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Naunyn-Schmiedeberg's Archives of... Jun 2024The endocannabinoid system and prostaglandins are important modulators in the genitourinary system. This study aimed to investigate the possible interactions between the...
Interaction of endocannabinoid system and cyclooxygenase metabolites with fatty acid amide hydrolase and cyclooxygenase enzyme activities on contractile responses in rat vas deferens tissue.
The endocannabinoid system and prostaglandins are important modulators in the genitourinary system. This study aimed to investigate the possible interactions between the endocannabinoid system and the cyclooxygenase (COX) pathway on rat vas deferens. For this purpose, the concentration responses of the endocannabinoid anandamide, prostaglandin F analog latanoprost, and prostaglandin E analog misoprostol on the electrical field stimulation (EFS)-induced contractile responses were obtained. The concentration responses to anandamide were obtained again in the presence of nonselective COX inhibitor flurbiprofen and prostaglandin analogs, while the concentration responses of latanoprost and misoprostol were obtained in the presence of cannabinoid receptor antagonists and fatty acid amide hydrolase (FAAH) enzyme inhibitor URB597. FAAH, COX-1, and COX-2 enzyme levels in vas deferens tissue samples were also determined. The cumulative addition of anandamide was not different from the vehicle; however, the EFS-induced contractile responses were significantly increased with the incubation of latanoprost or flurbiprofen in the prostatic portion. Flurbiprofen and misoprostol decreased FAAH enzyme levels in both portions of the vas deferens, while latanoprost induced the inhibition in the prostatic portion. The cumulative administration of latanoprost and misoprostol significantly enhanced the contractile responses in the prostatic portion. This effect of latanoprost was significantly antagonized by URB597 and AM251. The enhancing effect of misoprostol was antagonized by anandamide, URB597, AM251, and AM630. Anandamide, AM251, AM630, and URB597 decreased enzyme levels of COX-1 and COX-2 in both portions of the vas deferens. These results demonstrate an intricate crosstalk between endocannabinoids and prostaglandins in modulation of the vas deferens contractility.
Topics: Male; Animals; Vas Deferens; Amidohydrolases; Endocannabinoids; Muscle Contraction; Cyclooxygenase 1; Rats, Wistar; Cyclooxygenase 2; Polyunsaturated Alkamides; Cyclooxygenase Inhibitors; Benzamides; Arachidonic Acids; Prostaglandin-Endoperoxide Synthases; Rats; Electric Stimulation; In Vitro Techniques; Carbamates; Misoprostol; Membrane Proteins
PubMed: 38032490
DOI: 10.1007/s00210-023-02861-3 -
Animals : An Open Access Journal From... May 2024The advances in Assisted Reproductive Technologies (ARTs) applied in South American camelid species are still scarce. The aim of this study was to compare the effects of...
The advances in Assisted Reproductive Technologies (ARTs) applied in South American camelid species are still scarce. The aim of this study was to compare the effects of three semen extenders, before and after the cryopreservation of spermatozoa obtained from the vas deferens, on sperm quality parameters and in vitro fertilization rates of llama () oocytes. Mature fertile llama males (; = 6; age: 48-60 mo.; BCS: ~2.7) were included in the study. Sperm samples were collected from each male using the surgical technique of the vas deferens deviation. Then, the sperm samples were pooled and diluted with the Tris-EY, Andromed, or BioxCell extender in order to subsequently carry out the sperm cryopreservation process. The sperm quality assessment related to each extender was performed before and after cryopreservation with regard to sperm morphological abnormalities, acrosome integrity, sperm viability, membrane permeability, and sperm motility traits. Moreover, in vitro fertilization (IVF) procedures were carried out to evaluate the in vitro fertility of the cryopreserved sperm samples using each extender. Overall, significant differences were observed before and after cryopreservation regarding acrosome integrity, sperm viability, membrane permeability, and sperm motility traits among the extenders used, where Tris-EY and Andromed were better than BioxCell ( < 0.05); however, no differences were observed regarding the sperm morphological abnormalities among extenders ( > 0.05). Moreover, multiple differences were observed with regard to the velocity and linearity kinematic parameters obtained by computerized analysis before and after the cryopreservation process, irrespective of the extender used ( < 0.05). Finally, differences were observed regarding the in vitro fertilization rates among the different extender-derived samples ( < 0.05). In conclusion, the sperm quality using Tris-EY and Andromed was better before and after cryopreservation compared to that using BioxCell. Although the number of fertilized oocytes obtained after the IVF process between Tris-EY and Andromed was similar, Andromed-derived samples showed the best sperm quality results before and after cryopreservation. This indicates that the cryopreservation extender is a determining factor in significantly improving in vitro fertilization rates when using sperm samples obtained from vas deferens in llama () males.
PubMed: 38891621
DOI: 10.3390/ani14111573 -
Frontiers in Endocrinology 2024As an important gas signaling molecule, hydrogen sulfide (HS) affects multiple organ systems, including the nervous, cardiovascular, digestive, and genitourinary,... (Review)
Review
As an important gas signaling molecule, hydrogen sulfide (HS) affects multiple organ systems, including the nervous, cardiovascular, digestive, and genitourinary, reproductive systems. In particular, HS not only regulates female reproductive function but also holds great promise in the treatment of male reproductive diseases and disorders, such as erectile dysfunction, prostate cancer, varicocele, and infertility. In this review, we summarize the relationship between HS and male reproductive organs, including the penis, testis, prostate, vas deferens, and epididymis. As lower urinary tract symptoms have a significant impact on penile erection disorders, we also address the potential ameliorative effects of HS in erectile dysfunction resulting from bladder disease. Additionally, we discuss the regulatory role of HS in cavernous smooth muscle relaxation, which involves the NO/cGMP pathway, the RhoA/Rho-kinase pathway, and K channel activation. Recently, various compounds that can alleviate erectile dysfunction have been reported to be at least partly dependent on HS. Therefore, understanding the role of HS in the male reproductive system may help develop novel strategies for the clinical treatment of male reproductive system diseases.
Topics: Hydrogen Sulfide; Humans; Male; Genitalia, Male; Animals; Erectile Dysfunction; Signal Transduction
PubMed: 38883604
DOI: 10.3389/fendo.2024.1377090 -
Animal Reproduction 2023The objectives of the study were to (1) describe the kinematic parameters of spermatozoa (2) compare methods of evaluating sperm viability (3) validate assays of...
The objectives of the study were to (1) describe the kinematic parameters of spermatozoa (2) compare methods of evaluating sperm viability (3) validate assays of functionality and integrity of the sperm membrane and (4) evaluate possible changes between spermatozoa from the epididymis and the vas deferens of the greater rhea. Semen samples were recovered from 7 adult individuals. Sperm motility was characterized by adjusting the set-up for Computer-assisted semen analysis (CASA) to that new species. For sperm viability evaluation, smears of bromophenol blue and eosin-nigrosine dyes were used. Five solutions of different osmolarities were then tested for the hypoosmotic swelling test (HOST). The combination of fluorescent probes (propidium iodide - IP and Hoechst 33342) was also used to assess plasma membrane integrity. Data were presented as mean ± SEM. Rhea spermatozoa from the vas deferens had an overall motility of 14.6 ± 2.5%. The bromophenol blue staining technique revealed that 64.6 ± 5.2% sperm were viable, while that proportion was 72.1 ± 2.5% using eosin-nigrosine. An average of 77.6 ± 4.8% of spermatozoa reacted to the HOST with distilled water at 0 mOsm/l. Fluorescent probes indicated that 65.3 ± 2.6% of spermatozoa had intact membranes. Interestingly, no statistical differences were observed between the parameters analyzed in the epididymal spermatozoa and the vas deferens. These new assays set reference values that can now be used to further exploration of sperm handling conditions and freezing protocols in rheas.
PubMed: 38213765
DOI: 10.1590/1984-3143-AR2023-0113 -
Purinergic Signalling Feb 2024Stimulation of sympathetic nerves in the vas deferens yields biphasic contractions consisting of a rapid transient component resulting from activation of P2X1 receptors...
Stimulation of sympathetic nerves in the vas deferens yields biphasic contractions consisting of a rapid transient component resulting from activation of P2X1 receptors by ATP and a secondary sustained component mediated by activation of α-adrenoceptors by noradrenaline. Noradrenaline can also potentiate the ATP-dependent contractions of the vas deferens, but the mechanisms underlying this effect are unclear. The purpose of the present study was to investigate the mechanisms underlying potentiation of transient contractions of the vas deferens induced by activation of α-adrenoceptors. Contractions of the mouse vas deferens were induced by electric field stimulation (EFS). Delivery of brief (1s duration) pulses (4 Hz) yielded transient contractions that were inhibited tetrodotoxin (100 nM) and guanethidine (10 µM). α,β-meATP (10 µM), a P2X1R desensitising agent, reduced the amplitude of these responses by 65% and prazosin (100 nM), an α-adrenoceptor antagonist, decreased mean contraction amplitude by 69%. Stimulation of α-adrenoceptors with phenylephrine (3 µM) enhanced EFS and ATP-induced contractions and these effects were mimicked by the phorbol ester PDBu (1 µM), which activates PKC. The PKC inhibitor GF109203X (1 µM) prevented the stimulatory effects of PDBu on ATP-induced contractions of the vas deferens but only reduced the stimulatory effects of phenylephrine by 40%. PDBu increased the amplitude of ATP-induced currents recorded from freshly isolated vas deferens myocytes and HEK-293 cells expressing human P2X1Rs by 93%. This study indicates that: (1) potentiation of ATP-evoked contractions of the mouse vas deferens by α-adrenoceptor activation were not fully blocked by the PKC inhibitor GF109203X and (2) that the stimulatory effect of PKC on ATP-induced contractions of the vas deferens is associated with enhanced P2X1R currents in vas deferens myocytes.
PubMed: 38374492
DOI: 10.1007/s11302-024-09993-y -
Biological Research Oct 2023In vitro incubation of epididymal and vas deferens sperm with Mn induces Sperm Chromatin Fragmentation (SCF), a mechanism that causes double-stranded breaks in...
BACKGROUND
In vitro incubation of epididymal and vas deferens sperm with Mn induces Sperm Chromatin Fragmentation (SCF), a mechanism that causes double-stranded breaks in toroid-linker regions (TLRs). Whether this mechanism, thought to require the participation of topoisomerases and/or DNAses and thus far only described in epididymal mouse sperm, can be triggered in ejaculated sperm is yet to be elucidated. The current study aimed to determine if exposure of pig ejaculated sperm to divalent ions (Mn and Mg) activates SCF, and whether this has any impact on sperm function and survival. For this purpose, sperm DNA integrity was evaluated through the Comet assay and Pulsed Field Gel Electrophoresis (PFGE); sperm motility and agglutination were assessed with computer assisted sperm analysis (CASA); and sperm viability and levels of total reactive oxygen species (ROS) and superoxides were determined through flow cytometry.
RESULTS
Incubation with Mn/Ca activated SCF in a dose-dependent (P < 0.05) albeit not time-dependent manner (P > 0.05); in contrast, Mg/Ca only triggered SCF at high concentrations (50 mM). The PFGE revealed that, when activated by Mn/Ca or Mg/Ca, SCF generated DNA fragments of 33-194 Kb, compatible with the size of one or multiple toroids. Besides, Mn/Ca affected sperm motility in a dose-dependent manner (P < 0.05), whereas Mg/Ca only impaired this variable at high concentrations (P < 0.05). While this effect on motility was concomitant with an increase of agglutination, neither viability nor ROS levels were affected by Mn/Ca or Mg/Ca treatments.
CONCLUSION
Mn/Ca and Mn/Ca were observed to induce SCF in ejaculated sperm, resulting in DNA cleavage at TLRs. The activation of this mechanism by an intracellular, non-oxidative factor sheds light on the events taking place during sperm cell death.
Topics: Male; Mice; Animals; Swine; Chromatin; Reactive Oxygen Species; Semen; Sperm Motility; Spermatozoa; DNA; DNA Fragmentation
PubMed: 37876007
DOI: 10.1186/s40659-023-00467-w -
Genes Jul 2023(1) Introduction: Pathogenic variants in the (Cystic Fibrosis Transmembrane conductance Regulator, OMIM: 602421) gene cause Cystic Fibrosis (CF, OMIM: 219700) and...
(1) Introduction: Pathogenic variants in the (Cystic Fibrosis Transmembrane conductance Regulator, OMIM: 602421) gene cause Cystic Fibrosis (CF, OMIM: 219700) and CF-related disorders (CF-RD), often accompanied by obstructive azoospermia due to congenital bilateral aplasia of vas deferens (CBAVD, OMIM: 277180) in male patients. The L138ins (c.413_415dup; p. (Leu138dup)) is a mild variant in the gene that is relatively common among CF-patients in Slavic populations. The frequency of this variant in Russian infertile men has not been sufficiently studied; (2) Materials and Methods: The sample consisted of 6033 Russian infertile men. The patients were tested for 22 common in Russian populations pathogenic variants of the gene and the IVS9Tn-polymorphic locus of the intron 9. Molecular-genetic studies were performed using amplified fragment length polymorphism (AFLP-PCR), multiplex ligation-dependent probe amplification (MLPA), and nested PCR (for analysis of the IVS9Tn-polymorphic locus); (3) Results: Pathogenic variants in the were detected in 3.9% of patients. The most frequent variants were F508del and CFTRdele2.3(21kb), accounted for 61.0% and 7.1% of detected variants, respectively. The L138ins variant was detected in 17 (0.28%) individuals: one of them was homozygous, 10 patients were heterozygous, and 6 patients were compound-heterozygous (F508del/L138ins, = 4; L138ins/N1303K, = 1; L138ins/5T, = 1). Two pathogenic CF-causing variants in the gene were detected in 8 patients, including 7 compound heterozygous (F508del/L138ins, = 4; F508del/N1303K, = 1; 2184insA/E92K, = 1; 3849+10kbC>T/E92K, = 1) and one homozygous (L138ins/L138ins). The L138ins variant was found in 7 out of 16 (43.75%) chromosomes in six of these patients. The most common pathogenic variant, F508del, was identified in five out of them, in 5 of 16 (31.25%) chromosomes. The allele frequency (AF) of the L138ins variant in the sample has been found to be 0.0014.; (4) Conclusions: The L138ins variant of the gene is the third most common variant after F508del and CFTRdele2.3(kb) among Russian infertile men.
Topics: Humans; Male; Amplified Fragment Length Polymorphism Analysis; Cystic Fibrosis Transmembrane Conductance Regulator; Gene Frequency; Mutation; Russia; Infertility, Male
PubMed: 37510311
DOI: 10.3390/genes14071407 -
Nature Communications Jan 2024Breeding bulls are well suited to investigate inherited variation in male fertility because they are genotyped and their reproductive success is monitored through semen...
Breeding bulls are well suited to investigate inherited variation in male fertility because they are genotyped and their reproductive success is monitored through semen analyses and thousands of artificial inseminations. However, functional data from relevant tissues are lacking in cattle, which prevents fine-mapping fertility-associated genomic regions. Here, we characterize gene expression and splicing variation in testis, epididymis, and vas deferens transcriptomes of 118 mature bulls and conduct association tests between 414,667 molecular phenotypes and 21,501,032 genome-wide variants to identify 41,156 regulatory loci. We show broad consensus in tissue-specific and tissue-enriched gene expression between the three bovine tissues and their human and murine counterparts. Expression- and splicing-mediating variants are more than three times as frequent in testis than epididymis and vas deferens, highlighting the transcriptional complexity of testis. Finally, we identify genes (WDR19, SPATA16, KCTD19, ZDHHC1) and molecular phenotypes that are associated with quantitative variation in male fertility through transcriptome-wide association and colocalization analyses.
Topics: Humans; Cattle; Animals; Male; Mice; Quantitative Trait Loci; Epididymis; Testis; Consensus; Fertility
PubMed: 38253538
DOI: 10.1038/s41467-024-44935-7 -
Urology Case Reports Nov 2023We report an exceedingly rare case of Klippel-Feil syndrome (KFS), compounded by ipsilateral absence of the vas deferens, renal agenesis, and diaphragmatic hernia....
We report an exceedingly rare case of Klippel-Feil syndrome (KFS), compounded by ipsilateral absence of the vas deferens, renal agenesis, and diaphragmatic hernia. Unilateral absence of the vas deferens was found incidentally during orchidopexy. To the best of our knowledge, no case of unilateral absence of the kidney and vas deferens has been reported in children with KFS.
PubMed: 37965125
DOI: 10.1016/j.eucr.2023.102600 -
Andrology Sep 2023The development of single-cell sequencing technology has expanded the understanding of cell heterogeneity and disease progression in the male genitourinary system....
BACKGROUND
The development of single-cell sequencing technology has expanded the understanding of cell heterogeneity and disease progression in the male genitourinary system. However, complex processing and unprofessional analytical annotations limit the daily use and widely sharing of published datasets.
OBJECTIVES
Single-cell sequencing data of male-specific tissues and organs.
MATERIALS AND METHODS
The data were downloaded from published studies and were processed based on the Seurat R package, including quality control, cell clustering, reduction and graph generation, and cell type annotation were differentiated by referring to the related paper or recognized cell markers. Input and visual results output through the Shiny package, which was loaded into the remote server.
RESULTS
The current version of the Male Health Atlas database includes two species (human and mouse), five male-specific tissues and organs (testis, epididymis, vas deferens, corpus cavernosum, and prostate), and eight major cell types, with a total of 57 samples and 258,428 single-cell profiles. The results were divided into two main parts: Cell Clustering and Gene Display. In Cell Clustering section, visitors are free to change cell dimensionality reduction (t-distributed stochastic neighbor embedding, or uniform manifold approximation and projection), color palette, and annotation (cell type or sample type). The Gene Display section includes a reduced dimension scatter plot, violin plot, and bubble plot. Visitors can easily view the expression characteristics of single or multiple genes, and compare the expression differences between different cell types or groups.
DISCUSSION AND CONCLUSION
Male Health Atlas is the first single-cell database website in the field of andrology and male reproduction, providing researchers with single-cell sequencing resources and an accessible tool. Male Health Atlas is freely available at http://malehealthatlas.cn/.
Topics: Male; Humans; Animals; Mice; Sequence Analysis, RNA; Single-Cell Gene Expression Analysis; Single-Cell Analysis; Cluster Analysis; Databases, Factual; Gene Expression Profiling
PubMed: 36710661
DOI: 10.1111/andr.13402