Japanese Lamprey Research Articles

Onset of Apoptosis in the Cystic Duct During Metamorphosis of a Japanese Lamprey, Lethenteron reissneri

A nonparasitic lamprey in Japan, Lethenteron reissneri, stops feeding prior to the commencement of metamorphosis. Resumption of feeding cannot take place due to major alterations in the digestive system, including loss of the gall bladder (GB) and biliary tree in the liver. This degeneration of bile ducts is considered to depend on programmed cell death or apoptosis, but molecular evidence of apoptosis remains lacking. Using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining and immunohistochemistry with an antibody against active caspase-3, we showed that epithelial cells of the cystic duct (CD) and GB became TUNEL-positive by the early metamorphosing stage. Immunohistochemical staining of active caspase-3, a key mediator in the apoptotic cascade, showed that the apoptotic signal was initiated in the region around the CD in the late larval phase. In later stages, active caspase-3-positive epithelial cells were also observed in the large intrahepatic bile duct (IHBD) and peripheral small IHBDs. At the early metamorphosing stage, bile canaliculi between hepatocytes were dilated and displayed features resembling canaliculi in cholestasis. Onset of apoptosis around the CD, which is the pathway for the storage of bile juice, and progression of apoptosis towards the large IHBD, which is the pathway for the secretion of bile juice, may lead to temporary intrahepatic cholestasis. The present study represents the first precise spatial and temporal analysis of apoptosis in epithelial cells of the biliary tract system during metamorphosis of any lamprey species.

A novel CD29-like protein expressed in Japanese lamprey ( Lethenteron japonicum) and involved in immune response

CD29 is the integrin beta1 subunit. Integrin family members are membrane receptors involved in cell adhesion and recognition in a variety of processes including embryogenesis, tissue repair and immune response. In this study, a novel CD29-like gene ( LCD29) was identified and characterized in Japanese lamprey ( Lethenteron japonicum), an agnathan that occupies a critical phylogenic position between cephalochordates and gnathostomes. After a partial cDNA sequence of LCD29 was found from the leucocyte cDNA library, the full-length cDNA was obtained by means of 3′ and 5′ RACE, respectively. LCD29 encodes 780 amino acids and shares high sequence homology with other vertebrates. Both real-time PCR and immunohistochemical assays have demonstrated the wide distribution of the LCD29 in lamprey tissues, and FACS analysis has shown that the expression level of this protein is higher in granulocytes than in lymphocytes. Furthermore, after the lampreys were stimulated with lipopolysaccharide, the levels of LCD29 mRNA were obviously up-regulated in the leucocytes, intestine, heart and gill tissues. Most importantly, an interaction between LCD29 and LCD9 in the intestine has been found with co-immunoprecipitation assays for the first time. The results obtained in the present study will help us to understand the function of LCD29 in immune response in jawless vertebrates.

Phylogenetic and expression analysis of lamprey toll-like receptors

Toll-like receptors (TLRs) have been identified as pivotal sensors recognizing microbial pattern molecules in vertebrates. Whole genome analysis of the teleost Takifugu rubripes supports the existence of a fundamental family of TLR genes in fish. However, the role of the innate immune system in the context of raising acquired immunity in jawless fish remains unclear. In this study, we annotated 16 lamprey TLR genes predicted from the latest genome assembly of lamprey on the basis of homology, and identified their cDNAs from Japanese lamprey, Lethenteron japonicum. Phylogenetic analyses indicated that the repertoire of lamprey TLRs consisted of both fish (F)- and mammalian (M)-type TLRs, and it was also demonstrated that lamprey TLRs are constitutively expressed in various organs. Our results suggest that lampreys protect against microorganisms using the innate system consisting of a similar set of M- and F-type TLRs, despite possessing a unique acquired immune system. In addition, type I interferon ( IFN), interferon regulatory factor ( IRF)- 3, and IRF7 were not identified in the lamprey genome although TLR adaptor and signal transduction genes were highly conserved upstream of (IRF)-3/7 and type I IFN in most vertebrates. This is the first report to describe the TLR repertoire and IFN system in one of the most primitive vertebrates, the lamprey.

Cloning and analysis of <I>Lyb</I> gene from <I>Lampetra japon-ica</I>

In the present study, three Y-box cDNAs, named as Lyb1, Lyb2, and Lyb3, were cloned from the liver and oral gland of Japanese lamprey (Lampetra japonica) using reverse transcript-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. Further bioinformatical analysis indicated that the three Y-box genes of lamprey, i.e., Lyb1, Lyb2, and Lyb3, encoded 331, 181, and 171 amino acids, respectively. The three amino acid sequences were homologically aligned with those from Ciona intestinalis, Danio rerio, Oryzias latipes, Takifugu rubripes, and Mus musculus by DNAMAN software. The Lyb genes were novel members of Y-box gene family containing a conserved cold-shock domain with two RNA binding motifs (RNP-1 and RNP-2). Moreover, phylogenetic analysis revealed that there was only one type of Y-box gene before the emergence of gnathostomata. By contrast, the Y-box gene were divided into three groups, YB1, YB2, and CSDA during evolution of jawed vertebrates.

Characterization of the neurohypophysial hormone gene loci in elephant shark and the Japanese lamprey: origin of the vertebrate neurohypophysial hormone genes

BackgroundVasopressin and oxytocin are mammalian neurohypophysial hormones with distinct functions. Vasopressin is involved mainly in osmoregulation and oxytocin is involved primarily in parturition and lactation. Jawed vertebrates contain at least one homolog each of vasopressin and oxytocin, whereas only a vasopressin-family hormone, vasotocin, has been identified in jawless vertebrates. The genes encoding vasopressin and oxytocin are closely linked tail-to-tail in eutherian mammals whereas their homologs in chicken, Xenopus and coelacanth (vasotocin and mesotocin) are linked tail-to-head. In contrast, their pufferfish homologs, vasotocin and isotocin, are located on the same strand of DNA with isotocin located upstream of vasotocin and separated by five genes. These differences in the arrangement of the two genes in different bony vertebrate lineages raise questions about their origin and ancestral arrangement. To trace the origin of these genes, we have sequenced BAC clones from the neurohypophysial gene loci in a cartilaginous fish, the elephant shark (Callorhinchus milii), and in a jawless vertebrate, the Japanese lamprey (Lethenteron japonicum). We have also analyzed the neurohypophysial hormone gene locus in an invertebrate chordate, the amphioxus (Branchiostoma floridae).ResultsThe elephant shark neurohypophysial hormone genes encode vasotocin and oxytocin, and are linked tail-to-head like their homologs in coelacanth and non-eutherian tetrapods. Besides the hypothalamus, the two genes are also expressed in the ovary. In addition, the vasotocin gene is expressed in the kidney, rectal gland and intestine. These expression profiles indicate a paracrine role for the two hormones. The lamprey locus contains a single neurohypophysial hormone gene, the vasotocin. The synteny of genes in the lamprey locus is conserved in elephant shark, coelacanth and tetrapods but disrupted in teleost fishes. The amphioxus locus encodes a single neurohypophysial hormone, designated as [Ile4]vasotocin.ConclusionThe vasopressin- and oxytocin-family of neurohypophysial hormones evolved in a common ancestor of jawed vertebrates through tandem duplication of the ancestral vasotocin gene. The duplicated genes were linked tail-to-head like their homologs in elephant shark, coelacanth and non-eutherian tetrapods. In contrast to the conserved linkage of the neurohypophysial genes in these vertebrates, the neurohypophysial hormone gene locus has experienced extensive rearrangements in the teleost lineage.

Identification of four Engrailed genes in the Japanese lamprey, Lethenteron japonicum

We have isolated four homologs of Engrailed genes from the Japanese lamprey, Lethenteron japonicum, an agnathan that occupies a critical phylogenic position between cephalochordates and gnathostomes. We named these four genes LjEngrailedA, LjEngrailedB, LjEngrailedC, and LjEngrailedD. LjEngrailedA, LjEngrailedB, and LjEngrailedD share a major expression domain in the presumptive midbrain-hindbrain boundary region of the central nervous system, although their levels and timing of expression differed. On the other hand, LjEngrailedC transcripts were in the pharyngeal ectoderm and the ventral ectoderm of the body wall. In addition, LjEngrailedA was expressed in the ventral side of the epibranchial muscle precursors. LjEngrailedD transcripts were seen in the mesodermal cells of the mandibular arch and later in a group of cells responsible for the formation of the upper lip, lower lip, and velum. Our results provide clues to the evolution of these structures as well as a possible scenario for duplication events of Engrailed genes.

Noncanonical role of Hox14 revealed by its expression patterns in lamprey and shark

Hox genes are arranged in uninterrupted clusters in vertebrate genomes, and the nested patterns of their expression define spatial identities in multiple embryonic tissues. The ancestral Hox cluster of vertebrates has long been thought to consist of, maximally, 13 Hox genes. However, recently, Hox14 genes were discovered in three chordate lineages, the coelacanth, cartilaginous fishes, and amphioxus, but their expression patterns have not yet been analyzed. We isolated Hox14 cDNAs from the Japanese lamprey and cloudy catshark. These genes were not expressed in the central nervous systems, somites, or fin buds/folds but were expressed in a restricted cell population surrounding the hindgut. The lack of Hox14 expression in most of the embryonic axial elements, where nested Hox expressions define spatial identities, suggests a decoupling of Hox14 genes' regulation from the ancestral regulatory mechanism. The relaxation of preexisting constraint for collinear expression may have permitted the secondary losses of this Hox member in the tetrapod and teleost lineages.

Identification and developmental expression of two Tbx1/10-related genes in the agnathan Lethenteron japonicum

We have identified two Tbx1/10-related genes, LjTbx1/10A and LjTbx1/10B, from the Japanese river lamprey Lethenteron japonicum. We used in situ hybridization to characterize their expression pattern during embryonic development. LjTbx1/10A and LjTbx1/10B shared common expression in the pharyngeal arches and otic vesicle, although their levels and timing of expression differed markedly. LjTbx1/10A was highly expressed in the mesodermal core of pharyngeal arches and the adjacent endoderm throughout pharyngeal arch development, whereas LjTbx1/10B expression was only transiently upregulated in forming pharyngeal pouches. LjTbx1/10A transcripts first appeared at stage 25 in otic vesicles, whereas LjTbx1/10B transcripts could already be detected in the developing otic placode at stage 20. These results suggest that lamprey LjTbx1/10A and LjTbx1/10B may play distinct roles in the patterning and development of the pharyngeal apparatus. It appears that lamprey Tbx1/10 genes have undergone subfunctionalization independent from gnathostomes, with regard to both regulation and function.

Molecular cloning, characterization and expression analysis of a putative C-type lectin (Fclectin) gene in Chinese shrimp Fenneropenaeus chinensis

Lectin is regarded as a potential molecule involved in immune recognition and phagocytosis through opsonization in crustacean. Knowledge on lectin at molecular level would help us to understand its regulation mechanism in crustacean immune system. A novel C-type lectin gene ( Fclectin) was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3′ and 5′ rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 1482 bp with an 861 bp open reading frame, encoding 287 amino acids. The deduced amino acid sequence contains a putative signal peptide of 19 amino acids. It also contains two carbohydrate recognition domains/C-type lectin-like domains (CRD1 and CRD2), which share 78% identity with each other. CRD1 and CRD2 showed 34% and 30% identity with that of mannose-binding lectin from Japanese lamprey ( Lethenteron japonicum), respectively. Both CRD1 and CRD2 of Fclectin have 11 amino acids residues, which are relatively invariant in animals’ C-type lectin CRDs. Five residues at Ca 2+ binding site 1 are conserved in Fclectin. The potential Ca 2+/carbohydrate-binding (site 2) motif QPD, E, NP (Gln-Pro-Asp, Glu, Asn-Pro) presented in the two CRDs of Fclectin may support its ability to bind galactose-type sugars. It could be deduced that Fclectin is a member of C-type lectin superfamily. Transcripts of Fclectin were found only in hemocytes by Northern blotting and RNA in situ hybridization. The variation of mRNA transcription level in hemocytes during artificial infection with bacteria and white spot syndrome virus (WSSV) was quantitated by capillary electrophoresis after RT-PCR. An exploration of mRNA expression variation after LPS stimulation was carried out in primarily cultured hemocytes in vitro. Expression profiles of Fclectin gene were greatly modified after bacteria, LPS or WSSV challenge. The above-stated data can provide us clues to understand the probable role of C-type lectin in innate immunity of shrimp and would be helpful to shrimp disease control.

Lampreys Rely on a Molecular Switch to Detect UV Light

By applying a combination of biochemical and biophysical techniques, researchers have identified a UV-sensitive photoreceptor in the pineal gland of the Japanese river lamprey.

Segmental development of reticulospinal and branchiomotor neurons in lamprey: insights into the evolution of the vertebrate hindbrain.

During development, the vertebrate hindbrain is subdivided along its anteroposterior axis into a series of segmental bulges called rhombomeres. These segments in turn generate a repeated pattern of rhombomere-specific neurons, including reticular and branchiomotor neurons. In amphioxus (Cephalochordata), the sister group of the vertebrates, a bona fide segmented hindbrain is lacking, although the embryonic brain vesicle shows molecular anteroposterior regionalization. Therefore, evaluation of the segmental patterning of the central nervous system of agnathan embryos is relevant to our understanding of the origin of the developmental plan of the vertebrate hindbrain. To investigate the neuronal organization of the hindbrain of the Japanese lamprey, Lethenteron japonicum, we retrogradely labeled the reticulospinal and branchial motoneurons. By combining this analysis with a study of the expression patterns of genes identifying specific rhombomeric territories such as LjKrox20, LjPax6, LjEphC and LjHox3, we found that the reticular neurons in the lamprey hindbrain, including isthmic, bulbar and Mauthner cells, develop in conserved rhombomere-specific positions, similar to those in the zebrafish. By contrast, lamprey trigeminal and facial motor nuclei are not in register with rhombomere boundaries, unlike those of gnathostomes. The trigeminal-facial boundary corresponds to the rostral border of LjHox3 expression in the middle of rhombomere 4. Exogenous application of retinoic acid (RA) induced a rostral shift of both the LjHox3 expression domain and branchiomotor nuclei with no obvious repatterning of rhombomeric segmentation and reticular neurons. Therefore, whereas subtype variations of motoneuron identity along the anteroposterior axis may rely on Hox-dependent positional values, as in gnathostomes, such variations in the lamprey are not constrained by hindbrain segmentation. We hypothesize that the registering of hindbrain segmentation and neuronal patterning may have been acquired through successive and independent stepwise patterning changes during evolution.

Expression of foreign genes in lamprey embryos: an approach to study evolutionary changes in gene regulation.

Evolution in development can be viewed as a sequence of changes in gene regulation. To investigate the cross-species compatibility of 5' upstream regulatory regions, we introduced exogenous gene constructs derived from a gnathostome genome into fertilized eggs of the Japanese lamprey, Lampetra japonica, a sister group of the gnathostomes. Eggs were injected with gene constructs in which a sequence encoding the green fluorescent protein (GFP) had been located downstream of either a virus promoter or 5' regulatory regions of medaka actin genes. Reporter gene expression was recorded for more than a month starting two days after injection. Although the expression patterns were highly mosaic and differed among individuals, GFP was expressed predominantly in the striated muscles of lamprey embryos when driven by the 5' upstream regions of the medaka muscle actin genes. This implies that a pan-vertebrate muscle-specific gene regulatory mechanism may have evolved before the agnathan/gnathostome divergence. This gene-transfer technique potentially facilitates the visualization of cells in various differentiating tissues throughout development. The introduction of developmental genes of the lamprey or other animals into lamprey embryos is another potentially important application, one that could provide us with information on the evolutionary changes in functions of genes or gene cascades.

Compartments in the lamprey embryonic brain as revealed by regulatory gene expression and the distribution of reticulospinal neurons

The vertebrate neural tube consists of a series of neuromeres along its anteroposterior axis. Between amphioxus that possesses no neuromeres and gnathostomes, the lamprey occupies a critical position in the phylogeny for the origin of the segmented brain. To clarify the rhombomeric configuration of the Japanese lamprey, Lampetra japonica, we injected rhodamine- and fluorescein-labeled dextrans into the larval spinal cord, and retrogradely labeled the reticulospinal neurons. We also isolated prosomere marker genes from the embryonic cDNA library of L. japonica, and performed in situ hybridization on the embryonic brain. Of the genes examined, LjOtxA, LjPax6, LjPax2/5/8, LjDlx1/6, and LjTTF-1 were expressed in clearly demarcated polygonal domains. In the telencephalon, LjDlx1/6, LjPax6, and a putative paralogue of LjEmx were expressed in different domains; the LjEmx paralogue was expressed in the dorsal region, and LjDlx1/6 and LjPax6 in a complimentary fashion of the middle part. These expression patterns implied existence of a tripartite configuration of the lamprey telencephalon similar to that in gnathostomes. All these evidences strongly suggest that the segmental and compartmental architecture of the vertebrate brain was already established before the divergence of agnathans and gnathostomes.

Embryology of the lamprey and evolution of the vertebrate jaw: insights from molecular and developmental perspectives.

Evolution of the vertebrate jaw has been reviewed and discussed based on the developmental pattern of the Japanese marine lamprey, Lampetra japonica. Though it never forms a jointed jaw apparatus, the L. japonica embryo exhibits the typical embryonic structure as well as the conserved regulatory gene expression patterns of vertebrates. The lamprey therefore shares the phylotype of vertebrates, the conserved embryonic pattern that appears at pharyngula stage, rather than representing an intermediate evolutionary state. Both gnathostomes and lampreys exhibit a tripartite configuration of the rostral-most crest-derived ectomesenchyme, each part occupying an anatomically equivalent site. Differentiated oral structure becomes apparent in post-pharyngula development. Due to the solid nasohypophyseal plate, the post-optic ectomesenchyme of the lamprey fails to grow rostromedially to form the medial nasal septum as in gnathostomes, but forms the upper lip instead. The gnathostome jaw may thus have arisen through a process of ontogenetic repatterning, in which a heterotopic shift of mesenchyme-epithelial relationships would have been involved. Further identification of shifts in tissue interaction and expression of regulatory genes are necessary to describe the evolution of the jaw fully from the standpoint of evolutionary developmental biology.

Isolation of Dlx and Emx gene cognates in an agnathan species, Lampetra japonica, and their expression patterns during embryonic and larval development: Conserved and diversified regulatory patterns of homeobox genes in vertebrate head evolution

Agnathan cognates of vertebrate homeobox genes, Emx and Dlx, were isolated from embryonic cDNA of a Japanese marine lamprey, Lampetra japonica. Analyses of amino acid sequences indicated that the Dlx cognate was closely related to the common ancestor of gnathostome Dlx1 and Dlx6 groups and termed LjDlx1/6. Southern blot analyses could not rule out the possibility that L. japonica possesses more than one paralog for both LjDlx1/6 and LjEmx, the lamprey cognate of Emx. Expression of LjDlx1/6 was regulated spatially as well as developmentally, and its transcripts were mainly found in the craniofacial and pharyngeal mesenchyme and in the forebrain. The expression pattern of LjEmx changed dramatically during embryogenesis; expression was seen initially in the entire neural tube and mesoderm, which were secondarily downregulated, and secondarily in cranial nerve ganglia and in the craniofacial mesenchyme. No specific expression of LjEmx was seen in the telencephalon. Comparisons of Dlx and Otx gene expression patterns suggested a shared neuromeric pattern of the vertebrate brain. Absence of Emx expression implied that the patterning of the lamprey telencephalon is not based on the tripartite plan that has been presumed in gnathostomes. Expression domains of LjDlx1/6 in the upper lip and of LjEmx in the craniofacial mesenchyme were peculiar features that have not been known in gnathostomes. Such differences in expression pattern may underlie distinct morphogenetic pathway of the mandibular arch between the agnathans and gnathostomes.

Pax1/Pax9-Related Genes in an Agnathan Vertebrate, Lampetra japonica: Expression Pattern of LjPax9 Implies Sequential Evolutionary Events toward the Gnathostome Body Plan

Among the transcription factor gene families, Pax genes play important and unique roles in morphological patterning of animal body plans. Of these, Group I Pax genes (Pax1 and Pax9) are expressed in the endodermal pharyngeal pouches in many groups of deuterostomes, and vertebrates seem to have acquired more extensive expression domains in embryos. To understand the evolution of Pax1/Pax9-related genes in basal groups of vertebrates, their cognates were isolated from the Japanese marine lamprey, Lampetra japonica. RT-PCR of larval lamprey cDNA yielded two different fragments containing vertebrate Pax1- and Pax9-like paired domains. The Pax9 orthologue was isolated and named LjPax9. Whole-mount in situ hybridization revealed that this gene was expressed in endodermal pharyngeal pouches, mesenchyme of the velum (the oral pumping apparatus) and the hyoid arch, and the nasohypophysial plate, but not in the somitic mesoderm of the lamprey embryo. These expression patterns could be regarded as a link between the basal chordates and the gnathostomes and are consistent with the phylogenetic position of the lamprey. Especially, the appearance of neural crest seemed to be the basis of velar expression. Homology of the velum and the jaw is also discussed based on the LjPax9 expression in the first pharyngeal pouch and in the velar mesenchyme. We conclude that Pax9 genes have sequentially expanded into new expression domains through evolution as more complicated body plans emerged.

Otx cognates in a lamprey, Lampetra japonica.

Gnathostomes have two lineages of Otx genes, Otx1 and Otx2, as cognates of a Drosophila head gap gene, orthodenticle. Previous studies with mutant mice have demonstrated that they play essential roles in the development of rostral head. To shed lights on the evolution of the rostral head in vertebrates we isolated their cognates in the Japanese marine lamprey, Lampetra japonica. The lamprey genome appeared to have two Otx cognantes, LjOtxA and LjOtxB. Phylogenetic analyses suggest that LjOtxA clusters with gnathostome Otx2 genes, but LjOtxB does not belong to either the Otx1 or Otx2 lineage. LjOtxA was expressed in the forebrain and midbrain with the caudal limit possibly at the midbrain/hindbrain junction as gnathostome Otx cognates are, but LjOtxB was not expressed in the brain. No Otx1 or Otx2 cognates are known in gnathostomes that are not expressed in the brain. Both LjOtxA and LjOtxB were expressed in the olfactory placode, epiphysis, optic stalks, and lower and upper lips. LjOtxB was also expressed in the eyes, where no LjOtxA transcripts were detected. Thus, Otx1 and Otx2 functions for the development of forebrain and midbrain in gnathostomes appear to be shouldered by LjOtxA alone in the lamprey. LjOtxB may have diverged from the stem of the Otx1 and Otx2 lineages and evolved independently.

Peripheral development of cranial nerves in a cyclostome,Lampetra japonica: morphological distribution of nerve branches and the vertebrate body plan

The development of peripheral nerves was studied in a Japanese marine lamprey, Lampetra japonica, in whole-mount and sectioned embryos from hatching until the earliest ammocoete. Nerve fibers were immunohistochemically stained with a monoclonal antibody against acetylated tubulin. Branchiomeric nerves first developed in a simple metamerical pattern, each associated with a single pharyngeal arch. Of those, the ophthalmicus profundus, maxillomandibular, and facial nerves later developed a highly modified branching pattern, whereas postotic nerves were less specialized and showed the stereotypical branching pattern of post-trematic nerves. The early distribution of melanocytes in myotome-free space largely overlapped with the morphology of the cranial nerve and ganglion anlage, and resembled the cephalic crest cell distribution pattern in the early chick embryo. It was suggested that the cephalic crest cell distribution, which is also inhibited by myotomes in the lamprey, would be the common basis for branchiomeric nerve patterning. In later development of the lamprey embryo, myotomes 1 through 3, which had originated in the postotic region, grew rostrally into the preotic region, laterally covering all of the branchiomeric nerves. This results in a deep position of the cranial nerves, which is not observed in gnathostomes.

Peripheral development of cranial nerves in a cyclostome, Lampetra japonica: morphological distribution of nerve branches and the vertebrate body plan

The development of peripheral nerves was studied in a Japanese marine lamprey, Lampetra japonica, in whole-mount and sectioned embryos from hatching until the earliest ammocoete. Nerve fibers were immunohistochemically stained with a monoclonal antibody against acetylated tubulin. Branchiomeric nerves first developed in a simple metamerical pattern, each associated with a single pharyngeal arch. Of those, the ophthalmicus profundus, maxillomandibular, and facial nerves later developed a highly modified branching pattern, whereas postotic nerves were less specialized and showed the stereotypical branching pattern of post-trematic nerves. The early distribution of melanocytes in myotome-free space largely overlapped with the morphology of the cranial nerve and ganglion anlage, and resembled the cephalic crest cell distribution pattern in the early chick embryo. It was suggested that the cephalic crest cell distribution, which is also inhibited by myotomes in the lamprey, would be the common basis for branchiomeric nerve patterning. In later development of the lamprey embryo, myotomes 1 through 3, which had originated in the postotic region, grew rostrally into the preotic region, laterally covering all of the branchiomeric nerves. This results in a deep position of the cranial nerves, which is not observed in gnathostomes. J. Comp. Neurol. 384:483–500, 1997. © 1997 Wiley-Liss, Inc.

Demonstration of rod and cone photoreceptors in the lamprey retina by freeze-replication and immunofluorescence.

In common with other cyclostomata, the Japanese river lamprey (Lampetra japonica) has a retina consisting of distinct types of photoreceptor cells called long and short photoreceptor cells. After freeze-fracture, disc membranes of these photoreceptor cells were characterized in common by a homogeneous distribution of intramembrane particles on the protoplasmic fracture faces, in contrast to those of the myeloid bodies bearing scattering particles. Immunofluorescent examination was applied to the retina with monoclonal antibodies raised against bovine and chicken rhodopsins. Positive immunoreactivity was found to be limited to outer segments of the short cell, leaving the entire body of the long cell and all other components of the retina negative. The results suggest that the short cell is more closely related to a rod-type photoreceptor cell characterized by rhodopsin as its visual pigment.