The opisthonephric kidney of the sea lamprey of the great lakes,Petromyzon marinus L. I. The renal corpuscle

The renal corpuscle of the opisthonephric kidney of the Great Lakes lamprey, Petromyzon marinus L., is described. In the larvae (ammocoetes) several renal corpuscles, each consisting of lobed glomeruli and Bowman's capsules, are placed behind one another and there is a progressive lengthening of the posterior corpuscles as the animal ages, apparently by the addition of new glomeruli posteriorly. Bowman's capsule surrounds a single loop and consists of parietal and visceral layers which are similar to those seen in higher vertebrates. The kidney of the adult has a single complex renal corpuscle which extends almost its entire length. Present evidence indicates that the adult kidney is formed during transformation and that the larval renal corpuscles do not contribute to the single elongate corpuscle of the adult. A large number of lobed glomeruli, lying one behind the other, form the elongate glomus. Interposed between each pair of lobes is the dilated end of a nephron. There are no typical Bowman's capsules in the adult although where these nephric capsules invest the blood vessels of a lobe they develop podocytes similar to those of the visceral layer of Bowman's capsule and elsewhere they consist of a simple squamous epithelium similar to that of the parietal layer.     

The opisthonephric kidney of the sea lamprey of the great lakes,Petromyzon marinus L. II. Neck and proximal segments of the tubular nephron

The spatial arrangement of the five basic segments of the tubular nephron of the sea lamprey is reversed between ammocoetes and adults due to differences in the position of the glomus. The initial segment, the ciliated neck segment, resembles the pronephric nephrostome. The proximal segment is divided into two distinct regions, the pars convoluta followed by the pars recta, and has a brush border which reacts positively with the periodic acid-Schiff stain. Ultrastructurally the cells of the proximal segment resemble the corresponding cells of other vertebrates except that they lack the usual infoldings of the basal plasma membrane and they often show intercellular spaces. The mitochondria of these cells often contain filamentous and crystalline material and are usually associated with “flocculent bodies” resembling “non-nucleoid” renal microbodies. The structure of the cells of the proximal segment suggest that they are involved in endocytosis.     

The opisthonephric kidney of the sea lamprey of the great lakes,Petromyzon marinus L. III. Intermediate,distal, and collecting segments of the ammocoete

The presence of microvilli and well-developed organelles in the cells of the non-ciliated intermediate segment of the ammocoete nephron suggests that this is not simply a conducting tubule but may be involved in the formation of urine. This region more closely resembles the thin limb of the loop of Henle of mammals than the intermediate segment of lower vertebrates. The distal segment consists of dark and light cells possessing varying amounts of glycogen, mitochondria, and smooth endoplasmic reticulum. Basal infoldings of the plasma membrane are lacking in the distal cells but wide intercellular spaces are present. Angular prismatic mitochondrial cristae are found within the glycogen-rich cells of the intermediate and distal segments. The collecting segment also possesses narrow intercellular spaces as well as an abundance of organelles, particularly smooth endoplasmic reticulum and mitochondria, suggesting that this region may also be involved in urine formation.     

The opisthonephric kidney of the sea lamprey of the great lakes,Petromyzon marinus L. IV. Intermediate,distal, and collecting segments of the adult

Both short and long intermediate segments are found in the opisthonephric kidney of the adult sea lamprey and they possess morphologically distinct cell types. Wide lateral intercellular spaces appear between both the cuboidal cells of short segments and the squamous cells of long segments. Periodically the epithelium of the long segment is reduced in thickness providing for a close relationship between the interstitium and the tubular fluid. These features indicate that the segment may be involved in urine formation. The distal segment is divisible into two distinct portions: a straight pars recta and a twisted pars convoluta. The cuboidal cells of the pars recta and the columnar cells of the pars convoluta are characterized by abundant smooth endoplasmic reticulum and mitochondria, features reminiscent of cells involved in ion transport. Multivesicular bodies, coated vesicles, and dense apical tubules suggest that cells of both regions may also be involved in endocytosis. The cells of the collecting segment have few organelles but they possess a cytoskeleton of coarse filaments and wide intercellular spaces. The parallel arrangement of the intermediate, distal, and collecting segments and of the blood vessels within the adult kidney is suggestive of a functional unit similar to that found in the medulla of the mammalian kidney.     

Intertubular circulation in the opisthonephric kidneys of adult and larval sea lamprey,Petromyzon marinus L.

The opisthonephric kidneys of both larval and adult Petromyzon marinus lack a renal portal system. The tubules are supplied with arterial blood from the glomus through a system of efferent arterioles, capillaries, and sinusoids with the latter draining into the venous system. The adult kidney possesses a more complex and larger system of sinusoids than ammocoetes and these sinusoids reach the venous system through large subcapsular sinuses. Paralleling of tubules with sinusoids and arterioles in the ventral portion of the adult kidney resembles the association of the vasa recta with the tubules of the renal medulla in the mammalian kidney. This may suggest a mechanism important to tubular transport in adult lampreys. The intertubular tissue of ammocoetes is infiltrated with haemopoietic tissue while large numbers of phagocytes within the adult sinusoids may be involved in the destruction of blood cells.     

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 prolcphios 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.     

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

The fine structure of mucocartilage, a tissue unique to larval lampreys, was examined in Petromyzon marinus L. This tissue is surrounded by a perichondrium of vascularized, dense connective tissue composed of fibroblasts, collagen fibrils, and elastic-like microfibrils, but it is avascular itself and consists of elastic-like microfibrils, ground substance, and a few diffusely scattered fibroblasts. Fibroblasts possess rough endoplasmic reticulum, many free ribosomes, a well-developed Golgi apparatus, a tubulo-vesicular network, and a number of secondary lysosomes containing crystalline material. The appearance of the organelles suggests the involvement of the cell in the synthesis and secretion of the ground substance and microfibrils. Tubular microfibrils, 11–13 nm in diameter, comprise the major portion of the matrix, and they are similar to those described in developing mammalian elastic tissue (Ross and Bornstein, 1969). The retention of the microfibrils may represent either a primitive form of elastic fiber in this “primitive” vertebrate or reflect the larval condition of the lampreys under examination. Scattered spherical to polyhedral-shaped matrix granules and inter-granular filaments make up the remainder of the matrix. It was concluded that mucocartilage in larval lampreys is not a conventional type of vertebrate connective tissue.     

Immunocytochemical localization of thyroglobulin in the endostyle of the anadromous sea lamprey,Petromyzon marinus L.

Thyroglobulin (TG) was localized in the endostyle of the anadromous sea lamprey, Petromyzon marinus L. by means of the unlabeled antibody peroxidase-antiperoxidase immunocytochemical method. TG was found localized on the apical surface and within the cytoplasm of type 2c and 3 cells and in some type 5 cells. By identifying the cells of the endostyle immunocytochemically it may be possible to study more readily the events of endostylar transformation during metamorphosis.     

Chondrogenesis of the branchial skeleton in embryonic sea lamprey, Petromyzon marinus

This study provides concise temporal and spatial characteristics of branchial chondrogenesis in embryonic sea lamprey, Petromyzon marinus, using high resolution light microscopy, transmission electron, and immunoelectron microscopy. Prechondrogenic condensations representing the first branchial arch appeared first in the mid-region of the third pharyngeal arch at 13 days post-fertilization (pf). Cartilage differentiation, defined by the presence of the unique, fibrillar, non-collagenous matrix protein characteristic of branchial cartilage, was first observed at 14 days pf. Development of lamprey branchial cartilage appeared unusual compared to that in jawed fishes, in that precartilage condensations appear as a one-cell wide orderly stack of flattened cells that extend by the addition of one dorsal and one ventral condensation. Development of lamprey gill arches from three condensations that fuse to form a single skeletal element differs from the developing gill arches of jawed fishes, where more than one skeletal element forms from a single condensation. The initial orderly arrangement of cells in the lamprey branchial prechondrogenic condensations remains throughout development. Once chondrification of the condensations begins, the branchial arches start to grow. Initially, growth occurs as a result of matrix secretion and cell migration. Later in development, the arches grow mainly by cell proliferation and enlargement. This study defines the morphology and timing of lamprey branchial chondrogenesis. Studies of lamprey chondrogenesis provide not only insight into the developmental biology of a unique non-collagenous cartilage in a primitive vertebrate but also into the general evolution of the skeletal system in vertebrates.     

Corneal splitting in the developing lamprey Petromyzon marinus L. eye

The cornea of the adult lamprey has both dermal (spectacle) and scleral components. These are separated by a thin mucoid layer that allows free movement of the globe. This study has shown that during the larval (ammocoete) stage, the lamprey cornea develops in a manner similar to that of other lower vertebrates. Just prior to the period of transformation to the adult parasite, the outer dermal portion of the ammocoete cornea (spectacle) consists of an anterior stratified columnar epithelium with goblet cells at the surface. The stroma of the dermal cornea consists of a thick outer layer of orthogonally oriented collagen with branching fibroblasts and a thin, loosely organized inner layer with slender elongated fibroblasts. The scleral cornea is lined internally by a flattened monolayer of mesodermal cells, the corneal endothelium. Its narrow stroma is composed entirely of thin, orthogonally aranged, collagen-fiber lamellae, and is bounded externally by a thin continuous mesothelial layer of cells that abuts directly onto the loose stromal component of the dermal cornea. During the early stages of transformation, the anterior epithelium of the dermal cornea becomes stratified squamous in type. Later, the inner loose stroma of the dermal cornea (spectacle) begins to separate from the scleral cornea components, and a third complete mesothelial layer forms a distinct inner border for the dermal cornea. A mucoid layer is formed between the dermal (spectacle) and scleral corneas and remains throughout the adult life.     

Development of the adult endocrine pancreas during metamorphosis in the sea lamprey,Petromyzon marinus L. II. Electron microscopy and immunocytochemistry

The development of the adult endocrine pancreas was followed throughout metamorphosis in the sea lamprey using electron microscopy and immunocytochemistry. It was discovered that the caudal pancreas develops from the larval extrahepatic common bile duct through the process of transdifferentiation (dedifferentiation/redifferentiation). Early in metamorphosis the bile duct epithelial cells possess large vacuoles, resembling autophagic vacuoles, containing recognizable cell material. There is a loss of the large bundles of intermediate filaments characteristic of the larval bile duct epithelium. These same cells are then seen to contain granules immunoreactive for insulin. Pancreatic islets develop within the base of the bile duct epithelium from these transdifferentiated cells and migrate into the surrounding connective tissue to form the caudal pancreas. The cranial pancreas was found to develop from the epithelia lining the developing adult diverticulum and anterior intestine in a similar fashion as those in the larva. The second cell type to appear in either portion of the developing pancreas is similar to the third cell type of the adult: cells immunoreactive for somatostatin do not appear until late in metamorphosis in either region. © 1993 Wiley-Liss Inc.     

Convoluted bile ducts in the liver of the larval lamprey, Petromyzon marinus L

Summary The three-dimensional structure of the bile ducts and their relationship to the blood vessels were studied in the larval lamprey by scanning electron microscopy of the intact tissue and of biliary and vascular casts. The intrahepatic gall bladder is situated in the cephalic portion of the liver and a cystic duct is connected to a straight intrahepatic common bile duct, which extends to the extrahepatic bile duct at the caudal end of the liver. Several smaller intrahepatic common bile ducts are connected directly to the intrahepatic common bile duct, are convoluted or serpiginous and are surrounded intimately by sinusoids. This arrangement enables the bile ducts to have increased surface area exposed to blood vessels. The functional significance of this arrangement is discussed with respect to the modification of bile through the transport of solutes and the similarity of this bilio-vascular relationship to the peribiliary vascular plexus of the mammalian liver.     

The anionic charge barrier in the renal corpuscle of the pronephros in the lamprey, Petromyzon marinus L.

The charge barrier within the renal corpuscle of the pronephric kidney of the lamprey, Petromyzon marinus, was investigated at two life cycle intervals using cationized ferritin and polyethyleneimine. In the larval renal corpuscle the endothelium of the glomerular capillaries and the laminae rarae externa and interna of the glomerular basement membrane show regularly-spaced deposits of the tracers. The lamina densa remains unstained. Concomitant with a loss of major processes of the visceral epithelial podocytes and development of an extensive mesangial matrix in late adult life are alterations in the distribution of the anionic sites. The lamina rara interna is no longer a distinct entity and the mesangium contains irregularly-distributed anionic sites surrounding electron-dense deposits. The results indicate that the distribution of the anionic sites during adult life most likely affects the ability of the renal corpuscle to act as an efficient filtration device. This charge distribution is consistent with that seen during some renal pathologies of higher vertebrates.     

Ultrastructure of cartilage from young adult sea lamprey,Petromyzon marinus L: A new type of vertebrate cartilage

Ultrastructural observations of cartilage from adult sea lamprey, Petromyzon marinus, reveal a highly cellular cartilage with an unusual extracellular matrix. The avascular cartilage is surrounded by a vascular perichondrium, which consists of dense connective tissue containing fibroblasts, collagen fibrils, and microfibrils. The cells (chondrocytes) vary in morphology in different parts of the cartilage in a way that may reflect their state of activity. Chondrocytes within the peripheral cartilage contain tubulo-vesicular structures along the cell surface, an extensive lamellar rough endoplasmic reticulum, and a well-developed Golgi complex with associated vesicles and vacuoles. The presence of material within the Golgi elements that resembles components of the extracellular matrix suggests the involvement of the peripheral chondrocytes in the synthesis and secretion of the matrix components. Chondrocytes within the central cartilage are hypertrophied and contain a pale cytoplasm with a reduced number of organells that are widely spaced throughout the cell. The appearance of the organelles within these cells suggests that they are not as actively involved in the production of the matrix as those of the peripheral cartilage. The extracellular matrix consists of a dense network of randomly arranged, branched, noncollagenous matrix fibrils 15–40 nm in diameter and varying amounts of electron-dense matrix granules. Due to the unique nature of its extracellular matrix, the cartilage of the lamprey cannot be likened to any of the known vertebrate cartilages and, therefore, must be considered a new type of vertebrate cartilage.     

Retinal development in the lamprey (Petromyzon marinus L.): Premetamorphic ammocoete eye

Development of the retina of the ammocoete begins early in embryogenesis, with the formation of the optic vesicle, but development of the rudimentary eye is suspended and remains arrested during larval life. Prior to the onset of metamorphosis, the retina of the ammocoete is completely undifferentiated, with the exception of a small area (Zone II) surrounding the optic nerve head, where all of the adult retinal layers are found. The photoreceptors in this area have developed to include synaptic contacts as well as inner and outer segments. The pigment epithelium in this area, too, has differentiated to include well-formed melanin granules, myeloid bodies and endoplasmic reticulum and is closely associated with the receptor cell outer segments. With the approach of metamorphosis, differentiation of the remainder of the retina (Zone I) begins, taking place in a radial fashion from the optic nerve head. Differentiating pigment epithelial cells adjacent to the differentiated retinal zone begin to accumulate melanin granules. In the neural retina, junctional complexes are established in the form of an external limiting membrane, and connecting cilia project into the optic ventricle. Photoreceptor differentiation begins with the formation of a mitochondria-filled ellipsoid within the inner segment. Development and differentiation of the ammocoete retina is unique to vertebrates in that only a small area of differentiated retina is present during the larval stage. The remainder of the retina differentiates and becomes functional during metamorphosis.