Reproduction, Fertility, And Development[JOURNAL]
Sun
200 papers
Understanding placentation in ruminants: a review focusing on cows and sheep.
Johnson GA, Bazer FW, Seo H … +4 more , Burghardt RC, Wu G, Pohler KG, Cain JW
Reprod Fertil Dev · 2023 Dec · PMID 38064193 · Publisher ↗
Mammals differ regarding their placentae, but in all species placental trophoblasts interact intimately with the uterine endometrium to mediate the transfer of nutrients from the mother to the embryo/fetus through the cl...
Mammals differ regarding their placentae, but in all species placental trophoblasts interact intimately with the uterine endometrium to mediate the transfer of nutrients from the mother to the embryo/fetus through the closely juxtaposed microcirculatory systems of the uterus and placenta. Placentation in ruminants is intermediate between the non-invasive type, as observed in the epitheliochorial placenta of pigs, and the invasive type, as observed in the haemochorial placentae of mice and humans. In ruminants, placental trophoblast cells invade uterine endometrial tissue, but invasion is believed to be limited to the endometrial luminal epithelium (LE). In the LE there are varying degrees of syncytialisation among species, with syncytialisation being more extensive in sheep than cows. The hallmarks of placentation in ruminants include: (1) an extended period in which conceptuses (embryos and associated placental membranes) elongate and must be supported by secretions (histotroph) from the uterus; (2) a cascade involving an array of adhesion molecules that includes integrin-mediated attachment of the conceptus trophoblast to the endometrial LE for implantation; (3) syncytialisation of the developing early placenta, a process for which there is currently limited understanding; and (4) development of placentomes that define the cotyledonary placentae of cows and sheep, and provide haemotrophic support of fetal development.
Production of light-coloured, low heat-absorbing Holstein Friesian cattle by precise embryo-mediated genome editing.
Wei J, Brophy B, Cole SA … +5 more , Leath S, Oback B, Boch J, Wells DN, Laible G
Reprod Fertil Dev · 2023 Dec · PMID 38064192 · Publisher ↗
CONTEXT: Genome editing enables the introduction of beneficial sequence variants into the genomes of animals with high genetic merit in a single generation. This can be achieved by introducing variants into primary cells...
CONTEXT: Genome editing enables the introduction of beneficial sequence variants into the genomes of animals with high genetic merit in a single generation. This can be achieved by introducing variants into primary cells followed by producing a live animal from these cells by somatic cell nuclear transfer cloning. The latter step is associated with low efficiencies and developmental problems due to incorrect reprogramming of the donor cells, causing animal welfare concerns. Direct editing of fertilised one-cell embryos could circumvent this issue and might better integrate with genetic improvement strategies implemented by the industry. METHODS: In vitro fertilised zygotes were injected with TALEN editors and repair template to introduce a known coat colour dilution mutation in the PMEL gene. Embryo biopsies of injected embryos were screened by polymerase chain reaction and sequencing for intended biallelic edits before transferring verified embryos into recipients for development to term. Calves were genotyped and their coats scanned with visible and hyperspectral cameras to assess thermal energy absorption. KEY RESULTS: Multiple non-mosaic calves with precision edited genotypes were produced, including calves from high genetic merit parents. Compared to controls, the edited calves showed a strong coat colour dilution which was associated with lower thermal energy absorbance. CONCLUSIONS: Although biopsy screening was not absolutely accurate, non-mosaic, precisely edited calves can be readily produced by embryo-mediated editing. The lighter coat colouring caused by the PMEL mutation can lower radiative heat gain which might help to reduce heat stress. IMPLICATIONS: The study validates putative causative sequence variants to rapidly adapt grazing cattle to changing environmental conditions.
How I overcame problems in fertilisation of livestock animals.
Nagai T
Reprod Fertil Dev · 2023 Dec · PMID 38064191 · Publisher ↗
In my research life of 35years, growing with IETS as a researcher of in vitro maturation and fertilisation (IVM/IVF) of porcine and cattle oocytes, I suffered from hard times related to solving problems that prevented th...
In my research life of 35years, growing with IETS as a researcher of in vitro maturation and fertilisation (IVM/IVF) of porcine and cattle oocytes, I suffered from hard times related to solving problems that prevented the progress of my research and conferment of my degrees. Many researchers may have similar problems. Thus, I would like to address a few examples of how I overcame these problems related to IVF and help young researchers with similar troubles. There were four main problems to be solved in my experiments. Problem 1: Establishment of IVF using only defined medium in pigs. Problem 2: Establishment of successful in vitro culture (IVC) of IVM/IVF bovine oocytes in defined medium. Problem 3: Low rate of male pronucleus formation in IVM porcine oocytes after IVF. Problem 4: Sedimentation of Ca2+ in the sperm capacitation solution for IVF in pigs. Problem 1 was solved by a lucky accident, in which a sperm suspension that would have otherwise been discarded happened to be successfully used for IVF in pigs. Problems 2, 3 and 4 were solved by communication with scientists whose fields were different from mine, where similar problems had been solved already. Young researchers are encouraged to transcend the boundaries of their research fields and solve problems by interacting with researchers in different fields. There are many good connections or answers around us that may be effective in resolving the problems that are hindering the progress of pending research.
Paternal determinants of early embryo development.
Lockhart KN, Fallon LC, Ortega MS
Reprod Fertil Dev · 2023 Dec · PMID 38064190 · Publisher ↗
Existing research has primarily focused on investigating the impacts of the maternal environment, female fertility phenotype, and genetics on pregnancy loss in dairy cattle. Recently, attention has been directed toward u...
Existing research has primarily focused on investigating the impacts of the maternal environment, female fertility phenotype, and genetics on pregnancy loss in dairy cattle. Recently, attention has been directed toward understanding the role the sire has on embryo quality and viability. Studies have shown there is a paternal influence on early pregnancy loss, but the specific mechanisms impacting pregnancy establishment and maintenance remain unclear. Despite clear differences that sires have on pregnancy outcomes, there is a lack of evidence regarding specifically how sires influence pregnancy. Sperm characteristics, such as motility, concentration, and morphology, have been extensively studied, but further research is needed to understand what makes one sire more or less fertile than another sire and how this affects pregnancy. To effectively address pregnancy loss, a deeper understanding of the processes involved from fertilisation to blastocyst formation is essential, particularly for understanding early pregnancy loss.
The oocyte: the key player in the success of assisted reproduction technologies.
Fair T, Lonergan P
Reprod Fertil Dev · 2023 Dec · PMID 38064189 · Publisher ↗
The ovulation of a mature oocyte at metaphase II of meiosis, with optimal potential to undergo fertilisation by a sperm cell, complete meiosis and sustain the switch to mitotic division, and support early embryo developm...
The ovulation of a mature oocyte at metaphase II of meiosis, with optimal potential to undergo fertilisation by a sperm cell, complete meiosis and sustain the switch to mitotic division, and support early embryo development, involves a protracted and disrupted/delayed series of processes. Many of these are targeted for exploitation in vivo , or recapitulation in vitro , by the livestock industry. Reproductive technologies, including AI, multiple ovulation embryo transfer, ovum pick-up, in vitro embryo production, and oestrus and ovulation synchronisation, offer practitioners and producers the opportunity to produce offspring from genetically valuable dams in much greater numbers than they would normally have in their lifetime, while in vitro oocyte and follicle culture are important platforms for researchers to interrogate the physiological mechanisms driving fertility. The majority of these technologies target the ovarian follicle and the oocyte within; thus, the quality and capability of the recovered oocyte determine the success of the reproductive intervention. Molecular and microscopical technologies have grown exponentially, providing powerful platforms to interrogate the molecular mechanisms which are integral to or affected by ART. The development of the bovine oocyte from its differentiation in the ovary to ovulation is described in the light of its relevance to key aspects of individual interventions, while highlighting the historical timeline.
From fertilised oocyte to cultivated meat - harnessing bovine embryonic stem cells in the cultivated meat industry.
Zehorai E, Maor-Shoshani A, Molotski N … +4 more , Dorojkin A, Marelly N, Dvash T, Lavon N
Reprod Fertil Dev · 2023 Dec · PMID 38064188 · Publisher ↗
Global demand for animal protein is on the rise, but many practices common in conventional production are no longer scalable due to environmental impact, public health concerns, and fragility of food systems. For these r...
Global demand for animal protein is on the rise, but many practices common in conventional production are no longer scalable due to environmental impact, public health concerns, and fragility of food systems. For these reasons and more, a pressing need has arisen for sustainable, nutritious, and animal welfare-conscious sources of protein, spurring research dedicated to the production of cultivated meat. Meat mainly consists of muscle, fat, and connective tissue, all of which can be sourced and differentiated from pluripotent stem cells to resemble their nutritional values in muscle tissue. In this paper, we outline the approach that we took to derive bovine embryonic stem cell lines (bESCs) and to characterise them using FACS (fluorescence-activated cell sorting), real-time PCR and immunofluorescence staining. We show their cell growth profile and genetic stability and demonstrate their induced differentiation to mesoderm committed cells. In addition, we discuss our strategy for preparation of master and working cell banks, by which we can expand and grow cells in suspension in quantities suitable for mass production. Consequently, we demonstrate the potential benefits of harnessing bESCs in the production of cultivated meat.
Mammalian embryo culture media: now and into the future.
Zander-Fox DL, Pacella-Ince L, Morgan DK … +1 more , Green MP
Reprod Fertil Dev · 2023 Dec · PMID 38064187 · Publisher ↗
For over 70years, since the culture of the first mammalian embryo in vitro , scientists have undertaken studies to devise and optimise media to support the manipulation and culture of gametes and embryos. This area of re...
For over 70years, since the culture of the first mammalian embryo in vitro , scientists have undertaken studies to devise and optimise media to support the manipulation and culture of gametes and embryos. This area of research became especially active in the late 1970s onwards following the successful birth of the first human in vitro fertilised embryo. This review summarises some of the key advances in mammalian embryo culture media over time based on a greater understanding of the biochemical milieu of the reproductive tract. It highlights how learnings from studies in mice and agricultural species have informed human culture media compositions, in particular the inclusion of albumin, growth factors, cytokines, and antioxidants into contemporary culture media formulations, and how these advances may then in turn help to inform and guide development of in vitro culture systems used in other arenas, in particular agriculture. Additionally, it will highlight how the introduction of new technologies, such as timelapse, can influence current trends in media composition and usage that may see a return to a single step medium.
Understanding conceptus-maternal interactions: what tools do we need to develop?
Butt Z, Tinning H, O'Connell MJ … +3 more , Fenn J, Alberio R, Forde N
Reprod Fertil Dev · 2023 Dec · PMID 38064186 · Publisher ↗
Communication between the maternal endometrium and developing embryo/conceptus is critical to support successful pregnancy to term. Studying the peri-implantation period of pregnancy is critical as this is when most preg...
Communication between the maternal endometrium and developing embryo/conceptus is critical to support successful pregnancy to term. Studying the peri-implantation period of pregnancy is critical as this is when most pregnancy loss occurs in cattle. Our current understanding of these interactions is limited, due to the lack of appropriate in vitro models to assess these interactions. The endometrium is a complex and heterogeneous tissue that is regulated in a transcriptional and translational manner throughout the oestrous cycle. While there are in vitro models to study endometrial function, they are static and 2D in nature or explant models and are limited in how well they recapitulate the in vivo endometrium. Recent developments in organoid systems, microfluidic approaches, extracellular matrix biology, and in silico approaches provide a new opportunity to develop in vitro systems that better model the in vivo scenario. This will allow us to investigate in a more high-throughput manner the fundamental molecular interactions that are required for successful pregnancy in cattle.
Looking back at five decades of embryo technology in practice.
Hasler JF
Reprod Fertil Dev · 2023 Dec · PMID 38064185 · Publisher ↗
This paper covers developments from International Embryo Technology Society (IETS) meetings over the past 50years. The IETS was officially 'born' at a meeting in Denver, Colorado on Sunday 26 May 1974. There have been 51...
This paper covers developments from International Embryo Technology Society (IETS) meetings over the past 50years. The IETS was officially 'born' at a meeting in Denver, Colorado on Sunday 26 May 1974. There have been 51 IETS meetings (the first was in May 1974), and the first conference at which papers were presented was in 1975. The name of the IETS was changed from International Embryo Transfer Society to International Embryo Technology Society in 2016. The annual IETS conferences are held once a year in January. Embryo technology advanced from the laborious and slow techniques of surgical recovery and transfer of bovine embryos to non-surgical recovery and transfer in many species. Cryopreservation of embryos was initially a slow process that met with only moderate technical success before the development of technology that resulted in high embryo survival rates. The polymerase chain reaction was used to determine the sex of embryos following laminar flow cytology, which came into use to separate X- and Y-bearing spermatozoa. IVF has grown to become widely used, and several factors make it more useful than superovulation and embryo recovery/transfer. Perhaps the most important tool of all was the system that allowed the genetic 'mapping' of the genomic sequence of Bos taurus cattle. Charting key DNA differences by scientists, 'haplotypes' are now used routinely to identify animals with desirable traits of economic importance, discover new genetic disorders, and track carrier status of genotyped animals. Future technology is discussed.
Reproduction in the era of genomics and automation.
Chebel RC, Bisinotto RS, Giordano J … +2 more , Maggiolino A, de Palo P
Reprod Fertil Dev · 2023 Dec · PMID 38064184 · Publisher ↗
Much progress has been made in the reproductive efficiency of lactating dairy cows across the USA in the past 20years. The standardisation of evaluation of reproductive efficiency, particularly with greater focus on metr...
Much progress has been made in the reproductive efficiency of lactating dairy cows across the USA in the past 20years. The standardisation of evaluation of reproductive efficiency, particularly with greater focus on metrics with lesser momentum and less lag-time such as 21-day pregnancy rates (21-day PR), and the recognition that subpar reproductive efficiency negatively impacted profitability were major drivers for the changes that resulted in such progress. Once it became evident that the genetic selection of cattle for milk yield regardless of fertility traits was associated with reduced fertility, geneticists raced to identify fertility traits that could be incorporated in genetic selection programs with the hopes of improving fertility of lactating cows. Concurrently, reproductive physiologists developed ovulation synchronisation protocols such that after sequential treatment with exogenous hormones, cows could be inseminated at fixed time and without detection of oestrus and still achieve acceptable pregnancy per service. These genetic and reproductive management innovations, concurrently with improved husbandry and nutrition of periparturient cows, quickly started to move reproductive efficiency of lactating dairy cows to an upward trend that continues today. Automation has been adopted in Israel and European countries for decades, but only recently have these automated systems been more widely adopted in the USA. The selection of dairy cattle based on genetic indexes that result in positive fertility traits (e.g. daughter pregnancy rate) is positively associated with follicular growth, resumption of ovarian cycles postpartum, body condition score and insulin-like growth factor 1 concentration postpartum, and intensity of oestrus. Collectively, these positive physiological characteristics result in improved reproductive performance. Through the use of automated monitoring devices (AMD), it is possible to identify cows that resume cyclicity sooner after calving and have more intense oestrus postpartum, which are generally cows that have a more successful periparturient period. Recent experiments have demonstrated that it may be possible to adopt targeted reproductive management, utilising ovulation synchronisation protocols for cows that do not have intense oestrus postpartum and relying more heavily on insemination at AMD-detected oestrus for cows that display an intense oestrus postpartum. This strategy is likely to result in tailored hormonal therapy that will be better accepted by the public, will increase the reliance on oestrus for insemination, will improve comfort and reduce labour by reducing the number of injections cows receive in a lactation, and will allow for faster decisions about cows that should not be eligible for insemination.
Pregabalin alters reproductive performance in male mice and causes congenital anomalies in offspring.
Mestre VF, Martins CCN, Brito LV … +3 more , Zeffa AC, Sestário CS, Salles MJS
Reprod Fertil Dev · 2023 Nov · PMID 37995339 · Publisher ↗
CONTEXT: Pregabalin is an anticonvulsant drug with analgesic activity for the treatment of neuropathic pain. AIMS: To valuate the toxicity of pregabalin in reproductive parameters, spermatogenesis, and teratogenicity in...
CONTEXT: Pregabalin is an anticonvulsant drug with analgesic activity for the treatment of neuropathic pain. AIMS: To valuate the toxicity of pregabalin in reproductive parameters, spermatogenesis, and teratogenicity in the offspring of mice. METHODS: Twenty male mice were randomly distributed into two groups: PGB group and group C (n =10 per group). The animals in the PGB group received, via gavage, 200mg/kg of pregabalin diluted in distilled water daily, for a period of 45days. Group C received distilled water under the same experimental design. KEY RESULTS: In the paternal parameters of the PGB group, there was a significant increase in the size of the testicles, morphological alterations in the spermatozoa, a decrease in the Johnsen score, an increase in the Leydig cells, and a decrease in the serum level of testosterone. In the intrauterine development parameters of females mated with males from the PGB group, a significant decrease in placental weight, weight and length of fetuses, and fetal viability rate was observed. There was a significant increase in the number of resorptions and post-implantation losses. The significant anomalies observed in the offspring were alteration in the size of the kidneys, absent metacarpals and phalanges, alteration in the sternum, and supernumerary thoracic vertebrae. CONCLUSION: Results suggest that pregabalin had toxic effects on the reproductive function of male mice and teratogenic potential. IMPLICATIONS: The findings of this study may provide new hypotheses, taking into account the risk-benefit ratio for male reproduction and offspring health.
Optimisation of hormonal treatment to improve follicular development in one-day-old mice ovaries cultured under condition.
Torkzadeh T, Asadi Z, Jafari Atrabi M … +5 more , Eivazkhani F, Khodadi M, Hajiaghalou S, Akbarinejad V, Fathi R
Reprod Fertil Dev · 2023 Nov · PMID 37995332 · Publisher ↗
CONTEXT: Base medium containing knock-out serum replacement (KSR) has been found to support formation and maintenance of follicles in one-day-old mice ovaries, but has not been shown to properly support activation and gr...
CONTEXT: Base medium containing knock-out serum replacement (KSR) has been found to support formation and maintenance of follicles in one-day-old mice ovaries, but has not been shown to properly support activation and growth of primordial follicles. AIMS: The present study was conducted to tailor the hormonal content of base medium containing KSR to enhance development of primordial follicles in neonatal ovaries. METHODS: One-day-old mice ovaries were initially cultured with base medium for four days, and then, different hormonal treatments were added to the culture media and the culture was proceeded for four additional days until day eight. Ovaries were collected for histological and molecular assessments on days four and eight. KEY RESULTS: In experiment I, the main and interactive effects of FSH and testosterone were investigated and FSH promoted activation of primordial follicles and development of primary and preantral follicles, and upregulated genes of phosphoinositide 3-kinase (Pi3k ), KIT ligand (Kitl ), growth differentiation factor 9 (Gdf9 ) and follicle stimulating hormone receptor (Fshr ) (P Bmp15 ), Connexin-43 (Cx43 ) and luteinising hormone and choriogonadotropin receptor (Lhcgr ) (P P Lhcgr (P P >0.05). CONCLUSIONS: Supplementation of culture medium containing KSR with gonadotropins, particularly hMG, could improve follicular growth and expression of factors regulating follicular development. IMPLICATIONS: This study was a step forward in formulating an optimal medium for development of follicles in cultured one-day-old mice ovaries.
Immunolocalisation of aquaporins 3, 7, 9 and 10 in the epididymis of three wild ruminant species (Iberian ibex, mouflon and chamois) and sperm cryoresistance.
Martinez-Madrid B, Martínez-Cáceres C, Pequeño B … +7 more , Castaño C, Toledano-Díaz A, Bóveda P, Prieto P, Alvarez-Rodriguez M, Rodriguez-Martinez H, Santiago-Moreno J
Reprod Fertil Dev · 2023 Nov · PMID 37968880 · Publisher ↗
CONTEXT: In the epididymis, epithelial cells manage changes in the luminal environment for proper sperm maturation. Moreover, aquaglyceroporins, a subgroup of aquaporins (AQP), modulate the transport of water, glycerol a...
CONTEXT: In the epididymis, epithelial cells manage changes in the luminal environment for proper sperm maturation. Moreover, aquaglyceroporins, a subgroup of aquaporins (AQP), modulate the transport of water, glycerol and other small molecules in epithelial cells. AIMS: We aim to characterise the lining epithelium, quantify its cell composition and immunolocalise the aquaglyceroporins AQP3, AQP7, AQP9 and AQP10 alongside the epididymal ductus of three wild ruminant species, and to determine if species-specific differences could be associated with cauda sperm cryoresistance variations. METHODS: Epididymides from Iberian ibex (n =5), mouflon (n =5) and chamois (n =6) were obtained. Cauda spermatozoa were collected and sperm parameters were analysed before and after freezing. Histology and immunohistochemistry of AQP3, 7, 9, 10 and T-CD3 were performed in the caput, corpus and cauda epididymal regions. KEY RESULTS: This work first describes the lining epithelium in Iberian ibex, mouflon and chamois epididymis along the three anatomical regions, consisting of principal, basal, apical, clear and halo cells. However, the percentage of each cell type differed in ibex compared to mouflon and chamois. The positive T-CD3 immunolabeling of all the halo cells confirmed their T-lymphocyte nature. Aquaglyceroporin expression patterns were similar among species, except for differences in AQP7 and AQP10 immunolocalisation in ibex. Species-specific differences in epididymal sperm cryoresistance were confirmed. CONCLUSIONS: The epididymal epithelium of the three wild ruminants differ in their relative number of cell types and AQP immunolocalisation, which ultimately appears to affect cauda epidydimal spermatozoa cryoresistance. IMPLICATIONS: Our study provides information on the relevance of the quantitative composition and AQP pattern expression in epididymal lining epithelium on sperm cryoresistance.
RGMb expression in goat uterine tissues: possible role of RGMb in the proliferation and apoptosis of endometrial epithelial cells.
Xue Y, Pei X, Xia Y … +4 more , Chen H, Yu H, Wang W, Mao D
Reprod Fertil Dev · 2023 Nov · PMID 37967584 · Publisher ↗
Context Bone morphogenetic proteins (BMPs) play an important role in the uteri. Repulsive guidance molecule b (RGMb; a.k.a. Dragon) has been confirmed as the coreceptor of BMPs to function through drosophila mothers agai...
Context Bone morphogenetic proteins (BMPs) play an important role in the uteri. Repulsive guidance molecule b (RGMb; a.k.a. Dragon) has been confirmed as the coreceptor of BMPs to function through drosophila mothers against decapentaplegic protein (Smads) and mitogen-activated protein kinases (MAPK) pathways. We hypothesise that RGMb regulates the uterine function through the Smads and MAPK pathways. Aims This study aimed to investigate the expression of RGMb in goat uteri and the potential role of RGMb in the endometrial epithelial cells (EECs). Methods The localisation of RGMb in goat uterine tissues was detected by immunohistochemistry (IHC), EECs were isolated and transfected with siRNA to investigate the role of RGMb in proliferation, and apoptosis. The expression levels of Smads and MAPK members was measured by western blot (WB) and real-time PCR (RT-PCR). Key results IHC showed that RGMb was localised in goat endometrial luminal cells, glandular epithelial cells, and circular muscle fibres, but not in stromal cells. RT-PCR results showed that treatment with RGMb siRNA suppressed the expressions of proliferation-related genes cyclin D1 (CCND1 , P =0.0291), cyclin-dependent kinase 2 (CDK2 P =0.0107), and proliferating cell nuclear antigen (PCNA, P =0.0508), leading to the reduced viability of EECs (P =0.0010). WB results showed that the expression ratio of cleaved-caspase 3/caspase 3 (P =0.0013) was markedly increased after RGMb siRNA transfection. Likewise, the level of phospho-extracellular signal-regulated kinase 1/2 (p-ERK1/2, P =0.0068) and p-Smad1/5/8 (P =0.0011) decreased significantly, while there were no appreciable differences in the level of p-P38 MAPK expression (P >0.05). Conclusions RGMb might participate in the regulation of cell proliferation and apoptosis through Smads and ERK signalling pathways in goat EECs. Implications RGMb is involved in regulating the proliferation and apoptosis in goat endometrial epithelial cells.
Chemerin affects the expression of angiogenesis-related factors in the porcine endometrium during early pregnancy and the oestrous cycle: an study.
Kiezun M, Dobrzyn K, Kiezun J … +2 more , Kaminski T, Smolinska N
Reprod Fertil Dev · 2023 Nov · PMID 37955271 · Publisher ↗
CONTEXT: The appropriate course of angiogenesis in the endometrium is crucial for pregnancy establishment and maintenance. Very little is known about the factors linking vessel formation and immune system functioning. AI...
CONTEXT: The appropriate course of angiogenesis in the endometrium is crucial for pregnancy establishment and maintenance. Very little is known about the factors linking vessel formation and immune system functioning. AIMS: We hypothesised that chemerin, an adipokine known for its involvement in the regulation of energy balance and immunological functions, may act as a potent regulator of endometrial angiogenesis during early pregnancy in pigs. METHODS: Porcine endometrial tissue explants were obtained from pregnant pigs on days 10-11, 12-13, 15-16 and 27-28, and on days 10-12 of the oestrous cycle. The explants were in vitro cultured for 24h in the presence of chemerin (100, 200ng/mL) or in medium alone (control). We evaluated the in vitro effect of chemerin on the secretion of vascular endothelial growth factors A-D (VEGF-A-D), placental growth factor (PlGF), basic fibroblast growth factor (bFGF) and angiopoietin 1 and 2 (ANG-1, ANG-2) with the ELISA method. The protein abundance of angiogenesis-related factor receptors, VEGF receptors 1-3 (VEGFR1-3), FGF receptors 1 and 2 (FGFR1-2) and ANG receptor (TIE2) was evaluated with the Western blot (WB) method. We also analysed the influence of chemerin on the phosphorylation of AMPK using WB. KEY RESULTS: We found that in the studied endometrial samples, chemerin up-regulated the secretion of VEGF-A, VEGF-B and PlGF, and protein expression of VEGFR3. The adipokine caused a decrease in VEGF-C, VEGF-D and ANG-1 release. Chemerin effect on bFGF and ANG-2 secretion, and protein content of VEGFR1, VEGFR2, FGFR1, FGFR2 and TIE2 were dependent on the stage of pregnancy. Chemerin was found to down-regulate AMPK phosphorylation. CONCLUSIONS: The obtained in vitro results suggest that chemerin could be an important factor in the early pregnant uterus by its influence on angiogenic factors' secretion and signalling. IMPLICATIONS: The obtained results on the role of chemerin in the process of endometrial angiogenesis may, in the long term perspective, contribute to the elaboration of more effective methods of modifying reproductive processes and maintaining energy homeostasis in farm animals.
Construction of prognostic risk markers for cervical cancer combined with anoikis-related genes and their clinical significance.
Zhang J, Tian Y
Reprod Fertil Dev · 2023 Nov · PMID 37899003 · Publisher ↗
CONTEXT: Several studies have demonstrated that anoikis affects the development, metastasis and prognosis of cancer. AIMS: This study aimed to identify anoikis-related marker genes in cervical cancer (CC). METHODS: Least...
CONTEXT: Several studies have demonstrated that anoikis affects the development, metastasis and prognosis of cancer. AIMS: This study aimed to identify anoikis-related marker genes in cervical cancer (CC). METHODS: Least absolute shrinkage and selection operator (LASSO) combined with Cox regression analysis was used to construct a prognostic model and analyse the independent prognostic ability of riskscore. Receiver operating characteristic curve (ROC) and survival curves were used to evaluate and verify the performance and accuracy of the model. The nomogram of CC prognostic model was drawn using riskscore combined with clinical information. We analysed the relationship between prognostic riskscore and immune infiltration level and analysed immunophenoscore. Finally, qRT-PCR assay was used to verify the feature genes. KEY RESULTS: By Cox analysis, we found that the prognostic risk model could effectively predict the risk of CC in patients independently of other clinical factors. Both the levels of immune infiltration and the immunophenoscore were significantly lower in high-risk CC patients than those in low-risk patients, revealing that high-risk patients were likely to have bad response to immunotherapy. The qRT-PCR results of the feature genes were consistent with the results of gene expression in the database. CONCLUSIONS: The prognostic model constructed, based on anoikis-related genes in CC, could predict the prognosis of CC patients. IMPLICATIONS: The model described here can provide effective support for assessing prognostic risk and devising personalised protocols during clinical treatment.
Aquaporin 7 is upregulated through the PI3K-Akt pathway and modulates decidualisation of endometrial stromal cells.
Liu M, Mi YJ, Dai J
Reprod Fertil Dev · 2023 Nov · PMID 37879294 · Publisher ↗
CONTEXT: Aquaporin 7 (AQP7) is selectively expressed in decidualised endometrial stromal cells (ESCs) of mice surrounding the embryonic implantation sites. However, the roles of AQP7 and the underlying mechanism that reg...
CONTEXT: Aquaporin 7 (AQP7) is selectively expressed in decidualised endometrial stromal cells (ESCs) of mice surrounding the embryonic implantation sites. However, the roles of AQP7 and the underlying mechanism that regulates AQP7 expression in endometrial decidualisation after implantation are still unclear. AIMS: This study aimed to investigate the role of the PI3K-Akt pathway in regulating the expression of AQP7 in ESCs and decidualisation. METHODS: Primary ESCs of pregnant mice were isolated to establish in vitro decidualisation models. PI3K inhibitor LY294002 was added to the decidualisation models, then AQP7 expression, changes in decidualised ESC morphology and expression of decidualisation marker molecules were examined. KEY RESULTS: AQP7 knockdown reduced the proliferation and differentiation of ESCs with in vitro induced decidualisation. Furthermore, when the activity of PI3K was inhibited by LY294002, the expression of AQP7 in decidualised ESCs was decreased and both the proliferation and differentiation of ESCs were significantly reduced. CONCLUSIONS: This indicates that AQP7 is a key molecule involved in endometrial decidualisation and the expression of AQP7 is upregulated through activation of the PI3K-Akt pathways, which promotes the proliferation and differentiation of the ESCs, thus affecting occurrence of decidualisation. IMPLICATIONS: This study may provide a new biomarker for the diagnosis of infertility and a new drug target for the prevention and treatment of infertility.
Classical cadherins in the testis: how are they regulated?
Nguyen HT, Martin LJ
Reprod Fertil Dev · 2023 Nov · PMID 37717581 · Publisher ↗
Cadherins (CDH) are crucial intercellular adhesion molecules, contributing to morphogenesis and creating tissue barriers by regulating cells' movement, clustering and differentiation. In the testis, classical cadherins s...
Cadherins (CDH) are crucial intercellular adhesion molecules, contributing to morphogenesis and creating tissue barriers by regulating cells' movement, clustering and differentiation. In the testis, classical cadherins such as CDH1, CDH2 and CDH3 are critical to gonadogenesis by promoting the migration and the subsequent clustering of primordial germ cells with somatic cells. While CDH2 is present in both Sertoli and germ cells in rodents, CDH1 is primarily detected in undifferentiated spermatogonia. As for CDH3, its expression is mainly found in germ and pre-Sertoli cells in developing gonads until the establishment of the blood-testis barrier (BTB). This barrier is made of Sertoli cells forming intercellular junctional complexes. The restructuring of the BTB allows the movement of early spermatocytes toward the apical compartment as they differentiate during a process called spermatogenesis. CDH2 is among many junctional proteins participating in this process and is regulated by several pathways. While cytokines promote the disassembly of the BTB by enhancing junctional protein endocytosis for degradation, testosterone facilitates the assembly of the BTB by increasing the recycling of endocytosed junctional proteins. Mitogen-activated protein kinases (MAPKs) are also mediators of the BTB kinetics in many chemically induced damages in the testis. In addition to regulating Sertoli cell functions, follicle stimulating hormone can also regulate the expression of CDH2. In this review, we discuss the current knowledge on regulatory mechanisms of cadherin localisation and expression in the testis.
Use of sensitivity-enhanced nuclear magnetic resonance spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe in identifying human sperm intracellular metabolites.
Cheredath A, Uppangala S, Jijo A … +4 more , Lakshmi RV, Gowda GAN, Kalthur G, Adiga SK
Reprod Fertil Dev · 2023 Nov · PMID 37643634 · Publisher ↗
CONTEXT: The clinical value of human sperm metabolites has not been established due to the technical complexity in detecting these metabolites when sperm numbers are low. AIMS: To detect endogenous intracellular metaboli...
CONTEXT: The clinical value of human sperm metabolites has not been established due to the technical complexity in detecting these metabolites when sperm numbers are low. AIMS: To detect endogenous intracellular metabolites in fresh and post-thaw human spermatozoa using 800MHz nuclear magnetic resonance (NMR) spectroscopy equipped with a 1.7-mm cryo-probe. METHODS: Processed spermatozoa from 25 normozoospermic ejaculates were subjected to extraction of intracellular metabolites and then profiled by sensitivity-enhanced NMR spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe. In parallel, some of the processed sperm fractions were subjected to freeze-thawing and were then analysed for intracellular metabolites. KEY RESULTS: Twenty-three metabolites were profiled from only 1.25million sperm cells. Comparison of the metabolomic signature of pre-freeze and post-thaw sperm cells did not show significant changes in the levels of metabolites. CONCLUSIONS: Sensitivity-enhanced NMR spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe is a potential tool for identifying intracellular metabolites when sperm number is low. IMPLICATIONS: Use of sensitivity-enhanced NMR spectroscopy opens up the opportunity to test for endogenous metabolites in samples with a limited number of spermatozoa, to understand the patho-physiology of infertility.
Dynamic changes and importance of plasma concentrations of ether phospholipids, of which the majority are plasmalogens, in postpartum Holstein dairy cows.
Saito R, Kubo T, Wakatsuki T … +6 more , Asato Y, Tanigawa T, Kotaniguchi M, Hashimoto M, Kitamura S, Kadokawa H
Reprod Fertil Dev · 2023 Jul · PMID 37463606 · Publisher ↗
CONTEXT: Ethanolamine plasmalogens (EPls) and choline plasmalogens (CPls) are classes of ethanolamine ether phospholipids (ePE) and choline ether phospholipids (ePC), respectively. EPls play crucial roles in maternal and...
CONTEXT: Ethanolamine plasmalogens (EPls) and choline plasmalogens (CPls) are classes of ethanolamine ether phospholipids (ePE) and choline ether phospholipids (ePC), respectively. EPls play crucial roles in maternal and breastfed infant bodies and stimulate gonadotropin secretion by gonadotrophs. AIMS: To estimate changes in and importance of plasma concentrations of EPls and CPls, utilising newly developed enzymatic fluorometric assays for ePE and ePC in postpartum Holstein cows. METHODS: Plasma samples were collected from 3weeks before expected parturition until approximately 8weeks after parturition (16 primiparous and 38 multiparous cows) for analysis. KEY RESULTS: Plasma concentrations of ePE and ePC, most of which are plasmalogens, declined before and increased after parturition and stabilised near the day of the first postpartum ovulation (1stOV). From weeks 2 to 3 after parturition, third-parity cows exhibited ePE concentrations that were higher than those of other parity cows. The days from parturition to 1stOV correlated with days from parturition to conception. On the day of 1stOV, milk yield correlated with plasma concentration of both ePE and ePC, while ePC concentration correlated negatively with milk fat percentage. At the early luteal phase after 1stOV, plasma ePE concentration correlated with plasma anti-Müllerian hormone concentration (r =0.39, P <0.01), and plasma ePC concentration correlated with plasma follicle-stimulating hormone concentration (r =0.43, P <0.01). CONCLUSION: The concentrations of ePE and ePC changed dramatically around parturition and 1stOV, and the concentrations correlated with important parameters for milk production and reproduction. IMPLICATIONS: The blood plasmalogen may play important roles in postpartum dairy cows.
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