Document Actions

Microarray, Super-SAGE & RNA-seq

While several studies focused on the pairing-dependent development of the female reproductive organs as well as the involved genes and signaling processes, the influence of pairing on the male was neglected so far. Although male morphology seems to be unaffected upon pairing, actual transcriptome studies demonstrated a significant influence of pairing at the level of gene regulation including the gonads2-5.     

Aim of this project is the characterization of pairing-dependently transcribed genes in the male with a focus on the identification of competence factors. These are genes by definition whose expression is essential for the induction of pairing and maturation processes in the female.


 

SH Micr_SupSage_jpg.jpg

Comparative analyses of RNA obtained from pairing-experienced and pairing-unexperienced worms led to the identification of pairing-dependently transcribed genes in S. mansoni males. In total, transcripts of 6326 genes were detected by techniques such as SuperSAGE and Microarray (A, overlay), of which 29 were defined as significantly and differentially transcribed in accordance to statistical analyses of both methods (B, dark orange) 2.  

Overview of the RNAseq Data from Analysis of paired and unpairedS.mansoni, as well as

 their gonads. (a) Multidimesional scaling (Replcates are shown in the same colour. (b)

Sample distance matrix; colour code is replicated as in (a,c). Hierarchical

clustering based on RPKM. Horizontal lines represent single genes. Yellow

indicates higher transkript amount, while blue stands for lower amount. bM,

paired Males (darkblue in a,b); sM, single Male (in a,b: dark blue); bT, Testes

of bM, (in a,b: light blue); sT, testes from sM(in a,b light blue) bF, paired

female (in a,b:red) sF, unpaired female(in a,b:rose); bO, ovarie from bF(a,b:

yellow); sO, ovarie from sF (in a,b;okker)
SH qPCRs_jpg.jpg

Verification of the pairing-dependent transcription of different genes by qRT-PCR (grey bars are biological replicates; the mean value is shown in black). Differences in the transcript rates between pairing-experienced and pairing-unexperienced males are presented as log2ratio. Colored bars represent the results of former transcriptome analyses2,4. Positive values indicate genes with higher transcript levels in S. mansoni males after pairing. Genes whose transcription is lowered after pairing in males (and thus elevated in pairing-unexperienced males) show negative values.     

 

1Beckmann S, Quack T, Burmeister C, Buro C, Long T, Dissous C, Grevelding CG (2010). Schistosoma mansoni: Signal transduction processes during the development of the reproductive organs. Parasitology 137(3):497-20.

 

2Leutner S, Oliveira KC, Rotter B,  Beckmann S, Buro C, Hahnel S, Kitajima JP, Verjovski-Almeida S, Winter P, Grevelding CG (2013). Combinatory microarray and SuperSAGE analyses identify pairing-dependently transcribed genes in Schistosoma mansoni males, including follistatin. PLoS Negl Trop Dis. 7:e2532.

 

3Hahnel S, Quack T, Parker-Manuel SJ, Lu Z, Vanderstraete M, Morel M, Dissous C, Cailliau K, Grevelding CG (2014). Gonad RNA-specific qRT-PCR analyses identify genes with potential functions in schistosome reproduction such as SmFz1 and SmFGFRs. Front Genet. 5:170.

 

4LuZ, Sessler F, HolroydN, Hahnel S, Quack T, Berriman M, Grevelding CG (2016). Tissue-specific and pairing-dependent gene expression in the gonads of the blood fluke Schistosoma mansoni. Sci Rep. 6:31150.

 

5Lu, Z., Spänig, S., Weth, O., Grevelding, CG. (2019). Males, the Wrongly Neglected Partners of the Biologically Unprecedented Male-Female Interaction of Schistosomes. Front Genet. 10:796.