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Selected Publication:

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Type of publication: Master Thesis
Type of document:

Year: 2013

Authors: Hanna, Amgad

Title: An Approach to Analyze Proteins in Uterine Flushing Fluids of Mares.

Other title: Analyse des von Pferdeembryonen zur Zeit der maternalen Erkennung der Trächtigkeit sezernierten protein- bzw. Peptidmusters

Source: Master Thesis, Vet. Med. Univ. Wien, pp. 59.


Advisor(s):

Razzazi-Fazeli Ebrahim

Reviewer(s):
Böhm Josef

Vetmed Research Units:
VetCore


Graduation date: 24.09.13


Abstract:
The maternal recognition signal of mare has not been identified. Generally, embryo-derived pregnancy signal is released by the embryo and expected to be found in the uterine flushing fluid of the mother. This pilot study tried to find the proper way to analyze the uterine flushing fluid of mares, and the nature of the embryo-derived pregnancy signal. The uterus of pregnant mares was flushed with ringer-lactate solution. These samples of flushing fluid of mares were ultrafiltrated and then analyzed by gel electrophoresis. The main techniques which were used are one- and two-dimensional gel electrophoresis. The two dimension gel electrophoresis was optimized for separation of proteins. However, one dimension gel electrophoresis was used for identification of proteins. In the one dimensional gel electrophoresis, four different proteins were identified. These proteins are uterocalin (P19), Beta-2-microglobulin, putative non-heme bromoperoxidase and vascular non-inflammatory molecule 3-like. In two dimension gel electrophoresis technique, parameters were optimized such as rehydration process in IEF procedures, running buffer concentration, protein precipitation using TCA, using different size of IPG strips and adjust volt / hour in IEF procedures. It could be shown that passive rehydration process in IEF was more powerful than the active one. Also, longer focusing time provided a better separation according to the pI of the proteins. Finally, the higher SDS concentration in the running buffer led to a better resolution in the gel. In conclusions, one- and two-dimensional gel electrophoresis are reliable methods to detect proteins in the flushing fluids of mares. It is recommended to use flushing fluids from the same animal to avoid the biological variations.


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