Biology subjects

Best, C.

Publications and source records attributed to Best, C..

12 recordsLinked to original sources

bioRxiv2026

Stressor- and tissue-specific regulation of the corticotropin-releasing factor system across epithelial tissues in rainbow trout

The corticotropin-releasing factor (CRF) system is a major neural regulator of stress responses in vertebrates. However, stress-related roles for the CRF system in other tissues--and whether these roles vary between stressor types--remain unclear. To address this gap, we first characterized the CRF system in the gills and intestine of rainbow trout (Oncorhynchus mykiss) and then evaluated how it is transcriptionally regulated following either an immune (vaccination) or osmotic (seawater transfer) stressor. Additionally, since the CRF system is involved in food intake regulation, we also evaluated whether feeding state affects the intestinal CRF system. Vaccination against Vibrio anguillarum reduced CRF system activity in the intestine--as indicated by elevations in CRF binding protein transcripts paired with reductions in ligand (crfa2) and receptor (crfr1b) transcripts--but did not affect the gill CRF system. In contrast, seawater transfer caused the abundance of most CRF system transcripts to increase in the middle (but not posterior) portion of the intestine, while transcript levels of CRF binding proteins and receptors in the gills declined. Finally, levels of CRF system transcripts in the intestine varied with feeding state in a region-specific manner. In the middle intestine, transcript levels of most components declined with fasting and increased when feeding was resumed, whereas the opposite pattern occurred in the posterior intestine. Overall, our results implicate the peripheral CRF system as a stressor- and epithelial tissue-specific modulator of immune and osmoregulatory functions in teleosts.

physiology↗

bioRxiv2025

Myelin Decompaction in Mice Given Anesthetics during Magnetic Resonance Imaging

AbstractThe objective of this secondary analysis of a prior investigation was to determine if prolonged exposure to the anesthetics isoflurane and dexmedetomidine during MRI was associated with a higher proportion of axons with myelin decompaction. 16 mice underwent an MRI protocol in which they had prolonged exposure to isoflurane and dexmedetomidine, while 10 mice did not undergo this protocol. All mice were sacrificed and electron microscope images were taken of various brain regions including the right prefrontal cortex (anterior cingulate and prelimbic area), the nucleus accumbens, the amygdala, and the ventral hippocampus.. Proportion of decompacted axons was calculated for each mouse, and an inter-rater reliability score of 80% was achieved. Welchs t-tests were used to test the hypothesis that mice undergoing MRI with prolonged anesthesia had greater levels of myelin decompaction than mice that did not experience prolonged anesthesia. Mice with prolonged anesthetic exposure during MRI had significantly higher proportions of decompacted axons than mice that did not experience prolonged anesthesia (p-value of 0.00003642). Prolonged exposure to anesthetics, particularly isoflurane, may be associated with myelin decompaction. These findings, if replicated, have potential to impact future anesthesia use in clinical work and scientific research. Significance StatementProlonged exposure to the anesthetics isoflurane and dexmedetomidine during brain imaging may lead to myelin decompaction in adult rodents. Myelin is a protective sheath around nerve fibers that ensures efficient transmission of electrical signals in the nervous system. Decompaction of myelin can disrupt these signals, potentially leading to neurological issues. This discovery is significant because it highlights potential risks associated with these anesthetics, which are commonly used in fMRI studies of rodents and in veterinary and medical procedures.

neuroscience↗

bioRxiv2024

Transcriptional profiling provides insights on sublethal thermal stress thresholds in juvenile bull trout

Bull trout (Salvelinus confluentus) inhabit mainly cold-water streams in their Western North American range. Multiple anthropogenic stressors, including climate change, have caused population declines and range reduction, leading to classification of bull trout as threatened. Characterizing how fish respond to increasing temperature can identify key thresholds beyond which fish health is impacted, which is useful for developing species recovery strategies. Juvenile bull trout were acclimated to a range of relevant temperatures (6-21 {degrees}C), after which mRNA transcripts involved in responding to thermal stress were measured using high- throughput qPCR, and their upper thermal tolerance (critical thermal maximum, CTmax) was determined. Beyond 18 {degrees}C appears to be a critical sublethal threshold for bull trout: capacity to increase CTmax through acclimation was lost, growth and survival decreased, and transcriptional data suggest widespread activation of cellular stress and growth suppression. Below 18 {degrees}C, transcriptional data suggest subtler metabolic adjustments in response to acclimation temperature, preceding changes in whole-animal performance. Altogether, these data provide insight into thermal responses in bull trout, and their capacity to cope with climate-change related warming.

physiology↗

bioRxiv2023

Effect of Human Infant Gut Microbiota on Mouse Behavior, Dendritic Complexity, and Myelination

The mammalian gut microbiome influences numerous developmental processes. In human infants it has been linked with cognition, social skills, hormonal responses to stress, and brain connectivity. Yet, these associations are not necessarily causal. The present study tested whether two microbial stool communities, common in human infants, affected behavior, myelination, dendritic morphology, and spine density when used to colonize mouse models. Humanized animals were more like specific-pathogen free mice than germ-free mice for most phenotypes, although in males, both humanized groups were less social. Both humanized groups had thinner myelin sheaths in the hippocampus, than did germ-free animals. Humanized animals were similar to each other except for dendritic morphology and spine density where one group had greater dendritic length in the prefrontal cortex, greater dendritic volume in the nucleus accumbens, and greater spine density in both regions, compared to the other. Results add to a body of literature suggesting the gut microbiome impacts brain development. TeaserFecal transplants from human infants with highly abundant Bifidobacterium, an important inhabitant of the intestinal tract of breastfed newborns, may promote brain connectivity in mice.

neuroscience↗

bioRxiv2022

MSP1 encodes an essential RNA-binding PPR factor required for nad1 maturation and complex I biogenesis in Arabidopsis mitochondria

SummaryMitochondria are semi-autonomous organelles that serve as hubs for aerobic energy metabolism. The biogenesis of the respiratory (OXPHOS) system relies on nuclear-encoded factors, which regulate the transcription, processing and translation of mitochondrial (mt)RNAs. These include proteins of primordial origin, as well as eukaryotic-type RNA-binding families recruited from the host genomes to function in mitogenome expression. Pentatricopeptide repeat (PPR) proteins constitute a major gene-family in angiosperms that is pivotal in many aspects of mtRNA metabolism, such as editing, splicing or stability. Here, we report the analysis of MITOCHONDRIA STABILITY/PROCESSING PPR FACTOR1 (MSP1, At4g20090), a canonical mitochondria-localized PPR protein that is necessary for mitochondrial biogenesis and embryo-development. Functional complementation confirmed that the phenotypes result from a disruption of the MSP1 gene. As a loss-of-function allele of Arabidopsis MSP1 leads to seed abortion, we employed an embryo-rescue method for the molecular characterization of msp1 mutants. Our data show that msp1 embryo-development fails to proceed beyond the heart-torpedo transition stage as a consequence of a severe nad1 pre-RNA processing-defect, resulting in the loss of respiratory complex I (CI) activity. The maturation of nad1 involves the processing of three RNA-fragments, nad1.1, nad1.2 and nad1.3. Based on biochemical analyses and the mtRNA profiles in wild-type and msp1 plants, we concluded that through its association with a specific site in nad1.1, MSP1 facilitates the generation of its 3-terminus and stabilizes it -a prerequisite for nad1 exons a-b splicing. Our data substantiate the importance of mtRNA metabolism for the biogenesis of the respiratory machinery during early-plant development.

plant biology↗

bioRxiv2020

Insights into the mitochondrial transcriptome landscapes of two Brassicales plant species, Arabidopsis thaliana (var. Col-0) and Brassica oleracea (var. botrytis)

Mitochondria play key roles in cellular energy metabolism within eukaryotic cells. As relics of endosymbiotic bacteria, most (but not all) mitochondria contain their own genome (mitogenome, mtDNA), as well as intrinsic biosynthetic machinery for making organelle RNAs and proteins. The expression of the mtDNA requires regulated metabolism of its transcriptome by nuclear-encoded factors. Post-transcriptional mtRNA modifications play a central role in the expression of the plant mitogenomes, and hence in organellar biogenesis and plant physiology. Despite extensive investigations, a full map of angiosperm mitochondrial transcriptomes, a prerequisite for the elucidation of the basic RNA biology of mitochondria, has not been reported yet. Using RNA-seq data, RT-PCR and bioinformatics, we sought to explore the gene expression profiles of land plant mitochondria. Here, we present the mitochondrial transcriptomic maps of two key Brassicaceae species, Arabidopsis thaliana (var Col-0) and cauliflower (Brassica oleracea var. botrytis). The revised transcriptome landscapes of Arabidopsis and cauliflower mitogenomes provide with more detail into mtRNA biology and processing in angiosperm mitochondria, and we expect that they would serve as a valuable resource for the plant organellar community. Accession numbersSequences are available at the Sequence Read Archive (accession no. PRJNA472433), for both Arabidopsis thaliana var. Col-0 mtRNA (SRA no. SRX4110179) and Brassica oleracea var. botrytis mtRNA (SRA no. SRX4110177).

plant biology↗

bioRxiv2020

nMAT3 is an essential maturase splicing factor required for holo-complex I biogenesis and embryo-development in Arabidopsis thaliana plants

Group II introns are large catalytic RNAs that are particularly prevalent in the organelles of terrestrial plants. In angiosperm mitochondria, group II introns reside in the coding-regions of many critical genes, and their excision is essential for respiratory-mediated functions. Canonical group II introns are self-splicing and mobile genetic elements, consisting of the catalytic intron-RNA and its cognate intron-encoded endonuclease factor (i.e. maturase, Pfam-PF01348). Plant organellar introns are extremely degenerate, and lack many regions that are critical for splicing, including their related maturase-ORFs. The high degeneracy of plant mitochondrial introns was accompanied during evolution by the acquisition of host-acting protein cofactors. These include several nuclear encoded maturases (nMATs) and various other splicing-cofactors that belong to a diverse set of RNA-binding families, e.g. RNA helicases (Pfam-PF00910), Mitochondrial Transcription Termination Factors (mTERF, Pfam-PF02536), Plant Organelle RNA Recognition (PORR, Pfam-PF11955), and Pentatricopeptide repeat (PPR, Pfam-PF13812) proteins. Previously, we established the roles of MatR and three nuclear-maturases, nMAT1, nMAT2, and nMAT4, in the splicing of different subsets of mitochondrial introns in Arabidopsis. The function of nMAT3 (AT5G04050) was found to be essential during early embryogenesis. Using a modified embryo-rescue method, we show that nMAT3-knockout plants are strongly affected in the splicing of nad1 introns i1, i3 and i4 in Arabidopsis mitochondria. The embryo-defect phenotype is tightly associated with complex I biogenesis defects. Functional complementation of nMAT3 restored the splicing defects and altered embryogenesis phenotypes associated with the nmat3 mutant-line.

plant biology↗

bioRxiv2020

High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells

We present high-resolution, high-speed fluorescence lifetime imaging microscopy (FLIM) of live cells based on a compressed sensing scheme. By leveraging the compressibility of biological scenes in a specific domain, we simultaneously record the time-lapse fluorescence decay upon pulsed laser excitation within a large field of view. The resultant system, referred to as compressed FLIM, can acquire a widefield fluorescence lifetime image within a single camera exposure, eliminating the motion artifact and minimizing the photobleaching and phototoxicity. The imaging speed, limited only by the readout speed of the camera, is up to 100 Hz. We demonstrated the utility of compressed FLIM in imaging various transient dynamics at the microscopic scale.

bioengineering↗

bioRxiv2020

Quantifying myelin content in brain tissue using color spatial light interference microscopy (cSLIM)

Deficient myelination of the brain is associated with neurodevelopmental delays, particularly in high-risk infants, such as those born small in relation to their gestational age (SGA). New methods are needed to further study this condition. Here, we employ Color Spatial Light Interference Microscopy (cSLIM), which uses a brightfield objective and RGB camera to generate pathlength-maps with nanoscale sensitivity in conjunction with a regular brightfield image. Using tissue sections stained with Luxol Fast Blue, the myelin structures were segmented from a brightfield image. Using a binary mask, those portions were quantitatively analyzed in the corresponding phase maps. We first used the CLARITY method to remove tissue lipids and validate the sensitivity of cSLIM to lipid content. We then applied cSLIM to brain histology slices. These specimens are from a previous MRI study, which demonstrated that appropriate for gestational age (AGA) piglets have increased internal capsule myelination (ICM) compared to small for gestational age (SGA) piglets and that a hydrolyzed fat diet improved ICM in both. The identity of samples was blinded until after statistical analyses.

neuroscience↗

bioRxiv2019

Early life stress impairs social function through AVP-dependent mechanisms

Impaired social function is a core feature of many psychiatric illnesses. Adverse experiences during childhood increase risk for mental illness, however it is currently unclear whether stress early in life plays a direct role in the development of social difficulties. Using an animal model of pre-pubertal stress (PPS), we investigated effects on social behaviour, oxytocin and arginine vasopressin (AVP). We also explored social performance and AVP expression in participants with borderline personality disorder (BPD) who experienced a high incidence of childhood stress. Social behaviour was impaired and AVP expression increased in animals experiencing PPS and participants with BPD. Behavioural deficits in animals were rescued through administration of the AVP receptor 1a antagonist Relcovaptan (SR49059). AVP levels and recognition of negative emotions were significantly correlated in BPD participants only. In conclusion, early life stress plays a profound role in the precipitation of social dysfunction, and AVP mediates at least part of this effect.

neuroscience↗

bioRxiv2019

The unique topologies of N6-Adenosine methylation (m6A) in land-plant mitochondria and their putative effects on organellar gene-expression

Mitochondria are the main source of ATP production and also contribute to many other processes central to cellular function. Mitochondrial activities have been linked with growth, differentiation and aging. As relicts of bacterial endosymbionts, these organelles contain their own genetic system (i.e., mitogenome or mtDNA). The expression of the mtDNA in plants is complex, particularly at the posttranscriptional level. Following transcription, the polycistronic pre-RNAs in plant mitochondria are processed into individual RNAs, which then undergo extensive modifications, as trimming, splicing and C[->]U editing, before being translated by organellar ribosomes. Our study focuses on N6-methylation of Adenosine ribonucleotides (m6A-RNA) in plant mitochondria. m6A is the most common modification in eukaryotic mRNAs. The biological significance of this highly dynamic modification is under investigation, but its widely accepted that m6A mediates structural switches that affect RNA stability and activity. By performing m6A-pulldown/RNA-seq (m6A-RIP-seq) analyses of Arabidopsis and cauliflower mitochondrial transcripts (mtRNAs), we provide with detail information on the m6A landscapes in angiosperms mitochondria. The results indicate that m6A targets different types of mtRNAs, including coding sequences, UTRs, introns and non-coding RNA species. While introns and noncoding-RNAs undergo multiple m6A modifications along the transcript, in mRNAs m6A-modifications are preferably positioned near start-codons, and may modulate the translatability of the m6A-modified transcripts.

molecular biology↗

bioRxiv2019

The PPR-related splicing cofactor MSP1/EMB1025 protein, encoded by At4g20090, encode an essential protein that is required for the splicing of nad1 intron 1 and for the biogenesis of complex I in Arabidopsis mitochondria

Group II introns are particularly plentiful within plant mitochondrial genomes (mtDNAs), where they interrupt the coding-regions of many organellar genes, especialy within complex I (CI) subunits. Their splicing is essential for the biogenesis of the respiratory system and is facilitated by various protein-cofactors that belong to a diverse set of RNA-binding cofactors. These including maturases, which co-evolved with their host-introns, and various trans-acting factors, such as members of the pentatricopeptide-repeat (PPR) protein family. The genomes of angiosperms contain hundreds of PPR-related genes that are postulated to reside within the organelles and affect diverse posttranscriptional steps, such as editing, RNA-stability and processing or translation. Here, we report the characterization of MSP1 (Mitochondria Splicing PPR-factor 1; also denoted as EMB1025), which plays a key role in the processing of nad1 pre-RNAs in Arabidopsis mitochondria. Mutations in MSP1 gene-locus (At4g20090) result in early embryonic arrest. To analyze the putative roles of MSP1 in organellar RNA-metabolism we used a modified embryo-rescue method, which allowed us to obtain sufficient plant tissue for the analysis of the RNA and protein profiles associated with msp1 mutants. Our data indicate that MSP1 is essential for the trans-splicing of nad1 intron 1 in Arabidopsis mitochondria. Accordingly, msp1 mutants show CI biogenesis defects and reduced respiratory-mediated functions. These results provide with important insights into the roles of nuclear-encoded factors during early plant development, and contribute to our limited understanding of the importance of RNA-maturation and splicing in plant mitochondria during early embryogenesis.

plant biology↗