Single-neuron analysis of aging-associated changes in learning reveals impairments in transcriptional plasticity.

The molecular mechanisms underlying age-related declines in learning and long-term memory are still not fully understood. To address this gap, our study focused on investigating the transcriptional landscape of a singularly identified motor neuron L7 in Aplysia, which is pivotal in a specific type of nonassociative learning known as sensitization of the siphon-withdraw reflex. Employing total RNAseq analysis on a single isolated L7 motor neuron after short-term or long-term sensitization (LTS) training of Aplysia at 8, 10, and 12 months (representing mature, late mature, and senescent stages), we uncovered aberrant changes in transcriptional plasticity during the aging process. Our findings specifically highlight changes in the expression of messenger RNAs (mRNAs) that encode transcription factors, translation regulators, RNA methylation participants, and contributors to cytoskeletal rearrangements during learning and long noncoding RNAs (lncRNAs). Furthermore, our comparative gene expression analysis identified distinct transcriptional alterations in two other neurons, namely the motor neuron L11 and the giant cholinergic neuron R2, whose roles in LTS are not yet fully elucidated. Taken together, our analyses underscore cell type-specific impairments in the expression of key components related to learning and memory within the transcriptome as organisms age, shedding light on the complex molecular mechanisms driving cognitive decline during aging.

Antibiotics reduce Pocillopora coral-associated bacteria diversity, decrease holobiont oxygen consumption and activate immune gene expression.

Corals are important models for understanding invertebrate host-microbe interactions; however, to fully discern mechanisms involved in these relationships, experimental approaches for manipulating coral-bacteria associations are needed. Coral-associated bacteria affect holobiont health via nutrient cycling, metabolic exchanges and pathogen exclusion, yet it is not fully understood how bacterial community shifts affect holobiont health and physiology. In this study, a combination of antibiotics (ampicillin, streptomycin and ciprofloxacin) was used to disrupt the bacterial communities of 14 colonies of the reef framework-building corals Pocillopora meandrina and P. verrucosa, originally collected from Panama and hosting diverse algal symbionts (family Symbiodiniaceae). Symbiodiniaceae photochemical efficiencies and holobiont oxygen consumption (as proxies for coral health) were measured throughout a 5-day exposure. Antibiotics altered bacterial community composition and reduced alpha and beta diversity, however, several bacteria persisted, leading to the hypothesis that these bacteria are either antibiotics resistant or occupy internal niches that are shielded from antibiotics. While antibiotics did not affect Symbiodiniaceae photochemical efficiency, antibiotics-treated corals had lower oxygen consumption rates. RNAseq revealed that antibiotics increased expression of Pocillopora immunity and stress response genes at the expense of cellular maintenance and metabolism functions. Together, these results reveal that antibiotic disruption of corals' native bacteria negatively impacts holobiont health by decreasing oxygen consumption and activating host immunity without directly impairing Symbiodiniaceae photosynthesis, underscoring the critical role of coral-associated bacteria in holobiont health. They also provide a baseline for future experiments that manipulate Pocillopora corals' symbioses by first reducing the diversity and complexity of coral-associated bacteria.

A tropical eastern Pacific invasive brittle star species (Echinodermata: Ophiuroidea) reaches southeastern Florida.

The invasive brittle star Ophiothela mirabilis (family Ophiotrichidae), a tropical Indo-Pacific endemic species, first reported in Atlantic waters off southern Brazil in 2000, has extended its range northward to the Caribbean Sea, to the Lesser Antilles in 2011, and was first reported in south Florida in January 2019. Its occurrence in southeast Florida extends along nearly 70km of coastline, from near the Port of Miami, Miami-Dade County, northward to Deerfield Beach, Broward County. It occurs abundantly as an epizoite on octocorals, attaining population densities of 25 individuals and more per 10-cm long octocoral stem. The surface texture of octocoral hosts (rough, smooth) did not affect the densities of the ophiuroid epizoites, and there were significantly greater abundances on octocorals during two winter sampling periods than in the summer. Beige and orange-coloured morphs are sometimes present on the same octocoral stem. Gut content analysis supported a suspension feeding mode, revealing essentially identical ingested items in both colour morphs with a preponderance of amorphous detritus and filamentous algae. Molecular genetic evidence (COI & 16s) has established the identity of O. mirabilis and its relationship to invasive Brazilian populations. The orange and beige morphs form two distinct, but closely related lineages that may represent two separate introductions. The orange morph shares haplotypes with Brazilian and Caribbean specimens suggesting a further range expansion of the 'original' invasion. The beige morph, however, shares haplotypes with specimens from the Mexican Pacific and Peru and potentially represents a secondary introduction. Traits promoting dispersal and establishment of this species in new habitats are manifold: vagility and ability to cling tightly to diverse host taxa (e.g. sponges, cnidarians, bryozoans, and echinoderms), frequent asexual reproduction (fissiparity), suspension feeding, including a wide range of dietary items, possession of integument-covered ossicles and arm spines offering protection from predators, and an effective competitive edge over associated microbiota for substrate space.

A comparison of hatchery-rearing in exercise to wild animal physiology and reflex behavior in Aplysia californica.

Aplysia californica was hatchery-reared in two turbulence protocols intended to imitate the intermittent turbulence of the native habitat and to promote development of the foot muscle from the exercise of adhering to the substrate. Hatchery-reared animals in turbulence regimes were compared to siblings reared in quiet water, and to wild animals, using noninvasive assessments of the development of the foot muscle. The objective was to learn if the turbulence-reared phenotype mimicked laboratory-targeted aspects of the wild phenotype, that is, reflex behavior, swim tunnel performance, and resting oxygen consumption (MO2). No group exhibited different MO2. MO2 values for all of the compared groups of animals followed the power law, with an exponent of 0.69, consistent with this relationship throughout the animal kingdom. Turbulence-induced exercise did not affect the righting reflex or the tail withdrawal reflex, standard behavioral tests that involve the foot muscle, compared to quiet water-reared siblings. Wild individuals had significantly shorter time-to-right than all hatchery reared animals, however, wild animals did not perform better in flume tests. That turbulence-reared hatchery- or wild animals lacked superior flume performance suggests that this species may shelter from intertidal wave energy to remain near its optimal feeding areas.

The effects of Symbiodinium (Pyrrhophyta) identity on growth, survivorship, and thermal tolerance of newly settled coral recruits.

For many coral species, the obligate association with phylogenetically diverse algal endosymbiont species is dynamic in time and space. Here, we used controlled laboratory inoculations of newly settled, aposymbiotic corals (Orbicella faveolata) with two cultured species of algal symbiont (Symbiodinium microadriaticum and S. minutum) to examine the role of symbiont identity on growth, survivorship, and thermal tolerance of the coral holobiont. We evaluated these data in the context of Symbiodinium photophysiology for 9 months post-settlement and also during a 5-d period of elevated temperatures Our data show that recruits that were inoculated with S. minutum grew significantly slower than those inoculated with S. microadriaticum (occasionally co-occurring with S. minutum), but that there was no difference in survivorship of O. faveolata polyps infected with Symbiodinium. However, photophysiological metrics (∆Fv/F'm, the efficiency with which available light is used to drive photosynthesis and α, the maximum light utilization coefficient) were higher in those slower growing recruits containing S. minutum. These findings suggest that light use (i.e., photophysiology) and carbon acquisition by the coral host (i.e., host growth) are decoupled, but did not distinguish the source of this difference. Neither Symbiodinium treatment demonstrated a significant negative effect of a 5-d exposure to temperatures as high as 32°C under low light conditions similar to those measured at settlement habitats.

Juvenile growth of the tropical sea urchin Lytechinus variegatus exposed to near-future ocean acidification scenarios.

To evaluate the effect of elevated pCO(2) exposure on the juvenile growth of the sea urchin Lytechinus variegatus, we reared individuals for three months in one of three target pCO(2) levels: ambient seawater (380 µatm) and two scenarios that are projected to occur by the middle (560 µatm) and end (800 µatm) of this century. At the end of 89 days, urchins reared at ambient pCO(2) weighed 12% more than those reared at 560 µatm and 28% more than those reared at 800 µatm. Skeletons were analyzed using scanning electron miscroscopy, revealing degradation of spines in urchins reared at elevated pCO(2) (800 µatm). Our results indicate that elevated pCO(2) levels projected to occur this century may adversely affect the development of juvenile sea urchins. Acidification-induced changes to juvenile urchin development would likely impair performance and functioning of juvenile stages with implications for adult populations.

The effects of dietary silver on larval growth in the echinoderm Lytechinus variegatus.

Previous studies have demonstrated that the euryhaline copepod Acartia tonsa is extremely sensitive to dietborne silver (Ag) exposure, with a 20 % inhibition (EC(20)) of survival occurring when copepods are fed algae with 1.6 µg g(-1) dry weight (dw) Ag, corresponding to a waterborne Ag concentration of 0.46 µg l(-1) Ag. In contrast, 43 µg l(-1) Ag is required to elicit similar effects in copepods exposed to Ag by way of water. In the current study, we investigated whether another planktonic marine organism might also be sensitive to dietary Ag. Specifically, we tested larvae of the echinoderm, Lytechinus variegatus in an 18-day study in which larvae were continuously exposed to Ag-laden algae (Isochrysis galbana). After 7 days of exposure, no significant effects were observed on larval growth up to the highest concentration tested (10.68 µg g(-1) dw Ag in algae after exposure to 3.88 µg l(-1) waterborne Ag). After 18 days, significant effects were observed in all Ag treatments resulting in a lowest-observable effect concentration of 0.68 µg g(-1) dw Ag in algae and corresponding waterborne Ag concentration of 0.05-0.07 µg l(-1) Ag (depending on background Ag [see Results]). However, the dose-response relationship was quite flat with a similar level of growth inhibition (approximately 15 %) in all Ag treatments, resulting in an EC(20) of >10.68 µg g(-1) dw Ag in algae (>3.88 µg l(-1) Ag in water). This flat dose-response relationship is characteristic of dietary metal (silver, copper, cadmium, nickel, and zinc) toxicity to copepods as well, although the effect is slightly more robust (approximately 20-30 % inhibition of survival or reproduction). We conclude that echinoderm larvae may be similar to copepods in their sensitivity to dietary Ag, although a better understanding of the mechanisms underlying the apparent flat dose-response relationships is clearly needed.

Larval growth, development, and survival of laboratory-reared Aplysia californica: effects of diet and veliger density.

Over the last three decades, the California sea hare, Aplysia californica, has played an increasingly important role as a model organism in the neurosciences. Since 1995, the National Resource for Aplysia has supported a growing research community by providing a consistent supply of laboratory-reared individuals of known age, reproductive status, and environmental history. The purpose of the present study was to resolve the key biological factors necessary for successful culture of large numbers of high quality larval Aplysia. Data from a sequence of five experiments demonstrated that algal diet, food concentration, and veliger density significantly affected growth, attainment of metamorphic competency, and survival of Aplysia larvae. The highest growth and survival were achieved with a mixed algal diet of 1:1 Isochrysis sp (TISO) and Chaetoceros muelleri (CHGRA) at a total concentration of 250 x 10(3) cells/mL and a larval density of 0.5-1.0 per mL. Rapid growth was always correlated with faster attainment of developmental milestones and increased survival, indicating that the more rapidly growing larvae were healthier. Trials conducted with our improved protocol resulted in larval growth rates of >14 microm/day, which yielded metamorphically competent animals within 21 days with survival rates in excess of 90%. These data indicate the important effects of biotic factors on the critical larval growth period in the laboratory and show the advantages of developing optimized protocols for culture of such marine invertebrates.