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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

(A & B) This is an indirect immuno-fluorescence preparation using anti-keratin 7. The areas sampled for the intensity distribution graphs are shown on the left and the pixels of highest grey scale value (210–255) equivalent to immunofluorescence intensity have been banded red. The red banding corresponds to the location of EVT and is only infrequently seen in chorionic villous trophoblast. The graph on the right shows the pixel intensity distribution of high greyscale pixels within this brightness band over areas of chorionic villous trophoblast (pink line) and extravillous trophoblast (blue line).
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fig03: (A & B) This is an indirect immuno-fluorescence preparation using anti-keratin 7. The areas sampled for the intensity distribution graphs are shown on the left and the pixels of highest grey scale value (210–255) equivalent to immunofluorescence intensity have been banded red. The red banding corresponds to the location of EVT and is only infrequently seen in chorionic villous trophoblast. The graph on the right shows the pixel intensity distribution of high greyscale pixels within this brightness band over areas of chorionic villous trophoblast (pink line) and extravillous trophoblast (blue line).

Mentions: The degree of difference in immunofluorescence intensity can be highlighted by a banding procedure (Figs 2 and 3, left panels), where a threshold of immunofluorescence intensity is set and all pixels in the image above that level of intensity are colour-coded (here in red). A plot of the incidence of high-intensity pixels in the range 210–255 comparing areas of the image occupied by EVT (blue) and CVT (pink) shows a consistent excess of high greyscale value pixels in the EVT areas. The pattern of expression of specific keratin immunofluorescence is described in Figs 4–7 and the statistical analysis is shown in Tables 2–5. The overall picture is summarised in Table 6.


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)

(A & B) This is an indirect immuno-fluorescence preparation using anti-keratin 7. The areas sampled for the intensity distribution graphs are shown on the left and the pixels of highest grey scale value (210–255) equivalent to immunofluorescence intensity have been banded red. The red banding corresponds to the location of EVT and is only infrequently seen in chorionic villous trophoblast. The graph on the right shows the pixel intensity distribution of high greyscale pixels within this brightness band over areas of chorionic villous trophoblast (pink line) and extravillous trophoblast (blue line).
© Copyright Policy
Related In: Results  -  Collection

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

fig03: (A & B) This is an indirect immuno-fluorescence preparation using anti-keratin 7. The areas sampled for the intensity distribution graphs are shown on the left and the pixels of highest grey scale value (210–255) equivalent to immunofluorescence intensity have been banded red. The red banding corresponds to the location of EVT and is only infrequently seen in chorionic villous trophoblast. The graph on the right shows the pixel intensity distribution of high greyscale pixels within this brightness band over areas of chorionic villous trophoblast (pink line) and extravillous trophoblast (blue line).
Mentions: The degree of difference in immunofluorescence intensity can be highlighted by a banding procedure (Figs 2 and 3, left panels), where a threshold of immunofluorescence intensity is set and all pixels in the image above that level of intensity are colour-coded (here in red). A plot of the incidence of high-intensity pixels in the range 210–255 comparing areas of the image occupied by EVT (blue) and CVT (pink) shows a consistent excess of high greyscale value pixels in the EVT areas. The pattern of expression of specific keratin immunofluorescence is described in Figs 4–7 and the statistical analysis is shown in Tables 2–5. The overall picture is summarised in Table 6.

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