INTRODUCTION: In this study we detail the effect of different fixation agents and the duration of storage has on the immunohistochemical staining positivity of samples of archival embryonic and fetal tissues. MATERIALS AND METHODS: The samples were stained by indirect two-step immunohistochemistry (IHC) method for Ki-67, cyclin A and β-actin. RESULTS: Irrespective of the length of tissue archiving, tissue fixation with 10% neutral buffered formalin had better IHC intensity results in all cases when compared to methacarn-fixed tissues. In the case of β-actin, this difference was statistically significant, while differences in Ki-67 and cyclin A were not. The second aspect studied was which effect tissue block archiving duration has on the IHC reactivity. We demonstrated a statistically significant decrease in IHC positivity for all studied antigens between the samples that were archived for 10-19 or 20-45 years, regardless the fixative solution. CONCLUSION: To the best of our knowledge, the influence that the duration of tissue block archiving has on IHC positivity in human embryo and fetal tissue material has not yet been studied. Although the causes of the IHC positivity decline in archived tissue blocks are not well understood, a possible decrease in IHC over time should be considered, particularly in retrospective studies.
- MeSH
- aktiny analýza MeSH
- antigen Ki-67 analýza MeSH
- barvení a značení normy MeSH
- časové faktory MeSH
- chloroform MeSH
- cyklin A analýza MeSH
- fixace tkání metody MeSH
- fixativa MeSH
- formaldehyd MeSH
- gestační stáří MeSH
- imunohistochemie normy MeSH
- játra embryologie MeSH
- králíci MeSH
- kyselina octová MeSH
- lidé MeSH
- methanol MeSH
- myši MeSH
- placenta embryologie MeSH
- střeva embryologie MeSH
- těhotenství MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- lidé MeSH
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
CYP2C and CYP2 J enzymes, commonly named as cytochrome P450 (CYP) epoxygenases, convert arachidonic acid to four regioisomeric epoxyeicosatrienoic acids (EETs), biologically active eicosanoids with many functions in organism. EETs are rapidly hydrolysed to less active dihydroxyeicosatrienoic acids (DHETs) by soluble epoxide hydrolase (sEH). We investigated spatio-temporal expression pattern of CYP2C8, CYP2C9, CYP2 J2 and sEH in normal human placenta by immunohistochemical method. In the villous trophoblast, CYP2C8 was the most abundant protein. Its expression is higher than the CYP2C9 and CYP2 J2 in the cytotrophoblast in the embryonic stage of development and remains higher in syncytiotrophoblast of term placenta. Unlike to CYP2C8, CYP2C9 and CYP2 J2 expression decrease in term placenta. sEH expression increases with gestation age and is strictly limited to cytotrophoblast in embryonic and foetal stages of the development. Moreover, CYP2C8 shows more intensive staining than the other protein monitored in Hofbauer cells in villous stroma. Specific information regarding the exact role of EETs and DHETs functions in a normal placenta is still unknown. Based on CYP epoxygenases and sEH localization and well known information about the functions of placental structures during development, we suggest that these enzymes could play different roles in various cell populations in the placenta. As the placenta is absolutely crucial for prenatal development, arachidonic acid is essential part of human nutrient and CYP epoxygenases expression can be affected by xenobiotics, further investigation of the exact role of CYP epoxygenases, sEH, and their metabolites in normal pregnancy and under pathological conditions is needed.
- MeSH
- cytochrom P450 CYP2C8 biosyntéza MeSH
- cytochrom P450 CYP2C9 biosyntéza MeSH
- lidé MeSH
- placenta cytologie enzymologie MeSH
- regulace genové exprese enzymů fyziologie MeSH
- systém (enzymů) cytochromů P-450 biosyntéza MeSH
- těhotenské proteiny biosyntéza MeSH
- těhotenství metabolismus MeSH
- Check Tag
- lidé MeSH
- těhotenství metabolismus MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- biologické markery MeSH
- C-reaktivní protein * MeSH
- lidé MeSH
- míra přežití MeSH
- nemalobuněčný karcinom plic * diagnóza farmakoterapie MeSH
- pemetrexed * aplikace a dávkování farmakologie terapeutické užití MeSH
- prognóza MeSH
- progrese nemoci MeSH
- retrospektivní studie MeSH
- stupeň závažnosti nemoci MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
Three ABC transporters (MDR1, MRP1, BCRP), belonging to the family of multidrug resistance (MDR) proteins, play a crucial role in the protection mechanisms during embryogenesis and mediate drug resistance in cancer cells. The distribution of these transporters in the series of human embryonal/fetal intestine, liver and kidneys of various stages of intrauterine development (IUD) by indirect two-step immunohistochemical method was investigated. The organ- and age-specific expression patterns of these transporters were depicted and compared with the expression in adult organs. The evaluation of intestine and liver samples demonstrate differences in expression pattern of ABC transporters during IUD. On the contrary, in kidneys the age-specific localization was not observed. However, the increasing positivity from the kidney surface towards deeper, more differentiated parts was found. Hopefully, our study may contribute to elucidation of the role of multidrug resistance (MDR) pathways during IUD in man.
- MeSH
- ABC transportéry biosyntéza genetika metabolismus MeSH
- embryonální vývoj genetika fyziologie MeSH
- exprese genu MeSH
- játra embryologie MeSH
- ledviny embryologie MeSH
- lidé MeSH
- mnohočetná léková rezistence MeSH
- P-glykoproteiny biosyntéza genetika metabolismus MeSH
- střeva embryologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
