Validation of reference genes in leaf-cutting ant Atta sexdens rubropilosa in different developmental stages and tissues

Moreira, Ariele C.; Santos, Adriana Miranda dos; Carneiro, Renato Lajarim; Bueno, Odair Corrêa; Souza, Dulce Helena Ferreira de

doi:10.22161/ijeab/2.2.23

Cited by 8 publications

(7 citation statements)

References 39 publications

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“…Two AsChEs were purified from worker heads of A. sexdens . Medium worker ants were selected considering our previous studies, in which we identified higher AChE expression levels in this developmental stage, compared to larva and pupa [40].…”

Section: Resultsmentioning

confidence: 99%

Acetylcholinesterases from Leaf-Cutting antAtta sexdens: Purification, Characterization, and Capillary Reactors for On-Flow Assays

et al. 2019

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Acetylcholinesterase (AChE) is responsible for catalyzing the hydrolysis of the neurotransmitter acetylcholine (ACh) leading to acetate and choline (Ch) release. The inhibition of AChE produces a generalized synaptic collapse that can lead to insect death. Herein we report for the first time the isolation of two AChEs from Atta sexdens which were purified by sulphate ammonium precipitation followed by ion exchange chromatography. AsAChE-A and AsAChE-B enzymes have optimum pH of 9.5 and 9.0 and higher activities in 30/50°C and 20°C, respectively, using acetylthiocholine (ATCh) as substrate. Immobilized capillary enzyme reactors (ICERs) were obtained for both enzymes (AsAChE-A-ICER and AsAChE-B-ICER) and their activities were measured by LC-MS/MS through hydrolysis product quantification of the natural substrate ACh. The comparison of activities by LC-MS/MS of both AChEs using ACh as substrate showed that AsAChE-B (free or immobilized) had the highest affinity. The inverse result was observed when the colorimetric assay (Elman method) was used for ATCh as substrate. Moreover, by mass spectrometry and phylogenetic studies, AsAChE-A and AsAChE-B were classified as belonging to AChE-2 and AChE-1 classes, respectively.

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Section: Resultsmentioning

confidence: 99%

Acetylcholinesterases from Leaf-Cutting antAtta sexdens: Purification, Characterization, and Capillary Reactors for On-Flow Assays

et al. 2019

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“…These results do not differ from those found in the reference gene evaluation in A. sexdens rubropilosa , on which RPL18 was the most stable gene in tissues under development stage. [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Gene Expression Profile Analysis is Directly Affected by the Selected Reference Gene: The Case of Leaf-Cutting Atta Sexdens

Livramento

Freitas

Máximo

et al. 2018

Insects

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Although several ant species are important targets for the development of molecular control strategies, only a few studies focus on identifying and validating reference genes for quantitative reverse transcription polymerase chain reaction (RT-qPCR) data normalization. We provide here an extensive study to identify and validate suitable reference genes for gene expression analysis in the ant Atta sexdens, a threatening agricultural pest in South America. The optimal number of reference genes varies according to each sample and the result generated by RefFinder differed about which is the most suitable reference gene. Results suggest that the RPS16, NADH and SDHB genes were the best reference genes in the sample pool according to stability values. The SNF7 gene expression pattern was stable in all evaluated sample set. In contrast, when using less stable reference genes for normalization a large variability in SNF7 gene expression was recorded. There is no universal reference gene suitable for all conditions under analysis, since these genes can also participate in different cellular functions, thus requiring a systematic validation of possible reference genes for each specific condition. The choice of reference genes on SNF7 gene normalization confirmed that unstable reference genes might drastically change the expression profile analysis of target candidate genes.

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“…Neuropeptide expression levels were determined relative to transcripts of elongation factor‐1 alpha (EF1 α) using the ΔΔCt method. EF1 α was shown to be a suitable reference gene in brains of honeybees (Lourenço et al, 2008; Reim et al, 2013) and several ant species (Moreira et al, 2017; Ratzka et al, 2011). Therefore, we decided to use this gene in C .…”

Section: Methodsmentioning

confidence: 99%

Neuropeptides as potential modulators of behavioral transitions in the ant Cataglyphis nodus

Habenstein

Thamm

Rößler

2021

J of Comparative Neurology

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Age‐related behavioral plasticity is a major prerequisite for the ecological success of insect societies. Although ecological aspects of behavioral flexibility have been targeted in many studies, the underlying intrinsic mechanisms controlling the diverse changes in behavior along the individual life history of social insects are not completely understood. Recently, the neuropeptides allatostatin‐A, corazonin, and tachykinin have been associated with the regulation of behavioral transitions in social insects. Here, we investigated changes in brain localization and expression of these neuropeptides following major behavioral transitions in Cataglyphis nodus ants. Our immunohistochemical analyses in the brain revealed that the overall branching pattern of neurons immunoreactive (ir) for the three neuropeptides is largely independent of the behavioral stages. Numerous allatostatin‐A‐ and tachykinin‐ir neurons innervate primary sensory neuropils and high‐order integration centers of the brain. In contrast, the number of corazonergic neurons is restricted to only four neurons per brain hemisphere with cell bodies located in the pars lateralis and axons extending to the medial protocerebrum and the retrocerebral complex. Most interestingly, the cell‐body volumes of these neurons are significantly increased in foragers compared to freshly eclosed ants and interior workers. Quantification of mRNA expression levels revealed a stage‐related change in the expression of allatostatin‐A and corazonin mRNA in the brain. Given the presence of the neuropeptides in major control centers of the brain and the neurohemal organs, these mRNA‐changes strongly suggest an important modulatory role of both neuropeptides in the behavioral maturation of Cataglyphis ants.

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Validation of reference genes in leaf-cutting ant Atta sexdens rubropilosa in different developmental stages and tissues

Abstract: Abstract-Atta sexdens rubropilosa is an important leafcutting ant species considered as a pest in agricultural

Cited by 8 publications

References 39 publications

Acetylcholinesterases from Leaf-Cutting antAtta sexdens: Purification, Characterization, and Capillary Reactors for On-Flow Assays

Acetylcholinesterases from Leaf-Cutting antAtta sexdens: Purification, Characterization, and Capillary Reactors for On-Flow Assays

Gene Expression Profile Analysis is Directly Affected by the Selected Reference Gene: The Case of Leaf-Cutting Atta Sexdens

Neuropeptides as potential modulators of behavioral transitions in the ant Cataglyphis nodus

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