Entorhinal cortex vulnerability to human APP expression promotes hyperexcitability and tau pathology.

Preventative treatment for Alzheimer's Disease is of dire importance, and yet, cellular mechanisms underlying early regional vulnerability in Alzheimer's Disease remain unknown. In human patients with Alzheimer's Disease, one of the earliest observed pathophysiological correlates to cognitive decline is hyperexcitability. In mouse models, early hyperexcitability has been shown in the entorhinal cortex, the first cortical region impacted by Alzheimer's Disease. The origin of hyperexcitability in early-stage disease and why it preferentially emerges in specific regions is unclear. Using cortical-region and cell-type-specific proteomics coupled with ex vivo and in vivo electrophysiology, we uncovered differential susceptibility to human-specific amyloid precursor protein (hAPP) in a model of sporadic Alzheimer's. Unexpectedly, our findings reveal that early entorhinal hyperexcitability may result from intrinsic vulnerability of parvalbumin (PV) interneurons, rather than the suspected layer II excitatory neurons. This vulnerability of entorhinal PV interneurons is specific to hAPP, as it could not be recapitulated with increased murine APP expression. However, partial replication of the findings could be seen after introduction of a murine APP chimera containing a humanized amyloid-beta sequence. Surprisingly, neurons in the Somatosensory Cortex showed no such vulnerability to adult-onset hAPP expression. hAPP-induced hyperexcitability in entorhinal cortex could be ameliorated by enhancing PV interneuron excitability in vivo. Co-expression of human Tau with hAPP decreased circuit hyperexcitability, but at the expense of increased pathological tau species. This study suggests early disease interventions targeting non-excitatory cell types may protect regions with early vulnerability to pathological symptoms of Alzheimer's Disease and downstream cognitive decline.

Comparative Phytotoxicity of Metallic Elements on Duckweed Lemna gibba L. Using Growth- and Chlorophyll Fluorescence Induction-Based Endpoints.

In this study, we exposed a commonly used duckweed species-Lemna gibba L.-to twelve environmentally relevant metals and metalloids under laboratory conditions. The phytotoxic effects were evaluated in a multi-well-plate-based experimental setup by means of the chlorophyll fluorescence imaging method. This technique allowed the simultaneous measuring of the growth and photosynthetic parameters in the same samples. The inhibition of relative growth rates (based on frond number and area) and photochemical efficiency (Fv/Fo and Y(II)) were both calculated from the obtained chlorophyll fluorescence images. In the applied test system, growth-inhibition-based phytotoxicity endpoints proved to be more sensitive than chlorophyll-fluorescence-based ones. Frond area growth inhibition was the most responsive parameter with a median EC50 of 1.75 mg L-1, while Fv/Fo, the more responsive chlorophyll-fluorescence-based endpoint, resulted in a 5.34 mg L-1 median EC50 for the tested metals. Ag (EC50 0.005-1.27 mg L-1), Hg (EC50 0.24-4.87 mg L-1) and Cu (EC50 0.37-1.86 mg L-1) were the most toxic elements among the tested ones, while As(V) (EC50 47.15-132.18 mg L-1), Cr(III) (EC50 6.22-19.92 mg L-1), Se(VI) (EC50 1.73-10.39 mg L-1) and Zn (EC50 3.88-350.56 mg L-1) were the least toxic ones. The results highlighted that multi-well-plate-based duckweed phytotoxicity assays may reduce space, time and sample volume requirements compared to the standard duckweed growth inhibition tests. These benefits, however, come with lowered test sensitivity. Our multi-well-plate-based test setup resulted in considerably higher median EC50 (3.21 mg L-1) for frond-number-based growth inhibition than the 0.683 mg L-1 median EC50 derived from corresponding data from the literature with standardized Lemna-tests. Under strong acute phytotoxicity, frond parts with impaired photochemical functionality may become undetectable by chlorophyll fluorometers. Consequently, the plant parts that are still detectable display a virtually higher average photosynthetic performance, leading to an underestimation of phytotoxicity. Nevertheless, multi-well-plate-based duckweed phytotoxicity assays, combined with chlorophyll fluorescence imaging, offer definite advantages in the rapid screening of large sample series or multiple species/clones. As chlorophyll fluorescence images provide information both on the photochemical performance of the test plants and their morphology, a joint analysis of the two endpoint groups is recommended in multi-well-plate-based duckweed phytotoxicity assays to maximize the information gained from the tests.

An enhancer-AAV approach selectively targeting dentate granule cells of the mouse hippocampus.

The mammalian brain contains a diverse array of cell types, including dozens of neuronal subtypes with distinct anatomical and functional characteristics. The brain leverages these neuron-type specializations to perform diverse circuit operations and thus execute different behaviors properly. Through the use of Cre lines, access to specific neuron types has improved over past decades. Despite their extraordinary utility, development and cross-breeding of Cre lines is time consuming and expensive, presenting a significant barrier to entry for investigators. Furthermore, cell-based therapeutics developed in Cre mice are not clinically translatable. Recently, several adeno-associated virus (AAV) vectors utilizing neuron-type-specific regulatory transcriptional sequences (enhancer-AAVs) were developed that overcome these limitations. Using a publicly available RNA sequencing (RNA-seq) dataset, we evaluated the potential of several candidate enhancers for neuron-type-specific targeting in the hippocampus. Here, we demonstrate that a previously identified enhancer-AAV selectively targets dentate granule cells over other excitatory neuron types in the hippocampus of wild-type adult mice.

Entorhinal cortex vulnerability to human APP expression promotes hyperexcitability and tau pathology.

Preventative treatment for Alzheimer's Disease is of dire importance, and yet, cellular mechanisms underlying early regional vulnerability in Alzheimer's Disease remain unknown. In human patients with Alzheimer's Disease, one of the earliest observed pathophysiological correlates to cognitive decline is hyperexcitability1. In mouse models, early hyperexcitability has been shown in the entorhinal cortex, the first cortical region impacted by Alzheimer's Disease2-4. The origin of hyperexcitability in early-stage disease and why it preferentially emerges in specific regions is unclear. Using cortical-region and cell-type- specific proteomics and patch-clamp electrophysiology, we uncovered differential susceptibility to human-specific amyloid precursor protein (hAPP) in a model of sporadic Alzheimer's. Unexpectedly, our findings reveal that early entorhinal hyperexcitability may result from intrinsic vulnerability of parvalbumin interneurons, rather than the suspected layer II excitatory neurons. This vulnerability of entorhinal PV interneurons is specific to hAPP, as it could not be recapitulated with increased murine APP expression. Furthermore, the Somatosensory Cortex showed no such vulnerability to adult-onset hAPP expression, likely resulting from PV-interneuron variability between the two regions based on physiological and proteomic evaluations. Interestingly, entorhinal hAPP-induced hyperexcitability was quelled by co-expression of human Tau at the expense of increased pathological tau species. This study suggests early disease interventions targeting non-excitatory cell types may protect regions with early vulnerability to pathological symptoms of Alzheimer's Disease and downstream cognitive decline.

Duckweed: Research Meets Applications.

The Special Issue "Duckweed: Research Meets Applications" of the journal Plants (ISSN 2223-7747) presents a comprehensive update of the current progress in the field [...].

Phytoremediation, recovery and toxic effects of ionic gadolinium using the free-floating plant Lemna gibba.

The biosorption and recovery of ionic gadolinium (Gd) from contaminated water by the free-floating duckweed Lemna gibba was studied. The highest non-toxic concentration range was determined as (6.7 mg L-1). The concentration of Gd in the medium and in the plant biomass was monitored and a mass balance was established. Tissue Gd concentration of Lemna increased with increasing Gd concentration of the medium. The bioconcentration factor was up to 1134 and in nontoxic concentrations up to 2.5 g kg-1 Gd tissue concentration was reached. Lemna ash contained 23.2 g Gd kg-1. Gd removal efficiency from the medium was 95%, however, only 17-37% of the initial Gd content of the medium accumulated in Lemna biomass, an average of 5% remained in the water, and 60-79% was calculated as a precipitate. Gadolinium-exposed Lemna plants released ionic Gd into the nutrient solution when they were transferred to a Gd-free medium. The experimental results revealed that in constructed wetlands, L. gibba is able to remove ionic Gd from the water and can be suitable for bioremediation and recovery purposes.

Sixth International Conference on Duckweed Research and Applications Presents Lemnaceae as a Model Plant System in the Genomics and Postgenomics Era.

The 6th International Conference on Duckweed Research and Applications (6th ICDRA) was organized at the Institute of Plant Genetics and Crop Plant Research (IPK) located in Gatersleben, Germany, from 29 May to 1 June 2022. The growing community of duckweed research and application specialists was noted with participants from 21 different countries including an increased share of newly integrated young researchers. The four-day conference focused on diverse aspects of basic and applied research together with practical applications of these tiny aquatic plants that could have an enormous potential for biomass production.

Cryptogamic communities on flatroofs in the city of Debrecen (East Hungary).

Cryptogams of ten urban flatroofs, contrasting in their age and size, were studied between 2016 and 2018. Siliceous (bituminous felt, gravel, brick) and calcareous (concrete) substrata occurred at each site. Microclimate (T, RH) at two sites of contrasting shading was monitored from September 2016 to January 2017. Biomass of two differently aged, exposed flatroofs was sampled in October 2018. Taxa of Cladonia and Xanthoparmelia have been identified by spot tests and HPTLC. A total of 61 taxa (25 bryophytes, 36 lichens), mostly widespread synanthropic species, have been detected with an explicit difference of species composition between shaded and exposed sites. Floristically interesting species included acidophilous bryophytes (Hedwigia ciliata, Racomitrium canescens) and lichens (Xanthoparmelia conspersa, Stereocaulon tomentosum) of montane character. The most widespread lichen is Cladonia rei which accounted for a significant part of the biomass at selected sites. Species-area curves for bryophytes at exposed sites have become saturated at 100-150 m2. In contrast, saturation of lichen diversity has not been reached even at the largest sites. Flatroofs with traditional roofing techniques can harbour relatively diverse microhabitats and species-rich synanthropic vegetation. It is urgent to study these sites before renovation with modern roofing techniques eliminates them. Diversification of urban surroundings is possible in the future via application of various substrats in renovated and newly constructed roofs.

A novel enhancer-AAV approach selectively targeting dentate granule cells.

The mammalian brain contains the most diverse array of cell types of any organ, including dozens of neuronal subtypes with distinct anatomical and functional characteristics. The brain leverages these neuron-type-specializations to perform diverse circuit operations and thus execute different behaviors properly. Through the use of Cre lines, access to specific neuron types has steadily improved over past decades. Despite their extraordinary utility, development and cross-breeding of Cre lines is time-consuming and expensive, presenting a significant barrier to entry for many investigators. Furthermore, cell-based therapeutics developed in Cre mice are not clinically translatable. Recently, several AAV vectors utilizing neuron-type-specific regulatory transcriptional sequences (enhancer-AAVs) were developed which overcome these limitations. Using a publicly available RNAseq dataset, we evaluated the potential of several candidate enhancers for neuron-type-specific targeting in the hippocampus. Here we identified a promising enhancer-AAV for targeting dentate granule cells and validated its selectivity in wild-type adult mice.

Classifier for the Rapid Simultaneous Determination of Sleep-Wake States and Seizures in Mice.

Independent automated scoring of sleep-wake and seizures have recently been achieved; however, the combined scoring of both states has yet to be reported. Mouse models of epilepsy typically demonstrate an abnormal electroencephalographic (EEG) background with significant variability between mice, making combined scoring a more difficult classification problem for manual and automated scoring. Given the extensive EEG variability between epileptic mice, large group sizes are needed for most studies. As large datasets are unwieldy and impractical to score manually, automatic seizure and sleep-wake classification are warranted. To this end, we developed an accurate automated classifier of sleep-wake states, seizures, and the post-ictal state. Our benchmark was a classification accuracy at or above the 93% level of human inter-rater agreement. Given the failure of parametric scoring in the setting of altered baseline EEGs, we adopted a machine-learning approach. We created several multi-layer neural network architectures that were trained on human-scored training data from an extensive repository of continuous recordings of electrocorticogram (ECoG), left and right hippocampal local field potential (HPC-L and HPC-R), and electromyogram (EMG) in the murine intra-amygdala kainic acid model of medial temporal lobe epilepsy. We then compared different network models, finding a bidirectional long short-term memory (BiLSTM) design to show the best performance with validation and test portions of the dataset. The SWISC (sleep-wake and the ictal state classifier) achieved >93% scoring accuracy in all categories for epileptic and non-epileptic mice. Classification performance was principally dependent on hippocampal signals and performed well without EMG. Additionally, performance is within desirable limits for recording montages featuring only ECoG channels, expanding its potential scope. This accurate classifier will allow for rapid combined sleep-wake and seizure scoring in mouse models of epilepsy and other neurologic diseases with varying EEG abnormalities, thereby facilitating rigorous experiments with larger numbers of mice.

Species- and Metal-Specific Responses of the Ionome of Three Duckweed Species under Chromate and Nickel Treatments.

In this study, growth and ionomic responses of three duckweed species were analyzed, namely Lemna minor, Landoltia punctata, and Spirodela polyrhiza, were exposed for short-term periods to hexavalent chromium or nickel under laboratory conditions. It was found that different duckweed species had distinct ionomic patterns that can change considerably due to metal treatments. The results also show that, because of the stress-induced increase in leaf mass-to-area ratio, the studied species showed different order of metal uptake efficiency if plant area was used as unit of reference instead of the traditional dry weight-based approach. Furthermore, this study revealed that µXRF is applicable in mapping elemental distributions in duckweed fronds. By using this method, we found that within-frond and within-colony compartmentation of metallic ions were strongly metal- and in part species-specific. Analysis of duckweed ionomics is a valuable approach in exploring factors that affect bioaccumulation of trace pollutants by these plants. Apart from remediating industrial effluents, this aspect will gain relevance in food and feed safety when duckweed biomass is produced for nutritional purposes.

Ultrafast simulation of large-scale neocortical microcircuitry with biophysically realistic neurons.

Understanding the activity of the mammalian brain requires an integrative knowledge of circuits at distinct scales, ranging from ion channel gating to circuit connectomics. Computational models are regularly employed to understand how multiple parameters contribute synergistically to circuit behavior. However, traditional models of anatomically and biophysically realistic neurons are computationally demanding, especially when scaled to model local circuits. To overcome this limitation, we trained several artificial neural network (ANN) architectures to model the activity of realistic multicompartmental cortical neurons. We identified an ANN architecture that accurately predicted subthreshold activity and action potential firing. The ANN could correctly generalize to previously unobserved synaptic input, including in models containing nonlinear dendritic properties. When scaled, processing times were orders of magnitude faster compared with traditional approaches, allowing for rapid parameter-space mapping in a circuit model of Rett syndrome. Thus, we present a novel ANN approach allowing for rapid, detailed network experiments using inexpensive and commonly available computational resources.

Biophysical Kv3 channel alterations dampen excitability of cortical PV interneurons and contribute to network hyperexcitability in early Alzheimer's.

In Alzheimer's disease (AD), a multitude of genetic risk factors and early biomarkers are known. Nevertheless, the causal factors responsible for initiating cognitive decline in AD remain controversial. Toxic plaques and tangles correlate with progressive neuropathology, yet disruptions in circuit activity emerge before their deposition in AD models and patients. Parvalbumin (PV) interneurons are potential candidates for dysregulating cortical excitability as they display altered action potential (AP) firing before neighboring excitatory neurons in prodromal AD. Here, we report a novel mechanism responsible for PV hypoexcitability in young adult familial AD mice. We found that biophysical modulation of Kv3 channels, but not changes in their mRNA or protein expression, were responsible for dampened excitability in young 5xFAD mice. These K+ conductances could efficiently regulate near-threshold AP firing, resulting in gamma-frequency-specific network hyperexcitability. Thus, biophysical ion channel alterations alone may reshape cortical network activity prior to changes in their expression levels. Our findings demonstrate an opportunity to design a novel class of targeted therapies to ameliorate cortical circuit hyperexcitability in early AD.

Cell type-specific biotin labeling in vivo resolves regional neuronal and astrocyte proteomic differences in mouse brain.

Proteomic profiling of brain cell types using isolation-based strategies pose limitations in resolving cellular phenotypes representative of their native state. We describe a mouse line for cell type-specific expression of biotin ligase TurboID, for in vivo biotinylation of proteins. Using adenoviral and transgenic approaches to label neurons, we show robust protein biotinylation in neuronal soma and axons throughout the brain, allowing quantitation of over 2000 neuron-derived proteins spanning synaptic proteins, transporters, ion channels and disease-relevant druggable targets. Next, we contrast Camk2a-neuron and Aldh1l1-astrocyte proteomes and identify brain region-specific proteomic differences within both cell types, some of which might potentially underlie the selective vulnerability to neurological diseases. Leveraging the cellular specificity of proteomic labeling, we apply an antibody-based approach to uncover differences in neuron and astrocyte-derived signaling phospho-proteins and cytokines. This approach will facilitate the characterization of cell-type specific proteomes in a diverse number of tissues under both physiological and pathological states.

Chlorophyll Fluorescence Imaging-Based Duckweed Phenotyping to Assess Acute Phytotoxic Effects.

Duckweeds (Lemnaceae species) are extensively used models in ecotoxicology, and chlorophyll fluorescence imaging offers a sensitive and high throughput platform for phytotoxicity assays with these tiny plants. However, the vast number of potentially applicable chlorophyll fluorescence-based test endpoints makes comparison and generalization of results hard among different studies. The present study aimed to jointly measure and compare the sensitivity of various chlorophyll fluorescence parameters in Spirodela polyrhiza (giant duckweed) plants exposed to nickel, chromate (hexavalent chromium) and sodium chloride for 72 h, respectively. The photochemistry of Photosystem II in both dark- and light-adapted states of plants was assessed via in vivo chlorophyll fluorescence imaging method. Our results indicated that the studied parameters responded with very divergent sensitivity, highlighting the importance of parallelly assessing several chlorophyll fluorescence parameters. Generally, the light-adapted parameters were more sensitive than the dark-adapted ones. Thus, the former ones might be the preferred endpoints in phytotoxicity assays. Fv/Fm, i.e., the most extensively reported parameter literature-wise, proved to be the least sensitive endpoint; therefore, future studies might also consider reporting Fv/Fo, as its more responsive analogue. The tested toxicants induced different trends in the basic chlorophyll fluorescence parameters and, at least partly, in relative proportions of different quenching processes, suggesting that a basic distinction of water pollutants with different modes of action might be achievable by this method. We found definite hormetic patterns in responses to several endpoints. Hormesis occurred in the concentration ranges where the applied toxicants resulted in strong growth inhibition in longer-term exposures of the same duckweed clone in previous studies. These findings indicate that changes in the photochemical efficiency of plants do not necessarily go hand in hand with growth responses, and care should be taken when one exclusively interprets chlorophyll fluorescence-based endpoints as general proxies for phytotoxic effects.

Mitochondrial Proteostasis Requires Genes Encoded in a Neurodevelopmental Syndrome Locus.

Eukaryotic cells maintain proteostasis through mechanisms that require cytoplasmic and mitochondrial translation. Genetic defects affecting cytoplasmic translation perturb synapse development, neurotransmission, and are causative of neurodevelopmental disorders, such as Fragile X syndrome. In contrast, there is little indication that mitochondrial proteostasis, either in the form of mitochondrial protein translation and/or degradation, is required for synapse development and function. Here we focus on two genes deleted in a recurrent copy number variation causing neurodevelopmental disorders, the 22q11.2 microdeletion syndrome. We demonstrate that SLC25A1 and MRPL40, two genes present in the microdeleted segment and whose products localize to mitochondria, interact and are necessary for mitochondrial ribosomal integrity and proteostasis. Our Drosophila studies show that mitochondrial ribosome function is necessary for synapse neurodevelopment, function, and behavior. We propose that mitochondrial proteostasis perturbations, either by genetic or environmental factors, are a pathogenic mechanism for neurodevelopmental disorders.SIGNIFICANCE STATEMENT The balance between cytoplasmic protein synthesis and degradation, or cytoplasmic proteostasis, is required for normal synapse function and neurodevelopment. Cytoplasmic and mitochondrial ribosomes are necessary for two compartmentalized, yet interdependent, forms of proteostasis. Proteostasis dependent on cytoplasmic ribosomes is a well-established target of genetic defects that cause neurodevelopmental disorders, such as autism. Here we show that the mitochondrial ribosome is a neurodevelopmentally regulated organelle whose function is required for synapse development and function. We propose that defective mitochondrial proteostasis is a mechanism with the potential to contribute to neurodevelopmental disease.

Small Size of Recorded Neuronal Structures Confines the Accuracy in Direct Axonal Voltage Measurements.

Patch-clamp instruments including amplifier circuits and pipettes affect the recorded voltage signals. We hypothesized that realistic and complete in silico representation of recording instruments together with detailed morphology and biophysics of small recorded structures will reveal signal distortions and provide a tool that predicts native, instrument-free electrical signals from distorted voltage recordings. Therefore, we built a model that was verified by small axonal recordings. The model accurately recreated actual action potential (AP) measurements with typical recording artefacts and predicted the native electrical behavior. The simulations verified that recording instruments substantially filter voltage recordings. Moreover, we revealed that instrumentation directly interferes with local signal generation depending on the size of the recorded structures, which complicates the interpretation of recordings from smaller structures, such as axons. However, our model offers a straightforward approach that predicts the native waveforms of fast voltage signals and the underlying conductances even from the smallest neuronal structures.

FvmnSOD is involved in oxidative stress defence, mitochondrial stability and apoptosis prevention in Fusarium verticillioides.

Superoxide dismutases are key enzymes in elimination of the superoxide anion radical (O2 •- ) generated intracellularly or by exogenous oxidative stress eliciting agents, like menadione. In this study, we investigated the physiological role of the manganese superoxide dismutase-encoding gene in Fusarium verticillioides via the construction of a gene deletion mutant, ΔFvmnSOD and comparing its phenotype with that of the wild-type parental strain and a ΔFvmnSOD' C strain, complemented with the functional manganese superoxide dismutase gene. Deletion of FvmnSOD had no effect on the relative intracellular superoxide ratio but increased the sensitivity of the fungus to menadione sodium bisulphite on Czapek-Dox stress agar plates. The lack of FvmnSOD caused changes in mitochondrial morphology and physiology: The volumetric ratio of these cell organelles in the second hyphal segment, as well as the total, the KCN-sensitive cytochrome c-dependent and the KCN+SHAM (salicylhidroxamic acid)-resistant residual respiration rates, were higher in the mutant as compared to the wild-type and the complemented strains. Nevertheless, changes in the respiration rates were attributable to the higher volumetric ratio of mitochondria found in the gene deletion mutant. Changes in the mitochondrial functions also brought about higher sensitivity to apoptotic cell death elicited by the Penicillium chrysogenum antifungal protein. The gene deletion mutant developed significantly thinner hyphae in comparison to the wild-type strain. Deletion of FvmnSOD had no effect on fumonisin B1 and B2 production of the fungus grown in Myro medium as a static culture.

Functional specification of CCK+ interneurons by alternative isoforms of Kv4.3 auxiliary subunits.

CCK-expressing interneurons (CCK+INs) are crucial for controlling hippocampal activity. We found two firing phenotypes of CCK+INs in rat hippocampal CA3 area; either possessing a previously undetected membrane potential-dependent firing or regular firing phenotype, due to different low-voltage-activated potassium currents. These different excitability properties destine the two types for distinct functions, because the former is essentially silenced during realistic 8-15 Hz oscillations. By contrast, the general intrinsic excitability, morphology and gene-profiles of the two types were surprisingly similar. Even the expression of Kv4.3 channels were comparable, despite evidences showing that Kv4.3-mediated currents underlie the distinct firing properties. Instead, the firing phenotypes were correlated with the presence of distinct isoforms of Kv4 auxiliary subunits (KChIP1 vs. KChIP4e and DPP6S). Our results reveal the underlying mechanisms of two previously unknown types of CCK+INs and demonstrate that alternative splicing of few genes, which may be viewed as a minor change in the cells' whole transcriptome, can determine cell-type identity.

Retrospective analyses of archive phytotoxicity test data can help in assessing internal dynamics and stability of growth in laboratory duckweed cultures.

High growth potential of duckweed species (Lemnaceae family) has been utilized in wide range of research and practical applications. Based on literature data, however, it can be assumed that duckweed populations maintain constant growth rates only when short periods are considered but can vary over longer time scales. This intrinsic instability in growth can affect the interpretation of growth data. Duckweed phytotoxicity tests are usually performed according to highly standardized protocols. Therefore the archive data provide an opportunity for retrospective comparisons. In the present study we collected growth (frond number- and frond area-based relative growth rates) and morphology (average frond and colony sizes) data from control treatments of phytotoxicity tests. All the analyzed tests were carried out with the same Spirodela polyrhiza (L.) Schleid. (giant duckweed) clone (RDSC ID No. 5501) under the same experimental conditions over more than four years. We aimed to assess the overall variability of the above parameters and to test if intrinsic growth patterns affect growth data in short-term. In general, the results reflected high stability of the measured parameters in long term but also indicated that some temporal variability is inevitable which can bias the comparability of growth tests. The frond area-based relative growth rate resulted in smaller coefficient of variation than the usually preferred frond number-based one. The results also revealed a negative correlation between mean growth rates and their coefficients of variation. Therefore, it would be advisable to introduce higher minimal growth rates and/or maximized tolerable coefficients of variation for control cultures into the standard duckweed growth inhibition tests. Analyses of growth data aggregated on seasonal basis indicated faster growth and larger mean frond size in laboratory duckweed cultures from mid-autumn till mid-spring than during summer and early autumn. But, in shorter term (∼50 days) we did not observe distinct trends in growth suggesting that the successive frond generations have no effect on growth traits within this time-scale. Our results point to the importance of assessing intrinsic growth dynamics in duckweed cultures and also to the re-usability of the already collected phytotoxicity data in addressing new research questions.