF, ALB, HSP90B1, PIGR for seminal plasma. In conclusion, this study provides the first characterization of the proteome in cetacean sperm and seminal plasma, opening the way to future research into new biomarkers, the analysis of conservation capacity or possible additional applications in the field of assisted reproductive technologies.

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190 Effect of TCM-199 and synthetic oviductal fluid medium supplemented with varying hormone concentrations on invitro maturation of canine oocytes

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab190 ◽

2020 ◽

Vol 32 (2) ◽

pp. 223

Author(s):

M. Tscharke ◽

K. Kind ◽

J. Kelly ◽

J. Len

Keyword(s):

Oocyte Maturation ◽

Assisted Reproductive Technologies ◽

Germinal Vesicle ◽

Pairwise Comparison ◽

Reproductive Technologies ◽

Future Research ◽

Maturation Stage ◽

Fluid Medium ◽

Nuclear Maturation ◽

Oviductal Fluid

Invitro oocyte maturation (IVM) in the domestic canine is yet to be optimized, with low rates of cumulus-oocyte complexes (COCs) reaching MII. This limits the progression of assisted reproductive technologies, which could benefit breeding programs for assistance dogs and endangered Canidae. Canine oocyte maturation differs from that in other mammals, with the ovulation of a COC in the germinal vesicle stage and nuclear maturation occurring in the oviduct. Because of this, the environment in which a canine COC matures is unlike that of other mammals, meaning that IVM protocols cannot be readily adapted. The aim of the current work was to determine (1) the effects of varying concentrations of FSH, human chorionic gonadotrophin (hCG), and oestradiol (E2) during IVM on meiotic resumption and nuclear maturation of canine COCs; and (2) the optimal medium base, either synthetic oviductal fluid (SOF) or tissue culture medium-199 (TCM). Reproductive tracts of bitches (6 months to 7 years of age) were collected from veterinary clinics within 2h of routine spaying. Ovaries were sliced using a scalpel blade, releasing the COCs into aspiration medium. The COCs were randomly allocated to a maturation medium consisting of one of the hormones at two concentrations (FSH: 5 or 10µgmL−1; hCG: 5 or 10IUmL−1; E2: 1 or 5µgmL−1) and for both SOF and TCM base. Each hormone was tested individually for a replicate of eight animals per hormone (total of 12 experimental groups; 24 animals). The COCs were cultured for 72h in their allocated medium and then denuded and stained with Hoechst 33258. Fluorescence microscopy was used to determine nuclear maturation stage. Nuclear maturation rates to MII were analysed using a general linear model with pairwise comparison (SPSS version 25; SPSS Inc./IBM Corp.) with each individual animal acting as a replicate. Canine COCs matured in a SOF-based media had higher rates of meiotic resumption (MI and MII) (SOF: 38.68%, n=515; TCM: 25.78%, n=542; P<0.05) and number reaching MII (SOF: 7.54%; TCM: 4.39%; P<0.05) compared with TCM-based medium. Resumption of meiosis and nuclear maturation to MII did not differ between media with differing E2 or hCG concentrations. The use of FSH at 10µgmL−1 in SOF medium decreased resumption of meiosis (8.57%) and MII rates (0%) compared with 5µgmL−1 FSH in SOF (29.41% and 3.92%, respectively; P<0.05). In summary, our data indicated that higher concentrations of FSH during IVM have a negative effect on meiotic resumption and maturation to MII, whereas canine COCs resume meiosis and mature to MII in higher rates in a SOF-based medium compared with a TCM base. An IVM medium that replicates the invivo environment in which canine COCs mature is beneficial. However, rates of IVM canine oocytes reaching maturity are low, signifying that future research must investigate a greater range of hormone concentrations and combinations to better mimic invivo conditions to assess the possible benefits for canine IVM.

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From One Ejaculate to Another: Transference of Sperm Traits via Seminal Plasma Supplementation in the Ram

Biology ◽

10.3390/biology9020033 ◽

2020 ◽

Vol 9 (2) ◽

pp. 33

Author(s):

Christine Green ◽

Jessica P. Rickard ◽

Simon P. de Graaf ◽

Angela J. Crean

Keyword(s):

Sperm Motility ◽

Seminal Plasma ◽

Perceived Risk ◽

Positive Relationship ◽

Assisted Reproductive Technologies ◽

Sperm Quality ◽

Reproductive Technologies ◽

Before And After ◽

The Difference ◽

Sperm Traits

Males can adjust sperm motility instantaneously in response to the perceived risk of sperm competition. The speed of this response suggests that sperm motility is regulated by changes in seminal plasma rather than changes in the sperm cells themselves. Hence, here we test whether inter-ejaculate variation in seminal plasma can be used to alter sperm quality prior to use in assisted reproductive technologies. We supplemented fresh ejaculates of Merino rams with seminal plasma collected from previous ‘donor’ ejaculates to test whether changes in sperm kinetics were related to the relative quality of donor to focal ejaculates. We found a positive relationship between the change in sperm traits before and after supplementation, and the difference in sperm traits between the donor and focal ejaculate. Hence, sperm motility can be either increased or decreased through the addition of seminal plasma from a superior or inferior ejaculate, respectively. This positive relationship held true even when seminal plasma was added from a previous ejaculate of the same ram, although the slope of the relationship depended on the identity of both the donor and receiver ram. These findings indicate that seminal plasma plays a key role in the control and regulation of sperm kinetics, and that sperm kinetic traits can be transferred from one ejaculate to another through seminal plasma supplementation.

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Features of seminal plasma cytokine profile and an occurrence of pregnancy when carrying out a program of assisted reproductive technologies

GYNECOLOGY ◽

10.26442/20795696.2019.1.190233 ◽

2019 ◽

Vol 21 (4) ◽

pp. 9-13

Author(s):

Tatiana V Dontsova ◽

Alina A Babayan ◽

Marina A Nikolaeva ◽

Alla S Arefieva ◽

Elena A Kalinina ◽

...

Keyword(s):

Sexual Intercourse ◽

Seminal Plasma ◽

Transforming Growth Factor ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Female Reproductive Tract ◽

Cytokine Profile ◽

Sexual Life ◽

Operating Characteristics ◽

Diagnostic Significance

Aim. To asses an effectiveness of assisted reproductive technologies (ART) program given characteristics of the cytokine profile of seminal plasma (SP) entering the female reproductive tract during sexual intercourse. Outcomes and methods. 33 married couples who applied for a treatment of infertility by means of in vitro fertilization /ICSI (Intracytoplasmic Sperm Injection) were included in the prospective study. Patients were recommended to have sexual intercourse with no restrictions during treatment and to have the last sexual intercourse 3 days before an intended transvaginal puncture (ITP). Testing of cytokines (transforming growth factor (TGF)-b1, interferon (IFN)-g, interleukin (IL)-33, IL-6, IL-8, IL-23, IL-10, tumor necrosis factor (TNF)-a, IL-18, IL- 17A, IFN-a, IL-12, monocyte chemotactic protein-1) levels in samples of partners’ SP obtained on the day having ITP were carried out using multiplex analysis with LegendPlex kits (BioLegend, USA). Results. When comparing a cytokine profile of SP in couples who did not become pregnant (n=25) and couples who become pregnant (n=8) increased IL-18 and reduced IL-10 levels (p=0,017 and p=0, 01 respectively) were revealed in the group which got pregnant. To assess a clinical relevance of cytokine content in SP ROC (Receiver Operating Characteristics) curve was used. It was established that determining of IL-18 concentration in SP has the greatest diagnostic significance (the area under a curve was 0.792±0.107, test sensitivity - 62.5%, test specificity - 95.24% at threshold concentration>210.43 pg/ml). Incidence of pregnancy at IL-18 concentration above threshold levels was 83,3% while at lower concentrations of IL-18 pregnancy occurred only in 13.0% of women. Conclusions. Elevated IL-18 levels and decreased IL-10 levels in SP of female patients’ partners who don’t have restriction of sexual life when treating infertility with ART are favorable factors for a pregnancy to occur.

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Fertile Connections: Thinking across Assisted Reproductive Technologies and Parenting Culture Studies

Sociology ◽

10.1177/0038038517696219 ◽

2017 ◽

Vol 52 (5) ◽

pp. 983-1000 ◽

Cited By ~ 23

Author(s):

Charlotte Faircloth ◽

Zeynep B Gürtin

Keyword(s):

Social Science ◽

Assisted Reproductive Technologies ◽

Sociological Theory ◽

Holistic Approach ◽

Reproductive Technologies ◽

Future Research ◽

Culture Studies ◽

Contemporary Context ◽

Processual Approach

While studies of ‘parenting culture’ and ‘assisted reproductive technologies’ are now well-established areas of social science scholarship, so far, the potential connections between the two fields have not been significantly explored. Responding to calls for a more ‘processual’ approach to studying reproduction in order to make clearer contributions to sociological theory more broadly, we begin a dialogue between these mutually relevant bodies of literature, highlighting connections and crosscutting findings. We focus on four interlinked themes – Reflexivity, Gender, Expertise and Stratification – and promote a more holistic approach to understanding how children are conceived and cared for within the current ‘Euro-American’ reproductive landscape. By way of conclusion, we draw attention to the contemporary context of ‘anxious reproduction’ and propose directions for future research.

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An International Terminology for Endometriosis, 2021

Facts, Views and Vision in ObGyn ◽

10.52054/fvvo.13.4.036 ◽

2021 ◽

Vol 13 (4) ◽

Author(s):

C Tomassetti ◽

N.P. Johnson ◽

J Petrozza ◽

M.S. Abrao ◽

J.I. Einarsson ◽

...

Keyword(s):

Working Group ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

International Committee ◽

Classification Systems ◽

Current Paper ◽

Future Research ◽

Research Use ◽

System A ◽

Descriptive System

Background: Different classification systems have been developed for endometriosis, using different definitions for the disease, the different subtypes, symptoms and treatments. In addition, an International Glossary on Infertility and Fertility Care was published in 2017 by the International Committee for Monitoring Assisted Reproductive Technologies (ICMART) in collaboration with other organisations. An international working group convened over the development of a classification or descriptive system for endometriosis. As a basis for such system, a terminology for endometriosis was considered a condition sine qua non. Objectives: The aim of the current paper is to develop a set of terms and definitions on endometriosis that would be the basis for standardisation in disease description, classification and research. Materials and Methods: The working group listed a number of terms relevant to be included in the terminology, documented currently used and published definitions, and discussed and adapted them until consensus was reached within the working group. Following stakeholder review, further terms were added, and definitions further clarified. Although definitions were collected through published literature, the final set of terms and definitions is to be considered consensus-based. After finalisation of the first draft, the members of the international societies and other stakeholders were consulted for feedback and comments, which led to further adaptations. Results: A list of 49 terms and definitions in the field of endometriosis is presented, including a definition for endometriosis and its subtypes, different locations, interventions, symptoms and outcomes. Endometriosis is defined as a disease characterised by the presence of endometrium-like epithelium and/or stroma outside the endometrium and myometrium, usually with an associated inflammatory process. Conclusions: The current paper outlines a list of 49 terms and definitions in the field of endometriosis. The application of the defined terms aims to facilitate harmonisation in endometriosis research and clinical practice. Future research may require further refinement of the presented definitions. What is new? A consensus based international terminology for endometriosis for clinical and research use.

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Distribution and abundance estimates of bottlenose dolphins (Tursiops truncatus) around Lampedusa Island (Sicily Channel, Italy): implications for their management

Journal of the Marine Biological Association of the United Kingdom ◽

10.1017/s0025315413000842 ◽

2013 ◽

Vol 94 (6) ◽

pp. 1175-1184 ◽

Cited By ~ 7

Author(s):

Marina Pulcini ◽

Daniela Silvia Pace ◽

Gabriella La Manna ◽

Francesca Triossi ◽

Caterina Maria Fortuna

Keyword(s):

Tursiops Truncatus ◽

Marine Protected Area ◽

Bottlenose Dolphins ◽

Future Research ◽

Photo Identification ◽

Abundance Estimates ◽

Core Study ◽

Future Data ◽

Special Area ◽

Baseline Information

This paper represents the first quantitative assessment of the distribution and abundance of bottlenose dolphins (Tursiops truncatus) inhabiting the waters around Lampedusa Island, Italy. Eleven years of photo-identification data, collected from 1996 to 2006 by three different research groups, were brought together, reviewed and analysed to fulfil the following objectives: (i) to obtain baseline information on the abundance and residency of the local bottlenose dolphin putative population; (ii) to review the current Marine Protected Area (MPA) boundaries, especially those referred to waters around Lampedusa Island, with a view to establish a new Special Area of Conservation (SAC); and (iii) to explore the potential and limits of analysing heterogeneous datasets to improve future data collection methods. The most resident dolphins were regularly observed in six specific areas around Lampedusa Island. From a total of 148 photo-identified bottlenose dolphins, 102 were classified as well-marked. The capture histories and the distribution of sightings clearly show a number of dolphins regularly use the study area. Best estimates for the first period within the ‘core study area’ were obtained for 1998 data. The 2005 estimate was significantly larger than the 1998 estimates (z = 2.160;P< 0.05) compared to that of 1998. Implications of our results for the current MPA, for transboundary conservation initiative involving Italy, Malta and Tunisia and for directing future research within and outside the MPA are fully discussed.

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The Development of Echolocation in Bottlenose Dolphins (Tursiops truncatus)

International Journal of Comparative Psychology ◽

10.46867/ijcp.2016.29.00.17 ◽

2016 ◽

Vol 29 ◽

Author(s):

Jennifer H. Harder ◽

Heather M. Hill ◽

Kathleen M. Dudzinski ◽

Karissa T. Sanabria ◽

Sara Guarino ◽

...

Keyword(s):

Tursiops Truncatus ◽

Bottlenose Dolphins ◽

Future Research ◽

Anatomy And Physiology ◽

Behavioral Observations ◽

Sonar System ◽

Social Grouping ◽

The Social ◽

Click Train

This study aimed to expand on previous efforts to evaluate the ontogeny of echolocation in Atlantic bottlenose dolphins (Tursiops truncatus). Data consisted of echolocation recordings and concurrent behavioral observations taken from one calf in 2000 and from five additional dolphin calves and their mothers in 2002 housed at the U.S. Naval facility in San Diego, CA. A total of 361 echolocation click train samples from calves were recorded weekly over the first 6 months of the calves’ lives. The earliest calf echolocation train was recorded at 22 days postpartum and the number of echolocation attempts from calves increased steadily with age. Calf echolocation trains increased in duration and the number of clicks per train with age while train density (clicks/sec) and interclick interval values remained more consistent. Calves swimming independent of their mothers produced more click trains, especially when multiple calves were present in the social grouping. When considering these results in the context of possible maturation of a calf’s melon physiology, it seems very likely that the first two months of life are critical for the development of echolocation and related behaviors. While the first click train recorded in this sample was approximately 3 weeks of age by two different calves, it is possible that dolphin calves may innately produce functional sonar clicks immediately after birth, which were not captured in the current study. Future research will need to investigate this possibility using more controlled conditions and a better understanding of the anatomy and physiology of the sonar system of neonates as well as the possible role of the mother in echolocation development.

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Evolutionary consequences of environmental effects on gamete performance

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2020.0122 ◽

2021 ◽

Vol 376 (1826) ◽

Author(s):

Angela J. Crean ◽

Simone Immler

Keyword(s):

Reproductive Tract ◽

Assisted Reproductive Technologies ◽

Later Life ◽

Reproductive Technologies ◽

Female Reproductive Tract ◽

Future Research ◽

Male And Female ◽

Intrinsic Factors ◽

Gamete Interactions ◽

Evolutionary Consequences

Variation in pre- and post-release gamete environments can influence evolutionary processes by altering fertilization outcomes and offspring traits. It is now widely accepted that offspring inherit epigenetic information from both their mothers and fathers. Genetic and epigenetic alterations to eggs and sperm-acquired post-release may also persist post-fertilization with consequences for offspring developmental success and later-life fitness. In externally fertilizing species, gametes are directly exposed to anthropogenically induced environmental impacts including pollution, ocean acidification and climate change. When fertilization occurs within the female reproductive tract, although gametes are at least partially protected from external environmental variation, the selective environment is likely to vary among females. In both scenarios, gamete traits and selection on gametes can be influenced by environmental conditions such as temperature and pollution as well as intrinsic factors such as male and female reproductive fluids, which may be altered by changes in male and female health and physiology. Here, we highlight some of the pathways through which changes in gamete environments can affect fertilization dynamics, gamete interactions and ultimately offspring fitness. We hope that by drawing attention to this important yet often overlooked source of variation, we will inspire future research into the evolutionary implications of anthropogenic interference of gamete environments including the use of assisted reproductive technologies. This article is part of the theme issue ‘How does epigenetics influence the course of evolution?’

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Glycosaminoglycans in the accessory sex glands, testes and seminal plasma of alpaca and ram

Reproduction Fertility and Development ◽

10.1071/rd11152 ◽

2012 ◽

Vol 24 (2) ◽

pp. 362 ◽

Cited By ~ 10

Author(s):

Claire M. Kershaw-Young ◽

G. Evans ◽

W. M. C. Maxwell

Keyword(s):

Seminal Plasma ◽

Assisted Reproductive Technologies ◽

Prostate Gland ◽

Keratan Sulfate ◽

Reproductive Technologies ◽

Plasma Viscosity ◽

Accessory Sex Glands ◽

The Relationship

The viscous nature of alpaca semen limits its use in cryopreservation and other assisted reproductive technologies. The cause and source of this viscosity is unknown although it has been postulated, but never proven, that glycosaminoglycans (GAGs) secreted by the bulbourethral gland are responsible. The present study investigated the concentration and composition of GAGs in alpaca seminal plasma, testes, bulbourethral gland and prostate gland and compared them to those in the ram to determine the relationship between seminal plasma GAGs and viscosity and to identify the source of seminal plasma GAGs. Alpaca seminal plasma contained more GAGs than ram (P < 0.001) and the predominant GAG, keratan sulfate, was correlated with viscosity (P = 0.05, R2 = 0.2635). The alpaca bulbourethral gland contained most GAGs compared with prostate or testis (P < 0.001). In the ram, the prostate contained most GAGs. These findings suggest that GAGs, particularly keratan sulfate, may be the cause of seminal plasma viscosity in alpacas, and that the seminal plasma GAGs originate from the bulbourethral gland.

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Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates

Reproduction Fertility and Development ◽

10.1071/rd15261 ◽

2017 ◽

Vol 29 (6) ◽

pp. 1107 ◽

Cited By ~ 12

Author(s):

C. M. Kershaw ◽

G. Evans ◽

R. Rodney ◽

W. M. C. Maxwell

Keyword(s):

Sperm Motility ◽

Exposure Time ◽

Seminal Plasma ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Plasma Viscosity ◽

Dna Integrity ◽

Sperm Function ◽

Viscous Component ◽

Acrosome Integrity

In camelids, the development of assisted reproductive technologies is impaired by the viscous nature of the semen. The protease papain has shown promise in reducing viscosity, although its effect on sperm integrity is unknown. The present study determined the optimal papain concentration and exposure time to reduce seminal plasma viscosity and investigated the effect of papain and its inhibitor E-64 on sperm function and cryopreservation in alpacas. Papain (0.1 mg mL–1, 20 min, 37°C) eliminated alpaca semen viscosity while maintaining sperm motility, viability, acrosome integrity and DNA integrity. Furthermore E-64 (10 µM at 37°C for 5 min after 20 min papain) inhibited the papain without impairing sperm function. Cryopreserved, papain-treated alpaca spermatozoa exhibited higher total motility rates after chilling and 0 and 1 h after thawing compared with control (untreated) samples. Papain treatment, followed by inhibition of papain with E-64, is effective in reducing alpaca seminal plasma viscosity without impairing sperm integrity and improves post-thaw motility rates of cryopreserved alpaca spermatozoa. The use of the combination of papain and E-64 to eliminate the viscous component of camelid semen may aid the development of assisted reproductive technologies in camelids.

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