Comprehensive histological and immunological studies reveal a novel glycoprotein hormone and thyrostimulin expressing proto-glycotrope in the sea lamprey pituitary.

In the adenohypophysis (anterior pituitary) of all gnathostomes, there are six tropic cell types: corticotropes, melanotropes, somatotropes, lactotropes, gonadotropes and thyrotropes; each cell type produces specific tropic hormones. In contrast, we report in this study that there are only four tropic cell types in the sea lamprey (Petromyzon marinus) adenohypophysis. We specifically focused on the cell types that produce the glycoprotein hormones (GpHs).

Emergence of an Ancestral Glycoprotein Hormone in the Pituitary of the Sea Lamprey, a Basal Vertebrate.

The gnathostome (jawed vertebrates) classical pituitary glycoprotein hormones, FSH, LH, and TSH, consist of a common α-subunit (GpA1) and unique β-subunits (Gpβ1, -2, and -3), whereas a recently identified pituitary glycoprotein hormone, thyrostimulin, consists of GpA2 and GpB5. This paper reports the identification, expression, and function of an ancestral, nonclassical, pituitary heterodimeric glycoprotein hormone (GpH) consisting of the thyrostimulin A2 subunit with the classical β-subunit in the sea lamprey, Petromyzon marinus, a jawless basal vertebrate. Lamprey (l) GpA2, and lGpHβ were shown to form a heterodimer by coimmunoprecipitation of lGpA2 with FLAG-tagged lGpHβ after the overexpression in transiently transfected COS7 cells using a bipromoter vector.

Molecular cloning of cytochrome P450 side-chain cleavage and changes in its mRNA expression during gonadal development of brown hagfish, Paramyxine atami.

Since hagfishes are considered the most primitive vertebrate known, extant or extinct, studies on their reproduction are indispensable for understanding phylogenetic aspects of vertebrate reproduction. However, little information is available on the endocrine regulation of the gonadal function in the hagfish. Based on EST analysis of the testis of the brown hagfish (Paramyxine atami), P450 side chain cleavage (CYP11A), which is the first and essential enzyme for steroidogenesis in jawed vertebrates, was cloned.

Hypothalamic-pituitary-gonadal endocrine system in the hagfish.

The hypothalamic-pituitary system is considered to be a seminal event that emerged prior to or during the differentiation of the ancestral agnathans (jawless vertebrates). Hagfishes as one of the only two extant members of the class of agnathans are considered the most primitive vertebrates known, living or extinct. Accordingly, studies on their reproduction are important for understanding the evolution and phylogenetic aspects of the vertebrate reproductive endocrine system.

Relationships between plasma concentrations of sex steroid hormones and gonadal development in the brown hagfish, Paramyxine atami.

The relationship between sex steroid hormone profiles in plasma and gonadal function in hagfish is poorly understood. In the present study, plasma concentrations of estradiol, testosterone, and progesterone were examined with respect to gonadal development, sexual differences, and possible function of atretic follicles in the brown hagfish, Paramyxine atami, using a time-resolved fluoroimmunoassay. Plasma concentrations of these three hormones were low in juveniles of both sexes.

Effects of estradiol or testosterone treatment on expression of gonadotropin subunit mRNAs and proteins in the pituitary of juvenile brown hagfish, Paramyxine atami.

A single functional gonadotropin (GTH) comprising two subunits, α and β, was recently identified in the pituitary of brown hagfish (Paramyxine atami). Little is known about the feedback mechanisms that regulate these GTH subunits by sex steroids in the hagfish. The present study was designed to examine feedback effects of estradiol and testosterone on mRNA expression and protein expression of both GTHα and GTHβ subunits in the pituitary of the juvenile P.

Glycoprotein hormone in the pituitary of hagfish and its evolutionary implications.

The pituitary gland is present in all vertebrates, from agnathans (jawless vertebrates) to mammals, but not in invertebrates. Reproduction in gnathostomes (jawed vertebrates) is controlled by two pituitary gonadotropins (GTHs), luteinizing hormone and follicle-stimulating hormone, which are part of the pituitary glycoprotein hormone (GPH) family. Hagfishes, which lack both jaws and vertebrae, are considered the most primitive vertebrate known, living or extinct.

Evolutionary origin of the structure and function of gonadotropin-inhibitory hormone: insights from lampreys.

Gonadotropin (GTH)-inhibitory hormone (GnIH) is a novel hypothalamic neuropeptide that inhibits GTH secretion in mammals and birds by acting on gonadotropes and GnRH neurons within the hypothalamic-pituitary-gonadal axis. GnIH and its orthologs that have an LPXRFamide (X = L or Q) motif at the C terminus (LPXRFamide peptides) have been identified in representative species of gnathostomes. However, the identity of an LPXRFamide peptide had yet to be identified in agnathans, the most ancient lineage of vertebrates, leaving open the question of the evolutionary origin of GnIH and its ancestral function(s).

Characterization of novel RFamide peptides in the central nervous system of the brown hagfish: isolation, localization, and functional analysis.

RFamide (RFa) peptides play various important roles in the central nervous system in both invertebrates and vertebrates. However, there is no evidence of the existence of any RFamide peptide in the brain of hagfish, one of the oldest lineages of vertebrates. In this study, we sought to identify novel RFamide peptides from the brains of hagfish (Paramyxine atami).

Evolutionary origin of a functional gonadotropin in the pituitary of the most primitive vertebrate, hagfish.

Hagfish, which lack both jaws and vertebrae, are considered the most primitive vertebrate known, living or extinct. Hagfish have long been the enigma of vertebrate evolution not only because of their evolutionary position, but also because of our lack of knowledge on fundamental processes. Key elements of the reproductive endocrine system in hagfish have yet to be elucidated.

The hagfish pituitary gland and its putative adenohypophysial hormones.

The pituitary gland is present in all vertebrates, from agnathans (jawless fishes) to mammals, but not in invertebrates. Hagfishes, which lack both jaws and vertebrae, are considered the most primitive vertebrate known, living or extinct. Accordingly, studies on hagfishes are indispensable for understanding the origin and evolution of the pituitary hormones.

Gene structure and functional characterization of growth hormone in dogfish, Squalus acanthias.

Dogfish (Squalus acanthias) growth hormone (GH) was identified by cDNA cloning and protein purification from the pituitary gland. Dogfish GH cDNA encoded a prehormone of 210 amino acids (aa). Sequence analysis of purified GH revealed that the prehormone is composed of a signal peptide of 27 aa and a mature protein of 183 aa.

Origins of gonadotropin-releasing hormone (GnRH) in vertebrates: identification of a novel GnRH in a basal vertebrate, the sea lamprey.

We cloned a cDNA encoding a novel (GnRH), named lamprey GnRH-II, from the sea lamprey, a basal vertebrate. The deduced amino acid sequence of the newly identified lamprey GnRH-II is QHWSHGWFPG. The architecture of the precursor is similar to that reported for other GnRH precursors consisting of a signal peptide, decapeptide, a downstream processing site, and a GnRH-associated peptide; however, the gene for lamprey GnRH-II does not have introns in comparison with the gene organization for all other vertebrate GnRHs.

Identity and distribution of immunoreactive adenohypophysial cells in the pituitary during the life cycle of sea lampreys, Petromyzon marinus.

Adrenocorticotropin (ACTH), melanotropins (MSHs), growth hormone (GH) and gonadotropin (GTH) have been identified or cloned from the pituitary gland of sea lampreys (Petromyzon marinus). The present study was designed to gain insights into the functional significance of these hormones through a description of changes in the occurrence and distribution of cells immunoreactive to their antibodies at several different stages of the sea lamprey life cycle. ACTH-like cells and MSH-like cells were distributed in the rostral pars distalis and the pars intermedia, respectively, throughout the life cycle from ammocoetes (larvae) to pre-spawning adults.

Gonadotropin-like and adrenocorticotropin-like cells in the pituitary gland of hagfish, Paramyxine atami; immunohistochemistry in combination with lectin histochemistry.

The pituitary system of the hagfish remains an enigma. The present study has aimed to detect possible adenohypophysial hormones in the pituitary gland of the brown hagfish, Paramyxine atami, by means of immunohistochemistry in combination with lectin histochemistry. Rabbit antisera raised against ovine luteinizing hormone (LH)beta, proopiomelanocortin (POMC)-related peptides, and the growth hormone/prolactin family of tetrapod and fish species were used, and 25 kinds of lectins were tested.

Identification and cloning of a glycoprotein hormone receptor from sea lamprey, Petromyzon marinus.

A full-length transcript encoding a functional lamprey glycoprotein hormone receptor I (lGpH-R I, GenBank AY750688) was cloned from the testes of the sea lamprey, Petromyzon marinus, using the GpH-R protein fingerprint GLYCHORMONER from the PRINTS database. The present study is the first to identify a GpH-R transcript in an agnathan, which is one of the only two representatives of the oldest lineage of vertebrates. The 719-amino acid full-length cDNA encoding lGpH-R I is highly similar and is likely a homolog of the vertebrate GpH-Rs (including LH, FSH, and TSH receptors).

Identification of a natriuretic peptide (NP) in cyclostomes (lamprey and hagfish): CNP-4 is the ancestral gene of the NP family.

In bony fishes, natriuretic peptides (NPs) comprise a hormone family that is composed of seven subtypes; ANP, BNP, VNP that have an intramolecular ring and N- and C-terminal extensions, and four CNPs (CNP-1 to -4) that lack the C-terminal extension. To assess the ancestral molecule of the NP family, we determined the NP sequences in several species of two extant cyclosotome groups, lampreys and hagfishes. A cDNA encoding CNP was cloned from the heart and brain of three phylogenetically distant species of lampreys, Geotria australis, Lampetra japonica, and Petromyzon marinus.

Immunohistochemical detection of gonadotropin-like material in the pituitary of brown hagfish (Paramyxine atami) correlated with their gonadal functions and effect of estrogen treatment.

Since hagfish are members of the most primitive group of living vertebrates, studies on their reproduction are indispensable for understanding phylogenetic aspects of vertebrate reproductive system. Nevertheless, our knowledge of the reproductive physiology of the hagfish, especially of the pituitary-gonadal axis, is almost completely lacking. In the present study, the relationship between the amount of immunoreactive gonadotropin (GTH)-like material in the pituitary gland and gonadal conditions was examined in the brown hagfish, Paramyxine atami.

A comparative study on the anatomy and development of different shapes of domatia in Cinnamomum camphora (Lauraceae).

Background And Aims: Domatia are small organs usually found in the axils of major veins on the underside of leaves and, although they have received wide attention from ecologists, few detailed reports exist on their anatomy or development. This study is focused on the domatia of Cinnamomum camphora (Lauraceae) and is the first comparative study on the anatomy and development of the different shapes of domatia within a single plant.

Methods: Four types of domatia in C.

Identification of sea lamprey GTHbeta-like cDNA and its evolutionary implications.

We have identified the first and perhaps only gonadotropin beta-like protein by cDNA cloning in sea lamprey, a member of the oldest lineage of vertebrates, the agnathans. Two pituitary gonadotropins (GTHs: follicle-stimulating hormone (FSH) and luteinizing hormone (LH)) have been identified in representative species of all classes of vertebrates except the agnathans. The present study was undertaken to identify GTH in sea lamprey, Petromyzon marinus, to gain a further understanding of the origin and evolution of reproductive pituitary hormones and their respective genes in vertebrates.

Distribution of immunoreactivities for adenohypophysial hormones in the pituitary gland of the polypteriform fish, Polypterus endlicheri.

Polypteriform fish constitutes the most primitive living descendent of the ancient bony fish. In polypteriform fish, only proopiomelanocortin (POMC) has been identified so far in the adenohypophysis, which is surprising in view of their evolutionary importance. In the present study, distribution of immunoreactive adenohypophysial hormones was examined in juvenile individuals of Polypterus endlicheri.

Occurrence of two functionally distinct proopiomelanocortin genes in all modern lampreys.

The lampreys (family Petromyzontidae) are divided into three subfamilies, the Petromyzontinae in the Northern Hemisphere and the Geotriinae and Mordaciinae in the Southern Hemisphere. We previously found two proopiomelanocortin subtypes, proopiocortin (POC) and proopiomelanotropin (POM) in sea lamprey, Petromyzon marinus (Petromyzontinae). POC encoding adrenocorticotropic hormone (ACTH) and beta-endorphin (beta-END) is expressed in the pars distalis of the pituitary, while POM encoding melanophore-stimulating hormone (MSH)-A and B together with a different beta-END is expressed in the pars intermedia of the pituitary.

Distribution of immunoreactive adenohypophysial cell types in the pituitaries of the Atlantic and the Pacific hagfish, Myxine glutinosa and Eptatretus burgeri.

The hagfish is considered the most primitive vertebrate known, living or extinct. It remains an enigma whether adenohypophysial hormones similar to those of more advanced vertebrates are present in the hagfish pituitary gland or not. The present study aimed to detect immunoreactive adenohypophysial hormones in the hagfish pituitary gland, using antisera to tetrapod and fish adenohypophysial hormones as immunohistochemical probes.

Cloning and analysis of the lamprey GnRH-III cDNA from eight species of lamprey representing the three families of Petromyzoniformes.

The lamprey, which are divided into three families, including the Petromyzonidae, Geotriidae, and Mordaciidae, have been shown to regulate the reproductive axis through a functional hypothalamic-pituitary-gonadal axis. To date, two forms of gonadotropin-releasing hormone (GnRH) have been identified in the sea lamprey (Petromyzon marinus), lamprey GnRH-I (decapeptide and cDNA) and lamprey GnRH-III (decapeptide), both of which have been shown to be expressed in the preoptic-anterior hypothalamic region and both forms have been demonstrated to regulate reproductive function (i.e.