高通量质谱法用于小鼠脑部神经肽的鉴定

神经肽在参与调控人体各种生理功能上发挥着重要的作用,如痛觉、睡眠、情绪、学习与记忆等生理活动都受到神经肽的影响。神经肽主要存在于机体的神经组织内,其他体液和器官中也有少量的分布。目前对全脑组织神经肽高通量鉴定的研究仍不足,高通量检测这些神经肽对了解神经肽的组成和功能具有重要的意义。本研究通过对小鼠全脑组织内源性肽段的萃取,运用液相串联质谱(LC-MS/MS)技术对全脑组织的神经肽进行检测,共鉴定到1 830条内源性肽段和99条预测神经肽肽段。这些内源性肽段的鉴定在疾病的治疗和机制研究以及药物的研发方面提供了参考价值,也为研究新的神经肽及其功能奠定了基础。     

脑啡肽研究的新进展

随着实验方法的改进,特别是免疫组化技术的应用,脑内微量神经肽的测定和定位才成为可能。有关神经肽和肽能神经原的研究是当前神经生理学中最活跃的一个领域,最近已发表了不少专题论述。目前已发现的神经肽还不足脑内总数的10％,Snyder估计脑内肽类递质将在200种以上。已知的神经肽大多为偶然发现,多数未曾进行系统研究,唯有对脑啡肽(Enk)研究得最深入,有关Enk与内啡肽的分布、功能及生化特性等,已有综述。有关鸦片受体(OR)的近况也有介绍,本文仅就近年来有关Enk研究进展作一扼要介绍。     

稳定同位素技术在头足类摄食生态学研究中的应用

头足类在海洋食物网营养关系中占有重要地位,然而对其复杂的生活史过程,尤其是摄食生态学信息仍知之甚少.稳定同位素技术作为传统食物网科学研究方法的有力补充,可更深层次地分析头足类的摄食习性和栖息地等方面的重要信息.本文在比较分析国内外头足类摄食生态学研究方法的基础上,系统归纳总结了稳定同位素技术在头足类摄食生态学研究中的发展现状并介绍最新进展情况,着重分析稳定同位素技术在头足类生活史信息,尤其是在摄食生态学研究中的应用现状及发展前景,包括测定样品标准化、头足类生长发育过程中的食性转换和洄游分布等核心问题,以促进其在头足类生物学研究中的应用.     

安徽贵池早奥陶世晚期及中奥陶世的头足类

一、前言安徽贵池一带的奥陶系发育良好,含丰富的头足类化石。1984年夏,笔者等在贵池百安的方村剖面采获不少头足类化石,其中以下奥陶统的紫台组、牯牛潭组及中奥陶统的大田坝组、宝塔组所产的头足类最为丰富。这批化石经研究共有33种(其中10新种),分属15属8科。另有2未定属种。这些化石的发现增加了我国奥陶纪头足类的新材料,并对这个地区奥陶纪地层的划分以及与国内外的对比提供了可靠的依据。笔者在研究过程中,得到中国科学院南京     

基于质谱技术的神经肽研究进展

神经肽是一类重要的内源活性物质,在神经系统中发挥重要的作用,并连接大脑和其他神经器官。基于质谱技术的神经肽组学研究旨在对神经肽进行大规模研究,在分子水平上得到重要信息,进一步加深对神经系统调控机制以及神经疾病致病机理的理解。文中综述了利用质谱技术进行神经肽研究的基本策略,包括样品处理、定性定量方法以及质谱成像等研究进展。     

PBAN/pyrokinin神经肽类及其基因的研究进展

昆虫在其生长发育过程中,如胚胎发育、蜕皮变态、滞育、迁飞、代谢、生殖等都离不开神经肽的调控。信息素合成激活肽(pheromone biosynthesis activating neuropeptide,PBAN)和Pyrokinin神经肽是C端具有五肽FXPRL(X=S,V,T,G等)(苯丙-X-脯-精-亮氨酸)序列的一类神经肽,在昆虫的生长发育中起重要的生理功能,如性信息素的合成、控制表皮色素、促进胚胎滞育和刺激内脏肌肉收缩等重要的生理功能。因此近几年对PBAN/pyrokinin神经肽的鉴定、加工、作用和降解方式的研究成为研究的热点,为研制高效、低毒、专一性强、无公害的杀虫剂提供了思路。介绍了PBAN/pyrokinin神经肽类及其基因的研究进展,并对PBAN/pyrokinin神经肽在害虫防治中的应用进行了展望。     

《新疆石炭纪头足类》中国古生物志,新乙种,第27号

<正> 本书共分六部分:前言、含头足类地层剖面简介、新疆石炭系简介与含头足类地层的对比、新疆北部泥盆-石炭系的界线、新疆石炭纪头足类动物群的分析、系统描述。新疆的石炭系相当发育、产丰富的头足类,是我国研究石炭纪头足类的重要地区之一。本书系统描述了新疆地区石炭系12个剖面中所产的头足类标本,鹦鹉螺15属21种(含5新种)归10科3目,菊石27属(亚属)75种(含2新属1新亚属27新种)归15科2目。对新疆含头足类的石炭纪地层进行了综合研究,共建立     

头足类早期生长阶段的比较研究

本文首次系统描述了乌贼科、枪乌贼科、微鳍乌贼科和蛸科(章鱼科)浅海性种类早期生长阶段的形态特征,并与成体的形态特征相比较。对比了浅海性头足类与大洋性头足类的早期生长阶段。发现浅海性头足类在早期生长阶段中的形态变化较小,而大洋性的头足类在早期生长阶段中的形态变化较大。表明在系统演化中,浅海性头足类已走在大洋性头足类的前面。     

家蚕神经肽及其受体的功能和信号转导机制研究进展

神经肽是一类由神经分泌细胞分泌、用于调节生物胞间信号传递的信号分子,其信号分子的膜定位、相应胞内信使的激活以及一系列级联反应的引发,是由存在于细胞表面的特异性受体分子来完成的。神经肽及其受体能够调控昆虫的几乎所有生命活动,在昆虫生长发育中起着关键作用。家蚕Bombyx mori作为鳞翅目昆虫的模式物种,是昆虫生长发育与生理学研究的重要模型。特别是家蚕基因组测序完成后,越来越多的家蚕神经肽及其受体被鉴定,并发现其在家蚕的生长发育、取食消化、蜕皮、滞育、繁殖、吐丝结茧等各种生理活动中都发挥了重要的调节作用。本文综述了家蚕重要神经肽的种类及其对家蚕取食消化、蜕皮变态、生殖发育等的调控作用,探讨了神经肽通过结合特异性受体而激活细胞内ERK、TOR等下游信号通路的分子作用机制,以期为昆虫神经肽及其受体研究提供借鉴和参考,并以此推进家蚕功能基因的研究,促进蚕丝产业的发展。     

家蚕神经肽基因的筛查及成熟肽的预测

神经肽（neuropeptide）是家蚕Bombyx mori体内重要的调控物质。为了充分理解神经肽对家蚕发育的调控, 扩充家蚕神经肽的数量, 本研究利用BLAST的tblastn程序结合OpenOffice软件的查找程序, 基于其他昆虫和无脊椎动物神经肽的同源性和保守的结构特点, 在家蚕基因、理论蛋白质数据库及NCBI中进行全面而系统的基因筛查; 并利用各种在线工具对所筛查到基因和理论蛋白质的结构和成熟肽进行预测分析。结果共获得allatostatin-A （AST-A）, allatostatin-B （AST-B）, allatostatin-C （AST-C）, allatropin (AT), ecdysis-triggering hormone （ETH）, crustacean cardioactive peptide (CCAP)和FMRFamide等31个神经肽基因家族, 包括37个神经肽基因亚家族, 共计44个神经肽基因; 预测出193个成熟的神经肽, 其中73个根据家族同源性预测在C末端发生酰胺化, 而6个被预测在N末端发生了环化, 9个被预测酪氨酸发生了硫酸化。大部分成熟神经肽都具有明显的家族结构特征, 但proctolin, CCAP及CAPA-PK成熟肽结构上与其他昆虫相比有所扩展。结果提示, 家蚕神经和内分泌细胞产生了几乎在所有昆虫中具有的神经肽前体; 进化过程中大部分成熟神经肽的氨基酸序列在种间产生了差异, 但家族特征性基序高度保守。本研究为神经肽功能研究以及神经肽对家蚕发育尤其是蛹期发育调控的研究奠定了基础。     

Immunohistochemical Localization of Cardio-Active Neuropeptides in the Heart of a Living Fossil, Nautilus pompilius L. (Cephalopoda, Tetrabranchiata)

Neuropeptides play an important role in modulating the effects of neurotransmitters such as acetylcholine and noradrenaline in the heart and the vascular system of vertebrates and invertebrates. Various neuropeptides, including substance P (SP), vasoactive intestinal polypeptide (VIP) and FMRFamide, have been localized in the brain in cephalopods and the neurosecretory system of the vena cava. Previous studies involving cephalopods have mainly focussed on the modern, coleoid cephalopods, whereas little attention was paid to the living fossil Nautilus. In this study, the distributions of the peptides related to tachykinins (TKs) and the high affinity receptor for the best characterized TK substance P (tachykinin NK-1), VIP, as well as FMRFamide were investigated in the heart of Nautilus pompilius L. by immunohistochemistry. TK-like immunoreactivity (TK-LI) was seen associated to a sub-population of hemocytes, VIP-LI glial cells in larger nerves entering the heart, whereas FMRFamide immunoreactivity was distributed throughout the entire heart, including the semilunar atrioventricular valves. The pattern of FMRFamide immunoreactivity matched that of Bodian silver staining for nervous tissue. The NK-1-LI receptor was located on endothelial cells, which were also positive for endothelial nitric oxide synthase-LI (eNOS). The results indicate that neuropeptides may be involved in the regulation of the Nautilus heart via different mechanisms, (1) by direct interaction with myocardial receptors (FMRFamide), (2) by interacting with the nervus cardiacus (VIP-related peptides) and (3) indirectly by stimulating eNOS in the endothelium throughout the heart (TK-related peptides).     

The presence of APGWamide in Octopus vulgaris: a possible role in the reproductive behavior

The concerted action of many neuropeptides has been implicated in the nervous control of specific behaviors in many molluscs. In the present study, the presence of amidated tetrapeptide Ala-Pro-Gly-Trp-NH2 (APGWamide) in those lobes that are involved in the control of reproductive behavior in Octopus vulgaris has been investigated. APGWamide immunoreactivity was mainly confined to the posterior olfactory lobule and in the inferior frontal system. These areas are involved in Octopus in the processing of either chemotactile sense or olfaction. From these lobes, immunoreactive fibers reached other lobes of the central nervous system (CNS) which could be indirectly involved in the reproductive behavior. APGWamide immunoreactivity was also present in the glandular cells of the oviducal gland in the female reproductive system. These results constitute the first detailed immunolocalization of APGWamide in cephalopods and open a new insight into the possible effects that both distant and close chemical stimuli can exert on neuropeptidergic circuitries, which may affect the reproductive behavior of cephalopods.     

How intelligent is a cephalopod? Lessons from comparative cognition

The soft-bodied cephalopods including octopus, cuttlefish, and squid are broadly considered to be the most cognitively advanced group of invertebrates. Previous research has demonstrated that these large-brained molluscs possess a suite of cognitive attributes that are comparable to those found in some vertebrates, including highly developed perception, learning, and memory abilities. Cephalopods are also renowned for performing sophisticated feats of flexible behaviour, which have led to claims of complex cognition such as causal reasoning, future planning, and mental attribution. Hypotheses to explain why complex cognition might have emerged in cephalopods suggest that a combination of predation, foraging, and competitive pressures are likely to have driven cognitive complexity in this group of animals. Currently, it is difficult to gauge the extent to which cephalopod behaviours are underpinned by complex cognition because many of the recent claims are largely based on anecdotal evidence. In this review, we provide a general overview of cephalopod cognition with a particular focus on the cognitive attributes that are thought to be prerequisites for more complex cognitive abilities. We then discuss different types of behavioural flexibility exhibited by cephalopods and, using examples from other taxa, highlight that behavioural flexibility could be explained by putatively simpler mechanisms. Consequently, behavioural flexibility should not be used as evidence of complex cognition. Fortunately, the field of comparative cognition centres on designing methods to pinpoint the underlying mechanisms that drive behaviours. To illustrate the utility of the methods developed in comparative cognition research, we provide a series of experimental designs aimed at distinguishing between complex cognition and simpler alternative explanations. Finally, we discuss the advantages of using cephalopods to develop a more comprehensive reconstruction of cognitive evolution.     

Contrasting Responses to Harvesting and Environmental Drivers of Fast and Slow Life History Species

According to their main life history traits, organisms can be arranged in a continuum from fast (species with small body size, short lifespan and high fecundity) to slow (species with opposite characteristics). Life history determines the responses of organisms to natural and anthropogenic factors, as slow species are expected to be more sensitive than fast species to perturbations. Owing to their contrasting traits, cephalopods and elasmobranchs are typical examples of fast and slow strategies, respectively. We investigated the responses of these two contrasting strategies to fishing exploitation and environmental conditions (temperature, productivity and depth) using generalized additive models. Our results confirmed the foreseen contrasting responses of cephalopods and elasmobranchs to natural (environment) and anthropogenic (harvesting) influences. Even though a priori foreseen, we did expect neither the clear-cut differential responses between groups nor the homogeneous sensitivity to the same factors within the two taxonomic groups. Apart from depth, which affected both groups equally, cephalopods and elasmobranchs were exclusively affected by environmental conditions and fishing exploitation, respectively. Owing to its short, annual cycle, cephalopods do not have overlapping generations and consequently lack the buffering effects conferred by different age classes observed in multi-aged species such as elasmobranchs. We suggest that cephalopods are sensitive to short-term perturbations, such as seasonal environmental changes, because they lack this buffering effect but they are in turn not influenced by continuous, long-term moderate disturbances such as fishing because of its high population growth and turnover. The contrary would apply to elasmobranchs, whose multi-aged population structure would buffer the seasonal environmental effects, but they would display strong responses to uninterrupted harvesting due to its low population resilience. Besides providing empirical evidence to the theoretically predicted contrasting responses of cephalopods and elasmobranchs to disturbances, our results are useful for the sustainable exploitation of these resources.     

Cephalopods in neuroscience: regulations,research and the 3Rs

Cephalopods have been utilised in neuroscience research for more than 100 years particularly because of their phenotypic plasticity, complex and centralised nervous system, tractability for studies of learning and cellular mechanisms of memory (e.g. long-term potentiation) and anatomical features facilitating physiological studies (e.g. squid giant axon and synapse). On 1 January 2013, research using any of the about 700 extant species of “live cephalopods” became regulated within the European Union by Directive 2010/63/EU on the “Protection of Animals used for Scientific Purposes”, giving cephalopods the same EU legal protection as previously afforded only to vertebrates. The Directive has a number of implications, particularly for neuroscience research. These include: (1) projects will need justification, authorisation from local competent authorities, and be subject to review including a harm-benefit assessment and adherence to the 3Rs principles (Replacement, Refinement and Reduction). (2) To support project evaluation and compliance with the new EU law, guidelines specific to cephalopods will need to be developed, covering capture, transport, handling, housing, care, maintenance, health monitoring, humane anaesthesia, analgesia and euthanasia. (3) Objective criteria need to be developed to identify signs of pain, suffering, distress and lasting harm particularly in the context of their induction by an experimental procedure. Despite diversity of views existing on some of these topics, this paper reviews the above topics and describes the approaches being taken by the cephalopod research community (represented by the authorship) to produce “guidelines” and the potential contribution of neuroscience research to cephalopod welfare.     

The SepOvotropin: a new ovarian peptide regulating oocyte transport in Sepia officinalis

In the cuttlefish Sepia officinalis, the successive steps of egg laying are controlled by multiple neuropeptides. Recent experiments led us to suppose that there was possible involvement of a second regulation pathway by the release of ovarian regulatory peptides in the genital tract. Using HPLC fractionation and an in vitro biological test, a C-terminal amidated peptide modulating the motility of the Sepia officinalis oviduct was isolated from an extract of vitellogenic ovarian follicles. The mass of this peptide as determined by MALDI-TOF (1501.8 Da) and analysis by Edman degradation led to the following sequence: Pro-Lys-Asp-Ser-Met-Leu-Leu-Leu-Gln-Val-Pro-Val-Tyr-amide. The peptide mapping performed by LC/MS revealed a distribution restricted to the follicles, the full grown oocytes and the eggs. This new peptide, called SepOvotropin, modulated contractions of the whole genital tract in physiological conditions from a threshold concentration between 10(-20) and 10(-19) M, demonstrating for the first time the occurrence of a specific peptidergic control of egg-laying in cephalopods.     

Surface-feeding on cephalopods by procellariiform seabirds in the southern Benguela region, South Africa

Cephalopod beaks recovered from stomach samples taken from 14 seabird species in the southern Benguela region off Southern Africa and from one species at Sub-Antarctic Marion Island, were identified as far as possible, counted and the lower rostral lengths (LRLs) measured. Dorsal mantle lengths (DMLs) and body masses of the cephalopods eaten were estimated. The results of analyses by percentage frequency of occurrence and numerical abundance are discussed with reference to present knowledge of the distribution of cephalopods eaten by seabirds in the areas studied. Division of the cephalopod component of seabird diets into species which float, and species which sink, after death indicates that the birds forage on dead or moribund cephalopods on the surface, rather than catching live bioluminescent cephalopods at night.     

The ecological role of cephalopods and their representation in ecosystem models

Cephalopods, especially squids, are believed to have a structuring role in marine ecosystems as a link between different trophic levels, primarily due to their voracious prey consumption and high production rate. Cephalopod ecology, however, is still poorly understood as observational studies often give highly uncertain and variable results due to the peculiarities of cephalopod behaviour and biology, and their responsiveness to external drivers. This review evaluates our representation of cephalopods in ecosystem models and the insights given by these models on the role of cephalopods in our oceans. We examined ecosystem models from 13 regions to analyse the representation of cephalopods and compared their results to local trophic studies. Our analysis indicated that most ecosystem models inadequately include cephalopods in terms of model structure and parametrization; although some models still have the capacity to draw valuable conclusions regarding the impact and role of cephalopods within the system. Oceanic squid species have a major role linking trophic levels and food webs from different habitats. The importance of neritic species varies locally, but generally cephalopods have a substantial impact via their consumer role. To better understand the ecological role of cephalopods, improved representation of these species in ecosystem models is a critical requirement and could be achieved relatively easily to more accurately articulate the mechanisms regulating the ecological role of cephalopods.

     

Experimental fragmentation patterns of modern Nautilus shells and the implications for fossil cephalopod taphonomy

Shells of modern Nautilus pompilius from the Philippines were experimentally fragmented designed to mimic: (1) transport with sediment; (2) sediment loading; and (3) collision during floating. The breaking patterns by other mechanisms (predation and implosion by hydrostatic pressure) documented in the literature were also considered. The breaking patterns produced by various mechanisms are distinct and can therefore be differentiated. The results allow identification of the distinct mechanisms responsible for specific fragmentation not only in modern Nautilus but also in fossil cephalopods. This is a new approach for the more complete recognition of post-mortem transport of fossil cephalopods and their early taphonomic history, and contributes to our understanding of their palaeobiogeography and palaeoecology.     

CEPHALOPODS AND FISH: THE LIMITS OF CONVERGENCE

Resemblances between cephalopods and fish 1. Modern cephalopods (coleoids < 1000 species) resemble modern fish (30,000 species) more closely than any of their ancestors did. They have not been replaced by the more diversified group in geological time. 2. The main body of the article (pp. 245–283) reviews these resemblances. They are to be found at all levels of analysis. 3. Basic physiological mechanisms of molluscs (pp. 262–5) have been incorporated into systems with performances comparable to those of vertebrate systems. For instance the cephalopod locomotory system (pp. 249–56) and hydrostatic control system (pp. 256–60), structurally very different from their fish counterparts, have similar adaptive radiation. 4. Behaviour (p. 278) and growth of the brain (p. 265) are characteristically vertebrate-like. 5. Cephalopods and fish are considered as occupying the same broad adaptive zone though modal differences (pp. 283–5)-in reproductive habits, growth rate and light-dependent behaviour where extraocular photoreceptors appear to be important - mean that they occupy different areas within that zone. (ii) Evolution of convergence 6. Evidence is presented (pp. 287–293) for considering the convergence as due not merely to similar physical demands of the marine environment, but to dynamic interactions between cephalopods and vertebrates from the late Palaeozoic onwards. 7. The convergence was set on its way when the two groups, independently of each other, acquired locomotory methods that allowed them to increase in size. 8. It is argued that reduction and eventually complete loss of the chambered shell (in all but sepiids) was an evolutionary response to the needs of increased mobility and to the need to go deeper as vertebrate predators pushed out into oceanic waters. 9. The ammonites (pp. 291–2) present a partial model of the course that coleoid ancestors may have taken. 10. Coleoids subsequently reinvaded surface and coastal waters, competing successfully in a teleost habitat. Their most striking adaptations are ones that they share with teleosts. 11. Behavioural interactions in a vertebrate-dominated environment have probably been responsible for the vertebrate-type eye of cephalopods (p. 293). 12. The conclusion that the common adaptive zone shared by cephalopods and fish has been achieved by mutual interactions between the two groups evokes no special evolutionary principles. It assumes that all cephalopod species have (at some period of their evolutionary history) been in competition for food with some vertebrate species and that vertebrates are a source of selection pressures - largely operating through visual behaviour - that maintain and promote convergence upon the ‘fish’ modal type. 13. Selection pressures also operate within behaviour space to maintain and promote the special differences that separate any competitively successful species from all others. Coleoids as a group appear to have retained adaptations associated with such molluscan features as high growth rate and rapid turnover of the population. They are still characteristically crepuscular in habits and have extensive vertical mobility.