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Pulse of inflammatory proteins in the pregnant uterus of European polecats ( Mustela putorius ) leading to the time of implantation

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ABSTRACT

Uterine secretory proteins protect the uterus and conceptuses against infection, facilitate implantation, control cellular damage resulting from implantation, and supply pre-implantation embryos with nutrients. Unlike in humans, the early conceptus of the European polecat (Mustela putorius; ferret) grows and develops free in the uterus until implanting at about 12 days after mating. We found that the proteins appearing in polecat uteri changed dramatically with time leading to implantation. Several of these proteins have also been found in pregnant uteri of other eutherian mammals. However, we found a combination of two increasingly abundant proteins that have not been recorded before in pre-placentation uteri. First, the broad-spectrum proteinase inhibitor α2-macroglobulin rose to dominate the protein profile by the time of implantation. Its functions may be to limit damage caused by the release of proteinases during implantation or infection, and to control other processes around sites of implantation. Second, lipocalin-1 (also known as tear lipocalin) also increased substantially in concentration. This protein has not previously been recorded as a uterine secretion in pregnancy in any species. If polecat lipocalin-1 has similar biological properties to that of humans, then it may have a combined function in antimicrobial protection and transporting or scavenging lipids. The changes in the uterine secretory protein repertoire of European polecats is therefore unusual, and may be representative of pre-placentation supportive uterine secretions in mustelids (otters, weasels, badgers, mink, wolverines) in general.

No MeSH data available.


Changes in European polecat uterine secretory proteins with time after mating. Animals sampled on days 4 and 14 were non-pregnant. The sample volumes were adjusted to normalize the intensity of the strong band at approximately 65 kDa (serum albumin). See electronic supplementary material, figure S1, for SDS-PAGE of all of the samples collected and upon which the adjustments were based. Gel band codes are indicated by letters and are referred to in the text and in table 1. The proteins that most clearly increased in concentration with time were α2-macroglobulin (uniquely identified in bands A, B, J and K) and lipocalin-1 (uniquely identified in bands G and U). M, marker/calibration proteins with relative mobilities (Mr) as indicated in kilodaltons (kDa).
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RSOS161085F1: Changes in European polecat uterine secretory proteins with time after mating. Animals sampled on days 4 and 14 were non-pregnant. The sample volumes were adjusted to normalize the intensity of the strong band at approximately 65 kDa (serum albumin). See electronic supplementary material, figure S1, for SDS-PAGE of all of the samples collected and upon which the adjustments were based. Gel band codes are indicated by letters and are referred to in the text and in table 1. The proteins that most clearly increased in concentration with time were α2-macroglobulin (uniquely identified in bands A, B, J and K) and lipocalin-1 (uniquely identified in bands G and U). M, marker/calibration proteins with relative mobilities (Mr) as indicated in kilodaltons (kDa).

Mentions: The protein profiles of all the uterine flush samples collected from days 4 to 14 after mating are shown in the SDS-PAGE analysis in electronic supplementary material, figure S1. The disparities between overall protein concentrations among the samples could be due to differences in efficiency of flushing, changes in the total tissue volume, differences between individual animals' secretion volumes or changes in secretory activity. In order to improve comparability, selected samples were concentrated as described above and/or the volume of loaded sample adjusted to approximately equalize the intensity of the band at ca 65 kDa Mr (band C; figure 1), suspected (and subsequently confirmed by mass spectrometry), to be serum albumin from its size and slower migration under reduction.Figure 1.


Pulse of inflammatory proteins in the pregnant uterus of European polecats ( Mustela putorius ) leading to the time of implantation
Changes in European polecat uterine secretory proteins with time after mating. Animals sampled on days 4 and 14 were non-pregnant. The sample volumes were adjusted to normalize the intensity of the strong band at approximately 65 kDa (serum albumin). See electronic supplementary material, figure S1, for SDS-PAGE of all of the samples collected and upon which the adjustments were based. Gel band codes are indicated by letters and are referred to in the text and in table 1. The proteins that most clearly increased in concentration with time were α2-macroglobulin (uniquely identified in bands A, B, J and K) and lipocalin-1 (uniquely identified in bands G and U). M, marker/calibration proteins with relative mobilities (Mr) as indicated in kilodaltons (kDa).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383852&req=5

RSOS161085F1: Changes in European polecat uterine secretory proteins with time after mating. Animals sampled on days 4 and 14 were non-pregnant. The sample volumes were adjusted to normalize the intensity of the strong band at approximately 65 kDa (serum albumin). See electronic supplementary material, figure S1, for SDS-PAGE of all of the samples collected and upon which the adjustments were based. Gel band codes are indicated by letters and are referred to in the text and in table 1. The proteins that most clearly increased in concentration with time were α2-macroglobulin (uniquely identified in bands A, B, J and K) and lipocalin-1 (uniquely identified in bands G and U). M, marker/calibration proteins with relative mobilities (Mr) as indicated in kilodaltons (kDa).
Mentions: The protein profiles of all the uterine flush samples collected from days 4 to 14 after mating are shown in the SDS-PAGE analysis in electronic supplementary material, figure S1. The disparities between overall protein concentrations among the samples could be due to differences in efficiency of flushing, changes in the total tissue volume, differences between individual animals' secretion volumes or changes in secretory activity. In order to improve comparability, selected samples were concentrated as described above and/or the volume of loaded sample adjusted to approximately equalize the intensity of the band at ca 65 kDa Mr (band C; figure 1), suspected (and subsequently confirmed by mass spectrometry), to be serum albumin from its size and slower migration under reduction.Figure 1.

View Article: PubMed Central - PubMed

ABSTRACT

Uterine secretory proteins protect the uterus and conceptuses against infection, facilitate implantation, control cellular damage resulting from implantation, and supply pre-implantation embryos with nutrients. Unlike in humans, the early conceptus of the European polecat (Mustela putorius; ferret) grows and develops free in the uterus until implanting at about 12 days after mating. We found that the proteins appearing in polecat uteri changed dramatically with time leading to implantation. Several of these proteins have also been found in pregnant uteri of other eutherian mammals. However, we found a combination of two increasingly abundant proteins that have not been recorded before in pre-placentation uteri. First, the broad-spectrum proteinase inhibitor α2-macroglobulin rose to dominate the protein profile by the time of implantation. Its functions may be to limit damage caused by the release of proteinases during implantation or infection, and to control other processes around sites of implantation. Second, lipocalin-1 (also known as tear lipocalin) also increased substantially in concentration. This protein has not previously been recorded as a uterine secretion in pregnancy in any species. If polecat lipocalin-1 has similar biological properties to that of humans, then it may have a combined function in antimicrobial protection and transporting or scavenging lipids. The changes in the uterine secretory protein repertoire of European polecats is therefore unusual, and may be representative of pre-placentation supportive uterine secretions in mustelids (otters, weasels, badgers, mink, wolverines) in general.

No MeSH data available.