Neuroendocrinology of Teleosts

The nucleus preopticus has been shown to receive afferent inputfrom certain cranial nerves and the spinal cord. In addition,the nucleus preopticus and its tracts can synthesize and transporthormones about as rapidly as a mammal can. The nucleus preopticusis functionally involved in the spawning reflex behavior. The hypothalamic control of each of the adenohypophysial hormonesis discussed. There is conflicting and incomplete evidence forthe control of melanocyte-stimulating hormone (MSH), prolactin,and somatotropin. Secretion of prolactin and MSH may each becontrolled by an inhibitory factor. Corticotropin secretionhas been shown to be controlled by corticotropin releasing factor(CRF). There is a negative fedback effect by cortisol on thepituitary to suppress corticotropin secretion. Gonadotropinsecretion is controlled by gonadotropin releasing factor (GRF).A part of the nucleus lateralis tuberis is involved in the controlof gonadotropin secretion. A great deal of indirect evidenceindicates that a thyrotropin inhibitory factor (TIF) controlsthyrotropin secretion. There is a negative feedback effect bythyroxine on the pituitary to suppress thyrotropin secretionand a positive feedback effect on the hypothalamus to stimulateTIF secretion. The above findings are restricted to only one or two speciesin each instance. It is not known how general the above mechanismsof control are found throughout the teleosts.     

Expression Patterns of CREBs in Oocyte Growth and Maturation of Fish

In fish, oocyte meiotic maturation is regulated by 17α, 20β-dihydroxy-progesterone through cAMP. To study the role of cAMP response element binding protein (CREB) in meiotic maturation, we cloned and characterized the expression pattern of CREBs from two fish models, the Nile tilapia and catfish. In the Nile tilapia three different CREBs were identified where in CREB1 was found in many tissues including gonads with abundant expression in testis. CREB2, few amino acids shorter than CREB1, was expressed in several tissues with abundant expression in ovary. In addition, a 3’UTR variant form, CREB3 was exclusively found in ovary. During natural 14-day ovarian cycle of the Nile tilapia, CREB1 expression was stable throughout vitellogenesis with a sharp decrease on the day of spawning. In contrast, CREB2 remain unchanged throughout the ovarian cycle, however elevated in 11-day full-grown immature ovarian follicle and after hCG-induction. Interestingly, CREB3 expression was induced three folds on the day of spawning as well as during hCG-induced oocyte maturation. Based on the synergistic expression pattern, CREB1 is likely to control oocyte growth, whereas CREB 2 and 3 contribute to oocyte maturation in tilapia and the latter seems to be critical. In catfish, a single form of CREB showed a maximum expression during spawning phase and hCG-induced maturation both in vivo and in vitro augmented CREB expression. These results suggest that spatial and temporal expression of CREBs seems to be important for final oocyte maturation and may also regulate oocyte growth in fish.     

硬骨鱼中vasa基因的研究

vasa基因编码一个DEAD-box家族的ATP依赖性RNA解旋酶,最早在果蝇中发现.在绝大多数的脊椎动物和非脊椎动物中,vasa基因的杂交信号仅在生殖细胞系中特异性表达,因此它可以作为一种分子标签,广泛用于性腺发生、配子发生、原生殖细胞(primordial germ cells,PGCs)的起源、迁移、分化等方面的研究.综述了国内外有关硬骨鱼类vasa基因结构,表达与功能等方面的研究报道,并对其应用前景作了展望.     

Urokinase-type Plasminogen Activator-like Proteases in Teleosts Lack Genuine Receptor-binding Epidermal Growth Factor-like Domains

Plasminogen activation catalyzed by urokinase-type plasminogen activator (uPA) plays an important role in normal and pathological tissue remodeling processes. Since its discovery in the mid-1980s, the cell membrane-anchored urokinase-type plasminogen activator receptor (uPAR) has been believed to be central to the functions of uPA, as uPA-catalyzed plasminogen activation activity appeared to be confined to cell surfaces through the binding of uPA to uPAR. However, a functional uPAR has so far only been identified in mammals. We have now cloned, recombinantly produced, and characterized two zebrafish proteases, zfuPA-a and zfuPA-b, which by several criteria are the fish orthologs of mammalian uPA. Thus, both proteases catalyze the activation of fish plasminogen efficiently and both proteases are inhibited rapidly by plasminogen activator inhibitor-1 (PAI-1). But zfuPA-a differs from mammalian uPA by lacking the exon encoding the uPAR-binding epidermal growth factor-like domain; zfuPA-b differs from mammalian uPA by lacking two cysteines of the epidermal growth factor-like domain and a uPAR-binding sequence comparable with that found in mammalian uPA. Accordingly, no zfuPA-b binding activity could be found in fish white blood cells or fish cell lines. We therefore propose that the current consensus of uPA-catalyzed plasminogen activation taking place on cell surfaces, derived from observations with mammals, is too narrow. Fish uPAs appear incapable of receptor binding in the manner known from mammals and uPA-catalyzed plasminogen activation in fish may occur mainly in solution. Studies with nonmammalian vertebrate species are needed to obtain a comprehensive understanding of the mechanism of plasminogen activation.     

The Parapineal Organ of Teleosts

A parapineal organ was found to be present in 21 teleost fishes belonging to 20 different families, but was absent in poecilids and cyprinodontids. The parapineal organ was situated on the left side of the brain and sent a nerve tract to the left habenular nucleus, except in Gadus, where a “parapineal organ” appeared to send a nerve tract into the pineal stalk. The parapineal organ of adult Gasterosteus consisted of glial elements and parapinealocytes. The latter were small neurons which sent off the unmyelinated axons that formed the parapineal tract. A single photoreceptor cell was found in a stickleback parapineal organ.     

Endocrine Control of Calcium Metabolism in Teleosts

It is evident that fishes regulate their serum calcium efficientlybut that endocrine systems involved may be different from thosein tetrapods. A functional parathyroid gland has not yet beendemonstrated in fishes. The majority of evidence indicates thatcalcitonin has little or no effect on fish calcium regulation.Instead, the corpuscles of Stannius and the pituitary glandare necessary for maintaining fish serum calcium levels. Inthe killifish, Fundulus heteroclitus, the removal of the corpusclesproduces hypercalcemia in sea water but not in artificial seawater deficient in calcium. Transplants of the corpuscles orthe administration of corpuscle homogenate corrects the increasein calcium. On the other hand, hypophysectomy elicits hypocalcemiaunder calcium deficient conditions but not in calcium rich seawater. Replacement therapy with pituitary homogenate or hypophysialtransplant prevents the fall in calcium. It is postulated thatthe hypocalcemic corpuscles of Stannius and the hypercalcemicpituitary gland enable the euryhaline killifish to regulateits serum calcium levels in high calcium sea water and low calciumfresh water, respectively.     

Cellular Aspects of the Control of Physiological Color Changes in Amphibians

SYNOPSIS. The present paper is concerned mainly with the melanin-dispersingeffect of melanocyte-stimulating hormones (MSH's) on the skinmelanophores of amphibians. In addition, some of the more recentevidence for the unihumoral theory of the control of color changeis reviewed. The mechanism of dispersion of melanin is stillunknown, but evidence is accumulating that the action of MSHmay be mediated by an increase in the melanophoric content ofadenosine 3', 5'-monophosphate (cyclic AMP). For example, cyclicAMP has a specific, reversible melanin-dispersing effect onthe melanophores of the isolated skin of R. pipiens and Xenopuslaevis. It also has a reversible "melanophore—expanding"effect on the tissue—cultured embryonic melanophores ofthe spotted salamander, Ambystoma maculatum. The effect of cyclicAMP on melanophores of R. pipiens does not require sodium butis inhibited by hypertonicity. Finally, new evidence is presented that confirms that the melanin-dispersingeffect of catecholamines on melanophores of X. laevis is mediatedby beta adrenergic receptors,because it is blocked by the highlyspecific ß—blocking agent, propranolol. On theother hand, the melanin-aggregating effect of catecholamineson amphibian melanophores appears to be mediated by alpha adrenergicreceptors. There is even a possibility that the effects of catecholaminesare also mediated through a control of cyclic AMP levels inmelanophores, with beta adrenergic stimulation producing anincrease in cyclic AMP levels, followed by dispersion of melanin,and alpha adrenergic stimulation producing a decrease in cyclicAMP levels, followed by aggregation of melanin.     

Cellular Aspects of the Control of Physiological Color Changes in Crustaceans

SYNOPSIS. Red chromatophores(erythrophores) of the prawn, Palaemonetesvulgaris, are controlled by pigment—dispersing and -concentratinghormones. Recent experiments on the modes of action of thesehormones are described, followed by a theory which satisfactorilyexplains the data. Red pigment-concentrating hormone is dependentupon sodium ions for a strong response to occur. There is asimilar dependency of red pigment—dispersing hormone uponcalcium ions. Ouabain inhibits the response to red pigment—concentratinghormone; tetrodotoxin enhances it. Erythrophores with maximallydispersed pigment had a transmembrane potential of 55±15mv inside negative in one series of experiments and 56±4mv in another. No appreciable changes in permeability occurwhen depolarizing and hyperpolarizing currents are passed througha microelectrode within the chromatophore. Red pigmentconcentratinghormone causes hyperpolarization of the transmembrane potential.The magnitude of hyperpolarization is directly related to thedegree of pigment concentration. Adenosine 3`;, 5`-monophosphate(cyclic AMP) causes dispersion of the red pigment but has nopigment-concentrating effect. The primary action of red pigmentconcentratinghormone is most likely stimulation of a pump which exchangessodium ions from inside the chromatophore with potassium ionsfrom the outside, whereas red pigment-dispersing hormone quitelikely stimulates entry of calcium ions into the chromatophore.     

Cellular Aspects of the Control of Physiological Color Changes in Fishes

SYNOPSIS. Recent advances in the cellular aspects of chromatophoricactivities in fishes are reviewed, special emphasis being laidon the black pigment-containing cells, the melanophores. A fewrecent electron-microscopic studies have disclosed the finestructure of melanophores. They are enclosed with a single cellmembrane, within which melanosomes and other cell organellesare found. All observations favor the view that melanosomesare selectively moved through the cellular processes, leavingthe cell contour rather fixed. In regard to these findings,current ideas about the mechanisms of pigment movements arediscussed. Particular attention is directed to the possibleintervention of microtubules and the theory of migration ofpigment by intracellular electrophoresis. The regulatory mechanismsof pigment cells are then dealt with. The adrenergic natureof transmission is affirmed in the peripheral melanin-aggregatingnervous system. The mode of nervous supply to a melanophoreis also analyzed. Investigations of the antagonistic, melanin—dispersing,nervous system are also considered, with special reference torecent physiological studies and to the finding of synapticvesicles by electron microscopy. On the basis of these results,a new interpretation of the so-called Parker effect is proposed.     

Gonadal Hormones and Brain Development: Cellular Aspects of Sexual Differentiation

Sexual differentiation of the neural control of reproductivefunction with respect to both gonadotropin secretion and sexualbehavior is thought to result from exposure of the brain totesticular androgens during a very restricted or critical periodof CNS differentiation and development, when the tissue is competentto respond to the hormone, and after which it is refractoryor responds in a reversible manner. This paper reviews the cellularaspects of sexual differentiation with particular emphasis onthe morphological expression of the gonadal hormonal effectsin the adult brain. It presents experimental evidence for themorphogenetic basis for the observed steroid effects by showinghow the addition of steroid to undifferentiated hypothalamiccultures produces a selective neuritic response that is steroid-dependent.These results suggest that since afferent axonal input and temporalfactors are critical for dendritic and synaptic differentiation,steroid-induced variations in neuritic development could resultin gender-specific responses seen in sexual differentiationof reproductive function.     

Effects of Mechano-Gated Cation Channel Blockers on Xenopus Oocyte Growth and Development

The putative role(s) of a mechanically gated (MG) cation channel in Xenopus oocyte growth, maturation, fertilization and embryogenesis has been examined. Using a pharmacological approach, we have tested the effects of the MG channel blockers, gadolinium, gentamicin and amiloride on the above developmental events. Our results indicate that oocyte maturation, fertilization and early embryogenesis (up to the free-swimming stage 45) can proceed normally in the presence of concentrations of agents that either completely abolish (i.e., ≥10 μm Gd³⁺) or partially block (i.e., 1 mm gentamicin) single MG channel activity as measured by patch-clamp recording. However, we also find that higher concentrations of Gd³⁺ (≥50 μm) can lead to an increased percentage (>20%) of axis-perturbed embryos compared with control (<1%) and that amiloride (0.5 mm) reduces the success of fertilization (from 100% to <50%) and increases mortality (by ∼75%) in developing embryos. Furthermore, we find that all three agents inhibit oocyte growth in vitro. However, their order of effectiveness (amiloride > gentamicin > Gd³⁺) is opposite to their order for blocking MG channels (Gd³⁺≫ gentamicin > amiloride). These discrepancies indicated that the drugs effects occur by mechanisms other than, or in addition to, MG channel block. Our results provide no compelling evidence for the idea that MG channel activity is critical for development in Xenopus. This could mean that there are other mechanisms in the oocyte that can compensate when MG channel activity is blocked or that the protein that forms the channel can undergo additional interactions that result in a function insensitive to MG channel blockers. Received: 27 March 1998/Revised: 10 June 1998     

Variability and Constancy in Cellular Growth of Arabidopsis Sepals

Growth of tissues is highly reproducible; yet, growth of individual cells in a tissue is highly variable, and neighboring cells can grow at different rates. We analyzed the growth of epidermal cell lineages in the Arabidopsis (Arabidopsis thaliana) sepal to determine how the growth curves of individual cell lineages relate to one another in a developing tissue. To identify underlying growth trends, we developed a continuous displacement field to predict spatially averaged growth rates. We showed that this displacement field accurately describes the growth of sepal cell lineages and reveals underlying trends within the variability of in vivo cellular growth. We found that the tissue, individual cell lineages, and cell walls all exhibit growth rates that are initially low, accelerate to a maximum, and decrease again. Accordingly, these growth curves can be represented by sigmoid functions. We examined the relationships among the cell lineage growth curves and surprisingly found that all lineages reach the same maximum growth rate relative to their size. However, the cell lineages are not synchronized; each cell lineage reaches this same maximum relative growth rate but at different times. The heterogeneity in observed growth results from shifting the same underlying sigmoid curve in time and scaling by size. Thus, despite the variability in growth observed in our study and others, individual cell lineages in the developing sepal follow similarly shaped growth curves.Cells undergo multiple rounds of growth and division to create reproducible tissues. In some plant tissues, such as expanding cotyledons, reproducibility can occur on a cellular level during specific intervals of development, where cotyledon cells exhibit uniform cellular growth (Zhang et al., 2011). However, several studies on cell division and growth in other developing plant tissues have demonstrated that plant cells exhibit considerable cell-to-cell variability during development (Meyer and Roeder, 2014). For example, in both the Arabidopsis (Arabidopsis thaliana) meristem and leaf epidermis, cells show spatiotemporal variation in individual cell growth rates (GRs; Asl et al., 2011; Elsner et al., 2012; Kierzkowski et al., 2012; Uyttewaal et al., 2012). Furthermore, cell divisions have been observed with marked randomness in their timing and orientation (Roeder et al., 2010; Besson and Dumais, 2011; Roeder, 2012). In this study, we identify a hidden, underlying pattern in the seemingly random GR (Box 1) of cells during the formation of sepals in Arabidopsis.Open in a separate windowBox 1.Definitions of GR terms. (For details on the calculations, see “Materials and Methods.”)Plant cell growth is defined as an increase in cell size due to an irreversible expansion of the cell wall. Neighboring cells physically accommodate one another during plant growth because their cell walls are glued together with a pectin-rich middle lamella, which prevents cell mobility. The cell wall is a thin, stiff layer composed of a polymer matrix including cellulose, hemicellulose, and pectin (Somerville et al., 2004; Cosgrove, 2005). Plant cells change their size and shape by modifying their turgor pressure and/or the mechanical properties of their walls, such as elasticity, plasticity, and extensibility. Growing plant cells exert forces on their neighbors through their walls, and cell wall stresses created by these forces feed back to alter the growth anisotropy (Hamant et al., 2008; Sampathkumar et al., 2014). Although these feedbacks can coordinate growth, they may also amplify differences in growth between neighboring cells (Uyttewaal et al., 2012).Two competing computational models have proposed explanations of the cellular heterogeneity observed in growing tissues by making different assumptions about how cells grow. In the first, it is assumed that relative growth rates (RGRs) of all cells are uniform in space and time, whereas variation in the timing of division causes the heterogeneity of cell sizes (Roeder et al., 2010). This model suggests that cell divisions cut the sepal into semiindependent cells, which grow uniformly within the expanding organ (Kaplan and Hagemann, 1991). The second model postulates the reverse process: timing of cell division is uniform, but cellular growth is variable and depends on the size of the cell (Asl et al., 2011). This model suggests that cells are autonomous. Currently, there is biological evidence for both models. Variability in cell division timing is observed in sepals and meristems, whereas variability in cellular GRs has been observed in leaves and meristem cells (Reddy et al., 2004; Roeder et al., 2010; Asl et al., 2011; Elsner et al., 2012; Kierzkowski et al., 2012; Uyttewaal et al., 2012). Thus, the debate on how the growth of individual cells within an organ relates to one another remains unresolved.The identification of underlying patterns in noisy cellular growth processes is challenging. Technical difficulties include the capability for cellular-resolution imaging of the tissue at sufficiently small time intervals. Previous studies (Zhang et al., 2011; Elsner et al., 2012; Kierzkowski et al., 2012) did not image and track individual cells, or they had a coarse time resolution, with 11- to 48-h intervals between images, which may have hidden important temporal dynamics. We studied growing cells in the Arabidopsis sepal, which allows for live imaging with cellular resolution at 6-h intervals (Roeder et al., 2010). The sepal is the leaf-like outermost floral organ of Arabidopsis (Fig. 1) with four sepals of stereotypical size produced per flower. Its accessibility for live imaging makes the sepal an excellent system for studying organogenesis (Roeder et al., 2010, 2011, 2012; Qu et al., 2014). Sepals exhibit high cellular variability in the timing of division and endoreduplication, an alternative cell cycle in which a cell replicates its DNA but fails to divide (Roeder et al., 2010). Furthermore, quantifying cell growth in sepals may shed light on growth mechanisms of other plant organs, such as leaves (Poethig and Sussex, 1985; Roeder et al., 2010).Open in a separate windowFigure 1.Diverse sizes of Arabidopsis sepal cells. A, Four sepals (s) are the outermost green leaf-like floral organs in Arabidopsis. B and C, Scanning electron micrographs of a mature Arabidopsis sepal show that the outer epidermal cells have a wide range of sizes. Asterisks mark some of the largest cells (giant cells) that can span 1/4 the length of the sepal. Scale = 100 µm.Another key challenge in analyzing cellular growth is the identification of trends in noisy data. Inaccuracies in data acquisition, such as segmentation errors, and noisy growth of individual cells can hide meaningful spatiotemporal trends in growth. GRs measured over longer time intervals will have reduced noise, but they may also obscure important temporal dynamics. Alternatively, previous studies have examined growth of the whole organ or its subregions to avoid cellular noise (De Veylder et al., 2001; Mündermann et al., 2005; Rolland-Lagan et al., 2005, 2014; Kuchen et al., 2012; Remmler and Rolland-Lagan, 2012). However, precise cellular patterns are not resolved. In our study, we use cellular resolution data to define spatially averaged kinematics while keeping the full temporal resolution to identify course-grained spatial trends in the dynamics of cellular growth (Box 1).We analyze the relationships among the growth of individual cell lineages in a developing Arabidopsis sepal by live imaging and computational analyses. We have developed continuous low-order displacement fields to represent the spatially averaged kinematics of the sepal (Box 1). We find that the growth of the tissue surface area, cell lineage area, and wall length follows S curves, suggesting that their GRs vary over time. Additionally, we find that there is a linear correlation between the maximum GR (i.e. size increase per hour) and the size of the cell. We furthermore find that each sepal cell lineage reaches the same maximum RGR (i.e. GR divided by size). However, each cell reaches the maximum RGR at a different time during its development, generating the observed heterogeneity. Thus, we find underlying similarities in the growth curves of sepal cells.     

The Development of Saccus vasculosus in Two Teleosts

The development and differentiation of the ventro-caudal part of the diencephalon, the saccus infundibuli, has been examined in salmon, Salmo salar L., and in pike, Esox lucius L. In salmon, the saccus infundibuli becomes the saccus vasculosus; but in pike, this does not occur. The question of saccus vasculosus and part of the neural lobe of the tetrapod pituitary being homologous is dealt with.