Paternal knockdown of tRNA (cytosine-5-)-methyltransferase (Dnmt2) increases offspring susceptibility to infection in red flour beetles

Schulz, Nora; Mfd, Buhr; Kurtz, Joachim

doi:10.1101/422063

“…More recent studies tend, however, to point out some role of epigenetic in TGIP. In T. castaneum , priming of fathers with B. thuringiensis combined with RNAi of a DNA methyltransferase ( Dnmt2 ), known to drive CpG methylation on tRNA (142), led to a low (~10%) although significant decrease in offspring survival to the same pathogen (143). However, the mechanism by which tRNA methylation could participate to TGIP is still unclear and further investigation is now required.…”

Section: The Many Roads To Tgip: Hypothetical Scenarios Based On Empimentioning

confidence: 99%

Trans-generational Immune Priming in Invertebrates: Current Knowledge and Future Prospects

Tetreau

¹

,

Dhinaut

²

,

Gourbal

³

et al. 2019

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Trans-generational immune priming (TGIP) refers to the transfer of the parental immunological experience to its progeny. This may result in offspring protection from repeated encounters with pathogens that persist across generations. Although extensively studied in vertebrates for over a century, this phenomenon has only been identified 20 years ago in invertebrates. Since then, invertebrate TGIP has been the focus of an increasing interest, with half of studies published during the last few years. TGIP has now been tested in several invertebrate systems using various experimental approaches and measures to study it at both functional and evolutionary levels. However, drawing an overall picture of TGIP from available studies still appears to be a difficult task. Here, we provide a comprehensive review of TGIP in invertebrates with the objective of confronting all the data generated to date to highlight the main features and mechanisms identified in the context of its ecology and evolution. To this purpose, we describe all the articles reporting experimental investigation of TGIP in invertebrates and propose a critical analysis of the experimental procedures performed to study this phenomenon. We then investigate the outcome of TGIP in the offspring and its ecological and evolutionary relevance before reviewing the potential molecular mechanisms identified to date. In the light of this review, we build hypothetical scenarios of the mechanisms through which TGIP might be achieved and propose guidelines for future investigations.

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“…Trizol was subsequently added to the homogenate. The RNA of each frozen larva was extracted following a combined TRIZol / spin column protocol developed by Schulz, Buhr, & Kurtz (2018). The quality and the concentration of the RNA were controlled on a 1.5 % agarose gel, and its concentration assessed with a nanophotometer (IMPLEN, München, Germany).…”

Section: Assessment Of Gene Expression By Qpcr and Verification Of Gene Knock-down Efficiencymentioning

confidence: 99%

A Gene Knock-Down Approach in Tribolium castaneum to Study Survival and Priming Towards Bacillus thuringiensis var. tenebrionis

Lindeza¹,

Barth²,

Kurtz³

et al. 2021

Preprint

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Insects possess an array of defense molecules allowing them to fight infections. They can also show a form of immune memory, named priming. However, the involvement of insect immune defense mechanisms in priming is unclear, since invertebrates lack the molecular machinery present in vertebrates to build an immune memory. In the red flour beetle Tribolium castaneum, larvae can be primed via the oral route with Bacillus thurigiensis var. tenebrionids (Btt). This results in changes in the expression of a large number of genes, among which some belong to families of ancient defense genes. In the present work, we tested whether three chosen candidate genes (a Thaumatin, a C-type Lectin and an Osiris-like gene) could be involved in the survival to a Btt exposure, as well as in the priming phenotype. We assessed changes in their expression over time and according to the priming treatment, knocked them down individually by RNA interference (RNAi), and observed how it affected survival upon challenge. The quantification of gene expression patterns in our larvae with RT-qPCR showed that up- and/or down-regulation of the genes, after the priming treatment, was quite volatile and time dependent. Upon knock-down, we did not observe the expected decrease in survival to Btt or the abolishment of the priming phenotype. We conclude that knocking down genes individually is probably insufficient to affect survival and priming in our system. This gives us insight into the complexity of the molecular processes underpinning priming.

show abstract

“…Trizol was subsequently added to the homogenate. The RNA of each frozen larva was extracted following a combined TRIZol / spin column protocol developed by Schulz, Buhr, & Kurtz (2018). The quality and the concentration of the RNA were controlled on a 1.5 % agarose gel, and its concentration assessed with a nanophotometer (IMPLEN, München, Germany).…”

Section: Assessment Of Gene Expression By Qpcr and Verification Of Gene Knockdown Efficiencymentioning

confidence: 99%

A Gene Knock-Down Approach in Tribolium castaneum to Study Survival and Priming Towards Bacillus thuringiensis var. tenebrionis

As¹,

Barth²,

Kurtz³

et al. 2021

Preprint

Self Cite

0

View full text Add to dashboard Cite

Insects possess an array of defense molecules allowing them to fight infections. They can also show a form of immune memory, named priming. However, the involvement of insect immune defense mechanisms in priming is unclear, since invertebrates lack the molecular machinery present in vertebrates to build an immune memory. In the red flour beetle Tribolium castaneum, larvae can be primed via the oral route with Bacillus thurigiensis var. tenebrionids (Btt). This results in changes in the expression of a large number of genes, among which some belong to families of ancient defense genes. In the present work, we tested whether three chosen candidate genes (a Thaumatin, a C-type Lectin and an Osiris-like gene) could be involved in the survival to a Btt exposure, as well as in the priming phenotype. We assessed changes in their expression over time and according to the priming treatment, knocked them down individually by RNA interference (RNAi), and observed how it affected survival upon challenge. The quantification of gene expression patterns in our larvae with RT-qPCR showed that up- and/or down-regulation of the genes, after the priming treatment, was quite volatile and time dependent. Upon knock-down, we did not observe the expected decrease in survival to Btt or the abolishment of the priming phenotype. We conclude that knocking down genes individually is probably insufficient to affect survival and priming in our system. This gives us insight into the complexity of the molecular processes underpinning priming.

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Paternal knockdown of tRNA (cytosine-5-)-methyltransferase (Dnmt2) increases offspring susceptibility to infection in red flour beetles

Cited by 5 publications

References 68 publications

Trans-generational Immune Priming in Invertebrates: Current Knowledge and Future Prospects

Trans-generational Immune Priming in Invertebrates: Current Knowledge and Future Prospects

A Gene Knock-Down Approach in <i>Tribolium castaneum</i> to Study Survival and Priming Towards <i>Bacillus thuringiensis</i> var. <i>tenebrionis</i>

A Gene Knock-Down Approach in <i>Tribolium castaneum</i> to Study Survival and Priming Towards <i>Bacillus thuringiensis</i> var. <i>tenebrionis</i>

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