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Motilin (MLN) is a peptide hormone originally isolated from the mucosa of the porcine intestine. Its orthologs have been identified in various vertebrates. Although MLN regulates gastrointestinal motility in tetrapods from amphibians to mammals, recent studies indicate that MLN is not involved in the regulation of isolated intestinal motility in zebrafish, at least in vitro. To determine the unknown function of MLN in teleosts, we examined the expression of MLN and the MLN receptor (MLNR) at the cellular level in Japanese medaka (Oryzias latipes). Quantitative PCR revealed that mln mRNA was limitedly expressed in the gut, whereas mlnr mRNA was not detected in the gut but was expressed in the brain and kidney. By in situ hybridization and immunohistochemistry, mlnr mRNA was detected in the dopaminergic neurons of the area postrema in the brain and the noradrenaline-producing cells in the interrenal gland of the kidney. Furthermore, we observed efferent projections of mlnr-expressing dopaminergic neurons in the lobus vagi (XL) and nucleus motorius nervi vagi (NXm) of the medulla oblongata by establishing a transgenic medaka expressing the enhanced green fluorescence protein driven by the mlnr promoter. The expression of dopamine receptor mRNAs in the XL and cholinergic neurons in NXm was confirmed by in situ hybridization. These results indicate novel sites of MLN activity other than the gastrointestinal tract. MLN may exert central and peripheral actions through the regulation of catecholamine release in medaka. [ABSTRACT FROM AUTHOR]

The larval period in anurans is highly plastic. Even though larval period length and larval size have been the most studied characteristics showing intraspecific variability, other features such as morphology, behavior, food preferences and physiology have also been described as plastic. Metamorphic changes are triggered by thyroid and interrenal hormones, and environmental signals modulate the onset of metamorphosis via the hypothalamic–pituitary–interrenal axis. The South American bufonid toad Rhinella arenarum from nearby Buenos Aires City has a larval period of 30–120 days under the same environmental conditions. The goal of this study was to evaluate life‐history traits in tadpoles from different localities and potential morphological differences among tadpoles with different larval period length. For this purpose, we reared under natural conditions tadpoles from two different localities nearby Buenos Aires City and in three different years measured their size at the end of metamorphosis and recorded their larval period length. The external appearance, histo‐morphology and morphometry of the skin and kidney/interrenal complex were studied in metamorphs with short and long larval periods (SLP and LLP, respectively). We observed a larval period lasting 30–120 days in tadpoles from different localities or years. SLP metamorphs were smaller, darker, had less protruding eyes without eyelids, a thinner epidermis and fewer skin glands than LLP metamorphs. The secretion of skin glands from SLP tadpoles had acidic glycoconjugates. Kidney size was similar between LLP and SLP metamorphs, but the former had an adult‐like histomorphology. Interrenal gland in LLP early and late climax tadpoles had cells with a high nucleus/cell diameter ratio, a sign of highly active steroidogenic cells. Overall, these results show that intraspecific variability in size, skin and interrenal gland development is associated with differences in larval period length. Distinct skin gland number and secretion, along with distinct interrenal histology at late climax, could have physiological consequences in the post‐metamorphic life. [ABSTRACT FROM AUTHOR]

The datasets of this article present the experimental parameters resulting from the assessment of adrenal gland as a potential biomarker of endocrine disruption mediated by graphene oxide (GO), a nanocarbon, using Japanese medaka fish as the model. These data sets support the article “Histopathological evaluation of the interrenal gland (adrenal homolog) of Japanese medaka (Oryzias latipes) exposed to graphene oxide”. The experiments were conducted on reproductively active adult fish maintained as a breeding pair (one male and one female) in 500 mL balanced salt solution (BSS) either by immersion in GO (20 mg/L in BSS) continuously for 96 h with refreshing of media once in every 24 h or by a single intraperitoneal (IP) injection of GO (100 µg/g) to both male and female fish. The experimental fish were allowed breeding and assessed after 21 days post-treatment. Moreover, one day-post hatch (dph) Japanese medaka fries (orange-red variety) were exposed to different concentrations of GO (2.5–20 mg/L) by immersion in embryo-rearing medium (ERM) for 96 h (1–5 dph) with refreshing of media every 24h. Food was given to the adults, however, the larvae remained fasting during the GO-exposure (0–5 dph) period. Control adults and larvae were identically maintained either in BSS (adults) or ERM (larvae), with no GO. After treatment, both adults and the larvae were maintained in BSS with feeding in a GO-free environment. After 21 days post-treatment, adults, and after six weeks post-treatment, larvae, were anaesthetized in MS-222, and the trunk region was preserved in 4% paraformaldehyde in PBS (20 mM) containing 0.05% Tween 20. Evaluation of interrenal gland (IRG) in kidneys were made in 5 µm thick sections stained on haematoxylin-eosin (HE). The phenotypic sex of adults was assessed by secondary sex characters (fin features) and gonad (testis and ovary) histology; in larvae, phenotypic sex was determined by gonad histology and the genotypic sex by genotyping dmy gene. The location of IRG in the kidney were determined by immunohistochemical technique using rabbit polyclonal antityrosine hydroxylase antibody as primary antibody. The digital images of sections were captured using an Olympus CKX53 inverted microscope with DP22 camera and CellSens software. Using imagej software, a minimum of 3 images of kidney consisting IRG were assessed for cell (separated as dark and pale stained nucleus after HE staining) sorting (cells/ mm2) and also measured the nuclear area (µm2). Counting of IRG cells, lined between the cardinal vein and the interstitial cells in the kidneys, were limited to maximum three layers in a given area. Numerical data, presented as means ± SEM, were analysed by one-way ANOVA followed by post-hoc Tukey's multiple comparison test or unpaired parametric ‘t’ test including Welch's correction, if distributed normally; or by Kruskal-Wallis test followed by Mann-Whitney's test as post hoc test, if the data did not meet the criteria of using a parametric test. Statistically significant difference were considered for p ≤ 0.05. The collected data on IRG of Japanese medaka fish will be used for the assessment of GO as an EDC disposed in the environment.

Due to unique physicochemical properties and wide industrial and biomedical applications, graphene oxide (GO) is ubiquitous in the aquatic ecosystem. Using Japanese medaka (Oryzias latipes) fish as a model, we previously demonstrated minimal endocrine disrupting (ED) effects of GO on reproductive organs, and thyroids. Current study investigated the ED‐effects of GO on the interrenal gland (IRG) of medaka. Breeding pairs of adult male and female fish were exposed to 0 mg/L (control) or 20 mg/L GO by continuous immersion for 96 h, or to 0 or 100 μg/g GO by intraperitoneal administration. Also, 1 day post‐hatch (dph) larvae were exposed to different concentrations of GO (2.5–20 mg/L) for 96 h. IRG was evaluated by immunohistochemical techniques after 21 days depuration in adults and 6 weeks in larvae. IRG cells were counted and the nuclear area was measured in hematoxylin–eosin stained sections using ImageJ software. We found that IRG is distributed adjacent to the posterior cardinal vein and its branches within the head kidney. Columnar/oval shaped periodic acid‐Schiff negative, tyrosine hydroxylase positive cells are arranged either in a single, or in groups, sometimes encircling a sinusoid, or in a straight chord, laying adjacent to the endothelium of the cardinal vein, and having eosinophilic cytoplasm with round/oval basophilic nuclei. GO effect on nuclei and cell population in IRG was inconsistent; depending on exposure route, sex, and/or age of the fish. Also, because of its high adsorptive property and sharp edges, GO probably agglomerated on IRG, and induced physical injury, and ED effects. [ABSTRACT FROM AUTHOR]

The present study examined the effects of maternal social status, with subordinate status being a chronic stressor, on development and activity of the stress axis in zebrafish embryos and larvae. Female zebrafish were confined in pairs for 48 h to establish dominant/subordinate hierarchies; their offspring were reared to 144 h post-fertilization (hpf) and sampled at five time points over development. No differences were detected in maternal cortisol contribution, which is thought to be an important programmer of offspring phenotype. However, once zebrafish offspring began to synthesize cortisol de novo (48 hpf), larvae of dominant females exhibited significantly lower baseline cortisol levels than offspring of subordinate females. These lower cortisol levels may reflect reduced hypothalamic-pituitary-interrenal (HPI) axis activity, because corticotropin-releasing factor (crf) and cytochrome p450 side chain cleavage enzyme (p450scc) mRNA levels also were lower in larvae from dominant females. Moreover, baseline mRNA levels of HPI axis genes continued to be affected by maternal social status beyond 48 hpf. At 144 hpf, stress-induced cortisol levels were significantly lower in offspring of subordinate females. These results suggest programming of stress axis function in zebrafish offspring by maternal social status, emphasizing the importance of maternal environment and experience on offspring stress axis activity. [ABSTRACT FROM AUTHOR]

Vitellogenin (Vtg) is an important precursor yolk protein in egg-laying vertebrates, including fish. The 17β-oestradiol (E2) plays a crucial role in the Vtg synthesis; moreover, certain hormones can stimulate Vtg synthesis. We investigated the possible role of E2, carp recombinant growth hormone (cr GH), insulin (Ins), progesterone (P4) and 11-deoxycortisol (11- DOC) hormones in Vtg synthesis on primary juvenile Chalcalburnus tarichi hepatocyte culture. The amount of Vtg in the medium was measured at 2-day intervals using enzyme-linked immunosorbent assay (ELISA). The hepatocytes were maintained in culture for more than 2 weeks without the addition of serum components. Vitellogenin localization was visualized with the immunofluorescence method in E2-supplemented hepatocytes. Among hormones applied to the culture, only E2 had an influence on Vtg synthesis in a time-dependent manner, while cr GH, Ins, P4 and 11- DOC had no effect. However, in hepatocytes stimulated with E2 in combination with P4, a lower Vtg production was seen compared with Vtg produced when hepatocytes were stimulated with E2 alone. P4 proved to have potentiating effects on co-treatment with E2-induced Vtg production. As a result, E2 and P4 are the most important hormones for Vtg synthesis in juvenile C. tarichi hepatocyte culture. [ABSTRACT FROM AUTHOR]

Indirect development is widespread in anurans and is considered an ancestral condition. The metamorphosis of larvae into juveniles involves highly coordinated morphological, physiological, biochemical, and behavioral changes, promoted by the thyroid hormone and interrenal corticosteroids. Stress response to environmental changes is also mediated by corticosteroids, affecting the timing and rate of metamorphosis and leading to great developmental plasticity in tadpoles. Given the potential effect of interrenal gland ontogeny alterations on metamorphosis and the lack of studies addressing both the morphology and endocrinology of this gland in tadpoles, we present corticosterone (CORT) production and histological changes through the ontogeny of interrenal gland in the generalized pond-type tadpole of Rhinella arenarum (Anura, Bufonidae). This species shows the highest concentration of whole-body CORT by the early climax when drastic metamorphic changes begin. This is coincident with the morphological differentiation of steroidogenic cells and the formation of interrenal cords. By this stage, steroidogenic cells have a shrunken cytoplasm, with a significantly higher nucleus-to-cell diameter ratio. The lowest CORT concentration during premetamorphosis and late climax is associated with small undifferentiated cells with lipid inclusions surrounding large blood vessels between kidneys, and with cords of differentiated steroidogenic cells with a significantly lower nucleus-to-cell diameter ratio, respectively. Our study characterizes the morphological and physiological pattern of interrenal gland development, showing an association between certain histological and morphometric characteristics and CORT levels. Variations in this morpho-physiological pattern should be considered when studying the phenotypic plasticity or variable growth rates of tadpoles., (© 2022 Anatomical Society.)

Highlights: [•] The interrenal gland was exclusively located in the posterior portion of the cephalic kidney. [•] Nuclear area of interrenal steroidogenic cells differed between males of two distinct social ranks. [•] Cortisol and 11-ketotestosterone plasma levels differed between males of two distinct social ranks. [•] Cortisol plasma levels correlated positively with the nuclear area of interrenal steroidogenic cells. [•] For the first time, interrenal gland, cortisol plasma levels and social rank were studied together. [Copyright &y& Elsevier]

Macrophage aggregates (MA) occur in various organs of fish as discrete aggregations of pigmented macrophages. The study presented herein investigates the quantitative modifications from normal anatomical condition, of interrenal gland (IG) and kidney MA in six treatment groups of adult rainbow trout submitted to either specific or aspecific immune stimulation and subsequently challenged with Yersinia ruckeri. Routinely stained tissue sections from both IG and kidney were analysed. The percentage of tissues occupied by MA and the MA density (number/mm2) were calculated on at least 10 randomly selected nonoverlapping fields taken from each tissue section. MA morphometric findings from challenged fish were compared to those from a control group. Results showed that fish from control group displayed a statistically significant (P < 0.05) higher percentage of tissue occupied by MA and MA density. Among different treatment groups, anti-Yersinia ruckeri immunized fish, which did not show clinical signs of disease after bacterial challenge, displayed higher values of morphometric parameters compared with symptomatic fish from other groups. Our study demonstrates that the quantification of the area occupied by MA might be an efficient parameter to evaluate the general condition of a salmonid population since it positively correlates with the health status and negatively with stress factor such as the acute bacterial infection. [ABSTRACT FROM AUTHOR]

Studies of the barfin plaice Liopsetta pinnifasciata inhabiting Amursky Bay (Peter the Great Bay, Sea of Japan) performed in 1999-2005 showed a twofold reduction in the area of cell nuclei of the interrenal and chromaffin glands after 2001, which indicates a significant weakening of the damaging effect of the environment on fish. It is assumed that one of the main causes of the hypertrophy of both glands in barfin plaices before 2001 was the phenolic pollution of the environment. The study points to the sensitivity of the adrenal system of the barfin plaice to changes in the quality of environment. [ABSTRACT FROM AUTHOR]

The datasets of this article present the experimental parameters resulting from the assessment of adrenal gland as a potential biomarker of endocrine disruption mediated by graphene oxide (GO), a nanocarbon, using Japanese medaka fish as the model. These data sets support the article "Histopathological evaluation of the interrenal gland (adrenal homolog) of Japanese medaka ( Oryzias latipes ) exposed to graphene oxide". The experiments were conducted on reproductively active adult fish maintained as a breeding pair (one male and one female) in 500 mL balanced salt solution (BSS) either by immersion in GO (20 mg/L in BSS) continuously for 96 h with refreshing of media once in every 24 h or by a single intraperitoneal (IP) injection of GO (100 µg/g) to both male and female fish. The experimental fish were allowed breeding and assessed after 21 days post-treatment. Moreover, one day-post hatch (dph) Japanese medaka fries (orange-red variety) were exposed to different concentrations of GO (2.5-20 mg/L) by immersion in embryo-rearing medium (ERM) for 96 h (1-5 dph) with refreshing of media every 24h. Food was given to the adults, however, the larvae remained fasting during the GO-exposure (0-5 dph) period. Control adults and larvae were identically maintained either in BSS (adults) or ERM (larvae), with no GO. After treatment, both adults and the larvae were maintained in BSS with feeding in a GO-free environment. After 21 days post-treatment, adults, and after six weeks post-treatment, larvae, were anaesthetized in MS-222, and the trunk region was preserved in 4% paraformaldehyde in PBS (20 mM) containing 0.05% Tween 20. Evaluation of interrenal gland (IRG) in kidneys were made in 5 µm thick sections stained on haematoxylin-eosin (HE). The phenotypic sex of adults was assessed by secondary sex characters (fin features) and gonad (testis and ovary) histology; in larvae, phenotypic sex was determined by gonad histology and the genotypic sex by genotyping dmy gene. The location of IRG in the kidney were determined by immunohistochemical technique using rabbit polyclonal antityrosine hydroxylase antibody as primary antibody. The digital images of sections were captured using an Olympus CKX53 inverted microscope with DP22 camera and CellSens software. Using imagej software, a minimum of 3 images of kidney consisting IRG were assessed for cell (separated as dark and pale stained nucleus after HE staining) sorting (cells/ mm 2 ) and also measured the nuclear area (µm 2 ). Counting of IRG cells, lined between the cardinal vein and the interstitial cells in the kidneys, were limited to maximum three layers in a given area. Numerical data, presented as means ± SEM, were analysed by one-way ANOVA followed by post-hoc Tukey's multiple comparison test or unpaired parametric 't' test including Welch's correction, if distributed normally; or by Kruskal-Wallis test followed by Mann-Whitney's test as post hoc test, if the data did not meet the criteria of using a parametric test. Statistically significant difference were considered for p  ≤ 0.05. The collected data on IRG of Japanese medaka fish will be used for the assessment of GO as an EDC disposed in the environment., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships which have or could be perceived to have influenced the work reported in this article., (© 2022 The Author(s).)

Abstract: We examined the effects of carbaryl (1-naphthyl methylcarbamate; sevin), a carbamate pesticide, on interrenal and thyroid activities and mitochondrial rich (MR) cell function in climbing perch to understand the physiological basis of toxicity acclimation in this fish to the chemical stressor. Carbaryl exposure (5–20mgL−1) for 48h increased cortisol and glucose, but decreased the T3 level without affecting T4 concentration in the plasma. These responses of the carbaryl-exposed fish were nullified and a rise in plasma T4 occurred in these fish when they were kept for 96h recovery in clean water. A tight plasma mineral control was indicated in the carbaryl-exposed fish as reflected by the unchanged plasma Na, K, Ca and inorganic phosphate levels. The ouabain-sensitive Na+, K+-ATPase activity showed an increase in the gills but the intestinal and renal tissues showed little response to carbaryl treatment. However, substantial increases in the intestinal and renal Na+, K+-ATPase activities occurred in the recovery fish. The MR cells in the branchial epithelia showed a strong Na+, K+-ATPase immunoreactivity to carbaryl treatment indicating an activated MR cell function. The numerical MR cell density remained unchanged, but stretching of secondary gill lamellae as part of gill remodeling occurred during carbaryl exposure. The increased surface of these lamellae with abundant MR cells as a result of its migration into the lamellar surface points to marked structural and functional modifications of these cells in the carbaryl-treated fish which is likely to a target for carbaryl action. The rise in plasma T4 and the restoration of normal branchial epithelia in the recovery fish indicate a thyroidal involvement in the recovery response and survival. Our data thus provide evidence that carbaryl exposure and its recovery evoke interrenal and thyroid disruption in this fish leading to a modified osmotic response including an altered MR cell function. [Copyright &y& Elsevier]

Abstract: The effects of chronic exposure to adrenocorticotropic hormone (ACTH) or the synthetic glucocorticoid dexamethasone (DEX) on the expression of genes involved in cortisol synthesis were examined using quantitative RT-PCR and immunohistochemistry. Juvenile Chinook salmon were treated with either ACTH via micro-osmotic pumps or with DEX via a lipid-based sustained release vehicle. Plasma cortisol levels were significantly elevated in ACTH-treated fish after 1day, with a significant reduction in this effect with increasing treatment duration. ACTH also appeared to cause progressive hyperplasia of interrenal cells. Steroidogenic acute regulatory protein (StAR) and cytochrome P450 side chain cleavage enzyme (P450scc) transcripts but not 3β-hydroxysteroid dehydrogenase-isomerase (3β-HSD) or cytochrome P450 11β-hydroxylase (P45011β) transcripts in head kidneys significantly increased after 5days of ACTH treatment. Significant linear relationships between plasma cortisol levels and transcript levels were identified at day 1 and day 5 for StAR, and day 5 for P450scc. Increased immunoreactivity for P450scc was observed in interrenal cells of ACTH-treated fish after 5 and 10days. No effect of ACTH on 3β-HSD immunoreactivity was apparent at any time point. P45011β immunoreactivity was more intense after 5days treatment with ACTH. DEX significantly reduced resting plasma cortisol levels and induced interrenal cell atrophy. Although no significant effect of treatment with DEX was found for any transcript, immunoreactivity for P450scc and P45011β appeared to be reduced. These results indicate that StAR and P450scc are subject to transcriptional regulation by chronic changes in ACTH levels. [Copyright &y& Elsevier]

The impact of hypophysectomy on the state of the interrenal gland and ultrastructure of chloride cells of gills is investigated in 18-month old juveniles of starred sturgeon Acipenser stellatus in the process of its adaptation to artificial sea water (14.6‰). Hypophysectomized juveniles, similarly to intact juveniles, are able to support a relative stability of osmolarity of blood serum in the course of adaptation to sea water by transition from hyperosmotic to hypoosmotic type of osmoregulation. Changes in the investigated parameters of cells of the interrenal gland (volume of nuclei, areas of cells and of lipophilic vacuoles) occurring in the hypophysectomized and in intact specimens in the process of adaptation to sea water are generally similar, but have different dynamics. In contrast to many teleostean species, in acipenserids the hypophyectomy does not cause atrophy of the interrenal gland. The latter is incorporated in the process of regulation in the course of adaptation of fish to sea water. Hypophyecotmy results in partial destruction of organoids in some chloride cells of gills. However, when the fish are transferred to sea water, the structural changes occur in chloride cells characteristic of their transition to the excretory state. This may happen only at activation of the transport enzyme Na/K-ATPase of these cells by cortisol produced by the interrenal gland. In the absence of hypophysis, the functional connection of organs of the axis hypothalamus (ACTH-immunopositive cells of tuberal nucleus) → the interrenal gland → chloride cells is realized in the fish. [ABSTRACT FROM AUTHOR]

Abstract: The rate-limiting and regulated step in steroidogenesis, the conversion of cholesterol to pregnenolone, is facilitated by the steroidogenic acute regulatory protein (StAR) and cytochrome P450 cholesterol side-chain cleavage (P450scc). We have isolated cDNAs encoding StAR and P450scc from the Atlantic stingray, Dasyatis sabina, and characterized the steroidogenic activity of the encoded proteins using a heterologous expression system. Green monkey kidney (COS-1) cells cotransfected with D. sabina StAR and human P450scc/adrenodoxin reductase/adrenodoxin fusion (F2) constructs produced significantly more pregnenolone than cells transfected with the F2 construct alone. COS-1 cells transfected with a modified F2 construct (F2DS) in which human P450scc is replaced by D. sabina P450scc had higher rates than cells transfected with D. sabina P450scc alone. In other vertebrates, the stress peptide adrenocorticotropic hormone (ACTH) elicits its effects on corticosteroidogenesis in part through regulation of StAR and P450scc mRNAs. In vitro incubation of D. sabina interrenal tissue with porcine ACTH significantly increased intracellular cAMP and corticosteroid production. As demonstrated by quantitative PCR, ACTH also induced significant increases in mRNA abundance of both StAR and P450scc. Our results suggest that, as in higher vertebrates, chronic ACTH-induced glucocorticoid synthesis in elasmobranchs is mediated by regulation of primary steroidogenic mRNAs. This study is the first to demonstrate steroidogenic activity of an elasmobranch P450scc protein and express a composite elasmobranch steroidogenic pathway in a heterologous cell line. Also, the regulation of StAR and P450scc mRNAs has not previously been demonstrated in elasmobranch fishes. [Copyright &y& Elsevier]

This work analyses the distribution, histology and ultrastructure of chromaffin cells (CCs) and interrenal tissue (It) in the head kidney of Epinephilus tauvina. Histological examination revealed that chromaffin cells are found in small groups under the endothelium of the posterior cardinal vein (PCV) and are mostly closely associated with the interrenal tissue. Ultrastructure examination confirmed the existence of two main chromaffin cell types, distinguished by different types of secretory granules. The first type was characterized by the presence of vesicles with round, strongly electron dense core granules, which were eccentrically located. Such cells were interpreted as being noradrenaline cells. Meanwhile, cells with vesicles that were completely electron lucent or that contained small less dense eccentric granules were identified as adrenaline cells. Nerve endings were invaginated into the chromaffin cells through synaptic junctions. Interrenal tissue consisted of nests, cords, or strands of cells in contact with the posterior cardinal vein (PCV) and interposed with haematopoietic tissue. Ultrastructure analysis revealed only one interrenal cell type, which contained abundant smooth endoplasmic reticulum (sER) and numerous mitochondria with tubulo-vesicular cristae, characteristics of steroid-producing cells. The interrenal tissue cells have different cytological aspects that can be linked to a steroidogenic cell cycle allowing a periodical renewal of organelles. [ABSTRACT FROM AUTHOR]

The annual cycle of morphological changes in the interrenal gland of the flounder Liopsetta pinnifasciata, a species which had been exposed over several decades to polluted waters of the Amur Gulf (Peter the Great bay of the Sea of Japan), is investigated. The curve of annual dynamics of the volume of nuclei of interrenal cells is U-form. The maximum size of nuclei is recorded in the period of perivitellogenous growth of oocytes and the minimum size is recorded at early stages of vitellogenesis. The area of epithelium of the interrenal gland in flounders is the lowest also in the beginning of the vitellogenous growth of oocytes. It is assumed that the uncommon annual cycles of changes in the interrenal gland of this flounder living in the Amur Gulf is an index of its stressed condition. [ABSTRACT FROM AUTHOR]

In teleostean fishes the hypothalamic–pituitary–thyroid axis (HPT axis) and the hypothalamic–pituitary–interrenal axis (HPI axis) regulate the release of thyroid hormones (THs) and cortisol respectively. Since many actions of both hormones are involved in the regulation of metabolic processes, communication between both signal pathways can be anticipated. In this study, we describe central and peripheral sites for direct interaction between mediators of both neuroendocrine axes in the common carp (Cyprinus carpio). Despite suggestions in the literature that CRH is thyrotropic in some fish; we were not able to establish stimulatory effects of CRH on the expression of the pituitary TSHβ subunit gene. In preoptic area tissue incubated with 10–7M thyroxine (T4) a 2.9-fold increase in the expression of CRH-binding protein (CRHBP) was observed. Thus, T4could reduce the bioavailable hypothalamic crh via the up regulation of crhbp expression and hence down regulate the HPI axis. At the peripheral level, cortisol (10–6M), ACTH (10–7M), and α-MSH (10–7M) stimulate the release of T4from kidney and head kidney fragments, which contain all functional thyroid follicles in carp, by two- to fourfold. The substantiation of three pituitary thyrotropic factors, viz. TSH, ACTH, and α-MSH, in common carp, allows for an integration of central thyrotropic signals. Clearly, two sites for interaction between the HPT axis, the HPI axis, and α-MSH are present in common carp. These interactions may be key to the proper regulation of general metabolism in this fish. [ABSTRACT FROM AUTHOR]

Abstract: The Hedgehog (Hh) signaling pathway plays critical instructional roles during embryonic development. Misregulation of Hh/Gli signaling is a major causative factor in human congenital disorders and in a variety of cancers. The zebrafish is a powerful genetic model for the study of Hh signaling during embryogenesis, as a large number of mutants that affect different components of the Hh/Gli signaling system have been identified. By performing global profiling of gene expression in different Hh/Gli gain- and loss-of-function scenarios we identified known (e.g., ptc1 and nkx2.2a) and novel Hh-regulated genes that are differentially expressed in embryos with altered Hh/Gli signaling function. By uncovering changes in tissue-specific gene expression, we revealed new embryological processes that are influenced by Hh signaling. We thus provide a comprehensive survey of Hh/Gli-regulated genes during embryogenesis and we identify new Hh-regulated genes that may be targets of misregulation during tumorigenesis. [Copyright &y& Elsevier]

Abstract: Adenosine deaminase (ADA) is responsible for cleaving the neuromodulator adenosine to inosine. Two members of ADA subfamilies, known as ADA1 and ADA2, were described and evidence demonstrated another similar protein group named ADAL (adenosine deaminase “like”). Although the identification of ADA members seems to be consistent, the expression profile of ADA1, ADA2 and ADAL genes in zebrafish has not yet been reported. The aim of the present study was to map the expression pattern of ADA-related genes in various tissues of adult zebrafish (Danio rerio). An extensive search on zebrafish genome followed by a phylogenetic analysis confirmed the presence of distinct ADA-related genes (ADA1, ADAL and two orthologous genes of ADA2). Specific primers for each ADA member were designed, optimized semi-quantitative RT-PCR experiments were conducted and the relative amount of transcripts was determined. The tissue samples (brain, gills, heart, liver, skeletal muscle and kidney) were collected and the expression of ADA1, ADAL and ADA2 genes was characterized. ADA1 had a similar expression pattern, whereas ADAL was less expressed in the heart. The highest relative amount of ADA2-1 transcripts was observed in the brain, liver and gills and it was less expressed in the heart. RT-PCR assays revealed that the other ADA2 form (ADA2-2) was expressed ubiquitously and at comparable levels in zebrafish tissues. The strategy adopted also allowed the identification of an ADA2-1 truncated alternative splice isoform (ADA2-1/T), which was expressed at different intensities. These findings demonstrated the existence of different ADA-related genes, their distinct expression pattern and a truncated ADA2-1 isoform, which suggest a high degree of complexity in zebrafish adenosinergic system. [Copyright &y& Elsevier]

Abstract: The ultrastructural aspect of degeneration and recovery processes involving the steroidogenic interrenal cells of the stickleback was studied. Together with the adrenergic cells, the interrenals constitute the adrenal homolog in teleosts. From our study it appears that a process of massive cell death may lead to temporary disappearance of the gland. Moreover, our E.M. observations suggest two main ways, each leading to morphological dedifferentiation of the cells, no longer recognizable as interrenals: the first way involves elimination of organelles and recovery of the nucleus surrounded by a thin rim of cytoplasm; the second involves fragmentation of the cytoplasm by other pyknotic star-shaped interrenals, together with autophagocytosis processes. Our E.M. observations also suggest that the subsequent reconstitution of the tissue can occur in two ways. In the first, the interrenals appear mainly to differentiate from mesenchymatic-like electron-light cells, while in the second, the new interrenals appear mainly raising from some macrophagic electron-dense cells. Some data obtained with Mallory''s trichrome staining of histological sections, and localization of the enzyme 3β hydroxysteroid dehydrogenase in thin sections, support the above-mentioned results. A hypothesis is advanced on the origin of the electron-dense differentiating interrenals, and a possible role of dedifferentiated cells in restoration of the interrenal gland is also discussed. [Copyright &y& Elsevier]

Abstract: The interrenal gland (adrenocortical homolog) of elasmobranchs produces a unique steroid, 1α-hydroxycorticosterone (1α-B). The synthesis of this and most other steroids requires both cholesterol side chain cleavage (CYP11A) and 3β-hydroxysteroid dehydrogenase (HSD3). To facilitate the study of elasmobranch steroidogenesis, we isolated complementary DNAs encoding CYP11A and HSD3 from the freshwater stingray Potamotrygon motoro. The P. motoro CYP11A (2182 bp total length) and HSD3 (2248 bp total length) cDNAs harbor open reading frames that encode proteins of 542 and 376 amino acids (respectively) that are similar (CYP11A: 39–61% identical; HSD3: 36–53% identical) to their homologs from other vertebrates. In molecular phylogenetic analysis, P. motoro CYP11A segregates with CYP11A proteins (and not with related CYP11B proteins) and P. motoro HSD3 segregates with steroidogenic HSD3 proteins from other fishes. CYP11A and HSD3 mRNA is found only in interrenal and gonadal tissues, indicating de novo steroidogenesis is restricted to these tissues. Because 1α-B is thought to act in the elasmobranch response to hydromineral disturbances, we examined the effect of adapting P. motoro to 10 ppt seawater on mRNAs encoding steroidogenic genes. The P. motoro response to this salinity challenge does not include interrenal hypertrophy or an increase in the levels of interrenal CYP11A, HSD3 or steroidogenic acute regulatory protein (StAR) mRNA. This study is the first to isolate full length cDNAs encoding elasmobranch CYP11A and HSD3 and the first to examine the regulation of steroidogenic genes in elasmobranch interrenal cells. [Copyright &y& Elsevier]

Abstract: Red drum settle into shallow seagrass meadows during the larval stage. Day–night cycles in these habitats result in marked diel temperature and dissolved oxygen (DO) cycles, and it is possible that extreme fluctuations influence endocrine development and growth of larvae. Here, we described red drum interrenal and thyroid ontogeny and determine responses to environmental stimuli with special emphasis on settlement to explore possible role of hormones as mediator of directive environmental factors. This study detected an early activation of thyroid and interrenal axis during the yolk-sac phase and a second activation of the thyroid starting at settlement size to the end of the larval period. Whole-body l-thyroxine (T4) and 3-5-3′-triiodo-l-thyronine (T3) showed a sharp decline at the juvenile stage. In contrast, cortisol steadily declines during the larval phase to a minimum before the end of the larval period. Older settlement-size larvae exposed to a strong stimulus increased whole body cortisol. In contrast, new settlers showed a minor cortisol rise suggesting changes on stress responsiveness during the ontogeny of the species. Additionally, settlement-size larvae exposed to various environmentally realistic temperature or DO fluctuations showed no difference in growth compared to fish grown under stable conditions (control). However, growth rate was significantly reduced in DO cycled fish with prolonged exposure to hypoxia. No differences were found in whole-body cortisol levels in the reduced growth treatment groups, suggesting that growth retardation was not related to a cortisol-mediated stress response. In moderate DO and temperature treatment groups, cortisol showed wider fluctuations than control groups during the night time that were not related to stress. [Copyright &y& Elsevier]

Investigates the interrenal stress responsiveness of marbled sole in Tokyo, Japan. Effects of air-exposure on plasma cortisol and glucose levels in the fish; Impact of ACTH treatment on cortisol release from the head kidney; Responsiveness of the fish to cortisol-producing interrenal cells.

This work presents the structure and ultrastructure of the interrenal gland and chromaffin cells, as well as the morphology of the head kidney of Brycon cephalus. The head kidney is composed of fused bilateral lobes located anterior to the swim bladder and ventrolateral to the spinal column. The parenchyma revealed lympho-haematopoietic tissue, melano-macrophage centres, interrenal gland and chromaffin cells. The interrenal gland consisted of cords or strands of cells grouped around the posterior cardinal vein and their branches. Chromaffin cells are found in small groups, closely associated with the interrenal gland and/or under the endothelium of the posterior cardinal vein. So far, the ultrastructural analysis has revealed only one interrenal cell type which contained abundant smooth endoplasmic reticulum and numerous mitochondria with tubulo-vesicular cristae, characteristic of steroid-producing cells. Two types of chromaffin cells were observed. The first type was characterized by the presence of vesicles with round, strongly electron-dense granules, which were eccentrically located. Such cells were interpreted as noradrenaline cells. Meanwhile, cells which contained smaller vesicles and electron-lucent granules, with a small halo separating the granule from the vesicular limiting membrane, were identified as adrenaline cells. [ABSTRACT FROM AUTHOR]

In nine cell types of the adenohypophysis in untreated adult rainbow trout, histologically different activity phases, seasonal changes in activity, and the relation between certain cell types and the interrenal gland, thyroid or gonads were investigated by light and, occasionally, by electron microscopy. Special attention was given to the effect of social rank on the synthetic activity in adenohypophysial cells of trout kept in small groups in which a social hierarchy with one (light) dominant and several (dark) submissives is established. Cell types in the rostral pars distalis were azocarminophil (I) or amphiphil (II). Proximal pars distalis cell types were slightly basophil (IV), orangeophil (V), strongly basophil (VI) or chromophobe (VII). In the pars intermedia, cell types were amphiphil (VIII) or very slightly basophil (IX). Type III was a non-secretory supporting (?) cell. Histologically different activity phases abounded in type IV cells, which mainly occurred in the proximal pars distalis but were also found dispersed in the rostral pars distalis, the pars intermedia and the neurohypophysis. Influences of social rank were pronounced in type IV cells. Phases with a high synthetic activity were exclusively found in submissive animals, phases with a low synthetic activity occurred in dominants. As a positive relation existed between type IV cell activity and the social rank dependent activity of the interrenal gland, it was suggested that type IV cells produce ACTH. In (dominant) male trout treated with DOCG or ACTH, colloid-containing type IV cell phases, reflecting accumulation of the secretory product, were found. This supported the earlier suggestion that ACTH in the trout is produced in the basophil type IV cells and not, as reported in the literature, in cells comparable to type II. [ABSTRACT FROM AUTHOR]

Immunocytochemistry was used to investigate the presence of corticotropin-releasing factor-like peptides in the interrenal (adrenal) glands of the bullfrog Rana catesbeiana by using specific antisera raised against synthetic nonconjugated rat/human corticotropin-releasing factor, urotensin I, and sauvagine. From these three antisera, covering a broad range of corticotropin-releasing factor-like immunoreactivities, only the sauvagine antiserum gave positive immunoreactivity. Sauvagine immunoreactivity was found in cortical cells grouped into cords in the renal zone of the interrenal gland. The central and subcapsular cords were less stained. Tyrosine hydroxylase-positive chromaffin cells were not sauvagine-immunoreactive. The immunoreactivity was abolished, in all cases, by previous immunoabsorption of the sauvagine antiserum with synthetic sauvagine (0.1 μM), but it was not eliminated by sucker ( Catostomus commersoni) urotensin I, sole ( Hippoglossoides elassodon) urotensin I, sucker corticotropin-releasing factor, rat/human corticotropin-releasing factor, or ovine corticotropin-releasing factor (0.1-10 μM). In a sauvagine radioimmunoassay, interrenal extracts displaced I-sauvagine from antiserum only partially, and not in parallel with the sauvagine standard curve. The results suggest that the sauvagine immunoreactivity in the R. catesbeiana interrenal gland may represent a novel sauvagine-like peptide. [ABSTRACT FROM AUTHOR]

The conversion of 3b-hydroxy-δ5 steroids into their respective biologically active 3-keto-δ4 steroid counterparts is catalyzed by a microsomal enzyme, 3β-hydroxysteroid dehydrogenase /5-ene-4-ene isomerase (3β-HSD). In order to study this enzyme in elasmobranchs, partial cDNA clones of 3β-HSD were isolated from steroidogenic tissue of the southern stingray (Dasyatis americana), spiny dogfish shark (Squalus acanthias), and the blacktip shark (Carcharhinus limbatus). The deduced amino acid sequences of the elasmobranchs were 40-44% identical to mammalian 3β-HSD sequences, and approximately 50% identical to the rainbow trout (Oncorhynchus mykiss) form. However, the highly conserved substrate-binding region was 80-90% identical among the various species. Northern blot analysis of interrenal gland RNA indicated a single 2.4 kb transcript for the stingray homolog and a 1.4 kb transcript for the blacktip shark homolog. Blacktip shark 3β-HSD activity was enriched in the microsomal fraction of the interrenal gland and was only detected in the presence of the oxidized forms of nicotinamide adenine dinucleotides. Maximal rates were observed near pH 7.5 and between 25 °C and 30 °C. Urea (400 mM) and trimethylamine oxide (200 mM), normally present in elasmobranch blood, had no effect upon 3β-HSD activity. Kinetic parameters of blacktip shark 3β-HSD were determined for pregnenolone (Km = 0.35 ± 0.06 μM; Vmax = 2.3 ± 0.11 nmol min-1 mg protein-1) and DHEA (Km = 0.12 ± 0.02 μM; Vmax = 1.1 ± 0.41 nmol min-1). The kinetic parameters of the stingray 3β-HSD for pregnenolone (Km = 0.13 ± 0.09 μM; Vmax = 4.0 ± 0.51 nmol min-1 mg protein-1) and DHEA (Km = 0.12 ± 0.08 μM; Vmax = 4.1 ± 0.53 nmol min-1 mg protein-1) were similar to those of the blackt [ABSTRACT FROM AUTHOR]

Cyanoketone administered via the food (0.1, 0.2 and 2 mg/g) for 8 weeks from the first feeding (day 46 after fertilization) or via the aquarium water (3 and 30 mg/100 l) for 4 weeks from day 41 does not influence the activity of 3 β-hydroxysteroid dehydrogenase (3 β-HSD) in the interstitial cells of the gonads or interrenal cells of juvenile trout in vivo. However, the number of 3 β-HSD-positive interrenal cells was strongly increased by administration of the highest dose of cyanoketone via both routes. These high doses furthermore affect the sex ratio in favor of males. It is concluded that interrenal tissue is responsible for the masculinizing effect of cyanoketone via increased production of androgens and/or corticosteroids. Cyanoketone at concentrations of 0.01 to 100 μg/ml causes a dose-response inhibition of 3 β-HSD activity in the interrenal cells, when the substance is administered to an incubation medium for demonstration of this enzyme in tissue sections. The controversial in-vivo and in-vitro effects of cyanoketone on 3 β-HSD activity are discussed. [ABSTRACT FROM AUTHOR]

Partial hepatectomy was carried out on Xenopus laevis to investigate its influence on the endocrine system. In addition to other endocrine effects, a marked hypertrophy and stimulation of the interrenal gland was observed. Activated cells contain mitochondria with extended and irregularly coiled tubules embedded in a low electron dense matrix. Hepatectomy induces two phases of proliferation [3 and 35 days postoperative (p.o.)]. After 106 days p.o. giant mitochondria possessing narrow and closely packed, parallel tubules surrounded by an electron dense matrix indicate a phase of inactivation. The smooth endoplasmic reticulum and the Golgi apparatus proliferate after hepatectomy. During activation the high lipid content seen in controls is decreased significantly. [ABSTRACT FROM AUTHOR]

The effects of gonadectomy on the cytological structure of the hypophysis and interrenal tissue of Clarias batrachus have been investigated. Forty days after gonadectomy remarkable hypertrophy of certain cyanophils, identified later as gonadotrops, has been observed followed by hyperplasia. The somatotrops also seem to undergo similar changes, and as a result the proximal pars distalis (PPD) increases enormously and encroaches upon the pars intermedia (PT). The gonadotrops undergo a gradual degranulation and become devoid of stainable cytoplasm by 90 days. No signet ring cells have been noticed at any stage. The cyanophils in the ventral region of the PPD, identified as thyrotrops, also show degranulation, but to a comparatively limited degree. The ACTH cells undergo gradual hypertrophy and degranulation. A progressive increase is noticed in the nuclear diameter. The interrenal cells are activated as indicated by the enhanced nuclear indices. It is suggested that a I all in gonadal steroids may result in ACTH release which in turn activates the interrenal cells. [ABSTRACT FROM AUTHOR]

The article studies the interrenal tissues of bony fishes especially Amia Calva. There are some confusions, that still reigns on the true significance of kidneys, in fishes. which are homologous with the cortex and the medulla of the mammalian adrenal, and, on the other hand, of those structures, which are known as Stannius corpuscles. The material for these investigations into Amia consisted of series of slides, made several years ago and used for the study of the gonad development in this fish. Entire sections had been made in the smaller specimens; portions of kidney or gonad had been prepared in the larger ones. Stains used were haematoxyline-eosine or Azan.

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Morphologic studies were performed on the summer cell of Stilling in the interrenal gland of the American bullfrog. Ultrastructural studies showed the cell to be packed with membrane-bound granules similar to those of the juxtaglomerular apparatus granules in the mammalian kidney. These granules reacted positively with several stains for granules of cells in the juxtaglomerular apparatus. The Stilling cells are intimately associated with the cortical cells and contain rough endoplasmic reticulum and granules in varying degrees of development associated with the Golgi apparatus. It is concluded that the Stilling cell may secrete a renin-like substance that may function in an aldosterone-stimulating reninangiotensin system. [ABSTRACT FROM AUTHOR]

Studies the effect of acute extracellular volume expansion, sodium status and renal perfusion pressure on prostaglandin E[sub2] (PGE[sub2]) synthesis in rat renal cortical and medullary interstitium using microdialysis. Modulatory role of prostaglandins (PG) in the control of hemodynamics and excretion; Factors influencing renal PG synthesis; Renal function during progressive alterations the renal medulla.