Reproduction, fertility, and development最新文献

Understanding of central nervous system mechanisms related to age-related infertility remains limited. Fibril α-synuclein, distinct from its monomer form, is implicated in age-related diseases and propagates among neurons akin to prions.

We compared α-synuclein expression in gonadotropin-releasing hormone-expressing neurons (GnRH neurons) in the pre-optic area, arcuate nucleus, and median eminence of healthy heifers and aged cows to determine its role in age-related infertility.

We analysed mRNA and protein expression, along with fluorescent immunohistochemistry for GnRH and α-synuclein, followed by Congo red staining to detect amyloid deposits, and confocal microscopy.

Both mRNA and protein expressions of α-synuclein were confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and western blots in bovine cortex, hippocampus, and anterior and posterior hypothalamus tissues. Significant differences in α-synuclein mRNA expression were observed in the cortex and hippocampus between young and old cows. Western blots showed five bands of α-synuclein, probably reflecting monomer, dimer, and oligomers, in the cortex, hippocampus, hypothalamus tissues, and there were significant differences in some bands between young and old cows. Bright-field and polarised light microscopy did not detect obvious amyloid deposition in aged hypothalami; however, higher-sensitive confocal microscopy unveiled strong positive signal of Congo red and α-synuclein in GnRH neurons in aged hypothalami. Additionally, α-synuclein expression was detected in immortalised GnRH neurons, GT1-7 cells.

Alpha-synuclein was expressed in GnRH neurons, and some differences were observed between young and old hypothalami.

Alpha-synuclein may play an important role in aging-related infertility.

In vitro embryo production in pigs is an important tool for advancing biomedical research. Intracytoplasmic sperm injection (ICSI) circumvents the polyspermy problems associated with conventional IVF in porcine. However, the suboptimal efficiency for ICSI in pigs requires new strategies to increase blastocyst formation rates.

To investigate novel methods for assisted activation using the zinc chelator 1,10-phenanthroline (PHEN), and to improve embryo developmental competence and quality of ICSI porcine blastocyst.

ICSI embryos were treated with PHEN after or before sperm injection, recording pronuclear formation, blastocyst rate and the expression of SMARCA4, OCT4, SOX2 and CDX2.

Neither electrical nor PHEN significantly improves pronuclear formation rates before or after ICSI. Following in vitro culture to the blastocyst stage, no significant differences were observed in developmental rates among the groups. Moreover, the use of PHEN did not alter the total cell number or the expression of OCT4, SOX2 and CDX2 in pig ICSI blastocysts.

Assisted oocyte activation with PHEN does not affect the preimplantation development of ICSI-derived pig embryos.

These results hold significance in refining and advancing the application of assisted oocyte activation techniques. They offer insights into addressing fertility issues and propelling advancements in human and animal reproductive medicine.

Sires differ in their ability to produce viable blastocysts, yet our understanding of the cellular mechanisms regulated by the sire during early embryo development is limited.

The first aim was to characterise autophagy and reactive oxygen species (ROS) in embryos produced by high and low performing sires under normal and stress culture conditions. The second aim was to evaluate DNA damage and lipid peroxidation as mechanisms that may be impacted by increased cellular stress, specifically oxidative stress.

Embryos were produced using four high and four low performing sires based on their ability to produce embryos. Autophagy and ROS were measured throughout development. To evaluate oxidative stress response, autophagy, and ROS were measured in 2–6 cell embryos exposed to heat stress. To understand how cellular stress impacts development, DNA damage and lipid peroxidation were assessed.

Under normal conditions, embryos from low performing sires had increased ROS and autophagy. Under heat stress, embryos from low performing sires had increased ROS, yet those from high performing sires had increased autophagy. There was no difference in DNA damage or lipid peroxidation.

Results suggest that embryos from low performing sires may begin development under increased cellular stress, and autophagy potentially increases to mitigate the impacts of stress.

There is potential for improving embryonic competence through selection of sires with lower stress-related markers.

Prostate-specific antigen (PSA) is important for semen liquefaction and sperm motility. Anti-PSA antibodies may lead to immune infertility.

The present study was conducted to evaluate the impact of seminal anti-PSA antibodies on semen parameters in fertile and infertile men.

A cross-sectional analytic study was conducted on 105 fertile men (≥21–50 years) having biological children (within last 2 years) with normal semen analysis as controls and 105 infertile men with abnormal semen analysis as cases. All semen samples were cryopreserved till 210 samples were collected, followed by estimation of anti-PSA antibodies using an indirect enzyme-linked immunosorbent assay technique.

Mean ± standard deviation (s.d.) age of 210 participants was 30.0 ± 4.65 years. Mean ± s.d. levels of seminal anti-PSA antibodies in infertile men were 27.82 ± 102.19 ng/mL and in fertile men −30.45 ± 49.49 ng/mL (P = 0.001). A significant negative correlation was observed between anti-PSA antibody levels and sperm concentration (P = 0.013), rapid progressive motility (P = 0.001), slow progressive motility (P = 0.006), progressive sperm motility (P = 0.001), and normal morphology (P = 0.001), and significant positive correlation was observed with immotile sperms (P = 0.001). The overall accuracy of anti-PSA antibody for differentiating infertile from fertile men was 63.33%.

Seminal anti-PSA antibodies were significantly correlated with semen parameters in fertile and infertile men with an accuracy of 63.33%. A negative correlation was observed between antibody levels and progressive sperm motility.

Seminal anti-PSA antibodies can be used as a biomarker for male infertility assessment.

Infertility affects approximately 15% of couples trying to conceive. Male-related causes account for roughly 50% of cases, with obesity emerging as a possible significant factor. Obesity, defined as a body mass index of 30.0 or higher, has become a widespread epidemic associated with numerous health issues, including a decrease of fertility. This review discusses the relationship between obesity and male infertility, particularly focusing on sperm quality and function. An overview of the literature suggests that obesity may influence the male reproductive system via disruptions in hormonal profiles, oxidative stress, and inflammation, leading to changes in sperm parameters. Several studies have discussed if obesity causes a decrease in sperm concentration, motility, and normal morphology, so far without a consensus being reached. However, available evidence suggests an impairment of sperm function in obese men, due to an increase in DNA damage and oxidative stress, impaired mitochondrial function and acrosome reaction in response to progesterone. Finally, the relationship between obesity and assisted reproductive technologies outcomes remains debatable, with conflicting evidence regarding the influence on fertilisation, pregnancy, and live birth rates. Therefore, the actual impact of obesity on human spermatozoa still needs to be clarified, due to the multiple factors potentially in play.

Context: In pigs, in vitro fertilisation (IVF) is associated with high polyspermy rates, and for this reason, in vitro embryo production (IVP) is still an inefficient biotechnology. Coculture with somatic cells is an alternative to improve suboptimal in vitro maturation (IVM) conditions.

Aim: This study was conducted to test a coculture system of porcine luteal cells (PLC) and cumulus-oocyte complexes (COC) to improve oocyte metabolism.

Methods: COC were matured in vitro with PLC. Oocyte lipid content, mitochondrial activity, zona pellucida (ZP) digestibility and pore size, cortical reaction and in vitro embryo development were assessed.

Key results: Coculture reduced cytoplasmic lipid content in the oocyte cytoplasm without increasing mitochondrial activity. Although ZP digestibility and ZP pore number were not different between culture systems, ZP pores were smaller in the coculture. Coculture impacted the distribution of cortical granules as they were found immediately under the oolemma, and more of them had released their content in the ZP. Coculture with porcine luteal cells during IVM increased monospermic penetration and embryo development after IVF.

Conclusions: The coculture of COC with PLC affects the metabolism of the oocyte and benefits monospermic penetration and embryo development.

Implications: The coculture system with PLC could be an alternative for the conventional maturation medium in pigs.

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.

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.

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.