Immunohistochemical investigation of the pituitary of the sturgeon (Acipenser baeri,Chondrostei)

An immunohistochemical study of the sturgeon (Acipenser baeri) pituitary was undertaken using antisera directed against hormones from various classes of vertebrates, including the only pituitary hormone available from sturgeon, gonadotrophin. A positive reaction was obtained after application of antisera towards the following hormones 1–24 synthetic ACTH (1-24 ACTH), melanophore stimulating hormone (MSH), ovine prolactin (oPRL), ovine growth hormone (oGH), salmon growth hormone (sGH), carp gonadotrophin (cGTH) and its beta subunit (cGTH), sturgeon gonadotrophin (aciGTH), carp thyrotrophin (cTSH) and subunit of the human thyrotrophin (hTSH). The results demonstrate that, in general, the sturgeon pituitary resembles that of teleosts as regards the distribution of the different cell types: ACTH and PRL cells in the rostral pars distalis, GTH, TSH and GH cells in the proximal pars distalis and MSH and PAS-cells in pars intermedia. In addition to the topographical organization of the sturgeon pituitary, this study provides data on the immunological relationships between sturgeon pituitary hormones and those of other vertebrates.     

Glucagon-like peptide-1 (GLP-1) immunoreactivity in the forebrain and pituitary of the teleost, Clarias batrachus (Linn.)

Glucagon-like peptide-1-immunoreactivity (GLP-ir) was seen in several olfactory receptor neurons in the olfactory epithelium and in the olfactory nerve fascicles that extend caudally to the bulb and innervate the glomerular layer. Thick GLP-ir fascicles were seen in the medial olfactory tracts (MOT); these probably represent the extrabulbar olfactory projections. GLP-ir fibers in the MOT penetrate into the telencephalon, swing dorsocaudally into the area ventralis telencephali/pars supracommissuralis (Vs) just above the anterior commissure. The immunoreactive fascicles continue caudally in the dorsal hypothalamus and are traceable as far as the lateral recesses. Scattered GLP-ir fibers were seen in the ventral telencephalon and preoptic area. While several immunoreactive cells were seen in the pars intermedia of the pituitary gland, few were encountered in the rostral pars distalis and proximal pars distalis. Role for the peptide in the processing of olfactory information and in the regulation of pituitary gland may be suggested.     

石鲽脑垂体组织生理学研究

石鲽(Kareius bicoloratus)脑垂体呈鸡心形,由神经垂体和腺垂体两部分组成。腺垂体又包括前外侧部(RPD)、中外侧部(PPD)和垂体中间部(PI)。神经纤维束延伸至腺垂体的各个部分。采用HE、Jafri、HA、Mallory和PAS染色法,结合免疫组织化学方法,对石鲽脑垂体的6种内分泌细胞进行鉴别和定位。RPD部位主要含有PRL和ACTH细胞,PI部位主要含有MSH细胞。PPD部位有4类细胞,分别为GH、Gt H、TSH和嫌色细胞。GH、Gt H和TSH细胞数量随鱼体生长发育而变化,Gt H在性腺成熟期达到高峰,GH细胞和TSH细胞在生长早期数量多。     

Close association of gonadotropin-releasing hormone fibers and gonadotropin, growth hormone, somatolactin and prolactin expressing cells in pejerrey, Odontesthes bonariensis

The purpose of this study was to determine if there is any association between immunoreactive (ir) gonadotropin-releasing hormone (GnRH) fibers with different pituitary endocrine cell types in the pejerrey, Odontesthes bonariensis. Using a monoclonal antibody raised against mammalian GnRH (mGnRH) (LRH13), ir-GnRH fibers were observed passing through the pituitary stalk and reaching the three areas of the pituitary gland: rostral (RPD) and proximal pars distalis (PPD) and pars intermedia (PI). Double labeled immunocytochemistry showed ir-GnRH fibers in close association with prolactin (PRL)-producing cells in the RPD, growth hormone (GH)-producing cells in the PPD, gonadotropin (GtH)-producing cells in the PPD and the external border of the PI, and with somatolactin (SL)-producing cells in the PI. Our results show, direct morphological evidences of a close association of GnRH fibers with GH, PRL, GtH and SL-expressing cells. These results would suggest that GnRH has a broad role in the regulation of the secretion of different pituitary hormones.     

Hypophyseal adenocarcinoma in a horse. A case study

The paper presents the case-history of a 5-year-old pony suffering from an adenocarcinoma which proceeded from the pars intermedia of the pituitary gland. Neurological and patho-morphological findings are reported.     

Immunocytochemical and ultrastructural identification of adenohypophyseal cells in Ctenopharyngodon idella (Cypriniformes: Cyprinidae) during gonadal differentiation

The adenohypophysis was studied by immunocytochemical and ultrastructural methods in juvenile grass carp (Ctenopharyngodon idella) from natural reproduction in Northern Italian rivers. The adenohypophysis included the rostral pars distalis (RPD), the proximal pars distalis (PPD) and the pars intermedia (PI), all deeply penetrated by branches of the neurohypophysis (Nh). The prolactin (PRL), adrenocorticotropic (ACTH), somatotropic (GH), thyrotropic (TSH), gonadotropic type I (GtH I) and type II (GtH II), somatolactin (SL), melanotropic (MSH) and endorphin (END) cells were identified with antisera raised against piscine and human pituitary hormones. In juveniles of 51–69 mm of total body length (TL) with undifferentiated gonads, the PRL cells, arranged in thick strands, occupied most of the RPD. The ACTH and GH cells organized in cords bordering Nh were, respectively, confined to RPD and PPD. The TSH cells were scattered among ACTH cells in RPD and among GH cells in PPD. Cells simultaneously immunoreactive to anti-follicle stimulating hormone and to anti-croaker gonadotropin were intermingled among GH and TSH cells, which were mostly in the dorsal PPD. The SL cells were detected in PI layers bordering the Nh. The MSH and END cells were intermingled in PI and, unlike what observed in other teleosts, their respective antisera did not cross-react. In individuals of 78–112 mm TL with gonads at the beginning of differentiation, the GtH II cells were detected in PPD; all other cell types increased in number. These results, supported by ultrastructural investigations, suggest that SL and GtH II cells are directly involved in gonadal differentiation in C. idella.     

Food-deprivation-induced suppression of pituitary–testicular-axis in the tilapia Oreochromis mossambicus

In the present study, the tilapia Oreochromis mossambicus were exposed to food deprivation for a period of 6 or 12 days and changes in the luteinizing hormone (LH) immunoreactivity in the proximal pars distalis (PPD) of the pituitary gland and the testicular activity were examined. Intensely immunoreactive LH content was noticed in the PPD of the pituitary gland in the initial controls, controls on days 6 and 12, and fasting fish on day 6, whereas the LH immunoreactivity was moderate or weak in fasting fish on day 12. In addition, although the mean gonadosomatic and hepatosomatic indices among different experimental groups did not show any statistically significant difference, the mean numbers of spermatogonia, spermatocytes, and spermatids were significantly lower in food-deprived fish on days 6 or 12 compared to those of controls. The inhibition of the spermatogenesis was accompanied by the presence of abundant spermatozoa in the lumen of seminiferous tubules of the testis in food-deprived fish, whereas the occurrence of spermatozoa was relatively infrequent in initial controls and controls. Furthermore, refeeding to food-deprived fish on day 6 onwards resulted in occurrence of few intensely stained LH secreting cells and significantly higher numbers of spermatocytes and spermatids concomitant with sparse spermatozoa in majority of tubules compared to those of food-deprived fish. These results suggest that prolonged exposure to food-deprivation causes suppression of the LH secretory activity in the pituitary gland and disruption in the spermatogenesis in O. mossambicus.

     

Development of a homologous radioimmunoassay for Mediterranean yellowtail (Seriola dumerilii, Risso 1810) LH

Purified Mediterranean (M.) yellowtail luteinizing hormone-like gonadotropin (MyLH) and its β-subunit (MyLHβ) served to develop a radioimmunoassay (RIA) for MyLH. The rabbit antisera against MyLH and MyLHβ used for this purpose were tested on pituitary sections by immunocytochemistry. Anti-MyLHβ specifically detected a single type of cells, which were located at the periphery of the proximal pars distalis (PPD) and surrounding the pars intermedia (PI). Anti-MyLH, however, also recognized two other cell types, thyrotropin β-immunoreactive (ir) cells and putative follicle-stimulating hormone-like (MyFSH)-producing cells. Labeling of the two latter cell types was prevented by preabsorption of anti-MyLH with M. yellowtail pituitary glycoprotein -subunit. The standard curve for the RIA was generated using purified MyLH, ¹²⁵I-labeled MyLHβ and anti-MyLHβ at a dilution of 1:70,000, which resulted in the binding of 30% of the tracer added. The standard curve ranged from 0.25 to 50 ng/ml. The midrange of the assay (ED50) was obtained with 5.48–7.87 ng LH/ml. The variation between assays was less than 15%. An average cross-reactivity of FSH in the LH RIA of 8.4% was found. Serial dilutions of M. yellowtail pituitary extracts displaced radiolabelled MyLHβ parallel to the MyLH standard. Application of the LH RIA to blood samples and pituitary cell culture medium provided physiological validation of the assay. Significant increases in LH levels were recorded after salmon GnRH treatments in vivo and in vitro. Serum LH levels from wild fish sampled at the spawning season were significantly higher than those from captive fish sampled in the same period.     

Gonadotropin-releasing hormones in the African catfish: molecular forms, localization, potency and receptors

Two gonadotropin releasing hormones (GnRHs) were identified in the African catfish: chicken GnRH-II (cGnRH-II) and catfish GnRH (cfGnRH). Immunological screening of HPLC fractions from pituitary extracts indicated a third GnRH which co-eluted with lamprey GnRH-III. However, mass determination and amino acid sequencing identified this material as isotocin. This underlines the risk of identifying multiple forms of GnRH in tissue extracts on the basis of immunoreactivity in HPLC fractions. In vivo and in vitro studies demonstrated that cGnRH-II is an over 100-fold more potent gonadotropin (GTH) secretagogue than cfGnRH. This correlates with the respective receptor affinities. The presence of both GnRHs in the pituitary gland suggests that they may modulate each other's GTH release activity. Sub-threshold or low doses of cGnRH-II partly inhibited cfGnRH-induced GTH II secretion. Conversely, combinations of sub-threshold or low doses of cfGnRH with effective doses of cGnRH-II led to increases in GTH II levels similar to those induced by cGnRH-II alone. Combinations of submaximally effective dose of the 2 peptides resulted in additive effects. Hence, both GnRHs participate in the regulation of GTH II release, and their relative concentrations may determine the overall effect. Immunocytochemistry, using anti-bodies against the respective recombinant GnRH associated peptides (GAPs), as well as in situ hybridization showed that cfGnRH neurones are scattered in the ventral forebrain and project into the pituitary gland, while cGnRH-II neurones are confined to the midbrain tegmentum and without projections to the pituitary gland. Transfection experiments with GnRH receptor cDNA shows ligand activation characteristics similar to those of the native GnRH-R. Autoradiographic studies and hormone release studies indicate that GnRH-Rs in the African catfish pituitary gland are restricted to the gonadotrophs.     

Naltrexone attenuates stress-induced suppression of LH secretion in the pituitary gland in the Cichlid fish Oreochromis mossambicus: evidence for the opioidergic mediation of reproductive stress response

Opioid peptide β-endorphin (β-EP) plays a modulatory role in vertebrate reproduction. However, the role of opioid peptides in reproductive stress response is least understood in fishes. The aim of the present study was to determine the effect of different doses of β-EP on luteinizing hormone (LH) secretion in normal and the opioid receptor antagonist naltrexone (NALT) in stressed female tilapia Oreochromis mossambicus. Administration of 4 μg β-EP, but not 0.5 or 1.5 μg β-EP, daily for 22 days caused suppression of LH-secreting cells at the proximal pars distalis of the pituitary gland, concomitant with a significant reduction in the mean GSI and HSI in 4 μg β-EP-treated fish compared to controls. On the other hand, exposure of the fish to mild acute stressors for 22 days caused changes in the LH-secreting cells similar to that of high dose of β-EP, whereas administration of NALT attenuated these effects. Taken together, the results indicate that increased concentration of β-EP as may occur during stressful conditions can cause suppression of LH secretion, leading to the inhibition of spawning, and that treatment of NALT attenuates the stress-induced inhibition of LH secretion in fish.     

Immunocytochemical identification of adenohypophyseal cells in the pirarucu (<Emphasis Type="Italic">Arapaima gigas</Emphasis>), an Amazonian basal teleost

The adenohypophysis (AH) of juvenile pirarucu (Arapaima gigas), a representative species of the Osteoglossomorpha (bonytongue fishes, one of the oldest living groups of the teleosts), was studied using histochemical and immunocytochemical methods. The AH is comprised of the pars distalis (PD), without a clear distinction between rostral pars distalis (RPD) and proximal pars distalis (PPD), and the pars intermedia (PI). The neurohypophysis (NH) is positioned on top of the PD and penetrates and branches into the PI. In the most rostral dorsal portion of the PD, adrenocorticotropic cells and fusiform gonadotropic cells were found. In the central PD, scarce prolactin-producing cells and growth-hormone-producing cells were located mainly in the dorsal part, whereas round gonadotropic cells were abundant in the ventral portion of this region. Human thyrotropin immunoreactive cells were not found in the entire AH. In the PI, melanotropic, some adrenocorticotropic, and somatolactin-producing cells were located intermingled surrounding the neurohypophyseal branches. Our results showed that the A. gigas pituitary has some basal characteristics between the ancient Actinopterygii and the more derived teleosts.     

Cytology of the cerebrospinal fluid in dogs with brain tumors and spinal cord compression. Part 4]

The results of cerebrospinal fluid (CSF) cytology of 9 dogs with brain tumors and 50 dogs with spinal cord compression are discussed. Of the 50 dogs with spinal cord compression, disc protrusion was diagnosed in 31, myelomalacia in 7, discospondylitis in 3 and spinal cord tumors in 9 dogs. In 4 of 9 dogs with brain tumors, tumor cells could be found by the sedimentation apparatus of K?lmel. Pleocytosis existed in 6 patients. In about 70% (29 of 41) of cases with disc protrusion, more than 200 cells could be evaluated in the CSF sediment, consisting mostly of transformed lymphocytes and activated monocytes. As the neurologic deficits increased, the amount of cells and especially cell complexes increased. This was especially evident in cases with myelomalacia of the spinal cord. Only in cases with discospondylitis or spinal cord neoplasia was the CSF cytology unchanged.     

长吻鮠不同时期脑垂体的超微结构

采用组织学方法和透射电镜技术对性周期内不同阶段和不同季节的雌性长吻鮠(Leiocassis longirostris)脑垂体的超微结构进行较详细的研究，结果表明：长吻鮠脑垂体包括腺垂体和神经垂体两个部分，腺垂体组织中有6种分泌细胞，即促性腺激素(GTH)分泌细胞、生长激素(STH)分泌细胞、促甲状腺激素(TSH)分泌细胞、促黑色素细胞刺激激素(MSH)分泌细胞、催乳激素(PRL)分泌细胞、促肾上腺皮质激素(ACTH)分泌细胞。神经垂体组织中存在A型(A1、T2)和B型神经分泌纤维，A型分泌纤维轴突中具有不同类型的分泌颗粒，B型分泌纤维轴突中含有许多透明小囊泡。产卵前GTH细胞胞质中充满了电子密度较高的大、小两种分泌颗粒；TSH细胞胞质中分泌颗粒排空，电子密度降低，出现大空泡；STH细胞处于活跃的合成和分泌状态；MSH细胞粗面内质网相当发达，成层围核分布，分泌颗粒极少；PRL细胞电子密度高，具圆球形被膜的分泌颗粒；ACTH细胞分泌物颗粒数量少。产卵后GTH细胞中的分泌颗粒排空；TSH细胞出现空泡；STH细胞中的粗面内质网围核环状排列；MSH细胞分泌颗粒多但比较小，粗面内质网呈扁状或网状；PRL细胞分泌颗粒极少，分布不均匀；ACTH细胞内质网近核分布。秋季GTH细胞中发现GTH1和GTH2两种细胞，GTH1细胞具有大量围核分布的分泌颗粒，GTH2细胞胞质中分泌颗粒数量较少；STH细胞胞质中粗面内质网极发达，围核分布，线粒体数量增加；TSH细胞内质网膨大成泡状；MSH细胞有明显的双核结构，其中一核具核仁，胞质内分泌颗粒较产后增多；PRL细胞具有呈不均匀分布的圆球形分泌颗粒；ACTH细胞线粒体与内质网围核分布，分泌颗粒较少，不均匀分布于细胞质中。冬季GTH细胞内有少量分泌颗粒散布在核周围；TSH细胞含有较多的小型分泌颗粒，胞核外周有大量长条形的粗面内质网；STH细胞中分泌颗粒数量较少，合成和分泌活动微弱；MSH细胞粗面内质网呈泡化状，绕核排列；PRL细胞、ACTH细胞和秋季时的超微结构无明显差异。神经垂体中的分泌纤维各阶段也有一定的变化，产前阶段具有较多球形的无被膜分泌颗粒，产后阶段分泌颗粒呈现排空，秋季部分可见到分泌颗粒，但数量相对较少，冬季变化不明显。     

Presence of infectious salmon anaemia virus (ISAV) in tissues of Atlantic salmon, Salmo salar L., collected during three separate outbreaks of the disease

Abstract. Infectious salmon anaemia (ISA) is a viral disease of farmed Atlantic salmon, Salmo salar L., in Norway. The enveloped virus particles (100nm) believed to be the causative agent of the disease have been observed budding from endothelial cells in heart blood vessels. However, it is not known if the virus propagates in endothelial cells in all tissues/organs, if other target cells exist or if material collected from different salmon farms with natural outbreaks of ISA contain the same virus particles. Salmon smolts from three hatcheries with no history of disease were taken into the laboratory and experimentally challenged with ISA collected from Atlantic salmon during natural outbreaks of the disease in three different fish farms outside Bergen. Norway. Tissues for TEM studies were Collected from: (1) organs that showed clinical signs of ISA (i.e. used in the diagnosis of the disease); (2) tissues believed to be important in transmission of the virus (integument, kidney, urinary bladder, gut and somatic muscle); and (3) hormone-producing tissues (pituitary gland, saccus vasculosus, thymus, thyroid, ultimobranehial gland, gonad, head kidney, heart and ventral aorta). The same virus as that believed to be the causative agent of ISA was found in all tissues examined from the challenged fish, i.e. a multiorgan infection with the same virus present in salmon from all three fish farms. The virus particles are about 100 nm in diameter, consisting of a slightly pleomorphic unit membrane envelope within which are a number of granules about 10–12nm in diameter. The granules seemed to be arranged in two concentric circles (spheres). The virus was seen budding from the surface of endothelial cells in blood vessels/sinus only. However, the virus was found intracellularly in both endothelial cells and in leucocytes.     

Neuronal nitric oxide synthase immunoreactivity in forebrain,pineal, and pituitary of <Emphasis Type="Italic">Oreochromis mossambicus</Emphasis> (Tilapia)

Ultrastructural localization of neuronal nitric oxide synthase (nNOS) in olfactory receptor neurons (ORNs) and immunohistochemical detection of the enzyme in forebrain, pituitary, and pineal were undertaken in the teleost Oreochromis mossambicus. Application of post-embedding immunoelectron microscopy revealed nNOS-labeled gold particles on the cilia, microvilli, mitochondria, and Golgi complex of the ORNs. Gold particles were also seen adhered to microtubules in the axons that extend to the olfactory nerve layer in the olfactory bulb. With light microscopy, nNOS-immunoreactive neurons were seen in preoptic area, nucleus entopeduncularis, and parvocellular, and magnocellular subdivisions of nucleus preopticus (NPO). Numerous cerebrospinal fluid-contacting cells lining the wall of the third ventricle at the level of the NPO showed intense immunoreactivity. Intense to moderate immunoreactivity was observed in the neurons of suprachiasmatic nucleus, nucleus lateralis tuberis pars lateralis, and nucleus recessus lateralis. While several immunoreactive fibers were detected in medial olfactory tract, suprachiasmatic area, and hypothalamo-hypophyseal tract, a few were seen throughout the telencephalon, in the optic chiasma, tuberal area, and inferior lobes. In the pituitary, nNOS-containing fibers were seen in the neurohypophysis, rostral pars distalis, proximal pars distalis, and pars intermedia. While intense immunoreactivity was noticed in some cells in the pineal, immunoreactive fibers were detected in the pineal stalk as well as parenchyma. We suggest that nitric oxide may play a role in processing olfactory and photic information, circadian rhythms, and neuroendocrine regulation in tilapia.     

Immunohistochemical localization of three GnRH systems in brain and pituitary of Japanese flounder

ABSTRACT:   To clarify the possible roles of gonadotropin-releasing hormone (GnRH) in the reproduction of Japanese flounder Paralichthys olivaceus , localization of salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II), and sea bream GnRH (sbGnRH) immunoreactive (ir) cell bodies and fibers in the brain and pituitary were examined together with follicle stimulating hormone (FSH) and luteinizing hormone (LH)-ir cells in the pituitary by immunohistochemistry. sGnRH-ir cell bodies were localized in the ventromedial part of the rostral olfactory bulb and cGnRH-II-ir cell bodies were restricted to the midbrain tegmentum, while sbGnRH-ir cell bodies were evident in the preoptic area. sGnRH-ir fibers were distributed throughout the brain, especially abundant in the forebrain. cGnRH-II-ir fibers were also scattered in many areas of the brain with abundance in the midbrain, but sbGnRH-ir fibers were observed in the preoptic–hypothalamic area and innervated the pituitary. In the pituitary, neither sGnRH-ir fibers nor cGnRH-II-ir fibers were found, but sbGnRH-ir fibers were profuse in the neurohypophysis and invaded the proximal pars distalis, targeting FSH and LH cells. These results suggest that three GnRH systems can play different physiological roles in the brain of Japanese flounder. Among them, sbGnRH is considered to be involved in reproduction by stimulating gonadotropin secretion, while sGnRH and cGnRH-II can function as a neurotransmitter and/or neuromodulator within the brain in this species.     

Role of neuropeptide Y (NPY) in the regulation of reproduction: study based on catfish model

Significance of NPY in the regulation of GnRH–LH axis was evaluated. Considerable NPY immunoreactivity was seen in the components like olfactory system, basal telencephalon, preoptic and tuberal areas, and the pituitary gland that serve as neuroanatomical substrates for processing reproductive information. Close anatomical association as well as colocalizations of NPY and GnRH were seen in the olfactory receptor neurons, olfactory nerve fibers and their terminals in the glomeruli, ganglion cells of nervus terminalis, medial olfactory tracts, fibers in the ventral telencephalon and pituitary. In the pituitary, NPY fibers seem to innervate the GnRH as well as LH cells. Intracranial administration of NPY resulted in significant increase in the GnRH immunoreactivity in all the components of the olfactory system. In the pituitary, NPY augmented the population of GnRH fibers and LH cells. HPLC analysis showed that salmon GnRH content in the olfactory organ, bulb, preoptic area+telencephalon and pituitary was also significantly increased following NPY treatment. NPY may play a role in positive regulation of GnRH throughout the neuraxis and also up-regulate the LH cells in the pituitary.     

Neuropeptides regulating the activity of goldfish corticotropes and melanotropes

The goldfish (Carassius auratus) has proven an advantageous model for investigations of the neuroendocrine regulation of pituitary hormone secretion in teleost fishes. Investigations examining the secretion of adrenocorticotropin (ACTH) and melanocyte-stimulating hormone (MSH) from pituitary cellsin vitro have been used to identify neuropeptides influencing goldfish corticotrope and melanotrope activity. Ovine CRF, urotensin I (UI), arginine vasotocin (AVT), isotocin and angiotensins I and II stimulate the release of ACTH from corticotropesin vitro. Thyrotropin-releasing hormone (TRH), oCRF, UI and neuropeptide Y stimulate the release of MSH from melanotropesin vitro. Immunocytochemical studies have revealed the presence of separate CRF- and UI-immunoreactive perikarya in the hypothalamus suggesting the existence of two structurally similar, yet distinct, hypothalamic CRF-UI-like peptides. Interactions of AVT and CRF in the regulation of ACTH secretion is suggested from studies demonstrating the co-localization of AVT- and CRF-immunoreactivities in perikarya of the preoptic-hypophyseal system. These investigations demonstrate that the secretory activity of goldfish corticotropes and melanotropes is influenced by a diversity of neuropeptides of hypothalamic origin.     

Immunohistochemical detection of gonadotropin-like material in the hagfish pituitary, correlated with their gonadal conditions

In the present study, immunoreactive gonadotropin (GTH)-like material was examined in the hagfish pituitary, in correlation with their gonadal conditions. For this purpose, three species of hagfish, Myxine glutinosa, Eptatretus burgeri and Paramyxine atami, were used. In all species, GTH-like cells were observed in the adenohypophysis. They were stained with several antibodies to LH-related GTHs, such as ovine LHβ, human LHβ, bullfrog LH, and salmon GTHIIβ. In the subsequent study, the relationship between the amount of GTH-like material and gonadal conditions was examined in Paramyxine atami. The adenohypophysis of adult animals exhibited well-developing features, and about half of the adenohypophysial cells were stained intensely with anti-ovine LHβ. On the other hand, the adenohypophysis of juveniles exhibited less developed features with few or no cells stained with anti-ovine LHβ. These results suggest that a GTH-gonadal system is truly present in the hagfish.