Ultrastructure of the adenohypophysis in the larval anadromous sea lamprey,Petromyzon marinus L.

The ultrastructure of the adenohypophysis (AH) in the larval anadromous sea lamprey, Petromyzon marinus L., was examined. The AH is subdivided into three regions, the pro-, meso-, and meta-AH. Cells of the nasopharyngeal stalk extend directly beneath the pro- and meso-AH to form the ventral surface of the gland. Some cells in the pro- and meso-AH are arranged into small follicles. Each region of the AH is characterized by a single granulated (secretory) cell type. Granulated cells constitute 80–90% of the pro-AH and contain secretory granules that range from 800 to 2400 Å in diameter. Only 10–20% of the cells in the meso-AH are granulated and they contain much smaller secretory granules (400 to 1250 Å diameter) than those in the pro-AH. Granulated cells constitute 80–90% of the meta-AH and contain only a few secretory granules, ranging from 1000 to 2500 Å in diameter, and many vesicles containing either a loose flocculent or dense granular material. Nongranulated (stellate) cells are found in all regions. They are characterized by their long cell processes, abundant cytoplasmic filaments, and variable electron density. The appearance of organelles in these cells suggests they are nonsecretory. They may play a role in maintaining the structural integrity of the gland and the regulation of granule release in the pro-AH. Two types of nongranulated cells make up 80–90% of the meso-AH. Type I are stellate cells, type II may be undifferentiated cells. The functional significance of the secretory cells in the larval AH is discussed.     

Morphology of the epithelium in the alimentary tract of the larval lamprey,Petromyzon marinus L.

The alimentary tract of the ammocoete of the lamprey, Petromyzon marinus L., is divisible into three morphologically distinct regions: the oesophagus, the anterior intestine, and the posterior intestine. The epithelium of the oesophagus possesses mucous, ciliated, and columnar cells and appears to be specialized for movement of food particles. The epithelium of the anterior intestine possesses secretory cells with numerous zymogen granules, ciliated cells, and columnar-absorptive cells. Although some absorption occurs in the anterior intestine, the main function of this region seems to be the release of digestive enzymes and the continued movement of food particles. The epithelium of the posterior intestine is entirely comprised of columnar absorptive cells, namely tall (light and dark) columnar and low columnar, and the primary function of this region is one of absorption. The epithelium of the hindgut resembles that of the archinephric duct (Youson and McMillan, '71). The morphology of the alimentary tract of ammocoetes suggests that some differentiation and renewal of cell types may occur in the epithelium of the three regions. Comparison of the alimentary tract of larval lamprey with that of other vertebrates indicates that the gut of the ammocoete represents a less specialized level of vertebrate development.     

Transformation of the intestinal epithelium of the larval anadromous sea lamprey Petromyzon marinus L. during metamorphosis

The events in the transformation of the intestine of the larval lamprey into the adult intestine were followed through the seven (1–7) stages of metamorphosis in anadromous Petromyzon marinus L. Light and electron-microscope observations demonstrated that the processes of degeneration, differentiation, and proliferation are involved in the transformation. In the anterior intestine, degeneration of cells and the extrusion of others into the lumen results in the disappearance of secretory (zymogen) cells and the decline in numbers of endocrine and ciliated cells. Larval absorptive cells, with a prominent brush border, are believed to dedifferentiate into unspecialized columnar cells with few microvilli. Degeneration and removal of cells occurs by both autophagy and heterography and cells extruded into the lumen in the anterior intestine are phagocytosed by epithelial cells of the posterior intestine. The loss of epithelial cells during transformation results in the folding and degradation of parts of the basal lamina and in an extensive widening of the lateral intercellular spaces in all parts of the intestine. As metamorphosis is a nontrophic period of the lamprey life cycle, the possible morphological effects of starvation on the intestinal epithelium are discussed. The development of longitudinal folds is a consequence of the events of metamorphic transformation of the intestinal mucosa. Although an interaction between the epithelium and the underlying tissues is believed to be importent, the actual mechanism of fold development is unknown. The intestinal epithelium of adult lampreys develops from surviving cells of the larval (primary) epithelium. Unlike the situation in amphibians, there does not appear to be a group (nest) of undifferentiated larval cells which differentiate into the adult (secondary) epithelium. Instead, in lampreys, columnar cells that persist through the degradative processes seem to be the source of absorptive and ciliated cells and probably are responsible for mucous and secretory cells. Preliminary observations indicate that the intestinal epithelium of feeding adults is specialized into an anterior region which liberates a secretion, absorbs lipid, and possesses the machinery for ion transport. A posterior region absorbs lipid, secretes mucus, and likely is involved in some protein absorption.     

Growth and intermediary metabolism of larval and metamorphosing stages of the landlocked sea lamprey,Petromyzon marinus L

Synopsis Seasonal changes in blood, liver and muscle substrate (glucose, glycogen and lipid) concentrations and enzyme (pyruvate kinase (PyK), fructose diphosphatase (FDPase), NADP-isocitrate dehydrogenase (ICDH), malic enzyme (ME) and the hexose monophosphate shunt dehydrogenases (HMSD)) activities were assessed in ammocoete and metamorphosing stages of a stream stock of the landlocked sea lamprey, Petromyzon marinus L. In all developmental stages studied, muscle rather than liver tissue served as the main site of carbohydrate and fat storage. Blood glucose and muscle lipid exhibited a positive relationship while liver HMSD and muscle ME activity, a negative relationship, with ammocoete weight. These responses were attributed to a proliferation of red fibers and adipocytes in the ammocoete muscle as the time of metamorphosis approched. Muscle lipid stores of ammocoetes in their last year of larval life increased dramatically during the fall and winter preceding metamorphosis. Changes in tissue enzyme activity of ammocoetes in their last year of larval life indicated that the liver was the site of amino acid incorporation into fat while muscle was the site of lipogenesis from glucose. During the non-trophic period of metamorphosis, stored material was catabolized to provide energy for protein synthesis.     

The hemoglobin of the sea lamprey,Petromyzon marinus

The blood hemoglobin of the sea lamprey presents a curious mixture of primitive and highly specialized properties. Like muscle hemoglobin, it has a molecular weight of about 17,000, and apparently contains a single heme. Its isoelectric point is like that of a typical invertebrate hemoglobin. Its amino acid composition is partly characteristic of invertebrate) partly of vertebrate hemoglobins (Pedersen; Roche and Fontaine). In the present experiments, the oxygen equilibrium curve of this pigment was measured at several pH's. As expected, it is a rectangular hyperbola, the first such function to be observed in a vertebrate blood hemoglobin. Other hemoglobins known to possess this type of oxygen dissociation curve—those of vertebrate muscle, the worm Nippostrongylus, and the bot-fly larva—appear to serve primarily the function of oxygen storage rather than transport. Lamprey hemoglobin on the contrary is an efficient oxygen-transporting agent. It achieves this status by having, unlike muscle hemoglobin, a relatively low oxygen affinity, and a very large Bohr effect. In these properties it rivals the most effective vertebrate blood hemoglobins.     

Ultrastructure of the pronephric kidney in upstream migrant sea lamprey, Petromyzon marinus L

The pronephric kidneys were examined in upstream migrant sea lampreys, Petromyzon marinus L., by transmission and scanning electron microscopy. Each pronephros consists of an enlarged renal corpuscle (glomus) and ciliated nephrostomes, but there are no renal tubules. The renal corpuscle contains an extensive mesangium, which consists of a highly fibrous extracellular matrix, numerous mesangial cells, granulocytes, and macrophages. The extracellular matrix contains microfibrils with a morphology similar to amyloid P microfibrils, fibrils with a periodicity similar to fibrin, and abundant collagen. Often these fibrillar components are aggregated in the region of the basement membrane, giving it a thickened appearance. Some podocytes of the visceral epithelium appear swollen, and their cytoplasm contains numerous vacuolar inclusions, and many have only primary major processes with only a few or no foot processes. The morphological features of the pronephric kidney of the lamprey at this time in the life cycle reflect the regression of this organ, but some features also resemble those seen in renal pathologies of higher vertebrates.     

The immune response in adult sea lamprey (Petromyzon marinus L.): the effect of temperature

Adult lamprey were shown to produce antigen-specific antibody in response to weekly injections of human O-type erythrocytes ('O'-RBC) at both 17 degrees C and 9 degrees C. At the higher temperature, high titre antisera were produced earlier and at a faster rate than in animals kept at 9 degrees C. Lamprey kept at 9 degrees C, after a long lag period, eventually produced titres comparable to those obtained at 17 degrees C. Lamprey immunized and maintained at 9 degrees C were capable of producing high titres of antibody in response to second injection of antigen after the primary response titre had declined.     

Immunohistochemical observations of the endocrine pancreas during metamorphosis of the sea lamprey,Petromyzon marinus L.

Summary Light-microscopic immunohistochemistry was used to localize insulin- and somatostatin-immunoreactive cells within developing endocrine pancreatic tissue of metamorphosing lampreys, Petromyzon marinus. The extrahepatic common bile duct and a portion of the intrahepatic bile duct develop into the caudal portion of the endocrine pancreas. The cranial pancreas is composed of follicles originating in the intestinal and diverticular epithelia, thus following the method of formation of pancreatic follicles from gut epithelium in larvae. In both the cranial and caudal portions, and in an intermediate cord of isolated follicles which connect these two major masses, insulin-immunoreactive cells appear first and are followed by cells showing somatostatin-immunoreactivity. In all stages of metamorphosis individual endocrine cells demonstrate immunoreactivity to a single hormone. Biliary atresia in lamprey may have some adaptive significance in providing cells that produce a caudal endocrine pancreas.Supported by NSERC of Canada grant No. A5945 and MRC of Canada grant No. MA8629 to JHY     

Cell renewal in the epithelium of the alimentary tract of the larval lamprey,Petromyzon marinus L.

Light microscopic autoradiography with ³H-thymidine demonstrates that the three regions of the alimentary tract in the larval (ammocoete) lamprey, Petromyzon marinus L., possess different patterns for renewing their epithelium. In the oesophagus, columnar and mucous cells originate from stem cells located at the bases of folds and migrate to the tops of the folds where they are apparently extruded. Ciliated cells, located only at the tops of the folds, seem to differentiate from migrating columnar cells. In the anterior intestine, stem cells are present throughout the epithelium so that there is limited migration of cells and their extrusion occurs randomly. In the posterior intestine, the stem cells located at the bases of the typhlosole provide a continuous population that differentiates and migrates to the top of the typhlosole and to the opposite epithelial wall where they are presumably extruded. The rates of cell renewal in all three epithelial regions of the alimentary tract are slower in animals maintained at 10 ± 1°C compared with those kept at 21 ± 1°C. Comparatively, ammocoetes have the least specialized system for cell renewal known in the alimentary tract of a vertebrate.     

Fruit of the forest - larval sea lamprey Petromyzon marinus are fuelled by allochthonous resources

We used stable isotopes of carbon, hydrogen and nitrogen to quantify the trophic position and resource use of larval sea lamprey Petromyzon marinus, four benthic macroinvertebrate functional feeding guilds (scraper, shredder, collector and predator) and other fishes in three rivers in eastern Canada. Larval lamprey and most invertebrate guilds foraged as primary consumers in all rivers whereas all other fishes predominantly foraged as secondary consumers. Larval lamprey obtained 75–85% of their resources from allochthonous derived material. This level exceeded all invertebrate guilds, which assimilated approximately 50% allochthonous and 50% autochthonous materials and fishes, which predominantly assimilated between 25% and 60% allochthonous material. Larval lamprey occupied a unique position within the river food webs analysed and show remarkable fidelity to a trophic niche specialising on terrestrially derived detritus.     

Migration and spawning energetics of the anadromous sea lamprey,Petromyzon marinus

Synopsis Energy expended in migration and reproduction was determined from measurements of caloric concentration and body and gonodal weight for nontrophic sea lampreys collected from different sites along the St. John River, New Brunswick. The estimated cost of locomotion in swimming the 140 km which separates the estuary from the spawning redds was 300 and 260 kcal for males and females respectively. Acutal distance which lampreys swam as well as mean swimming speed were estimated from a linear regression equation relating energy expenditure for locomotion and body weight. Energy expenditure for breeding was considerably greater than that catabolized throughout the upstream migration.     

Fine structure of the liver in the larval lamprey, Petromyzon marinus L.; hepatocytes and sinusoids

The ultrastructure of hepatocytes, bile canaliculi, and hepatic sinusoids of the larval lamprey, Petromyzon marinus, was examined using thin-sectioned and freeze-fractured tissues. The liver is a "tubular gland" with hepatocytes arranged in a tubular fashion around large bile canaliculi. Hepatocytes are roughly conical in shape, with their tapered apices facing a bile canalicular lumen. They possess extensive rough and smooth endoplasmic reticulum, a well-developed Golgi complex, abundant mitochondria, and varying numbers of large secondary lysosomes. Both secondary lysosomes and the Golgi complex are concentrated in the apical or peribiliary cytoplasm, indicating a possible role in bile secretion. The apical surfaces of the hepatocytes bear numerous elongate microvilli and occasional cilia, which project into the bile canaliculi. The hepatocytes are joined, apically, by junctional complexes composed of zonulae occludentes and adhaerentes. In freeze-fracture, the zonulae occludentes are of variable apicobasal depth and consist of honeycomb-like meshworks of fibrils. Spaces of variable width frequently appear in the P-face grooves, indicating that the zonulae occludentes are "leaky." Numerous communicating (gap) junctions join the hepatocytes laterally. Varying numbers of lateral microvilli project into the intercellular spaces and, basally, the plasma membrane is deeply infolded, resulting in the formation of apparently interdigitating basal processes resting upon a thin basal lamina. Sinusoids are composed of both a heavily-fenestrated, continuous endothelium, and phagocytic reticulo-endothelial (Kupffer) cells. Depsite the difference in arrangement of their hepatocytes, the mammalian and lamprey livers show similar ultrastructural features.     

Immunocytochemical demonstration of growth hormone,prolactin and somatostatin-like immunoreactivities in the brain of larval,young adult and upstream migrant adult sea lamprey,Petromyzon marinus

Summary Growth hormone, prolactin and somatostatinlike immunoreactivities were demonstrated in the brains of larval, young adult (parasitic) and upstream migrant adult sea lampreys, Petromyzon marinus, by means of immunoperoxidase techniques. Growth hormone (GH) and prolactin (PRL) were observed within separate perikarya in the nucleus praeopticus, within fibers in the commissura praeinfundibularis, and in nerve endings within the neurohypophysis of larval and adult-stage lampreys. Cell bodies demonstrating immunoreactive growth hormone were more numerous than those reactive for prolactin. Unlike in the upstream migrant adult lamprey, no GH or PRL was demonstrated in the adenohypophysis of larval or parasitic lamprey.Somatostatin (SRIF)-like immunoreactive neurons were demonstrated in the nucleus commissurae praeinfundibularis, anterior and posterior pars ventralis hypothalami, pars dorsalis thalami, and the tegmentum motorium rhombencephali of larval, parasitic and upstream migrant adult lampreys. Many of the SRIF containing neurons within the hypothalamus were cerebrospinal fluid (CSF)-contacting cells. SRIF fibers were found throughout most of the brain predominating within the nucleus praeopticus, pars ventralis hypothalami, and the nucleus interpeduncularis. No SRIF immunoreactivity was found within the neurophyophysis. The possible functions of these peptides within the brain of the lamprey are discussed.     

Changes in the proximate body composition of the landlocked sea lamprey Petromyzon marinus (L.) during larval life and metamorphosis

Estimates were made of the growth rates and proximate body composition of larval and metamorphosing P. marinus (L.) collected at various times of the year from Shelter Valley Creek, Lake Ontario. Analysis of length-frequency data indicates that the average duration of larval life was 6 years, with metamorphosis occurring predominantly in the length range above 13 cm. Increases in length were almost entirely restricted to the warmest months and did not take place during the final year of larval life. Three categories were thus recognized for the proximate analysis: ammocoetes <13 cm, ammocoetes > 13 cm and metamorphosing individuals. In ammocoetes <13 cm, seasonal differences were observed in the regression coefficients in the logarithmic relationships between wet weight and length and between each of water, lipid and ash and the wet weight. No such difference was found for the regressions between protein and wet weight. For a fixed length (9 cm), the wet weight varied only slightly during the year, although a small peak was seen in May. When considered on the basis of fixed weight (1 g), the relative amount of lipid deposited was greatest in May/July, coincident with a high diatom density. The water content followed an inverse pattern to that of lipid, while the protein and ash contents showed little seasonal variation and exhibited values lower than those normally found in teleosts. Throughout the last year of larval life, the animal stored a greater proportion of lipid, presumably to facilitate the energy demands of metamorphosis during which this food store underwent a marked reduction.     

Fine structure and immunocytochemistry of cells within the endocrine pancreas of larval and adult sea lampreys, Petromyzon marinus L

Immunocytochemistry with protein A-gold and routine electron microscopy were used to identify cell types within the endocrine pancreas of larvae, juvenile adults, and upstream-migrant adults of the sea lamprey, Petromyzon marinus. The larval pancreatic islets are composed only of insulin-immunoreactive B-cells, which are uniform in their fine structure. The cranial and caudal pancreatic tissue in both adult periods contains three cell types: B-cells, somatostatin-immunoreactive D-cells, and a third cell type of unknown content. No glucagon-immunoreactive cells are present in lampreys, but B- and D-cells exist in equal numbers in the pancreatic tissue of adults. The B-cells of adults have a fine structure similar to those in larvae. D-cells have secretory granules that are distinctly different from those both in B-cells and in the third cell type. Although B- and D-cells in lamprey pancreatic tissues have a basic morphological similarity to these cells in other vertebrates, their granules are generally of smaller dimensions. The inclusion of granules within large pleomorphic bodies in many D-cells indicates that granule turnover is common. Immunocytochemistry will be a useful tool for showing the relationship between the cells in the degenerating bile ducts and those of the developing adult pancreas.