Thermoadaptation Characteristics of European River Lamprey Lampetra fluviatilis Smolts

Journal of Ichthyology - Thermoadaptation characteristics of smolts of European river lamprey Lampetra fluviatilis are studied in experimental conditions for the first time. The final preferred...     

Electron Microscopic Analysis of the Inner Synaptic Layer in the River Lamprey (Lampetra fluviatilis)

Abstract Different types of synaptic contacts between bipolar, amacrine and ganglion cells were scored on random electron micrographs and on montages comprising the entire thickness of the inner synaptic layer. Currently accepted criteria were used when classifying the different cell processes. The percental distribution of dyads was estimated to 56 % amacrine-amacrine dyads, 34 % amacrine-ganglion dyads and 10 % ganglion-ganglion dyads. The ratio of amacrine conventional synapses to bipolar ribbon synapses was 6.8 : 1. The density per unit area of conventional synapses (0.035/μm²) and ribbon synapses (0.005/ μm²) was found markedly low as compared with other vertebrate species except the carp. The inner synaptic layer of the river lamprey is suggested to be of the intermediate type in which both simple and complex ganglion cell receptive fields may be expected.     

Aminergic and Indoleamine Accumulating Neurons in the Retina of the River Lamprey (Lampetra fluviatilis)

Tissues were processed for fluorescence microscopy of biogenic amines according to the method of Falck and Hillarp. Normal animals, and animals injected with α-methylnoradrenaline or 5,6-dihydroxytryptamine were used. Catecholamine containing neurons (junctional cells) occur in the innermost rows of cell bodies of the inner nuclear layer (INL) and close to the vitreous surface. Catecholamine containing fibers occur in three layers: (1) an outer layer around the innermost perikarya of the INL, which is a condition not found in retinas of gnathostome chordates; (2) a middle layer within the outer third of the inner synaptic layer (ISL), separated from the outer layer by ganglion cell axons; (3) a sparse inner layer within the innermost third of the ISL. A few catecholamine containing fibers were seen to extend from the innermost region of the INL to the outer synaptic layer. The position of the junctional cells in the lamprey corresponds to that in gnathostome chordates, but whereas all catecholamine containing fiber layers in gnathostomes are located sclerally to the optic fiber layer and within the ISL, the middle and the inner fiber layers in the lamprey occur vitreally to the optic fiber layer. Indoleamine accumulating neurons occur in the innermost row of perikarya of the INL and close to the vitreous surface. Those of the INL send fine, varicose branches to the ISL forming a network which is somewhat denser at the inner and outer borders of the ISL than in its middle. The indoleamine accumulating terminals do not ramify within the INL in contrast to the catecholamine containing terminals.     

Feeding Habits of Larval European River Lamprey Lampetra fluviatilis from the Chernaya River (Baltic Sea Basin)

Journal of Ichthyology - The results of the analysis of the bolus of the larval river lamprey Lampetra fluviatlis caught in the Chernaya River in May are presented. A quantitative analysis of the...     

Ultrastructure of the Pro-adenohypophysis of the River Lamprey,Lampetra fluviatilis,During Gonad Maturation

Abstract The pro-adenohypophysis (pro-AH) of migrating adult river lamprey, Lampetra fluviatilis, has been examined from October, when the gonads are developing slowly, until May, when the sexual maturation is completed. Two granulated cell types are dominating. One is chromophobic (C2, 95 nm), the other is basophilic (B1-B3) and probably gonadotropic. The mean diameter of the basophil secretory granules increases from 150 nm in October to 220 nm in April and May. The staining affinity of a rare, granulated cell type (D, 115 nm) has not been established. No acidophil cells are found in the pro-AH. The ultrastructural characteristics of the lead hematoxylin positive cells found sometimes in paraffin sections are unknown. Non-glandular stellate cells are common. They contain microfilaments, are probably contractile, and transform to phagocytes during the last months before spawning. This phagocytosis involves only the basophil cells. It is suggested that the stellate cells in this way destroy excess hormone. All cell types in the pituitary accumulate large irregular lipid droplets during the last month before spawning. These lipid droplets seem to be expelled from residual bodies as an end product of autolysis and phagocytosis.     

External Pathway of NADH Oxidation in the Liver of the River Lamprey Lampetra fluviatilis

A possibility of exogenous NADH oxidation via the external pathway has been shown on homogenates and isolated liver cells of the lamprey Lampetra fluviatilis in the presence of rotenone and antimycin A. The homogenates were incubated in isotonic and hypotonic sucrose media, while cells, in isotonic salt medium. At incubating the tissue preparations in isotonic media, digitonin was used to enhance membrane permeability to NADH and cytochrome c. In homogenates, the maximal rate of NADH oxidation via the external pathway in the presence of cytochrome c and digitonin was 5.3 nmol O₂/min/10 mg wet weight. This value in the cells amounted to 12.6, while without addition of exogenous NADH and cytochrome c, to 11.0 nmol O₂/min/10 million cells. Cyanide inhibited completely the NADH oxidation via the external pathway both in homogenates and in cells. The intact lamprey hepatocytes, unlike homogenates, are suggested to contain sufficient concentrations of cytochrome c and extramitochondrial NADH to provide maximal NADH oxidation rate in mitochondria through external pathway. This allows thinking that potential possibilities of NADH oxidation via the external pathway in Cyclostomata and mammals are qualitatively and quantitatively close.     

A Fluorescence Microscopic Study of the Retina of the River Lamprey (Lampetra fluviatilis) and the Atlantic Hagfish, (Myxine glutinosa)

During the last few years many facts have been revealed about the neural structure in the retina of several vertebrate groups, particularly with regard to the monoamine containing neurons. A number of investigations have been made on mammals, birds and teleost fishes by the Falck-Hillarp method for the cellular demonstration of such amines. The present study has paid attention to two cyclostomes, the river lamprey, Lampetra fluviatilis, and the atlantic hagfish, Myxine glutinosa.     

Activation of Na+,H+-Exchange in Erythrocytes of the River Lamprey Lampetra fluviatilis

To study H⁺ transport, the lamprey red blood cells were acidified to pH 6.0 by a pretreatment with an ionophore, nigericin. Incubation of the acidified cells in NaCl-medium at pH 8.0 was accompanied by a rapid H⁺ efflux from the erythrocytes. There was a tenfold decrease of the H⁺ efflux rate on addition to NaCl-medium of dimethylamiloride or on replacing Na⁺ in the medium (KCl-medium, pH 8.0). A high rate of Na+ influx into the acidified erythrocytes occurred only in the presence of H⁺ gradient (pH medium 8.0), but not in its absence (pH medium 6.0). The Na⁺-dependent H⁺ efflux from the cells and H⁺-dependent Na⁺ influx into the cells were quantitatively similar (about 700 mmol/l cells/h). A rapid elevation of the intracellular Na⁺ concentration as measured by flame photometry was also observed during incubation of the acidified cells in NaCl-medium (pH 8.0). The H⁺-dependent Na⁺ influx and an increase of the Na⁺ content in the acidified cells were significantly inhibited by amiloride. The data obtained for the first time prove with certainty the presence of the Na⁺/H⁺ exchanger in erythrocytes of the river lamprey.     

The Vertebrate 7S K RNA Separates Hagfish (Myxine glutinosa) and Lamprey (Lampetra fluviatilis)

7S RNA sequences from the hagfish (Myxiniformes) and lamprey (Petromyzontiformes) were cloned and analyzed. In both species, 7S L RNA (also designated SRP RNA, since it represents the RNA constituent of the signal recognition particle) was clearly detectable. The sequence similarity between the two species was 86%, compared with about 75% similarity between either of these species and mammals. 7S K RNA was also cloned from the lamprey. The similarity between the 7S K RNA of the lamprey and that of mammals was 68%. Interestingly, several interspersed elements were found with nearly 100% similarity compared with mammals. In contrast to the lamprey, no 7S K RNA-related sequences were detectable among hagfish RNA, neither in northern blots nor with the PCR assay. In view of the significant conservation between the 7S K RNA of lamprey and that of mammals (human), this unexpected result clearly separates lamprey and hagfish. In addition, the lack of detectable 7S K RNA sequences in an outgroup, such as amphioxus, indicates that these results do not reflect an autapomorphy of hagfish. Therefore, our data provide additional support to the notion of a sister group relationship between Petromyzontiformes and gnathostomous vertebrates to the exclusion of Myxiniformes. Received: 24 September 1999 / Accepted: 9 February 2000     

Transmitter Sensitivity of Primary Afferent Cells of the Lamprey Lampetra fluviatilis Spinal Cord

Using dorsal sensory cells (primary afferents) of the spinal cord of the lamprey Lampetra fluviatilis, isolated by an enzymatic-mechanical method, their responses have been studied to application of excitatory amino acids (EAA), glutamate, aspartate, kainate, N-methyl-D-aspartate (NMDA), and of inhibitory amino acids, GABA and glycine, as well as of neuromodulator serotonin (5-hydroxytryptamine, 5-HT). The patch-clamp method was applied to fix either potential or current through membranes of studied cells. It was shown that of all the above substances, only NMDA and kainate could produce in dorsal cells depolarization, input current with maximal amplitude up to 16 nA, and action potential. The main attention in this work was paid to NMDA-current properties. The dose–effect curves were obtained; a blocking effect of specific antagonist of NMDA-receptors, 2-amino-5-phosphonovalerian acid (APV), was shown; passage of Ba ions through Ca²⁺-channels of dorsal sensory cell membranes at NMDA application was proven. It has been established that in a half of the studied cells under effect of NMDA there is an increase of the maximal amplitude of the potential-activated current through Ca²⁺-channels, on average, by 22.5 ± 10.5% (n = 21). It was suggested that this variation might be essential for an increase of amount of the transmitter released in synapses formed by processes dorsal sensory cells, while NMDA could be considered a modulator facilitating synaptic activity of these cells.     

The Polarity of the Retinal Pigment Epithelium

The diversity of epithelia in the body permits a multitude of organ-specific functions. One of the foremost examples of this is the retinal pigment epithelium. Located between the photoreceptors of the retina and their principal blood supply, the choriocapillaris, the retinal pigment epithelium is critical for the survival and function of retinal photoreceptors. To serve this purpose, the retinal pigment epithelium cell has adapted the classic Golgi-to-cell-surface targeting pathways first described in such prototypic epithelial cell models as the Madin-Darby canine kidney cell, to arrive at a unique distribution of membrane and secreted proteins. More recent data suggest that the retinal pigment epithelium also takes advantage of its inherent asymmetry to augment the classical pathways of Golgi-to-cell-surface traffic. As retinal pigment epithelium transplants and gene therapy represent potential cures for retinal degenerative diseases, understanding the basis of the unique polarity properties of retinal pigment epithelium cells will be a critical issue for the development of future therapies.     

MicroRNA Expression Profiles of Human iPS Cells,Retinal Pigment Epithelium Derived From iPS,and Fetal Retinal Pigment Epithelium

The objective of this report is to describe the protocols for comparing the microRNA (miRNA) profiles of human induced-pluripotent stem (iPS) cells, retinal pigment epithelium (RPE) derived from human iPS cells (iPS-RPE), and fetal RPE. The protocols include collection of RNA for analysis by microarray, and the analysis of microarray data to identify miRNAs that are differentially expressed among three cell types. The methods for culture of iPS cells and fetal RPE are explained. The protocol used for differentiation of RPE from human iPS is also described. The RNA extraction technique we describe was selected to allow maximal recovery of very small RNA for use in a miRNA microarray. Finally, cellular pathway and network analysis of microarray data is explained. These techniques will facilitate the comparison of the miRNA profiles of three different cell types.     

The Ultrastructure of the Accessory Olfactory Organ in the River Lamprey (Lampetra Jiuviatilis)

The accessory olfactory organ of Lampetra fluviatilis was found to consist of clusters of interconnected vesicles in tenuous connection with the exterior medium via the cavity of the olfactory organ. The walls of the vesicles are composed of two types of cells. One type are primary sense cells that resemble the olfactory sense cells in that their nucleus is situated peripherally and their axons pass directly into the brain. They differ from the olfactory sense cells in the size and number of cilia they bear, and also in the internal structure of the cilia. The second cell type are supporting and/or secretory cells. It is concluded that this sense organ is capable of responding to a “special kind” of chemical stimulus and its possible function is discussed.     

The intestinal blood circulation in the River lamprey, Lampetra fluviatilis

Serial sections, injections with india ink and latex, and observations on fresh material, have been used to determine the pattern of blood circulation within the intestine of larval and adult lampreys. Attention has also been paid to resolving the variable terms previously applied to many of the blood vessels, and to the possible functional significance of the differences found between the two life cycle stages. In the larva, the main arterial supply to the intestine consists of a typhlosolar artery, while the venous return is comprised of a posterior and a left and right anterior intestinal vein that usually unite before entering the liver. Although a typhlosolar artery is also present in the adult, the main venous return of the ammocoete is replaced at metamorphosis by a newly formed typhlosolar vein. Moreover, in the ammocoete a considerable amount of blood is discharged into the haemopoietic sponge-work of the typhlosole and the arterial supply to the intestine is poorly developed. By contrast, the typhlosolar sponge-work is lost in the adult and a more efficient arterial supply is developed within the lamina propria of the various intestinal regions. Furthermore, vascular couples are developed in the adult which facilitate the flow of blood in opposite directions in the intestinal wall. Since, during both life cycle stages, the arterial blood passes into tissue spaces, there is no true capillary network in the intestine and no evidence was found for the presence of a lymphatic system. It is suggested that the changes which take place in the intestinal blood supply and the internal structure of the gut during metamorphosis result in improvements both to the vascular system and to the assimilation efficiency.     

日本七鳃鳗的二型体态

于国内首次揭示了日本七鳃鳗的二型体态现象,指出了国内文献资料对日本七鳃鳗特征描述的片面性,同时推出1个修正后的七鳃鳗种的检索表。     

Two Distinct K+ Channels in Lamprey (Lampetra fluviatilis) Erythrocyte Membrane Characterized by Single Channel Patch Clamp

Two channels, distinguished by using single-channel patch-clamp, carry out potassium transport across the red cell membrane of lamprey erythrocytes. A small-conductance, inwardly rectifying K⁺-selective channel was observed in both isotonic and hypotonic solutions (osmolarity decreased by 50%). The single-channel conductance was 26 ± 3 pS in isotonic (132 mm K⁺) solutions and 24 ± 2 pS in hypotonic (63 mm K⁺) solutions. No outward conductance was found for this channel, and the channel activity was completely inhibited by barium. Cell swelling activated another inwardly rectifying K⁺ channel with a larger inward conductance of 65 pS and outward conductance of 15 pS in the on-cell configuration. In this channel, rectification was due to the block of outward currents by Mg²⁺ and Ca²⁺ ions, since when both ions were removed from the cytosolic side in inside-out patches the conductance of the channel was nearly ohmic. In contrast to the small-conductance channel, the swelling-activated channel was observed also in the presence of barium in the pipette. Neither type of channel was dependent on the presence of Ca²⁺ ions on the cytosolic side for activity. Received: 18 July 1997/Revised: 30 January 1998