Differences in the strategies of mammalian and fish fertilization and the optimization of the protocol for immunofluorescence on fish spermatozoa
Koubek, Pavel; Králová, Alena; Pšenička, M.; Pěknicová, Jana
2008 - English
This lecture describes a special approach to handling fish spermatozoa in comparison with the symplex originating from mammals. This approach primarily emphasizes the concrete parts of immunofluorescence protocol determining accurate results. Vzhledem ke struktuře rybích spermií, která je jednoduchá a snadno poškoditelná, bylo třeba vypracovat protokol pro imunofluorescenční značení, který by minimalizoval jejich poškození a vznik nespecifických artefaktů. Cílem práce tak bylo především zamezit mylné interpretaci výsledků imunofluorescenčních experimentů na rybích spermiích a zvýšit vzájemnou porovnatelnost těchto experimentů.
Keywords:
fish spermatozoa; mammalian spermatozoa; immunofluorescence
Available at various institutes of the ASCR
Differences in the strategies of mammalian and fish fertilization and the optimization of the protocol for immunofluorescence on fish spermatozoa
This lecture describes a special approach to handling fish spermatozoa in comparison with the symplex originating from mammals. This approach primarily emphasizes the concrete parts of ...
Role povrchových proteinů v reprodukci
Jonáková, Věra; Postlerová, Pavla; Tichá, M.; Pěknicová, Jana
2008 - Czech
Během ejakulace se proteiny semenné plasmy navazují na povrch spermie a tvoří povrchové proteiny spermie, které mohou změnit vazebné vlastnosti spermie. DQH povrchový protein spermie vykazuje afinitu k oviduktálnímu epitelu a glykoproteinům zona pellucida (ZP). DQH mRNA byla detekována v semenných vacích (SV). DQH byl imunodetekován monoklonálními protilátkami (Mabs)v extraktu a tekutině i na tkáňových řezech SV vaků a na plasmatické membráně akrosomu ejakulovaných spermií. Výsledky potvrzují schopnost DQH proteinu se vázat na povrch spermie při ejakulaci, účast při tvorbě rezervoáru spermií v prasečím oviduktu. Spermie preinkubované s Mabs proti DQH proteinu před koinkubací s vajíčkem redukují vazbu spermie k vajíčku. Pro primární vazbu spermie s ZP glykoproteiny vajíčka je důležitý intaktní DQH protein na povrchu spermie. Seminal plasma proteins bind to the sperm surface at ejaculation and may modulate sperm properties during reproduction. DQH sperm surface protein shows affinity to oviductal epithelium and zona pellucida (ZP) glycoproteins. The mRNA transcript of DQH protein was found in seminal vesicles (SV). DQH was immunodetected by monoclonal antibodies (MAbs) in SV extract and fluid, on SV tissue sections and on the membrane-associated acrosome part of ejaculated spermatozoa. These results confirmed the ability of DQH protein to bind to the sperm surface at ejaculation and to participate in the formation of the sperm reservoir in the porcine oviduct. Moreover, monoclonal antibodies reduced binding of sperm to oocytes and proved the role of DQH protein in the sperm-ZP primary binding.
Keywords:
sperm surface protein; spermatozoa; speradhesin
Available at various institutes of the ASCR
Role povrchových proteinů v reprodukci
Během ejakulace se proteiny semenné plasmy navazují na povrch spermie a tvoří povrchové proteiny spermie, které mohou změnit vazebné vlastnosti spermie. DQH povrchový protein spermie vykazuje afinitu ...
Analysis of male sterility caused by T(16,17)43H chromosomal translocation
Čapková, Jana; Homolka, David; Ivánek, Robert; Jansa, Petr; Forejt, Jiří
2008 - English
Heterozygosity for certain mouse and human chromosomal rearrangements is characterized by male-limited sterility. Our model of a male-sterile chromosomal rearrangement represents mouse T(16;17)43H (T43) chromosomal translocation. The effect of the T43H translocation on spermatogenesis was studied. We analyzed meiosis in B10.T43/+ heterozygous male mice and we have obtained direct evidence on the transcriptional silencing of unsynapsed autosomal chromatin of the translocation quadrivalent and the subsequent failure of the X chromosome transcriptional inactivation at the mid-late pachytene of spermatogenesis. Po některých chromosomálních přestavbách vzniká heterozygosita, která vede ke sterilitě samců. Náš model chromosomální přestavby vedoucí samčí sterilitě představuje myší T(16,17)43H chromosomální translokace. Na tomto modelu jsme studovali účinek translokace na spermatogenezu. Analyzovali jsme meiosu u heterozygotních T43/+ samců a prokázali jsme transkripčním umlčení nesynaptického autosomálního chromatinu v translokačním kvadrivalentu a následnou neschopnost transkripční inaktivace X chromosomu v pachytenním stadiu spermatogeneze.
Keywords:
male sterility; chromosomal translocation; spermatogenesis
Available at various institutes of the ASCR
Analysis of male sterility caused by T(16,17)43H chromosomal translocation
Heterozygosity for certain mouse and human chromosomal rearrangements is characterized by male-limited sterility. Our model of a male-sterile chromosomal rearrangement represents mouse T(16;17)43H ...
Swim-up in assisted reproduction
Teplá, O.; Pěknicová, Jana; Mrázek, M.; Margaryan, Hasmik; Cibulková, E.; Králová, Alena; Strnadová, J.
2008 - English
To compare the effect on density-gradient centrifugation and swim-up on sperm preparation in assisted reproduction. Quality of sperm was evaluated by monoclonal antibodies.
Keywords:
swim-up; IVF; human spermatozoa
Available at various institutes of the ASCR
Swim-up in assisted reproduction
To compare the effect on density-gradient centrifugation and swim-up on sperm preparation in assisted reproduction. Quality of sperm was evaluated by monoclonal antibodies.
Expression and localization of acrosin inhibitor in the boar reproductive tract
Jonáková, Věra; Davidová, Nina; Maňásková, Pavla
2008 - English
Proteinase inhibitors present in seminal plasma inactivate prematurely released sperm acrosin and protect spermatozoa and reproductive epithelium of the male and female reproductive tracts against proteolytic degradation. Acrosin inhibitor mRNA was detectable in epididymis, seminal vesicles, prostate and Cowperś glands. By indirect immunofluorescence, acrosin inhibitors were detected in the secretory tissues of cauda epididymis, prostate and SV, and on epididymal and ejaculated spermatozoa in the acrosomal region. The presence of inhibitors of the proteolytic enzymes in epididymis is very important for the regulation of the sperm membrane protein processing. Inhibitory proteinas v semenné plasmě inhibují předčasně uvolněnou spermiovou proteinasu akrosin a ochraňují tak spermie a epitel samčího a samičího traktu proti proteolyse. Akrosin inhibitor mRNA byla nalezena v epididymu, v semených vacích, prostatě a Cowperových žlazách. Nepřímou imunofluorescencí inhibitor akrosinu byl detekován v sekreční tkáni cauda epididymu, prostatě a semených vacích a v akrosomální oblasti na epididymálních a ejakulovaných spermií. Přítomnost inhibitorů proteolytických enzymů v epididymu je důležitá pro processing spermiových membránových proteinů.
Keywords:
acrosin; acrosin inhibitor; boar reproductive tract
Available at various institutes of the ASCR
Expression and localization of acrosin inhibitor in the boar reproductive tract
Proteinase inhibitors present in seminal plasma inactivate prematurely released sperm acrosin and protect spermatozoa and reproductive epithelium of the male and female reproductive tracts against ...
Detection of transferrin in placentae from normal and abnormal pregnancies
Králová, Alena; Světlíková, M.; Madar, J.; Ulčová-Gallová, Z.; Bukovský, A.; Pěknicová, Jana
2008 - English
The placenta is one of the important sites for iron metabolism in human. It transfers the iron from the mother to the fetus. One of the major iron transport proteins is transferrin, which is a blood plasma protein crucial for iron uptake. Its localization and expression may be one of the markers to distinguish placental dysfunction. Differences of transferrin expression were foundbetween normal and abnormal placentae. Variations of its expression were detected in villous syncytiotrophoblast, which is in direct contact with maternal blood. In normal placentae, the expression of transferrin in syncytium was lower comparing to placentae with distinct pathologies. The results indicate that in the case of abnormal pregnancies, the fetus may require higher levels of transferrin in order to prevent iron depletion due to the stress from the placenta dysfunction Placenta je důležité místo pro metabolismus železa u člověka.Přenáší železo z matky pro plod. Jeden z hlavních transportních proteinů je transferrin, který jako protein krevní plazmy je rozhodující pro vstup železa. Jeho lokalizace a exprese může být jedním z ukazatelů dysfunkce placenty. Změny v expresi transferinu byly objeveny v klkatém syncitiotrofoblastu, který je v přímém kontaktu s krví matky. V placentě z normálního těhotenství, byla exprese transferrinu nižší nž u placenty z abnormálního těhotenství.Tyto výsledky ukazují na možnost, v případě abnormálních těhotenství, že plody mohou požadovat vyšší hladinu transferrinu k tomu, aby se zabránilo vyčerpání železa kvůli dysfunkci placenty.
Keywords:
transferrin; monoclonal antibody
Available at various institutes of the ASCR
Detection of transferrin in placentae from normal and abnormal pregnancies
The placenta is one of the important sites for iron metabolism in human. It transfers the iron from the mother to the fetus. One of the major iron transport proteins is transferrin, which is a blood ...
Sperm surface proteins: Their origin, biochemical properties and role in reproduction
Jonáková, Věra; Postlerová, Pavla; Tichá, M.; Pěknicová, Jana
2008 - English
Seminal plasma proteins bind to the sperm surface at ejaculation and may modulate sperm properties during reproduction. Porcine spermadhesins (AQN, AWN, PSP) are secreted mainly by the seminal vesicles (SV), but their mRNAs have been found also in the cauda epididymis and prostate. Using specific polyclonal antibodies, PSP-I and PSP-II proteins were immunodetected in tissue extracts from cauda epididymis, prostate, SV and Cowper´s glands on the blots, and in secretory tissues of cauda epididymis, prostate and SV by indirect immunofluorescence. We localized PSP spermadhesins on epididymal and ejaculated spermatozoa. PSP proteins are produced not only by SV and prostate, but also by epididymis. Characterization of seminal plasma proteins expressed in the individual reproductive organs might help to understand their role in the reproduction process. Proteiny semenné plasmy se váží na povrch spermie při ejakulaci a modulují vlastnosti spermie při reprodukci. Kančí spermadhesiny (AQN, AWN, PSP) jsou sekretovány hlavně semenými vaky (SV), ale jejich mRNA byla nalezena také v cauda epididymis a prostatě. Pomocí specifických protilátek PSP I a PSP II proteiny byly imunodetekovány na blotu v tkáňových extraktech cauda epididymis, prostatě, SV, a nepřímou imunofluorescencí v sekreční tkáni cauda epididymis, prostatě, SV. Lokalizovali jsme PSP spermadhesiny na epididymálnícj a ejakulovaných spermiích. PSP proteiny jsou produkovány SV, prostatou a také nadvarletem. Charakterizace proteinů semenné plasmy exprimovaných v jednotlivých reprodukčních orgánech napomůže pochopit jejich úlohu v reprodukci
Keywords:
sperm surface protein; spermadhesin; monoclonal antibody
Available at various institutes of the ASCR
Sperm surface proteins: Their origin, biochemical properties and role in reproduction
Seminal plasma proteins bind to the sperm surface at ejaculation and may modulate sperm properties during reproduction. Porcine spermadhesins (AQN, AWN, PSP) are secreted mainly by the seminal ...
The effect of low dose of vinclozolin on reproductive tract development in CD1 outbred mice
Pěknicová, Jana; Elzeinová, Fatima; Nováková, Vendula; Buckiová, Daniela; Kubátová, Alena
2008 - English
The effect of a low dose of vinclozolin within the development of the reproductive tract during gestation (VIN-GD 15-22) and puberty (VIN-PND 23-44) in CD1 mice was tested. We found a decrease in the anogenital distance, prostate weight and pathology of testes in both experimental groups. Sperm counts decreased to 46% (VIN-GD) and to 81% (VIN-PND), and also the acrosomal state (evaluated by antiacrosomal antibody) decreased in both groups to 89% in comparison to the control group (100%). Sperm head abnormalities increased by approximately 18% and 13%, respectively. In this connection, the expression of some genes was changed (arosome-related gene (Acr), apoptosis related genes (p53, p21)). In conclusion, a low dose of vinclozolin affected the reproductive tract, sperm parameters and expression of selected genes in both experimental groups.
Keywords:
vinclozolin; spermatozoa
Available at various institutes of the ASCR
The effect of low dose of vinclozolin on reproductive tract development in CD1 outbred mice
The effect of a low dose of vinclozolin within the development of the reproductive tract during gestation (VIN-GD 15-22) and puberty (VIN-PND 23-44) in CD1 mice was tested. We found a decrease in the ...
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