Microscopic identification of novel cell types in the integument of larval lake sturgeon,Acipenser fulvescens

Osmoregulation, respiration, nutrient/mineral transport, and defense mechanisms are all evident in the integument of fish. The role of the integument in these physiological processes is particularly important during early life history in larval fishes, as functional systems such as the gills and gastrointestinal tract are not fully developed. Using a variety of microscopy techniques, we describe the morphology of keratinocytes, mitochondria rich cells, ciliated cells and mucous cells of the skin, yolk sac, and gills. The cytology we observed was similar to previous studies describing the integument of larval fish, however, we have also identified two novel cell types on the integument of larval Lake Sturgeon, Acipenser fulvescens, between 9 and 34 days post fertilization. Our detailed analysis included a multifaceted microscopy approach using scanning electron, transmission electron, and light microscopy to elucidate the histology of the tissue and cellular morphology in addition to quantification and distribution of these novel cell types. The first cell type had a characteristic ampullary shape with a central cavity and a pore opening at the surface. The second, located on the free surface of the epidermis, had an uneven plasma membrane surface. Based on the abundance of secretory vesicles, organelles necessary for protein synthesis, and the lack of neural connection in both cell types, we propose these cells to be involved in the release of semiochemicals that may act as a pheromone, alarm substance, or chemical defense mechanism. J. Morphol. 277:86–95, 2016. © 2015 Wiley Periodicals, Inc.     

Isolation and characterization of microsatellites in Chinese sturgeon,Acipenser sinensis

From (GATA)_n and (AAAG)_n enriched genomic libraries for the Chinese sturgeon (Acipenser sinensis), 50 primer pairs were developed using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Forty‐six primer pairs exhibited highly polymorphic with two to 11 alleles per locus, while the rest four displayed monomorphic. These markers yielded 246 alleles in a survey of eight specimens of wild A. sinensis. Average observed heterozygosity ranged from 0.13 to 1.00. These loci should provide sufficient levels of genetic diversity to allow parentage analysis for artificial stocking management and delineation of fine‐scale population structure.     

水体中铜对中华鲟幼鱼血液生化指标的影响

通过检测中华鲟在不同铜离子浓度的水体中暴露60d后血液生化指标,研究Cu2+对幼鱼血浆生化成分和离子含量变化的影响及致毒效应.结果表明:中华鲟幼鱼血浆中血糖( Glu)、碱性磷酸酶(ALP)、尿素(Urea)、胆固醇(TC)、肌酐(CREA)含量随Cu2+浓度的增加而升高,低浓度组(0.40 μg· L-1)ALP含量与对照组有显著性差异(P＜0.05),Glu、Urea、TC、CREA含量无显著性差异;中浓度组(0.89 μg· L-1)和高浓度组(2.00 μg·L-1)含量与对照组均有显著性差异(P＜0.05).甘油三酯(TG)随着Cu2+浓度的增加而下降,低浓度组与对照组无显著性差异;中、高浓度组与对照组有显著差异(P＜0.05),总蛋白(TP)、谷草转氨酶(AST)、谷丙转氨酶(ALT)、乳酸脱氢酶(LDH-L)不受Cu2+的影响.Na+、Cl-、P含量和pH值随Cu2+浓度的增加而显著下降,Ca2+、Mg2+显著上升,其变化与Cu2+浓度存在相关性,K+含量不受Cu2+的影响.实验表明,Cu2+对中华鲟幼鱼血液生化指标的最低可观察效应浓度和最高无观察效应浓度分别为0.89和0.4 μg·L-1.血浆中ALP受Cu2+影响最明显,其含量除受Cu2+浓度影响外,随着时间的延长也显著升高,是Cu2+污染的敏感指标.     

Verification of meiotic gynogenesis in Siberian sturgeon (Acipenser baeri Brandt) using microsatellite DNA and cytogenetical markers

Diploid gynogenesis was induced in Siberian sturgeon Acipenser baeri using ultra violet (UV)-irradiated bester ( Huso huso × Acipenser ruthenus ) sperm. The higher survival rate of meiotic diploids was noted after 1350 ergs mm⁻² UV irradiation. A total of 80 meiotic diploids of known parentage from two different experimental treatments were screened using microsatellite DNA and cytogenetical analysis, and uniparental transmission in meiotic diploids was confirmed.     

From microsatellites to single nucleotide polymorphisms for the genetic monitoring of a critically endangered sturgeon

The use of genetic information is crucial in conservation programs for the establishment of breeding plans and for the evaluation of restocking success. Short tandem repeats (STRs) have been the most widely used molecular markers in such programs, but next‐generation sequencing approaches have prompted the transition to genome‐wide markers such as single nucleotide polymorphisms (SNPs). Until now, most sturgeon species have been monitored using STRs. The low diversity found in the critically endangered European sturgeon (Acipenser sturio), however, makes its future genetic monitoring challenging, and the current resolution needs to be increased. Here, we describe the discovery of a highly informative set of 79 SNPs using double‐digest restriction‐associated DNA (ddRAD) sequencing and its validation by genotyping using the MassARRAY system. Comparing with STRs, the SNP panel proved to be highly efficient and reproducible, allowing for more accurate parentage and kinship assignments' on 192 juveniles of known pedigree and 40 wild‐born adults. We explore the effectiveness of both markers to estimated relatedness and inbreeding, using simulated and empirical datasets. Interestingly, we found significant correlations between STRs and SNPs at individual heterozygosity and inbreeding that give support to a reasonable representation of whole genome diversity for both markers. These results are useful for the conservation program of A. sturio in building a comprehensive studbook, which will optimize conservation strategies. This approach also proves suitable for other case studies in which highly discriminatory genetic markers are needed to assess parentage and kinship.     

Repeat UCrit and endurance swimming in juvenile shortnose sturgeon (Acipenser brevirostrum)

Previous results show that juvenile shortnose sturgeon are steady swimmers and, compared with salmonids, generally have low critical swimming (UCrit) and endurance swimming capacities. Most studies on swimming capacities of sturgeon, and other fishes, include those where fish have only been swum once and the metrics of swimming performance are assessed (e.g., time swum, speed achieved). Under natural conditions, there are ample instances where fish undergo multiple swimming cycles when traversing fish ways, culverts and other sources of fast water flow. While some evidence exists for salmonids, the effects of repeat swimming are not well known for sturgeon. The current study consisted of two experiments. The first examined the UCrit of juvenile shortnose sturgeon following three consecutive swimming trials with a 30 min recovery period between subsequent tests. The second examined the endurance swimming capacities of juvenile shortnose sturgeon following three consecutive swimming trials with a 60 min recovery period between subsequent tests. Our findings indicate that (i) UCrit was consistent (~2 body lengths/s) among swimming trials; (ii) significant individual variation exists between individuals in the endurance swimming trials; and (iii) consistent results exist for individuals across swimming trials in both the UCrit and the endurance swimming tests. These results suggest that juvenile shortnose sturgeon have a high recovery capacity, and their behaviour and morphology likely reflect aspects of their swimming capacities.     

Annotation and analysis of expressed sequence tags (ESTs) from Chinese sturgeon (Acipenser sinensis) pituitary cDNA library

Chinese sturgeon Acipenser sinensis belongs to the family Acipenseridae, an ancient species of actinopterygian fishes. In order to advance molecular research on its reproduction, ontogenetic development, we were seeking for genomic information in the NCBI expressed sequence tag (EST database). We found 3384 indentified cDNA sequences which were assembled into 861 unigenes. Blast analysis revealed 301 unigenes shared high similarity with genes in the public databases, and these were classified into three groups: 202 known genes, 81 putative genes and 8 unknown genes. The remainder (560 genes) had no significant match to any protein sequence. Further, 255 unigenes and 333 unmatched unigenes were annotated with Gene Ontology (GO), which could be classified into cellular component, molecular function, and biological process. Among the known genes, the hormone genes pomc A (proopiomelanocortin), pomc B, GtH alpha I subunit (gonadotropin hormone), GtH alpha II subunit and GH (growth hormone) were present in this library. Comparison of the Chinese sturgeon proteins (GH, GtH alpha subunit and POMC) to proteins of other species showed higher levels of homology among sturgeon species. We performed five hormone related genes including GnRHRI (gonadotropin-releasing hormone receptor I), cpH (carboxypeptidase H), ppiB (peptidylprolyl isomerase B), stmn3 (stathmin-like 3), 7B2 (neuroendocrine protein 7B2), and four novel genes (contig 192, 177, 170 and 168) a semi-quantitative RT-PCR on different tissues from Chinese sturgeon.     

Comparative stable isotope analyses of green sturgeon (Acipenser medirostris) and white sturgeon (A. transmontanus) in the San Francisco estuary

Green sturgeon (Acipenser medirostris) and white sturgeon (A. transmontanus) are closely related, sympatric species that inhabit the San Francisco estuary. Green sturgeon have a more marine life history but both species spawn in the Sacramento River and reside for some duration in San Francisco Bay. These sturgeons are of conservation concern, yet little is known about their dietary competition when they overlap in space and time. To examine evidence of dietary differentiation, we collected whole blood and blood plasma from 26 green sturgeon and 35 white sturgeon in San Francisco Bay. Using carbon and nitrogen stable isotope analyses, we compared their relative trophic levels and foraging locations along the freshwater to marine gradient. Sampling blood plasma and whole blood allowed comparison of dietary integration over shorter and longer time scales, respectively. Plasma and whole blood δ¹³C values confirmed green sturgeon had more marine dietary sources than white sturgeon. Plasma δ¹⁵N values revealed white sturgeon fed at lower trophic levels than green sturgeon recently, however, whole blood δ¹⁵N values demonstrated the two species fed at the same trophic level over longer time scales. Larger individuals of both species had higher δ¹³C values than smaller individuals, reflecting more marine food sources in adulthood. Length did not affect δ¹⁵N values of either species. Isotope analyses supported the more marine life history of green than white sturgeon and potentially highlight a temporary trophic differentiation of diet between species during and preceding the overlapping life stage in San Francisco Bay.     

中华鲟铁蛋白基因cDNA全长克隆与组织表达分析

根据铁蛋白基因的保守序列,搜索GenBank数据库中华鲟的EST数据库得到一条同源序列.通过RT-PCR的方法对该序列进行扩增,修改其测序错误,获得中华鲟铁蛋白亚基cDNA全长,经过注释提交GenBank数据库,获取序列登录号EU348782.该cDNA长度为896bp,包含531bp的完整编码区,推测编码的蛋白质为176aa,分子量为20339.9Mr,理论等电点为5.66.它和大两洋鲑鱼铁蛋白序列同源性最高,达到82.9%.该基闪在中华鲟肝脏、胰脏、肌肉、脑、心脏、鳃和胃粘膜等多种组织表达,在胰脏和心脏中表达量较高,在肌肉组织中表达较低.根据同源模建的方法得到该蛋白质三维结构,其包括5个α螺旋和10个转角结构,和人、蛙和细菌的铁蛋白均能很好的叠合,表现了很高的相似性,表明该蛋白结构和功能在基因进化中的高度保守性.     

A bedform morphology hypothesis for spawning areas of Chinese sturgeon

Results from previous research suggested that geomorphic characteristics may be important controlling factors among other microhabitat variables for spawning fish. We investigated the bedform morphology of seven spawning areas (five historic and two present spawning areas) of Chinese sturgeon (Acipenser sinensis) by analysis of relief maps of the riverbed and by field surveys. We conclude that the topographic characteristics of the spawning areas include rocky or gravelly substrate river bend, large variations in water depth and river width and a slope with moderate length and slope. The turning structure and the adverse slope may be the most important topographic characteristics for the spawning areas. Combined with the reproductive occurrence of the fish in these spawning areas, we tentatively propose a bedform morphology hypothesis for the spawning areas. We consider that a spawning area of A. sinensis should consist of three functional areas: Mating area (M), Dispersal area (D) and Incubation area (I). M, D and I are distributed along the water current and their bedform characteristics are clearly different. M is the place for mature fish to mate and spawn. It is usually below a riffle with rolling terrain. D is the place for fertilizing and dispersing fertilized eggs. It is usually in a river bend with sharp variations in terrain, such as deep pools or steep slopes. I is the place for dispersal of fertilized eggs. It is usually in long point bars or broad shallow areas with rocky substrate. This hypothesis could serve as a guideline for improving the present spawning areas or constructing new spawning areas, so as to rehabilitate the wild stock of the sturgeon.