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Immunofluorescence confocal laser scanning microscopy and immuno-electron microscopic identification of keratins in human materno-foetal interaction zone.

Ahenkorah J, Hottor B, Byrne S, Bosio P, Ockleford CD - J. Cell. Mol. Med. (2008)

Bottom Line: This difference between healthy and pre-eclamptic chorionic villous trophoblast keratin expression was statistically significant in 4 out of the 5 keratins.This was not the case for the extravillous trophoblast at the immunofluorescence confocal level but significant differences were obtained using immunogold electron microscopy.We suggest that the villous trophoblast in pre-eclamptic placentae is cytoskeletally weaker with respect to the filaments made from these specific proteins and that this is one reason why, in pre-eclampsia, trophoblast is deported in greater quantity than in healthy placentae.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection Immunity and Inflammation, University of Leicester Medical School, UK.

ABSTRACT
We show here that at least 5 keratin proteins are present in villous trophoblast and the same 5 in extravillous trophoblast. A further 14 tested were undetectable in these tissues. In contrast, 10 of the 19 keratins tested were present in amniotic epithelium. The marking of amniotic epithelium on the one hand, as distinct from villous and extravillous trophoblast on the other, can be achieved using 5 keratins (K4, 6, 13, 14 and 17) with a mixture of positive and negative discrimination that is expected, in combination, to be highly sensitive. All the specific keratins identified in trophoblast were apparently up-regulated on the pathway to extravillous trophoblast. Co-ordinated differentiation at the molecular expression level is indicated by this finding. The relevant keratins are K5, 7, 8, 18 and 19. Specific keratins have been identified that are down-regulated in villous trophoblast in pre-eclamptic pregnancy. This difference between healthy and pre-eclamptic chorionic villous trophoblast keratin expression was statistically significant in 4 out of the 5 keratins. This was not the case for the extravillous trophoblast at the immunofluorescence confocal level but significant differences were obtained using immunogold electron microscopy. We suggest that the villous trophoblast in pre-eclamptic placentae is cytoskeletally weaker with respect to the filaments made from these specific proteins and that this is one reason why, in pre-eclampsia, trophoblast is deported in greater quantity than in healthy placentae.

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Related in: MedlinePlus

Boxplot showing interquartile range of the CLSM immunofluorescence data using the anti-keratin-7 antibody on pre-eclamptic chorionic villous trophoblast (CVTp) and extravillous trophoblast (EVTp), also healthy chorionic villous trophoblast (CVTh) and extravillous trophoblast (EVTh). The minimum, median and maximum values are indicated by the key and the whisker shows the range.
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fig04: Boxplot showing interquartile range of the CLSM immunofluorescence data using the anti-keratin-7 antibody on pre-eclamptic chorionic villous trophoblast (CVTp) and extravillous trophoblast (EVTp), also healthy chorionic villous trophoblast (CVTh) and extravillous trophoblast (EVTh). The minimum, median and maximum values are indicated by the key and the whisker shows the range.

Mentions: Using COMOS software, images of anti-keratin immunofluorescence were obtained for healthy and pre-eclamptic placentae, comprising both chorionic villous trophoblast (CVT) and extravillous trophoblast (EVT) of the basal plate tissue. Images were measured using the same rectangular boxed area of 340.80 × 226.60 μm (77,225.28 μm2). Percentage of pixels with intensity in the high intensity (Band 210–255) for both (CVT) and (EVT) were recorded. The criteria for taking measurements were to randomly select regions of EVTs or CVTs on the same basal plate tissue image and to ensure that each area selected to be measured did not overlap an area already chosen. Paired measurements of areas of EVT and CVT in each image from the same section were made. This was done to minimize the effects on the comparison of inter-preparation variation. The area to be measured contained only either EVT or CVT. Results were recorded in Excel spreadsheets and statistical analyses were carried out using STATA™ version 9.2 (StataCorp, College Station, TX, USA). The n values listed in Tables 2–5 and used for Figs 4–7 were the number of boxed areas scanned. Where quantitative data are presented all the antibodies were applied to the same range of tissues.


Immunofluorescence confocal laser scanning microscopy and immuno-electron microscopic identification of keratins in human materno-foetal interaction zone.

Ahenkorah J, Hottor B, Byrne S, Bosio P, Ockleford CD - J. Cell. Mol. Med. (2008)

Boxplot showing interquartile range of the CLSM immunofluorescence data using the anti-keratin-7 antibody on pre-eclamptic chorionic villous trophoblast (CVTp) and extravillous trophoblast (EVTp), also healthy chorionic villous trophoblast (CVTh) and extravillous trophoblast (EVTh). The minimum, median and maximum values are indicated by the key and the whisker shows the range.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822880&req=5

fig04: Boxplot showing interquartile range of the CLSM immunofluorescence data using the anti-keratin-7 antibody on pre-eclamptic chorionic villous trophoblast (CVTp) and extravillous trophoblast (EVTp), also healthy chorionic villous trophoblast (CVTh) and extravillous trophoblast (EVTh). The minimum, median and maximum values are indicated by the key and the whisker shows the range.
Mentions: Using COMOS software, images of anti-keratin immunofluorescence were obtained for healthy and pre-eclamptic placentae, comprising both chorionic villous trophoblast (CVT) and extravillous trophoblast (EVT) of the basal plate tissue. Images were measured using the same rectangular boxed area of 340.80 × 226.60 μm (77,225.28 μm2). Percentage of pixels with intensity in the high intensity (Band 210–255) for both (CVT) and (EVT) were recorded. The criteria for taking measurements were to randomly select regions of EVTs or CVTs on the same basal plate tissue image and to ensure that each area selected to be measured did not overlap an area already chosen. Paired measurements of areas of EVT and CVT in each image from the same section were made. This was done to minimize the effects on the comparison of inter-preparation variation. The area to be measured contained only either EVT or CVT. Results were recorded in Excel spreadsheets and statistical analyses were carried out using STATA™ version 9.2 (StataCorp, College Station, TX, USA). The n values listed in Tables 2–5 and used for Figs 4–7 were the number of boxed areas scanned. Where quantitative data are presented all the antibodies were applied to the same range of tissues.

Bottom Line: This difference between healthy and pre-eclamptic chorionic villous trophoblast keratin expression was statistically significant in 4 out of the 5 keratins.This was not the case for the extravillous trophoblast at the immunofluorescence confocal level but significant differences were obtained using immunogold electron microscopy.We suggest that the villous trophoblast in pre-eclamptic placentae is cytoskeletally weaker with respect to the filaments made from these specific proteins and that this is one reason why, in pre-eclampsia, trophoblast is deported in greater quantity than in healthy placentae.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection Immunity and Inflammation, University of Leicester Medical School, UK.

ABSTRACT
We show here that at least 5 keratin proteins are present in villous trophoblast and the same 5 in extravillous trophoblast. A further 14 tested were undetectable in these tissues. In contrast, 10 of the 19 keratins tested were present in amniotic epithelium. The marking of amniotic epithelium on the one hand, as distinct from villous and extravillous trophoblast on the other, can be achieved using 5 keratins (K4, 6, 13, 14 and 17) with a mixture of positive and negative discrimination that is expected, in combination, to be highly sensitive. All the specific keratins identified in trophoblast were apparently up-regulated on the pathway to extravillous trophoblast. Co-ordinated differentiation at the molecular expression level is indicated by this finding. The relevant keratins are K5, 7, 8, 18 and 19. Specific keratins have been identified that are down-regulated in villous trophoblast in pre-eclamptic pregnancy. This difference between healthy and pre-eclamptic chorionic villous trophoblast keratin expression was statistically significant in 4 out of the 5 keratins. This was not the case for the extravillous trophoblast at the immunofluorescence confocal level but significant differences were obtained using immunogold electron microscopy. We suggest that the villous trophoblast in pre-eclamptic placentae is cytoskeletally weaker with respect to the filaments made from these specific proteins and that this is one reason why, in pre-eclampsia, trophoblast is deported in greater quantity than in healthy placentae.

Show MeSH
Related in: MedlinePlus