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1.
Mol Cell Endocrinol ; 535: 111372, 2021 09 15.
Article in English | MEDLINE | ID: mdl-34175410

ABSTRACT

In this article we aim to provide an overview of the zebrafish interrenal development and function, as well as a review of its contribution to basic and translational research. A search of the PubMed database identified 41 relevant papers published over the last 20 years. Based on the common themes identified, we discuss the organogenesis of the interrenal gland and its functional development and we review what is known about the genes involved in zebrafish steroidogenesis. We also outline the consequences of specific defects in steroid biosynthesis, as revealed by evidence from genetically engineered zebrafish models, including cyp11a2, cyp21a2, hsd3b1, cyp11c1 and fdx1b deficiency. Finally, we summarise the impact of different chemicals and environmental factors on steroidogenesis. Our review highlights the utility of zebrafish as a research model for exploring important areas of basic science and human disease, especially in the current context of rapid technological progress in the field of Molecular Biology.


Subject(s)
Interrenal Gland/embryology , Steroids/biosynthesis , Zebrafish/growth & development , Animals , Animals, Genetically Modified , Genetic Engineering , Interrenal Gland/metabolism , Organogenesis , Zebrafish/genetics , Zebrafish/metabolism
2.
Endocrinology ; 158(12): 4206-4217, 2017 12 01.
Article in English | MEDLINE | ID: mdl-29029162

ABSTRACT

Although the zebrafish interrenal tissue has been used as a model for steroidogenesis and genesis of the adrenal gland, its specification and morphogenesis remains largely unclear. In the present study, we explored how the Wilms tumor 1 (WT1)-expressing cells are segregated from the SF-1-expressing steroidogenic cells in the zebrafish model. The interrenal tissue precursors expressing ff1b, the equivalent of mammalian SF-1, were derived from wt1-expressing pronephric primordia in the zebrafish embryo. Through histochemistry and in situ hybridization, we demonstrated that the size of functionally differentiated interrenal tissue was substantially increased on global inhibition of the Notch signaling pathway and was accompanied by a disrupted segregation between the wt1- and ff1b-expressing cells. As the Notch pathway was conditionally activated during interrenal specification, differentiation, but not ff1b expression, of interrenal tissue was drastically compromised. In embryos deficient for Notch ligands jagged 1b and 2b, transgenic reporter activity of wt1b promoter was detected within the steroidogenic interrenal tissue. In conclusion, our results indicate that Jagged-Notch signaling is required (1) for segregation between wt1-expressing cells and differentiated steroidogenic tissue; and (2) to modulate the extent of functional differentiation in the steroidogenic interrenal tissue.


Subject(s)
Jagged-1 Protein/genetics , Jagged-2 Protein/genetics , Receptors, Notch/genetics , Signal Transduction/genetics , WT1 Proteins/genetics , Zebrafish Proteins/genetics , Animals , Animals, Genetically Modified , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/embryology , Embryo, Nonmammalian/metabolism , Gene Expression Regulation, Developmental , Head Kidney/cytology , Head Kidney/embryology , Head Kidney/metabolism , In Situ Hybridization , Interrenal Gland/cytology , Interrenal Gland/embryology , Interrenal Gland/metabolism , Jagged-1 Protein/metabolism , Jagged-2 Protein/metabolism , Receptors, Notch/metabolism , Steroidogenic Factor 1/genetics , Steroidogenic Factor 1/metabolism , Steroids/biosynthesis , WT1 Proteins/metabolism , Zebrafish/embryology , Zebrafish/genetics , Zebrafish/metabolism , Zebrafish Proteins/metabolism
3.
Endocrinology ; 158(12): 4165-4173, 2017 12 01.
Article in English | MEDLINE | ID: mdl-28938470

ABSTRACT

Congenital adrenal hyperplasia is a group of common inherited disorders leading to glucocorticoid deficiency. Most cases are caused by 21-hydroxylase deficiency (21OHD). The systemic consequences of imbalanced steroid hormone biosynthesis due to severe 21OHD remains poorly understood. Therefore, we developed a zebrafish model for 21OHD, which focuses on the impairment of glucocorticoid biosynthesis. A single 21-hydroxylase gene (cyp21a2) is annotated in the zebrafish genome based on sequence homology. Our in silico analysis of the 21-hydroxylase (Cyp21a2) protein sequence suggests a sufficient degree of similarity for the usage of zebrafish cyp21a2 to model aspects of human 21OHD in vivo. We determined the spatiotemporal expression patterns of cyp21a2 by whole-mount in situ hybridization and reverse transcription polymerase chain reaction throughout early development. Early cyp21a2 expression is restricted to the interrenal gland (zebrafish adrenal counterpart) and the brain. To further explore the in vivo consequences of 21OHD we created several cyp21a2 null-allele zebrafish lines by using a transcription activator-like effector nuclease genomic engineering strategy. Homozygous mutant zebrafish larvae showed an upregulation of the hypothalamic-pituitary-interrenal (HPI) axis and interrenal hyperplasia. Furthermore, Cyp21a2-deficient larvae had a typical steroid profile, with reduced concentrations of cortisol and increased concentrations of 17-hydroxyprogesterone and 21-deoxycortisol. Affected larvae showed an upregulation of the HPI axis and interrenal hyperplasia. Downregulation of the glucocorticoid-responsive genes pck1 and fkbp5 indicated systemic glucocorticoid deficiency. Our work demonstrates the crucial role of Cyp21a2 in glucocorticoid biosynthesis in zebrafish larvae and establishes an in vivo model allowing studies of systemic consequences of altered steroid hormone synthesis.


Subject(s)
Adrenal Hyperplasia, Congenital/genetics , Interrenal Gland/metabolism , Steroid 21-Hydroxylase/genetics , Zebrafish Proteins/genetics , Adrenal Hyperplasia, Congenital/embryology , Adrenal Hyperplasia, Congenital/enzymology , Animals , Embryo, Nonmammalian/embryology , Embryo, Nonmammalian/enzymology , Embryo, Nonmammalian/metabolism , Fish Diseases/embryology , Fish Diseases/enzymology , Fish Diseases/genetics , Gene Expression Regulation, Developmental , Glucocorticoids/biosynthesis , Hyperplasia/enzymology , Hyperplasia/genetics , In Situ Hybridization , Interrenal Gland/embryology , Interrenal Gland/pathology , Larva/enzymology , Larva/genetics , Larva/metabolism , Mutation , Reverse Transcriptase Polymerase Chain Reaction , Steroid 21-Hydroxylase/metabolism , Zebrafish , Zebrafish Proteins/metabolism
4.
PLoS One ; 9(9): e107997, 2014.
Article in English | MEDLINE | ID: mdl-25248158

ABSTRACT

BACKGROUND: While the endothelium-organ interaction is critical for regulating cellular behaviors during development and disease, the role of blood flow in these processes is only partially understood. The dorsal aorta performs paracrine functions for the timely migration and differentiation of the sympatho-adrenal system. However, it is unclear how the adrenal cortex and medulla achieve and maintain specific integration and whether hemodynamic forces play a role. METHODOLOGY AND PRINCIPAL FINDINGS: In this study, the possible modulation of steroidogenic and chromaffin cell integration by blood flow was investigated in the teleostean counterpart of the adrenal gland, the interrenal gland, in the zebrafish (Danio rerio). Steroidogenic tissue migration and angiogenesis were suppressed by genetic or pharmacologic inhibition of blood flow, and enhanced by acceleration of blood flow upon norepinephrine treatment. Repressed steroidogenic tissue migration and angiogenesis due to flow deficiency were recoverable following restoration of flow. The regulation of interrenal morphogenesis by blood flow was found to be mediated through the vascular microenvironment and the Fibronectin-phosphorylated Focal Adhesion Kinase (Fn-pFak) signaling. Moreover, the knockdown of krüppel-like factor 2a (klf2a) or matrix metalloproteinase 2 (mmp2), two genes regulated by the hemodynamic force, phenocopied the defects in migration, angiogenesis, the vascular microenvironment, and pFak signaling of the steroidogenic tissue observed in flow-deficient embryos, indicating a direct requirement of mechanotransduction in these processes. Interestingly, epithelial-type steroidogenic cells assumed a mesenchymal-like character and downregulated ß-Catenin at cell-cell junctions during interaction with chromaffin cells, which was reversed by inhibiting blood flow or Fn-pFak signaling. Blood flow obstruction also affected the migration of chromaffin cells, but not through mechanosensitive or Fn-pFak dependent mechanisms. CONCLUSIONS AND SIGNIFICANCE: These results demonstrate that hemodynamically regulated Fn-pFak signaling promotes the migration of steroidogenic cells, ensuring their interaction with chromaffin cells along both sides of the midline during interrenal gland development.


Subject(s)
Camptothecin/administration & dosage , Chromaffin Cells/drug effects , Diacetyl/analogs & derivatives , Interrenal Gland/blood supply , Zebrafish Proteins/metabolism , Zebrafish/embryology , Animals , Cell Movement/drug effects , Cellular Microenvironment , Chromaffin Cells/physiology , Diacetyl/pharmacology , Gene Expression Regulation, Developmental/drug effects , Hemodynamics/drug effects , Interrenal Gland/cytology , Interrenal Gland/embryology , Neovascularization, Physiologic/drug effects , Norepinephrine/pharmacology , Signal Transduction/drug effects , Zebrafish/metabolism , Zebrafish Proteins/genetics
5.
Endocrinology ; 154(8): 2702-11, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23671259

ABSTRACT

Zebrafish are emerging as a model to study steroid hormone action and associated disease. However, steroidogenesis in zebrafish is not well characterized. Mammalian P450 side-chain cleavage enzyme (CYP11A1) catalyzes the first step of steroidogenesis, the conversion of cholesterol to pregnenolone. Previous studies describe an essential role for zebrafish Cyp11a1 during early development. Cyp11a1 has been suggested to be the functional equivalent of mammalian CYP11A1 in the zebrafish interrenal gland (equivalent to the mammalian adrenal), gonad, and brain. However, reported cyp11a1 expression is inconsistent in zebrafish larvae, after active cortisol synthesis commences. Recently a duplicated cyp11a gene, cyp11a2, has been described, which shares an 85% identity with cyp11a1. We aimed to elucidate the specific role of the two cyp11a paralogs. cyp11a1 was expressed from 0 to 48 hours post-fertilization (hpf), whereas cyp11a2 expression started after the development of the interrenal primordium (32 hpf) and was the only paralog in larvae. cyp11a2 is expressed in adult steroidogenic tissues, such as the interrenal, gonads, and brain. In contrast, cyp11a1 was mainly restricted to the gonads. Antisense morpholino knockdown studies confirmed abnormal gastrulation in cyp11a1 morphants. cyp11a2 morphants showed impaired steroidogenesis and a phenotype indicative of metabolic abnormalities. The phenotype was rescued by pregnenolone replacement in cyp11a2 morphants. Thus, we conclude that cyp11a1 is required for early development, whereas cyp11a2 is essential for the initiation and maintenance of zebrafish interrenal steroidogenesis. Importantly, this study highlights the need for a comprehensive characterization of steroidogenesis in zebrafish prior to its implementation as a model organism in translational research of adrenal disease.


Subject(s)
Cholesterol Side-Chain Cleavage Enzyme/genetics , Interrenal Gland/metabolism , Steroids/biosynthesis , Zebrafish Proteins/genetics , Zebrafish/genetics , Animals , Cholesterol Side-Chain Cleavage Enzyme/classification , Cholesterol Side-Chain Cleavage Enzyme/metabolism , Embryo, Nonmammalian/drug effects , Embryo, Nonmammalian/embryology , Embryo, Nonmammalian/metabolism , Gastrulation/drug effects , Gastrulation/genetics , Gene Expression Regulation, Developmental , Gene Expression Regulation, Enzymologic , Gene Knockdown Techniques , Interrenal Gland/embryology , Interrenal Gland/growth & development , Isoenzymes/genetics , Isoenzymes/metabolism , Larva/genetics , Larva/growth & development , Phenotype , Phylogeny , Pregnenolone/metabolism , Pregnenolone/pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Zebrafish/embryology , Zebrafish/growth & development , Zebrafish Proteins/metabolism
6.
Dev Dyn ; 242(5): 432-43, 2013 May.
Article in English | MEDLINE | ID: mdl-23362214

ABSTRACT

BACKGROUND: Fibronectin (Fn) forms a centripetal gradient during the fetal adrenal gland organogenesis, and modulates hormone responsiveness of adrenocortical cells in the primary culture. However, how Fn is involved in organ formation of the adrenal gland remains unclear. RESULTS: In this study, we found that Fn accumulates around migrating ff1b-expressing interrenal cells, which were marked by the ff1b promoter-driven transgenic fluorescence, during the course of interrenal organ assembly. The interrenal cells displaying the migratory phenotype were absent in the fn1 mutant, while specification and kidney association of the interrenal tissue remained normal. The Fn deposition in the interrenal microenvironment was severely reduced in the vessel-deficient ets1b morphant, implying its origin of synthesis from the peri-interrenal vasculature. In the fn1 mutant, early-migrating chromaffin cells were capable of interacting with steroidogenic interrenal cells, yet continuous migration and midline convergence of chromaffin cells were disrupted. Migration defects of both interrenal and chromaffin lineages, in the absence of Fn, thus led to incomplete interrenal organ assembly in aberrant positions. CONCLUSIONS: Our results indicate that Fn is essential for patterning interrenal organ formation, by modulating the migratory behavior of both steroidogenic interrenal and chromaffin cells.


Subject(s)
Body Patterning/genetics , Fibronectins/physiology , Interrenal Gland/embryology , Zebrafish/embryology , 3-Hydroxysteroid Dehydrogenases/genetics , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Animals, Genetically Modified , Cell Movement/genetics , Chromaffin Cells/metabolism , Chromaffin Cells/physiology , Embryo, Nonmammalian , Fibronectins/genetics , Fibronectins/metabolism , Gene Expression Regulation, Developmental , Gene Expression Regulation, Enzymologic , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Interrenal Gland/metabolism , Organogenesis/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Zebrafish/genetics , Zebrafish/metabolism , Zebrafish Proteins/genetics , Zebrafish Proteins/metabolism
7.
PLoS One ; 7(8): e43040, 2012.
Article in English | MEDLINE | ID: mdl-22937010

ABSTRACT

BACKGROUND: The early morphogenetic steps of zebrafish interrenal tissue, the teleostean counterpart of the mammalian adrenal gland, are modulated by the peri-interrenal angioblasts and blood vessels. While an organized distribution of intra-adrenal vessels and extracellular matrix is essential for the fetal adrenal cortex remodeling, whether and how an intra-interrenal buildup of vasculature and extracellular matrix forms and functions during interrenal organogenesis in teleosts remains unclear. METHODOLOGY AND PRINCIPAL FINDINGS: We characterized the process of interrenal gland vascularization by identifying the interrenal vessel (IRV); which develops from the axial artery through angiogenesis and is associated with highly enriched Fibronectin (Fn) accumulation at its microenvironment. The loss of Fn1 by either antisense morpholino (MO) knockdown or genetic mutation inhibited endothelial invasion and migration of the steroidogenic tissue. The accumulation of peri-IRV Fn requires Integrin α5 (Itga5), with its knockdown leading to interrenal and IRV morphologies phenocopying those in the fn1 morphant and mutant. fn1b, another known fn gene in zebrafish, is however not involved in the IRV formation. The distribution pattern of peri-IRV Fn could be modulated by the blood flow, while a lack of which altered angiogenic direction of the IRV as well as its ability to integrate with the steroidogenic tissue. The administration of Fn antagonist through microangiography exerted reducing effects on both interrenal vessel angiogenesis and steroidogenic cell migration. CONCLUSIONS AND SIGNIFICANCE: This work is the first to identify the zebrafish IRV and to characterize how its integration into the developing interrenal gland requires the Fn-enriched microenvironment, which leads to the possibility of using the IRV formation as a platform for exploring organ-specific angiogenesis. In the context of other developmental endocrinology studies, our results indicate a highly dynamic interrenal-vessel interaction immediately before the onset of stress response in the zebrafish embryo.


Subject(s)
Fibronectins/metabolism , Interrenal Gland/blood supply , Zebrafish Proteins/metabolism , Animals , Fibronectins/genetics , Immunohistochemistry , In Situ Hybridization , Interrenal Gland/embryology , Signal Transduction/genetics , Signal Transduction/physiology , Zebrafish , Zebrafish Proteins/genetics
8.
Mol Endocrinol ; 21(2): 472-85, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17082325

ABSTRACT

To further elucidate pituitary adrenal interactions during development, we studied the organogenesis of the interrenal organ, the teleost homolog of the mammalian adrenal gland, in zebrafish. To this end we compared wild-type zebrafish interrenal development with that of mutants lacking pituitary cell types including corticotrophs. In addition, we studied the effects of ACTH receptor (Mc2r) knockdown and dexamethasone (dex) on interrenal development and pituitary feedback. Until 2 d post fertilization (2 dpf) interrenal development assessed by transcripts of key steroidogenic genes (cyp11a1, mc2r, star) is independent of proopiomelanocortin (Pomc) as demonstrated in aal/eya1and lia/fgf3 mutants. However, at 5 dpf lack of pituitary cells leads to reduced expression of steroidogenic genes at both the transcriptional and the protein level. Pituitary control of interrenal development resides in corticotrophs, because pit1 mutants lacking pituitary cells except corticotrophs have a phenotype similar to that of wild-type controls. Furthermore, development in mc2r knockdown morphants does not differ from aal/eya1 and lia/fgf3 mutants. Inhibition of steroidogenesis by mc2r knockdown induces up-regulation of pomc expression in the anterior domain of pituitary corticotrophs. Accordingly, dex suppresses pomc in the anterior domain only, leading to impaired expression of steroidogenic genes commencing at 3 dpf and interrenal hypoplasia via reduced interrenal proliferation. In contrast, negative feedback on pituitary corticotrophs by dex is evident at 2 dpf and precedes effects of Pomc on the interrenal primordium. These data demonstrate a gradual transition from early pituitary-independent interrenal organogenesis to developmental control by the anterior domain of pituitary corticotrophs acting via Mc2 receptors.


Subject(s)
Interrenal Gland/embryology , Pituitary Gland/embryology , Zebrafish/embryology , Animals , Animals, Genetically Modified , Cell Proliferation , Cholesterol Side-Chain Cleavage Enzyme/metabolism , Corticotrophs/cytology , Corticotrophs/metabolism , Dexamethasone/pharmacology , Embryo, Nonmammalian/metabolism , Interrenal Gland/metabolism , Mutation , Phosphoproteins/metabolism , Pituitary Gland/cytology , Pituitary Gland/metabolism , Pro-Opiomelanocortin/metabolism , Receptors, Corticotropin/metabolism , Zebrafish/genetics , Zebrafish/metabolism , Zebrafish Proteins/metabolism
9.
Mol Endocrinol ; 20(11): 2630-40, 2006 Nov.
Article in English | MEDLINE | ID: mdl-16840536

ABSTRACT

Mutations in the human nuclear receptor, DAX1, cause X-linked adrenal hypoplasia congenita (AHC). We report the isolation and characterization of a DAX1 homolog, dax1, in zebrafish. The dax1 cDNA encodes a protein of 264 amino acids, including the conserved carboxy-terminal ligand binding-like motif; but the amino-terminal region lacks the unusual repeats of the DNA binding-like domain in mammals. Genomic sequence analysis indicates that the dax1 gene structure is conserved also. Whole-mount in situ hybridization revealed the onset of dax1 expression in the developing hypothalamus at approximately 26 h post fertilization (hpf). Later, at about 28 hpf, a novel expression domain for dax1 appeared in the trunk. This bilateral dax1-expressing structure was located immediately above the yolk sac, between the otic vesicle and the pronephros. Interestingly, weak and transient expression of dax1 was observed in the interrenal glands (adrenal cortical equivalents) at approximately 31 hpf. This gene was also expressed in the liver after 3 dpf in the zebrafish larvae. Disruption of dax1 function by morpholino oligonucleotides (MO) down-regulated expression of steroidogenic genes, cyp11a and star, and led to severe phenotypes similar to ff1b (SF1) MO-injected embryos. Injection of dax1 MO did not affect ff1b expression, whereas ff1b MO abolished dax1 expression in the interrenal organ. Based on these results, we propose that dax1 is the mammalian DAX1 ortholog, functions downstream of ff1b in the regulatory cascades, and is required for normal development and function of the zebrafish interrenal organ.


Subject(s)
Adrenal Cortex/embryology , DNA-Binding Proteins/physiology , Interrenal Gland/embryology , Receptors, Retinoic Acid/physiology , Repressor Proteins/physiology , Zebrafish/embryology , Amino Acid Sequence , Animals , Base Sequence , DAX-1 Orphan Nuclear Receptor , DNA-Binding Proteins/antagonists & inhibitors , DNA-Binding Proteins/metabolism , Embryo, Nonmammalian , Embryonic Development/physiology , Molecular Sequence Data , Oligonucleotides, Antisense/pharmacology , Phylogeny , Receptors, Retinoic Acid/antagonists & inhibitors , Receptors, Retinoic Acid/metabolism , Repressor Proteins/antagonists & inhibitors , Repressor Proteins/metabolism , Sequence Homology, Amino Acid , Transcription Factors/physiology , Water-Electrolyte Balance/physiology , Zebrafish Proteins/physiology
10.
Gen Comp Endocrinol ; 143(2): 99-103, 2005 Sep 01.
Article in English | MEDLINE | ID: mdl-16061067

ABSTRACT

ACTH and ACTH receptor-like molecules were found at the examined stages of development (2, 4, 8, 12, 18, and 24 days post-hatching) in yolk sac, pronephros tubules, interrenal tissue, thymus, liver, spleen, cardinal veins, and skin of the teleost fish Dicentrarchus labrax. ACTH and the related receptor-like molecules show a similar distribution. LPS treatment at two different stages (8 and 24 days post-hatching) provoked both a release and an induction of ACTH-like molecules, suggesting an important role of this peptide to control the modifications in body homeostasis during the first period of the sea bass' life, i.e., 30 days post-hatching, before the lymphoid cells have reached complete maturation.


Subject(s)
Adrenocorticotropic Hormone/metabolism , Bass/immunology , Interrenal Gland/immunology , Kidney/immunology , Lipopolysaccharides/immunology , Receptors, Corticotropin/metabolism , Adaptation, Physiological/immunology , Animals , Bass/embryology , Bass/metabolism , Homeostasis/immunology , Immune System/embryology , Immune System/immunology , Immune System/metabolism , Immunohistochemistry , Interrenal Gland/embryology , Interrenal Gland/metabolism , Kidney/embryology , Kidney/metabolism , Liver/embryology , Liver/immunology , Liver/metabolism , Neurosecretory Systems/embryology , Neurosecretory Systems/immunology , Neurosecretory Systems/metabolism , Signal Transduction/immunology , Thymus Gland/embryology , Thymus Gland/immunology , Thymus Gland/metabolism , Tissue Distribution , Yolk Sac/immunology , Yolk Sac/metabolism
11.
J Endocrinol ; 157(1): 127-37, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9614366

ABSTRACT

Whole-body levels of ACTH, alpha-MSH and cortisol in eggs and larvae of the common carp (Cyprinus carpio) were determined periodically up until 168 h after fertilisation. ACTH, alpha-MSH and cortisol immunoreactivity was detected in unfertilised eggs, and endogenous production of ACTH and alpha-MSH was observed 24 h after fertilisation and that of cortisol 36 h after fertilisation. ACTH immunoreactivity reached peak levels before hatching (56-72 h after fertilisation) and remained relatively stable thereafter, while alpha-MSH immunoreactivity started to increase after hatching. At 36 h after fertilisation, whole-body cortisol levels increased rapidly reaching peak levels at the end of hatching (72 h after fertilisation), remaining stable until the end of the experiment. From 50 h after fertilisation onwards, embryos and larvae increased their whole-body cortisol levels when subjected to handling (mechanical pressure during egg stage or netting during the larval stage). It is concluded that the pituitary-interrenal axis in carp is fully functional at the time of hatching. No indications of a stress non-responsive period after hatching were observed. To characterise ACTH and alpha-MSH immunoreactivities in carp larvae, whole-body homogenates were analysed by HPLC, with pituitary homogenates of adult carp serving as a reference. ACTH and alpha-MSH immunoreactivity in carp larvae homogenates consisted of three and two products respectively. HPLC of adult carp pituitaries revealed the presence of two ACTH immunoreactive products, which may represent a phosphorylated and a non-phosphorylated ACTH variant, while the three alpha-MSH peaks most likely represent des-acetylated, mono-acetylated and di-acetylated alpha-MSH, the latter being the predominant form. In carp larvae, however, one of the ACTH immunoreactive products co-eluted with the non-phosphorylated ACTH, while the two alpha-MSH products identified co-eluted with des-acetylated and mono-acetylated alpha-MSH, indicating that POMC processing at this stage of development is different from prohormone processing in adult fish.


Subject(s)
Adrenocorticotropic Hormone/metabolism , Carps/embryology , Hydrocortisone/metabolism , Interrenal Gland/embryology , Pituitary Gland/embryology , Stress, Physiological/physiopathology , Adrenocorticotropic Hormone/analysis , Animals , Binding, Competitive , Chromatography, High Pressure Liquid , Hydrocortisone/analysis , Immunohistochemistry , Interrenal Gland/metabolism , Pituitary Gland/metabolism , Radioimmunoassay , Statistics, Nonparametric , alpha-MSH/analysis , alpha-MSH/metabolism
12.
Biol Reprod ; 46(3): 458-63, 1992 Mar.
Article in English | MEDLINE | ID: mdl-1617018

ABSTRACT

Female sex determination can be induced in embryonic red-eared slider turtles (Trachemys scripta) by exogenous estrogen, as well as by incubation at warm temperature. In the present study, estrogen target areas were identified in embryos before (stage 15), during (stage 18), and after (stage 22) the critical period for sex determination. Both hyperfilm and emulsion autoradiography were used to localize tritium accumulation after the injection of radiolabeled 17 beta-estradiol. Site-specific tritium-labelling was found at all stages, notably in the mesonephros at stage 15, in the mesonephros and oviduct at stage 18, and in the mesonephros, oviduct, and the interrenal gland at stage 22. Few if any cells in the gonad were tritium-labeled at any stage. The large number of estrogen-concentrating cells in the mesonephros and interrenal and the lack of binding to gonadal tissues indicates that estrogen action on gonadal differentiation during the period of sex determination may be indirect.


Subject(s)
Estradiol/metabolism , Sex Determination Analysis , Turtles/embryology , Animals , Autoradiography , Gonads/embryology , Gonads/metabolism , Interrenal Gland/embryology , Interrenal Gland/metabolism , Mesonephros/metabolism , Oviducts/embryology , Oviducts/metabolism , Temperature , Time Factors , Tissue Distribution , Tritium
14.
Arch Anat Histol Embryol ; 68: 93-106, 1985.
Article in English | MEDLINE | ID: mdl-3024565

ABSTRACT

The ultrastructural features of the interrenal cells have been studied in 20-days-old chick embryos following the administration of adrenocorticotropin (ACTH), on the 8th or 15th day of incubation. The interrenal cells of normal embryos contain more SER than RER, mitochondria with tubular cristae, lipid droplets and Golgi complex. Adjacent cells had numerous regions of pentalaminar fusion and intermediate junctions. Neither a precise organization of medullary tissue in relation to interrenal tissue nor any structural differences between interrenal cells were found. Changes in the fine structure of the interrenal cells following ACTH treatment were extensive. The organelles that are known to be involved in the biosynthesis of steroids displayed structural modifications. These were mainly SER, mitochondria, and lipid droplets. Changes were also noticeable in the Golgi complexes, membrane-bound dense bodies (especially in 15 days treated embryos). These results indicate the well developed organelles in the interrenal cells of 20-days-old embryos, and their capacity to respond to ACTH stimulation as early as the 8th day of embryogenesis.


Subject(s)
Adrenal Glands/ultrastructure , Adrenocorticotropic Hormone/pharmacology , Chick Embryo/cytology , Interrenal Gland/ultrastructure , Animals , Cell Membrane/ultrastructure , Cell Nucleus/ultrastructure , Coated Pits, Cell-Membrane/ultrastructure , Cytoplasm/ultrastructure , Endoplasmic Reticulum/ultrastructure , Golgi Apparatus/ultrastructure , Interrenal Gland/drug effects , Interrenal Gland/embryology , Microscopy, Electron , Mitochondria/ultrastructure , Time Factors
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