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. 2010 Mar 9:11:160.
doi: 10.1186/1471-2164-11-160.

Transcriptome analysis of reproductive tissue and intrauterine developmental stages of the tsetse fly (Glossina morsitans morsitans)

Geoffrey M Attardo  1 , José Mc Ribeiro, Yineng Wu, Matthew Berriman, Serap Aksoy
Affiliations

Transcriptome analysis of reproductive tissue and intrauterine developmental stages of the tsetse fly (Glossina morsitans morsitans)

Geoffrey M Attardo et al. BMC Genomics. .

Abstract

Background: Tsetse flies, vectors of African trypanosomes, undergo viviparous reproduction (the deposition of live offspring). This reproductive strategy results in a large maternal investment and the deposition of a small number of progeny during a female's lifespan. The reproductive biology of tsetse has been studied on a physiological level; however the molecular analysis of tsetse reproduction requires deeper investigation. To build a foundation from which to base molecular studies of tsetse reproduction, a cDNA library was generated from female tsetse (Glossina morsitans morsitans) reproductive tissues and the intrauterine developmental stages. 3438 expressed sequence tags were sequenced and analyzed.

Results: Analysis of a nonredundant catalogue of 1391 contigs resulted in 520 predicted proteins. 475 of these proteins were full length. We predict that 412 of these represent cytoplasmic proteins while 57 are secreted. Comparison of these proteins with other tissue specific tsetse cDNA libraries (salivary gland, fat body/milk gland, and midgut) identified 51 that are unique to the reproductive/immature cDNA library. 11 unique proteins were homologous to uncharacterized putative proteins within the NR database suggesting the identification of novel genes associated with reproductive functions in other insects (hypothetical conserved). The analysis also yielded seven putative proteins without significant homology to sequences present in the public database (unknown genes). These proteins may represent unique functions associated with tsetse's viviparous reproductive cycle. RT-PCR analysis of hypothetical conserved and unknown contigs was performed to determine basic tissue and stage specificity of the expression of these genes.

Conclusion: This paper identifies 51 putative proteins specific to a tsetse reproductive/immature EST library. 11 of these proteins correspond to hypothetical conserved genes and 7 proteins are tsetse specific.

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Figures

Figure 1
Figure 1
Statistics on all predicted reproductive/immature proteins. This graph represents all putative proteins within the reproductive/immature cDNA library. Light bars represent the percentage of predicted proteins categorized by function from the reproductive/immature library. Dark bars represent the number of ESTs associated with the genes within each category.
Figure 2
Figure 2
Statistics on predicted proteins unique to the reproductive/immature library. This graph represents putative proteins unique to the reproductive/immature cDNA library relative to fat body, midgut and salivary gland specific cDNA libraries. Light bars represent the percentage of library specific predicted proteins categorized by function. Dark bars represent the number of ESTs associated with the genes within each category.
Figure 3
Figure 3
Phylogenetic and alignment based analysis and comparison of a gonad specific trypsin (EZ421932) with a midgut specific trypsin (EU589384). Orthologus sequences were identified by PSI-BLAST analysis. Sequences were then aligned with standalone ClustalX followed by manual adjustments. Phylogenetic tree construction and bootstrap analysis were performed with MEGA 3.1.
Figure 4
Figure 4
Phylogenetic and alignment based analysis and comparison of a library specific hexamerin protein (EZ421932). Orthologus sequences were identified by PSI-BLAST analysis. Sequences were then aligned with standalone ClustalX followed by manual adjustments. Phylogenetic tree construction and bootstrap analysis were performed with MEGA 3.1.
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