Infertility has recently emerged as a severe medical problem. The essential elements in male infertility are sperm morphology, sperm motility, and sperm density. In order to analyze sperm motility, density, and morphology, laboratory experts do a semen analysis. However, it is simple to err when using a subjective interpretation based on laboratory observation. In this work, a computer-aided sperm count estimation approach is suggested to lessen the impact of experts in semen analysis. Object detection techniques concentrating on sperm motility estimate the number of active sperm in the semen. This study provides an overview of other techniques that we can compare. The Visem dataset from the Association for Computing Machinery was used to test the proposed strategy. We created a labelled dataset to prove that our network can detect sperms in images. The best not-super tuned result is mAP 72.15.

• Klíčová slova
• computer-aided sperm analysis, small-object detection, sperm-cell detection, yolo,
• MeSH
• analýza spermatu MeSH
• lidé MeSH
• motilita spermií MeSH
• mužská infertilita * diagnóza   MeSH
• sperma * MeSH
• spermie MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Male infertility is a serious problem in an increasing number of couples. We report an infertile man with non-obstructive azoospermia and karyotype 45,XY,rob(14;22). The immunofluorescence analysis of his testicular tissue using antibodies to SYCP1, SYCP3, HORMAD2, MLH1, and centromeres showed delayed synapsis of the chromosomes involved in the translocation, a varying extent of trivalent asynapsis and its association with sex chromosomes. The mean frequency of meiotic recombination per cell was within the range of normal values. Fluorescence in situ hybridization (FISH) with probes for chromosomes 14 and 22 revealed 5.83% of chromosomally abnormal testicular spermatozoa. FISH with probes for chromosomes X, Y, and 21 showed frequencies of disomic and diploid testicular spermatozoa increased when compared to ejaculated sperm of healthy donors, but comparable with published results for azoospermic patients. PGD by FISH for the translocation and aneuploidy of chromosomes X, Y, 13, 18, and 21 showed a normal chromosomal complement in one out of three analyzed embryos. A healthy carrier girl was born after the embryo transfer. This study shows the benefits of preimplantation genetic diagnosis in a case of a rare Robertsonian translocation carrier with azoospermia and a relatively low frequency of chromosomally unbalanced testicular spermatozoa.

• Klíčová slova
• Aneuploidy, FISH, PGD, Robertsonian translocation, azoospermia, meiosis, testicular sperm,
• MeSH
• aneuploidie * MeSH
• azoospermie genetika   MeSH
• detekce genetických nosičů * MeSH
• karyotypizace MeSH
• lidé MeSH
• lidské chromozomy, pár 14 * MeSH
• lidské chromozomy, pár 22 * MeSH
• meióza genetika   MeSH
• spermie metabolismus   MeSH
• translokace genetická * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH

Complementary molecules on the surface of both gametes are responsible for the interaction of sperm protein receptors with zona pellucida (ZP) saccharide structures, and many primary sperm receptors for ZP glycoproteins have been disclosed in various mammals. For our study, proteins were obtained from the surface of ejaculated and in vitro capacitated boar sperm. The isolated proteins were characterized by 1D- and 2D-electrophoretic protein profiles, and by glycoprotein staining. Our results show quantitative and qualitative differences in protein and glycoprotein patterns between ejaculated and capacitated sperm. Far-western blotting with ZP glycoproteins identified 17 interactions in the subproteome of the ejaculated sperm and 14 interactions in the subproteome of the capacitated sperm. High-molecular-mass proteins, coincident with binding to ZP, were sequence-identified. Angiotensin-converting enzyme (ACE), polycystic kidney disease receptor and egg jelly receptor (PKDREJ), and acrosin precursor were successfully identified. This is the first time PKDREJ has been identified on the surface of boar spermatozoa.

• Klíčová slova
• PKDREJ protein, Sperm surface proteins, Zona pellucida-binding receptors,
• MeSH
• hmotnostní spektrometrie MeSH
• kapacitace spermií * MeSH
• membránové glykoproteiny metabolismus   MeSH
• membránové proteiny metabolismus   MeSH
• myši MeSH
• proteom MeSH
• proteomika metody   MeSH
• spermie metabolismus   MeSH
• Sus scrofa MeSH
• vazba proteinů MeSH
• zona pellucida metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• membránové glykoproteiny MeSH
• membránové proteiny MeSH
• proteom MeSH

Fertilization is a multiple step process leading to the fusion of female and male gametes and the formation of a zygote. Besides direct gamete membrane interaction via binding receptors localized on both oocyte and sperm surface, fertilization also involves gamete communication via chemical molecules triggering various signaling pathways. This work focuses on a mouse taste receptor, mTAS1R3, encoded by the Tas1r3 gene, as a potential receptor mediating chemical communication between gametes using the C57BL/6J lab mouse strain. In order to specify the role of mTAS1R3, we aimed to characterize its precise localization in testis and sperm using super resolution microscopy. The testis cryo-section, acrosome-intact sperm released from cauda epididymis and sperm which underwent the acrosome reaction (AR) were evaluated. The mTAS1R3 receptor was detected in late spermatids where the acrosome was being formed and in the acrosomal cap of acrosome intact sperm. AR is triggered in mice during sperm maturation in the female reproductive tract and by passing through the egg surroundings such as cumulus oophorus cells. This AR onset is independent of the extracellular matrix of the oocyte called zona pellucida. After AR, the relocation of mTAS1R3 to the equatorial segment was observed and the receptor remained exposed to the outer surroundings of the female reproductive tract, where its physiological ligand, the amino acid L-glutamate, naturally occurs. Therefore, we targeted the possible interaction in vitro between the mTAS1R3 and L-glutamate as a part of chemical communication between sperm and egg and used an anti-mTAS1R3-specific antibody to block it. We detected that the acrosome reacted spermatozoa showed a chemotactic response in the presence of L-glutamate during and after the AR, and it is likely that mTAS1R3 acted as its mediator.

• Klíčová slova
• L-glutamate, TAS1R family, acrosome reaction, chemoattractant, chemotaxis, gamete, mTAS1R3 receptor, mouse, sperm,
• MeSH
• buněčná diferenciace MeSH
• chemotaxe MeSH
• exprese genu MeSH
• glutamáty metabolismus   MeSH
• interakce spermie a vajíčka * MeSH
• messenger RNA genetika   MeSH
• mezibuněčná komunikace * MeSH
• myši MeSH
• receptory spřažené s G-proteiny genetika   metabolismus   MeSH
• spermie cytologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glutamáty MeSH
• messenger RNA MeSH
• receptory spřažené s G-proteiny MeSH
• taste receptors, type 1 MeSH Prohlížeč

OBJECTIVE: The transfer of good quality embryo in the program of assisted reproduction in the case of azoospermia, dg. Sertolli cells only syndrome (SCO sy) + maturation arrest (MA). Testes were assessed and found to have a high occurrence of Sertolli cells and very low occurrence of germinal cells, which were arrested at the round spermatid level. The histological evaluation was hypospermatogenesis gr. 3 (minimum 1 spermatid/sample). DESIGN: Case report. SETTING: Laboratory IVF, Iscare, a. s., Department of Biology and Biochemistry of Fertilization, Institute of Molecular Genetics, Czech Academy of Sciences, Prague. SUBJECT AND METHOD: The successful integration of three methods provides a solution for this case of azoospermia. Immunology and histology can more exactly diagnose the degree of azoospermia. Detection and visualisation of spermatids using monoclonal antibodies against sperm proteins predicts the eventual occurrence of spermatogenesis, and histological evaluation confirms these immunological findings. Using the information of both methods it is possible to use special in vitro cultivation of testicular cells and so obtain injectable spermatozoa, or precursors of sperm, for the ICSI method. CONCLUSION: The probability of acquisition of good-quality embryo in the program of assisted reproduction is higher when these three methods are applied in combination.

• MeSH
• dospělí MeSH
• intracytoplazmatické injekce spermie * MeSH
• lidé MeSH
• mužská infertilita patologie   terapie   MeSH
• oligospermie etiologie   patologie   MeSH
• Sertoliho buňky patologie   MeSH
• spermatidy patologie   MeSH
• těhotenství MeSH
• testis patologie   MeSH
• zrání spermie MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH

Primary binding of the sperm to the zona pellucida (ZP) is one of the many steps necessary for successful fertilization. Sperm bind ZP by means of membrane receptors which recognize carbohydrate moieties on ZP glycoproteins according to a well-defined sequential process. Primary binding receptors, many of which have been disclosed in various mammals, are localized throughout the acrosomal region of the sperm surface. A panel of monoclonal antibodies against proteins from the sperm surface was prepared. Antibodies were screened by immunofluorescence for protein localization and Western blotting. Proteins localized on the sperm head and simultaneously detected by Western blotting were further studied in terms of immunolocalization in reproductive tissues and fluids, binding to ZP, immunoprecipitation and sequencing. Of 17 prepared antibodies, 8 recognized proteins localized on the sperm head and also detected proteins of interest by Western blotting. Only three other antibodies recognized proteins that also coincided in binding to ZP. These three antibodies were used for immunoprecipitation, and further protein sequencing of immunoprecipitates revealed that these antibodies distinguished acrosin precursor, RAB-2A protein, and lactadherin P47. This is not the first time we have detected acrosin on the surface of ejaculated and capacitated sperm. However, to our knowledge, this is the first time RAB-2A has been detected on the sperm surface. Lactadherin P47 has already been characterized and its physiological function in reproduction has been proposed.

• MeSH
• fluorescenční protilátková technika MeSH
• interakce spermie a vajíčka * MeSH
• molekulová hmotnost MeSH
• monoklonální protilátky metabolismus   MeSH
• prasata MeSH
• receptory buněčného povrchu metabolismus   MeSH
• spermie metabolismus   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• monoklonální protilátky MeSH
• receptory buněčného povrchu MeSH

Polyspermically penetrated human zona-free eggs prepared from oocytes that had failed to be fertilized in an in-vitro fertilization programme were used. The pronuclear synthetic activity was evaluated by high-resolution autoradiography and correlated with the development of pronuclear structure. Incorporation of [3H]-thymidine, signalling the occurrence of a DNA synthetic phase, was only detected in structurally fully developed pronuclei previously shown to appear no sooner than 12 h after gamete union. However, [3H]adenosine was incorporated into very early pronuclei which had not yet completed the development of their nuclear envelopes and which first appeared about 4 h after sperm-egg fusion. In the absence of DNA synthesis (shown by the lack of thymidine incorporation), this early adenosine incorporation apparently reflects an early pronuclear RNA synthesis. Taken together, these results indicate that nucleic acid synthesis in human male pronuclei is tightly bound to the development of a corresponding pronuclear structure and that DNA synthesis, beginning about 12 h after fertilization, is preceded by a slight but evident RNA synthesis taking place during an early stage of human male pronuclear formation.

• MeSH
• adenosin metabolismus   MeSH
• blastocysta metabolismus   MeSH
• buněčné jádro metabolismus   ultrastruktura   MeSH
• DNA biosyntéza   MeSH
• elektronová mikroskopie MeSH
• interakce spermie a vajíčka * MeSH
• lidé MeSH
• RNA biosyntéza   MeSH
• spermie ultrastruktura   MeSH
• thymidin metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adenosin MeSH
• DNA MeSH
• RNA MeSH
• thymidin MeSH

PROBLEM: The role of Ala-Trp-Asn (AWN) and Ala-Gln-Asn (AQN) families of spermadhesive sperm proteins in fertilization. METHOD OF STUDY: The preparation and characterization of polyclonal antibodies against AWN and AQN spermadhesins and one monoclonal antibody (MAb), designated Bo.5, against AWN spermadhesin. The use of biochemical and immunocytochemical methods for characterization of spermadhesins on the sperm membrane of boar spermatozoa and in the cryostat sections of boar reproductive organs. RESULTS: Polyclonal anti-AWN and anti-AQN antibodies specifically reacted with AWN and AQN proteins, respectively. MAb Bo.5 detected the 17-, 16-, and 14-kDa protein members of AWN subfamily. The monoclonal, as well as the polyclonal, AWN antibodies remarkably decreased the sperm binding to the egg surface in an in vitro sperm zona pellucida binding assay. CONCLUSIONS: Presented results demonstrate that polyclonal antibodies and MAb Bo.5 against spermadhesins specifically recognize the membrane-associated antigens and inhibit the binding of sperm to oocytes. Reduced binding of sperm to oocytes, due to the antibodies, indicates the role of these spermadhesins in sperm-egg primary binding.

• MeSH
• fluorescenční protilátková technika nepřímá MeSH
• glykoproteiny imunologie   izolace a purifikace   fyziologie   MeSH
• interakce spermie a vajíčka imunologie   MeSH
• králíci MeSH
• lidé MeSH
• molekuly buněčné adheze imunologie   metabolismus   fyziologie   MeSH
• monoklonální protilátky biosyntéza   chemie   metabolismus   MeSH
• oocyty imunologie   metabolismus   fyziologie   MeSH
• prasata MeSH
• receptory buněčného povrchu MeSH
• spermie imunologie   metabolismus   fyziologie   MeSH
• vazebná místa protilátek * MeSH
• zona pellucida metabolismus   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• egg surface sperm receptor MeSH Prohlížeč
• glykoproteiny MeSH
• molekuly buněčné adheze MeSH
• monoklonální protilátky MeSH
• receptory buněčného povrchu MeSH

The developmental ability of hybrid zygotes, produced by in vitro fertilization of in vitro matured bovine oocytes with ram sperm, was evaluated by gross morphology, autoradiographic detection of (5-3H) uridine incorporation, and fine structure morphology. Fertilization was successful in 83% of bovine oocytes inseminated with bull sperm (control embryos) compared with 67% of bovine oocytes inseminated with ram sperm (hybrid embryos) and in both cases appeared two regularly developed pronuclei. Two-cell embryos were transferred to ewe oviducts and allowed to develop to the 8-cell stage. Although the ability of hybrid embryos to reach 8-cell stage was similar to that of control embryos, in nuclei of hybrid embryos the transition from maternal to embryonic genome control assessed according to the onset of RNA synthesis indicated the differences in the frequency of labelled nuclei and intensity of their labelling. In hybrid embryos these parameters were remarkably lower and may reflect the developmental failure of hybrid embryos. These observations are consistent with delay or inefficient reactivation of the embryonic genome in the hybrid embryos.

• MeSH
• buněčné jádro ultrastruktura   MeSH
• chiméra * genetika   MeSH
• embryo savčí cytologie   diagnostické zobrazování   ultrastruktura   MeSH
• fertilizace in vitro MeSH
• oocyty cytologie   MeSH
• ovce MeSH
• přenos embrya MeSH
• radiografie MeSH
• RNA analýza   MeSH
• skot MeSH
• spermie cytologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• RNA MeSH

A total of 271,547 records of semen collections were utilized to appraise sperm characteristics of 3319 boars belonging to eight breeds: Czech Large White (CLW), Czech Landrace (CLA), Prestice Black-Pied (PBP), Czech Meat Pig (CM), Hampshire (HA), Duroc (DC), Pietrain (PN), Large White (LW), and various crosses of these breeds. The data was collected over 8 years (1990-1997) from insemination stations for boars in the Czech Republic. The assessment of sperm output was based on semen volume, number of total spermatozoa and number of viable spermatozoa. A linear model was used for statistical analysis included fixed effects of breed or crossbred combinations, boar within breed or crossbred combinations, year-season, and linear and quadratic regression on age of boars at collection and on interval between collections. The average semen volume of boars ranged from 161 to 349 ml, number of total spermatozoa from 81x10(9) to 119x10(9) and number of viable spermatozoa from 60x10(9) to 86x10(9). The lowest values were detected in DC while the highest were observed in LW. In general, sperm output significantly differed across breeds and their crossbreeds. The highest heterosis effect for semen volume was 30.6% (HA x PN), for number of total spermatozoa 18.2% (HA x PN) and 10.4% for number of viable spermatozoa (CLA x DC). Sperm output varied with season, including high values in autumn and winter and low ones in spring and summer.

• MeSH
• chov MeSH
• druhová specificita MeSH
• hybridní efekt genetika   MeSH
• křížení genetické MeSH
• lineární modely MeSH
• počet spermií * MeSH
• prasata genetika   fyziologie   MeSH
• roční období MeSH
• sperma fyziologie   MeSH
• spermie fyziologie   MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH