Mutant analysis of Kcng4b reveals how the different functional states of the voltage-gated potassium channel regulate ear development.

The voltage gated (Kv) slow-inactivating delayed rectifier channel regulates the development of hollow organs of the zebrafish. The functional channel consists of the tetramer of electrically active Kcnb1 (Kv2.1) subunits and Kcng4b (Kv6.4) modulatory or electrically silent subunits. The two mutations in zebrafish kcng4b gene - kcng4b-C1 and kcng4b-C2 (Gasanov et al., 2021) - have been studied during ear development using electrophysiology, developmental biology and in silico structural modelling. kcng4b-C1 mutation causes a C-terminal truncation characterized by mild Kcng4b loss-of-function (LOF) manifested by failure of kinocilia to extend and formation of ectopic otoliths. In contrast, the kcng4b-C2-/- mutation causes the C-terminal domain to elongate and the ectopic seventh transmembrane (TM) domain to form, converting the intracellular C-terminus to an extracellular one. Kcng4b-C2 acts as a Kcng4b gain-of-function (GOF) allele. Otoliths fail to develop and kinocilia are reduced in kcng4b-C2-/-. These results show that different mutations of the silent subunit Kcng4 can affect the activity of the Kv channel and cause a wide range of developmental defects.

Immunohistochemical and ultrastructural characterization of the inner ear epithelial cells of splitnose rockfish (Sebastes diploproa).

The inner ear of teleost fish regulates the ionic and acid-base chemistry and secretes protein matrix into the endolymph to facilitate otolith biomineralization, which is used to maintain vestibular and auditory functions. The otolith is biomineralized in a concentric ring pattern corresponding to seasonal growth, and this calcium carbonate (CaCO3) polycrystal has become a vital aging and life-history tool for fishery managers, ecologists, and conservation biologists. Moreover, biomineralization patterns are sensitive to environmental variability including climate change, thereby threatening the accuracy and relevance of otolith-reliant toolkits. However, the cellular biology of the inner ear is poorly characterized, which is a hurdle for a mechanistic understanding of the underlying processes. This study provides a systematic characterization of the cell types in the inner ear of splitnose rockfish (Sebastes diploproa). Scanning electron microscopy revealed the apical morphologies of six inner ear cell types. In addition, immunostaining and confocal microscopy characterized the expression and subcellular localization of the proteins Na+-K+-ATPase, carbonic anhydrase, V-type H+-ATPase, Na+-K+-2Cl--cotransporter, otolith matrix protein 1, and otolin-1 in six inner ear cell types bordering the endolymph. This fundamental cytological characterization of the rockfish inner ear epithelium illustrates the intricate physiological processes involved in otolith biomineralization and highlights how greater mechanistic understanding is necessary to predict their multistressor responses to future climate change.

Gravity-Dependent Modulation of Downbeat Nystagmus and Subjective Visual Vertical in the Roll Plane.

Downbeat nystagmus (DBN) is the most common form of acquired central vestibular nystagmus. Gravity perception in patients with DBN has previously been investigated by means of subjective visual straight ahead (SVA) and subjective visual vertical (SVV) in the pitch and roll planes only during whole-body tilts. To our knowledge, the effect of head tilt in the roll plane on the SVV and on DBN has not yet been systematically studied in patients. In this study, we investigated static and dynamic graviceptive function in the roll-plane in patients with DBN (patients) and healthy-controls (controls) by assessment of the Subjective Visual Vertical (SVV) and the modulation of slow-phase-velocity (SPV) of DBN. SPV of DBN and SVV were tested at different head-on trunk-tilt positions in the roll-plane (0°,30° clockwise (cw) and 30° counterclockwise (ccw)) in 26 patients suffering from DBN and 13 controls. In patients, SPV of DBN did not show significant modulations at different head-tilt angles in the roll-plane. SVV ratings did not differ significantly between DBN patients vs. controls, however patients with DBN exhibited a higher variability in mean SVV estimates than controls. Our results show that the DBN does not exhibit any modulation in the roll-plane, in contrast to the pitch-plane. Furthermore, patients with DBN show a higher uncertainty in the perception of verticality in the roll-plane in form of a higher variability of responses.

Monsoonal wet season influences the migration tendency of a catadromous fish (barramundi Lates calcarifer).

Many animals exhibit partial migration, which occurs when populations contain coexisting contingents of migratory and resident individuals. This individual-level variation in migration behaviour may drive differences in growth, age at maturity and survival. Therefore, partial migration is widely considered to play a key role in shaping population demography. Otolith chemistry and microstructural analysis were used to identify the environmental- and individual-specific factors that influence migratory behaviour in the facultatively catadromous barramundi (Lates calcarifer) at two distinct life history stages: firstly, as juveniles migrating upstream into fresh water; and secondly, as adults or sub-adults returning to the estuarine/marine spawning habitat. Monsoonal climate played an important role in determining the migration propensity of juveniles: individuals born in the driest year examined (weak monsoon) were more than twice as likely to undergo migration to freshwater than those born in the wettest (strong monsoon) year. In contrast, the ontogenetic timing of return migrations to the estuary by adults and sub-adults was highly variable and not strongly associated with the environmental parameters examined. We propose that scarce resources within saline natal habitats during lower rainfall years may provide an ecological incentive for juveniles to migrate upstream, whereas more abundant resources in higher rainfall years may promote resident life histories within estuaries. We conclude that inter-annual climatic variation, here evidenced by monsoonal strength, likely plays an important role in driving the persistence of diversified life histories within wild barramundi populations.

Ciliopathy genes are required for apical secretion of Cochlin, an otolith crystallization factor.

Here, we report that important regulators of cilia formation and ciliary compartment-directed protein transport function in secretion polarity. Mutations in cilia genes cep290 and bbs2, involved in human ciliopathies, affect apical secretion of Cochlin, a major otolith component and a determinant of calcium carbonate crystallization form. We show that Cochlin, defective in human auditory and vestibular disorder, DFNA9, is secreted from small specialized regions of vestibular system epithelia. Cells of these regions secrete Cochlin both apically into the ear lumen and basally into the basal lamina. Basally secreted Cochlin diffuses along the basal surface of vestibular epithelia, while apically secreted Cochlin is incorporated into the otolith. Mutations in a subset of ciliopathy genes lead to defects in Cochlin apical secretion, causing abnormal otolith crystallization and behavioral defects. This study reveals a class of ciliary proteins that are important for the polarity of secretion and delineate a secretory pathway that regulates biomineralization.

Untangling the taxonomic ambiguities of the spotted seerfish Scomberomorus guttatus with the description of a new species from India.

Scomberomorus guttatus has been subjected to a series of synonym assignations over the years. Its taxonomy has been mired with ambiguities due to the greater-than-average morphological variations observed in samples from different regions. An integrated taxonomic revision with molecular support indicated that the species that was thought to be a single entity, in reality, is a complex of three distinct species. They are morphologically distinct with respect to the body depth and elongation, cephalic morphometry, and meristic characteristics. Otolith morphometry and phylogenetic evidence further established the taxonomic divergence within the spotted seerfish complex. The phylogenetic characteristic as indicated by the mitochondrial Cytochrome c Oxidase subunit I (COI) sequence unveiled that S. guttatus had a high intraspecific divergence of 11.1% from its two identical congeners and a divergence of 2.34% between the congeners, indicating scope for categorizing them as separate species. Based on the morphological and molecular evidence, S. guttatus is redescribed; the senior synonym Scomberomorus leopardus is resurrected as a valid species; and a new species Scomberomorus avirostrus n. sp. is described with keys for species distinction.

Age, growth, maturity and mortality of the tapetail anchovy Coilia brachygnathus (Engraulidae) in Lake Honghu, China.

The tapertail anchovy Coilia brachygnathus, a commercially important species mainly distributed along the mid-lower Chang-Jiang basin, is by far the most dominant species in Lake Honghu. To figure out its success in this semiclosed lake, some basic biological parameters of this anchovy were analysed based on samples seasonally collected from October 2020 to December 2021. The results demonstrated that the age classes of fished individuals varied from 0.5 to 3.5, with the majority (97.36%) being between 0.5 and 3 years old. The size at 50% maturity of 17.2 cm total length (TL) for females and 19.0 cm TL for males corresponded to 1 and 1.6 years, respectively. Coilia brachygnathus has a short life span, early sexual maturity and a relatively fast growth rate. The flourishing of the fish in the lake is mainly attributed to its short life span, early maturity, fast growth rate, closed fishing, pelagic spawners, the availability of plenty of food and low predation effect on it. Age 3.5 year occurs in an extremely small percentage of the total (<3%), indicating that a large number of larger-sized or older fish died after spawning, which is probably one of the major sources of water pollution if the closed fishing measure is adopted in Lake Honghu. Thus, individuals older than 2 years or more than 20.0 cm TL should be harvested. These findings have important management implications for the fish resources in Lake Honghu and beyond.

Otolith sectioning reveals higher maximum age in greater weever (Trachinus draco).

Growth and maximum age are two key parameters that inform resilience of fish populations to exploitation. Existing information on those for greater weever inhabiting the eastern North Sea is based on the analysis of whole otoliths. Here, we present a reanalysis using sectioned otoliths. The results reveal a different growth pattern and a higher maximum age than that previously reported. The higher maximum age makes greater weever populations more vulnerable to exploitation. Such information can serve as a basis for the estimation of the growth curve that can be used for future assessment of the species.

A life in the surf zone: Age and feeding habits of the pompano (Trachinotus ovatus, Linnaeus, 1758) on the beaches of the Gulf of Cádiz (south-west Iberian Peninsula).

Surf zones are crucial habitats for many fish species, where they spend the initial stages of their lives, finding food and shelter. One such species is the pompano (Trachinotus ovatus). The aim of this study was to examine the age and diet of the pompano within the surf zone, along with any potential variations in the environment. The average size of the specimens was 10.61 ± 4.91 cm. The length-weight relationship was total weight (TW) = 0.0136*total length (TL)2.8512 (parameter a: 95% C.I.: 0.0130-0.0142 and parameter b: 95% C.I.: 2.8318-2.8705). The most abundant age classes were 0+ and 1+, making up 97% of the captured specimens, with significant differences in the abundance of age classes depending on the time of the day and season. The pompano primarily fed on clupeiforms, copepods, and mysids, with significant variations in diet based on the time of the day, season, and size of the specimens. This study contributes new information about the use of the surf zone by T. ovatus in its early years of life, underscoring the importance of these areas and their role as an additional ecosystem service.

Unravelling the life-history patterns and habitat preferences of the Japanese eel (Anguilla japonica) in the Pearl River, China.

Eels have fascinated biologists for centuries due to their amazing long-distance migrations between freshwater habitats and very distant ocean spawning areas. The migratory life histories of the Japanese eel, Anguilla japonica, in the waters of south China are not very clear despite its ecological importance, and the need for fishery regulation and management. In this study, strontium (Sr) and calcium (Ca) microchemical profiles of the otoliths of silver eels were measured by X-ray electron probe microanalysis based on data collected from different habitats (including freshwater and brackish habitats), in the large subtropical Pearl River. The corresponding habitat preference characteristics were further analysed using redundancy analysis (RDA). A total of 195 Japanese eels were collected over 6 years. The collected individuals ranged from 180 to 771 mm in total length and from 8 to 612 g in body weight. Two-dimensional pictures of the Sr:Ca concentrations in otoliths revealed that the A. japonica in the Pearl River are almost entirely river eels, spending the majority of their lives in fresh water without exposure to salt water, while the catadromous migration time has delayed about 1 month in the Pearl River estuary in the past 20 years. RDA analysis further indicated that juveniles and adults preferred water with high salinity and high tide levels. Youth preferred habitats with high river fractals. Our findings contribute to a growing body of evidence showing that the eels are extremely scarce currently and conservation measures against them are imminent, including the protection of brackish and freshwater areas where they live in south China.

Staining protocols affect use of otolith to estimate the demography of the damselfish sergeant major (Abudefduf vaigiensis).

This study assessed the otolith (sagittae, lapilli, and asterisci) increment deposition rate in the range-shifting damselfish, A. vaigiensis, using different concentrations of Alizain Red S and evaluated the impact of staining on increment width. Daily increment deposition was verified in all otolith types and presented clearer fluorescent markings in the lapilli and sagittae than the asterisci, with high stain concentration showing the best clarity. Higher stain concentrations were found to decrease increment width, suggesting care is needed when using stained otoliths as a proxy for growth for this species.

Age estimates derived from hard parts of swordfish Xiphias gladius from the north-western Mediterranean Sea.

Accurate age estimates for fish are critical for properly understanding stock dynamics and health; this is particularly true for larger billfishes. Here we determined the most accurate aging estimation methods for swordfish (Xiphias gladius). We compared age estimates obtained from fin-ray sections, otolith sections, whole otoliths, and vertebrae collected from 87 swordfish off the east coast of Corsica. Age estimates from otolith sections were most consistently estimated across different readers (lowest average percentage error), followed by fin-ray sections, third vertebrae, and whole otoliths. When the age estimates from the otolith sections were compared with the other three age sclerochronological methods, we found the average percentage error to be lowest between the otolith section and fin-ray methods. However, age estimates from fin rays proved most useful for estimating swordfish younger than 6 years, as the fin ray-based age diverged from that of the otolith sections as the swordfish aged. Combining fin ray and otolith section techniques, we estimated the growth parameters of 1-12-year-old females (L∞ = 259.412, k = 0.113, t0 = -2.499) and 1-7-year-old males (L∞ = 175.543, k = 0.202, t0 = -2.239). We found that females grew significantly faster than males after 3 years and remained larger thereafter. Our calculated growth rates for this region of the north-western Mediterranean Sea were lower than those of the Atlantic, Pacific, and eastern Mediterranean Sea swordfish populations, and similar to growth rates recorded for the western Mediterranean Sea populations. Our study provides critical knowledge on biological-related parameters to serve as a guide for preserving the swordfish population in the Mediterranean Sea.

The incorporation of strontium and barium into the otoliths of the flounder Paralichthys olivaceus at early life stages demonstrates resilience to ocean acidification.

Ocean acidification could modify the bioavailability and chemical properties of trace elements in seawater, which could affect their incorporation into the calcareous structures of marine organisms. Fish otoliths, biomineralized ear stones made by aragonite, are suspended within the endolymph fluid of teleosts, indicating that the elemental incorporation of otoliths might also be susceptible to ocean acidification. In this study, we evaluated the combined effects of CO2-induced ocean acidification (pH 8.10, 7.70, and 7.30, corresponding to ocean acidification scenarios under the representative concentration pathway 8.5 model as projected by the Intergovernmental Panel on Climate Change) and water elemental concentrations of strontium (Sr) and barium (Ba; low, medium, and high) on elemental incorporation into otoliths of the flounder Paralichthys olivaceus at early life stages. Our results revealed that the elemental incorporation of Sr and Ba into otoliths was principally dependent on the corresponding water elemental concentrations rather than on ocean acidification. Moreover, the partition coefficients (DMe) of Sr and Ba may stabilize after dynamic equilibrium is reached as the water elemental concentration increases, but are not affected by ocean acidification. Therefore, the incorporation of Sr and Ba into otoliths of the flounder at early life stages may not serve as an effective indicator of ocean acidification. In other words, the findings suggest that ocean acidification does not impact the incorporation of Sr and Ba incorporation into otoliths when tracing the temperature or salinity experiences of the flounder. Our findings will provide new knowledge for understanding the potential ecological effects of ocean acidification on the recruitment dynamics of fish species.

Otolith morphogenesis during the early life stages of fish is temperature-dependent: Validation by experimental approach applied to European seabass (Dicentrarchus labrax).

Otolith shape is often used as a tool in fish stock identification. The goal of this study was to experimentally assess the influence of changing temperature and ontogenic evolution on the shape component of the European seabass (Dicentrarchus labrax) otolith during early-life stages. A total of 1079 individuals were reared in a water temperature of 16°C up to 232 days post hatch (dph). During this experiment, several specimens were transferred into tanks with a water temperature of 21°C to obtain at the end of this study four different temperature treatments, each with varying ratios between the number of days at 16 and 21°C. To evaluate the otolith morphogenesis, samples were examined at 43, 72, 86 and 100 dph. The evolution of normalized otolith shape from hatching up to 100 dph showed that there were two main successive changes. First, faster growth in the antero-posterior axis than in the dorso-ventral axis changed the circular-shaped otolith from that observed at hatching and, second, increasing the complexity relating to the area between the rostrum and the anti-rostrum. To test the effect of changing temperature, growing degree-day was used in three linear mixed-effect models. Otolith morphogenesis was positively correlated to growing degree-day, but was also dependent on temperature level. Otolith shape is influenced by environmental factors, particularly temperature, making it an efficient tool for fish stock identification.

Daily age, growth rate, and pelagic larval duration of commercially important snapper species in Abrolhos National Marine Park.

The age and daily growth of fish are registered through the deposition of increments in their otoliths, which are concretions formed by the precipitation of substances present in the endolymphatic fluid, mainly calcium carbonate (CaCO3). Faced with the need to fill some of the gaps in the knowledge on the occurrence and duration of the initial stages of snapper species' life cycles in the Abrolhos Bank, this study aimed to describe the growth rates, age, and period of pelagic larval duration (PLD) of three snapper species during the larval pre-settlement phase, in the Abrolhos Bank region. The post-larvae were captured using light traps. Otoliths were removed from 117 samples of snapper species; however, only 69 were viable for age estimation, of which 15 were Lutjanus analis, 25 were Lutjanus jocu, and 29 were Lutjanus synagris. Together, the samples presented individuals with total lengths ranging from 16.14 to 24.76 mm and ages from 21 to 39 days. Settlement marks were found for all three species, and the average PLD was ~25 days. The somatic growth of the snapper species was positively correlated with otolith growth. L. jocu presented the greatest daily growth compared to the other species. The three species use the Abrolhos Bank as a larval settlement site, demonstrating plasticity by using different habitats throughout their lives.

Post-settlement dynamics of Japanese flounder Paralichthys olivaceus in Tango Bay: Seasonal patterns in growth, mortality, and recruitment potential.

Japanese flounder Paralichthys olivaceus is one of the most valuable coastal flatfish species in East Asia. To investigate post-settlement growth and mortality, juveniles were sampled in Tango Bay (Japan) weekly throughout the settlement period in 2007 and 2008. Otolith (lapillus) microstructure analysis enabled the categorization of juveniles into six biweekly cohorts each year. Later cohorts exhibited higher growth rates possibly because of higher water temperatures. A key observation was the direct relationship between high mortality and high density in mid-season cohorts in both years, pointing to density-dependent mortality. This increased mortality may be attributed to predation, including cannibalism by earlier cohorts. Furthermore, growth-selective mortality was evident soon after settlement, underscoring the vulnerability of slow growers to predation during the early juvenile stage. Although earlier and later cohorts were less abundant but showed promising recruitment potential, the prospective contribution of mid-season cohorts to the adult population remained uncertain. The results clearly highlight the importance of density-dependent mortality in population regulation in post-settlement Japanese flounder.

Drivers of growth in strong year classes of the deepwater redfish (Sebastes mentella) population from the Gulf of St. Lawrence derived from otolith increment-based growth chronologies.

The case of the deepwater redfish (Sebastes mentella) in the Gulf of St. Lawrence (GSL) is a compelling example of drastic fluctuations in annual recruitment strength, characteristic of spasmodic stocks. After three decades of low abundance, the emergence of three consecutive strong year classes in 2011-2013 resulted in an unprecedented increase in biomass. In spasmodic stocks such as GSL redfish, strong year classes sustain both the biomass and catch for decades. Therefore, understanding the growth dynamics of these cohorts is essential. In the present study, we reconstructed the annual growth rates of redfish using otolith increment-based annual chronology and investigated the drivers of growth variation in redfish strong year classes of the early 2010s and early 1980s. Stock biomass was identified as the main extrinsic driver of redfish growth, suggesting intense competition for food at high conspecific density. Warming of deep waters in the GSL, where adult redfish settle, positively correlated with individual growth. However, recent warming of the cold intermediate layer showed a negative correlation with redfish growth, likely related to the shrinking of the habitat this water mass provides for various redfish cold-water prey rather than to a direct effect of temperature. Reconstruction of redfish annual growth trajectories from birth to capture emphasized the importance of carryover effects in the growth potential of strong year classes. This work provided an important first outlook of the factors driving growth variation in GSL redfish spasmodic stock and explored midterm consequences of density-dependent pressures on biological parameters of the population.

Recruitment of European sea bass (Dicentrarchus labrax) in northerly UK estuaries indicates a mismatch between spawning and fisheries closure periods.

European sea bass (Dicentrarchus labrax) is a species of high commercial and recreational value, but it exhibits highly variable recruitment rates and has been subject to recent declines. Emergency management measures put in place to protect spawning stocks include the annual closure of commercial and recreational fisheries over a 2-month, February-March, window. Whether this protection measure is having the desired outcome for this data-poor species remains unclear. Otolith microstructural analyses (counts and widths of daily growth rings and check marks indicative of settlement) were used to estimate (1) spawn timing, (2) pelagic larval duration and settlement timing, (3) growth rate and condition, and (4) the otolith-fish size relationship for juvenile European sea bass caught from two estuaries in Wales (Dwyryd, Y Foryd), located at the northern edge of the species range. We observed a significant mismatch between the timing of fisheries closures and the spawning, with 99.2% of recruits having been spawned after the fishery had reopened (back-calculated median spawn date = May 5 ± 17 days SD), suggesting that the closure may be too early to adequately protect this population. Further, we present the first empirically derived estimates of pelagic larval duration for sea bass recruits settling in UK habitats, which showed a strong negative relationship with spawn date. Finally, we found significant differences in fish condition between the two estuaries, suggesting local variation in habitat quality. The results suggest that the timing of current fisheries closures may not be adequately protecting the spawners supplying these northernmost estuaries, which are likely to become increasingly important as sea bass distributions shift northward in our climate future.

Juvenile downstream migration patterns of an anadromous fish, allis shad (Alosa alosa), before and after the population collapse in the Gironde system, France.

Diadromous fish have exhibited a dramatic decline since the end of the 20th century. The allis shad (Alosa alosa) population in the Gironde-Garonne-Dordogne (GGD) system, once considered as a reference in Europe, remains low despite a fishing ban in 2008. One hypothesis to explain this decline is that the downstream migration and growth dynamics of young stages have changed due to environmental modifications in the rivers and estuary. We retrospectively analysed juvenile growth and migration patterns using otoliths from adults caught in the GGD system 30 years apart during their spawning migration, in 1987 and 2016. We coupled otolith daily growth increments and laser ablation inductively-coupled plasma mass spectrometry measurements of Sr:Ca, Ba:Ca, and Mn:Ca ratios along the longest growth axis from hatching to an age of 100 days (i.e., during the juvenile stage). A back-calculation allowed us to estimate the size of juveniles at the entrance into the brackish estuary. Based on the geochemistry data, we distinguished four different zones that juveniles encountered during their downstream migration: freshwater, fluvial estuary, brackish estuary, and lower estuary. We identified three migration patterns during the first 100 days of their life: (a) Individuals that reached the lower estuary zone, (b) individuals that reached the brackish estuary zone, and (c) individuals that reached the fluvial estuary zone. On average, juveniles from the 1987 subsample stayed slightly longer in freshwater than juveniles from the 2016 subsample. In addition, juveniles from the 2016 subsample entered the brackish estuary at a smaller size. This result suggests that juveniles from the 2016 subsample might have encountered more difficult conditions during their downstream migration, which we attribute to a longer exposure to the turbid maximum zone. This assumption is supported by the microchemical analyses of the otoliths, which suggests based on wider Mn:Ca peaks that juveniles in 2010s experienced a longer period of physiological stress during their downstream migration than juveniles in 1980s. Finally, juveniles from the 2016 subsample took longer than 100 days to exit the lower estuary than we would have expected from previous studies. Adding a new marker (i.e., Ba:Ca) helped us refine the interpretation of the downstream migration for each individual.

[Practical instructions for recording vestibular evoked myogenic potentials]. / Praktische Hinweise zur Messung vestibulär evozierter myogener Potenziale.

Recording of vestibular evoked myogenic potentials (VEMPs) is a well-established method for functional diagnostics of the otolith organs. VEMPs are vestibular reflexes of the sacculus und utriculus to acoustic stimulation by air-conducted sound or bone-conducted vibration and are recorded by surface electrodes from the cervical (cVEMP) and ocular (oVEMP) muscles. The results of VEMP recordings are part of the neuro-otologic test battery and enable diagnosis of various vestibular disorders or differentiation between non-vestibular and peripheral vestibular vertigo. However, the methods for recording VEMPs vary substantially, although recording and stimulation parameters as well as methods of data analysis have a significant influence on the results. This article provides an overview of recommended parameters as well as practical instructions for the recording, analysis, and interpretation of VEMPs.