Reversible metabolic depression in lamprey hepatocytes during prespawning migration: dynamics of mitochondrial membrane potential

The lamprey (Lampetra fluviatilis L.) is an extant representative of the ancient vertebrate group of Agnathans. During the prespawning migration (the river period of life from autumn until spring) lamprey hepatocytes exhibit widely different energy states: a high-energy state in autumn and spring, corresponding to a normal physiological standard, and a low-energy state in winter, which is provoked by prolonged starvation and profound metabolic arrest. In spring the restoration of energy status (return to an active state) is associated with hormonally induced lipolysis of the lipid droplets stored in the cells. Lamprey hepatocytes demonstrate an aerobic metabolism based on oxidation of free fatty acids. The dynamics of mitochondrial membrane potential (MMP) were measured throughout the prespawning migration. Pharmacological inhibition of the electron transport chain decreased the MMP and caused extensive depletion of cellular ATP without loss of cell viability. The potential molecular mechanisms responsible for winter metabolic depression in lamprey hepatocytes are discussed.     

Anion movements across lamprey (Lampetra fluviatilis) red cell membrane

Chloride and bicarbonate movements across lamprey red cell membrane were investigated. The halftime for equilibration of radioactive chloride across the red cell membrane was 2.46 h, and apparent permeability for chloride-36 was approximately 10(-9) cm X s-1, a value similar to that observed for lipid bilayers. Chloride movements were not affected by the anion exchange inhibitor, 4,4'-diisothiocyano-stilbene-2,2'-disulfonic acid (DIDS). Furthermore, intracellular buffering is effectively isolated from the extracellular compartment, as shown by the fact that practically no pH recovery occurred in the unbuffered extracellular medium after either acidification or alkalinization. These observations show that lamprey red cell membrane is quite impermeable to bicarbonate and other acid/base equivalents.     

Red cell pH of lamprey (Lampetra fluviatilis) is actively regulated

Summary The red cell pH of lamprey (Lampetra fluviatilis) was measured using the DMO method (based on the passive distribution of the wead acid, DMO, across the red cell membrane). The measured red cell pH was higher than the pH of the incubation medium throughout the pH range (7.2–8.2) studied, and higher than the red cell pH calculated from the chloride distribution ratio. Treatment of cells with the metabolic inhibitors 2,4-dinitrophenol or KCN caused a drop in the red cell pH to values lower than the pH of incubation medium, and abolished the difference between the measured red cell pH and the pH calculated from the chloride distribution ratio. These data strongly suggest that the proton gradient across lamprey red cell membrane is actively maintained. Acid extrusion from lamprey red cells may require sodium, as indicated by the observation that when choline was substituted for sodium in the incubation medium, the intracellular pH decreased significantly.     

Locomotor activity of river lamprey Lampetra fluviatilis (L.) during the spawning season

At high latitudes (62–64°N) the river lamprey Lampetra fluviatilis (L.) is shown to have a 24-hr locomotor activity during its spawning period just before midsummer. By keeping lampreys in cold water, the spawning was delayed about one month. Analysis of event recorder graphs indicate that the 24-hr activity is caused by addition of diurnal spawning activity to the basic nocturnal activity and is not caused by weak Zeitgeber conditions during midsummer.Water temperature might indirectly determine time of spawning of the river lamprey.     

Population genetics of a cyclostome species pair, river lamprey (Lampetra fluviatilis L.) and brook lamprey (Lampetra planeri Bloch)

Horizontal agarose gel electrophoresis of 24 allozyme loci in four species of Central European lampreys (321 Lampetra planeri , 83 L. fluviatilis , 11 Eudontomyzon mariae and nine Petromyzon marinus ) was used to study the 'paired species' L. fluviatilis and L. planeri . The genetic differentiation of the anadromous river lamprey ( L. fluviatilis ) from the stationary brook lamprey ( L. planeri ) was within the range of ingroup differentiation of the latter, but L. fluviatilis exhibited much greater population cohesion over a more extended geographic range: G _ST = 0.0537 versus G _ST = 0.3398, N _em = 4.402 versus N _em = 0.4856, mean genetic among-stock distances D = 0.0047 versus D = 0.0257. L. planeri populations coexisting geographically with L. fluviatilis in the Rhine and Elbe river systems were genetically more cohesive than L. planeri stocks from the Danubian basin where L. fluviatilis is absent. Danubian L. planeri populations exhibit a lower degree of heterozygosity than brook lampreys from the Rhine river system, but comprise deeper genetic lineages ( G _ST = 0.4629 versus G _ST = 0.2434), despite being sampled from a much more restricted area. Isolation-by-distance is observed for L. planeri from the Danubian but not from the Atlantic drainage basins. Transspecific gene flow between L. planeri from Atlantic drainage basins and the long-distant migrating L. fluviatilis is inferred, raising doubt on the validity of two separate biospecies. E. mariae and P. marinus are clearly differentiated from Lampetra spp. at several allozyme loci.     

Phylogeography of the European brook lamprey (Lampetra planeri) and the European river lamprey (Lampetra fluviatilis) species pair based on mitochondrial data

The European river lamprey Lampetra fluviatilis and the European brook lamprey Lampetra planeri (Block 1784) are classified as a paired species, characterized by notably different life histories but morphological similarities. Previous work has further shown limited genetic differentiation between these two species at the mitochondrial DNA level. Here, we expand on this previous work, which focused on lamprey species from the Iberian Peninsula in the south and mainland Europe in the north, by sequencing three mitochondrial marker regions of Lampetra individuals from five river systems in Ireland and five in southern Italy. Our results corroborate the previously identified pattern of genetic diversity for the species pair. We also show significant genetic differentiation between Irish and mainland European lamprey populations, suggesting another ichthyogeographic district distinct from those previously defined. Finally, our results stress the importance of southern Italian L. planeri populations, which maintain several private alleles and notable genetic diversity.     

Cysts and tumour-like lesions in the endocrine pancreas of the lamprey (Lampetra fluviatilis)

Histological observations on the endocrine pancreas of the upstream migrant stage of the River lamprey, Lamperra fluviatilis , have revealed a relatively high incidence of cysts and other tumour-like lesions, remarkably similar to the hamartomas described in the Atlantic hagfish, Myxine glufinosa. The frequency of these lesions increases throughout the migratory period from autumn to spring. In the most common type the cysts are surrounded by an intensely granular, single-layered epithelium and both the epithelial cells and the cyst contents show a marked reaction to aldehyde-fuchsin staining. In more extreme forms the breakdown of the epithelial walls of neighbouring cysts leads to the formation of large multi-locular structures occupying almost the whole of the islet organ, but no consistent evidence of changes in blood sugar levels has been obtained in animals where the islet tissue is severely affected. A smaller proportion of the animals display neoplastic developments, frequently associated with hyperplasia and vacuolization of the foregut epithelium. In such cases, there is evidence of invasiveness and the development of epithelioid nodules within the adjoining islet tissue. Cavitation within these nodules leads to the formation of cyst-like structures containing cell debris which reacts to aldehyde-fuchsin staining. The development of cysts and hyperplasia is interpreted as a renewal in the sexually mature lamprey of inherent trends towards epithelial proliferation and vacuolization that are involved during metamorphosis in the morphogenetic processes that lead to the formation of the adult foregut and cranial islet tissue.     

Typology and distribution of ganglion cells in the retina of lamprey (Lampetra fluviatilis)

Retinal ganglion cells (GC) of the Lamprey were studied after in vitro labeling of these cells by iontophoretic deposition of horseradish peroxidase into the optic nerve. The majority of GC are located at the boundary between the inner nuclear layer and the inner plexiform layer; a small proportion (20%) lies in the vitreous portion of the inner plexiform layer. Four types of GC were identified.     

D-glucose permeability in river lamprey (Lampetra fluviatilis) and carp (Cyprinus carpio) erythrocytes.

1. The transport of 3-O-methyl-D-glucose (3-OMG) in lamprey and carp erythrocytes was studied. 2. In lamprey erythrocytes the half-time for the equilibration of 3-OMG was fast, approx. 8 min. In contrast, the erythrocytes of carp were almost impermeable to 3-OMG, with a half-time for equilibration of 14.2 hr. 3. 3-OMG was taken up by lamprey erythrocytes both by facilitated diffusion and simple diffusion. The presence of carrier-mediated transport was indicated by saturation kinetics and by inhibition by phloretin. The Km and Vm of the saturable component of 3-OMG transport were 1.6 mmol/l and 12.4 mmol/kg packed cells/hr. 4. Since the 3-OMG transport in carp erythrocytes showed no sign of saturation kinetics, it appears to proceed by simple diffusion only. 5. These results suggest that, as for most other teleost fish so far studied, low glucose permeability is insufficient to maintain normal energy metabolism in carp erythrocytes. In contrast, in agnathans facilitated glucose transport seems to be quite effective.     

Mitochondrial and lysosomal pathways of lamprey (Lampetra fluviatilis L.) hepatocyte death

Mechanisms of mitochondrial and lysosomal pathways of natural death of lamprey hepatocytes are described at the spring period of the prespawn migration. The mitochondrial pathway (release of cytochrome c from mitochondria into cytosol and activation of caspases) is realized by the classic scheme of apoptosis. Comparatively recently, the lysosomal pathway of cell death associated with cathepsin B activation has been revealed in cells in pathologies, specifically in obstruction of gallbladder and bile ducts. A peculiarity of lamprey hepatocytes consists in that in the adult animal liver there takes place biliary atresia (the absence of gallbladder and bile ducts. Thereby, the lamprey hepatocytes are an excellent object for study of this new pathway of cell death. We have revealed development of the mitochondrial and the lysosomal pathways of cell death of lamprey hepatocytes.     

Respiration rates, ATP content and ionic regulation in hepatocytes of starving lamprey during the pre-spawning period of their life cycle

During the winter pre-spawning migration, lampreys Lampetra fluviatilis stop feeding, and their liver metabolism is reduced substantially. Aerobic ATP production in hepatocytes decreased to one third and ATP content decreased by 50% as compared with the values in autumn. In spite of the decrease of endogenous and phosphorylating (oligomycin-sensitive) respiration in winter, the oxygen consumption used to drive sodium and potassium pumping through Na,K-ATPase activity (ouabain-sensitive respiration) remained virtually constant. Consequently its share in phosphorylating respiration increased from 16·3% in November to 54·2% in February. Potassium influx was similar within the range of ATP content between 2·5 and 1 nmol 10⁻⁶ cells and decreases only in hepatocytes which contained <0·8 nmol ATP 10⁻⁶ cells.     

Cell division in the olfactory epithelium of the lamprey,Lampetra fluviatilis

Summary The turnover of cells within the olfactory epithelium of the lamprey Lampetra fluviatilis was investigated using tritiated thymidine followed by autoradiography. It was found that cell division occurred in three distinct regions of the olfactory lamellae. Two of these regions — a distal lamellar region and a proximal lamellar region occurred outside the sensory area proper, but appeared to contribute cells to the sensory area as well as giving rise to secretory or ciliated cells outside the sensory area. A third region of division occurrred at the base of the sensory area. Division of specialised basal or blastema cells in this region gives rise to cells that are confined to the sensory region of the lamellae. These findings are discussed in the light of previous studies on cell replacement within the olfactory epithelium.