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Selected Publication:

Type of publication: Master Thesis
Type of document:

Year: 2010

Authors: Ondrovics, M

Title: Expression geschlechtsspezifischer und verschiedener Housekeeping-Gene in den parasitären Entwicklungsstadien von Oesophagostomum dentatum.

Other title: Expression of gender-specific and various housekeeping genes in different parasitic stages of Oesophagostomum dentatum

Source: Master Thesis, Vet. Med. Univ. Wien, pp. 92.


Advisor(s):

Joachim Anja

Reviewer(s):
Duvigneau Catharina

Vetmed Research Units:
Institute of Parasitology


Graduation date: 16.08.10


Abstract:
Oesophagostomum dentatum, the nodule worm, is one of the most common endoparasites of pigs. It also represents an ideal model organism for the order of the Strongylida, members of which are responsible for disease, fatalities and enormous economic losses. In order to investigate the expression of the two gender-specific genes vitellogenin and Major Sperm Protein (MSP) in O. dentatum RNA from different stages was extracted, reverse transcribed to cDNA and different levels of expression were investigated by real-time PCR using the dye SYBR® Green. Before gene expression could be detected the real-time protocol had to be established by testing the specificity of the primers and optimising the cDNA and primer concentrations. Furthermore it could be shown that the three employed housekeeping genes, ubiquitin-conjugating enzyme 2 (ubc-2), triosephosphate isomerase (TPI) and GTPcyclohydrolase (GTP-CH) which served as normalisers for the gender-specific genes were suitable as housekeeping genes for expression studies in O. dentatum. The relative quantification of vitellogenin and MSP with two different algorithms, the 􀇻􀇻Ct-method and the geNorm®-calculation, showed comparable results. Neither the immature larval stages nor worms from in vitro cultures (35 and 42 days) expressed vitellogenin or MSP, whereas adult females expressed vitellogenin and adult males MSP at high levels. It could also be shown that neither presence nor absence of the other sex, nor the duration of infection were pivotal for vitellogenin or MSP expression in O. dentatum. In small or “virgin” (also small) adults no or only low expression of vitellogenin and MSP was detectable. It can therefore be hypothesised that the expression of the gender-specific genes is related to worm size. This raises the question whether small size hampers the expression of vitellogenin and MSP, respectively, or whether lack of expression inhibits worm growth, for reasons not yet known. In many parasitic nematodes anthelmintic resistance has developed, so that new targets of intervention for parasitic control are highly important. A deeper understanding of the molecular mechanisms of the reproduction biology of parasitic nematodes could be of great importance for the development of new control measures. This work provides insight into the differential expression of the gender-specific genes vitellogenin and MSP during the development of O. dentatum. Inhibition of the expression could play a specific role for combating parasites.


Publication(s) resulting from University thesis:

Ondrovics, M; Gasser, RB; Ruttkowski, B; Nisbet, AJ; Joachim, A (2012): Transcription profiles for two key gender-specific gene families in Oesophagostomum dentatum during development in vivo and in vitro. Infect Genet Evol. 2012; 12(1):137-141

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