Mammalian GH Research Articles

The Role of the hGH Locus Control Region in Somatotrope Restriction of hGH-N Gene Expression

Expression of mammalian GH is normally restricted to somatotropes and somatolactotropes (somatotrope lineages) in the anterior pituitary. The basis for this restriction remains incompletely understood. Recent studies indicate that deoxyribonuclease I hypersensitive site I (HSI) of the hGH locus control region, located at -14.5 kb relative to the hGH-N promoter, acts as a potent long-range enhancer of hGH-N transcription. Here we report that HSI is also critical to somatotrope-restriction of hGH-N expression. Loss of HSI activity, either by direct inactivation of HSI or by interference with HSI-dependent downstream events, results in a relaxation of hGH-N cell-type specification with expansion of hGH-N expression to the full spectrum of Pit-1 positive pituitary cell types. These findings expand the defined roles for HSI of the hGH locus control region to include somatotrope lineage restriction as well as transcriptional enhancement of hGH-N gene expression.

CDNA cloning of growth hormone from giant panda ( Ailuropoda melanoleuca) and its expression in Escherichia coli

A cDNA encoding Ailuropoda melanoleuca growth hormone ( AmGH) was isolated from pituitary total RNA using RT-PCR and expressed in Escherichia coli. This is the first report of a GH nucleotide and amino acid (aa) sequence from giant panda. The open reading frame of AmGH (651 bp) encodes a precursor of 216 aa comprising a 26 aa signal peptide and a 190 aa mature protein with four cysteine residues similar to the typical primary structure of mammalian GH precursor. AmGH shares a high degree of identity (54–98.9%) with that of mammals, birds and amphibians, but a very low identity with bony fish GH (only 20–30%). The mature AmGH exhibits striking similarity to that of putative ancestral GH with a difference of only two residues, indicating a very slow basal rate of molecular evolution. The DNA fragment encoding mature AmGH was then subcloned into the pGEX-4T-1 expression vector and highly expressed in E. coli host BL21 with IPTG induction. The expressed proteins fused to GST were found to be sequestered into inclusion bodies and therefore the NaOH method was employed to solubilize the inclusion bodies; the proteins were further purified by Glutathione Sepharose 4B affinity chromatography. The production and purification of GST- AmGH reported here provide a basis for further studies on the biological activity of AmGH.

Toxic activity from cultures of Karlodinium micrum (= Gyrodinium galatheanum) (Dinophyceae)—a dinoflagellate associated with fish mortalities in an estuarine aquaculture facility

The goal of this study was to test for, and partially characterize, toxic activity associated with the dinoflagellate Karlodinium micrum. Since 1996, three fish kill events associated with blooms of K. micrum have occurred at HyRock Fish Farm, an estuarine pond aquaculture facility raising hybrid striped bass on the Chesapeake Bay, MD, USA. Using an assay based on the lysis of rainbow trout erythrocytes, cultures of a Chesapeake Bay isolate of K. micrum have been shown to produce toxic substances which are released upon cell disturbance or damage. The LC 50 for hemolysis of a sonicated cell suspension was 2.4×10 4 cells ml −1, well within the range of cell concentrations observed associated with fish kills. The toxic activity from K. micrum cells and culture filtrates was traced to two distinct fractions that co-elute with polar lipids. The LC 50 for hemolysis of the larger of these two fractions (Tox A) was 284 ng ml −1 while the LC 50 of the second, smaller, fraction (Tox B) was 600 ng ml −1. For comparison, the LC 50 for the standard hemolysin saponin was 3203 ng ml −1. At concentrations of 800 and 2000 ng ml −1, respectively, Tox A was further shown to be ichthyotoxic to zebrafish ( Danio rerio) larvae (80% mortality), and cytotoxic to a mammalian GH(4)C(1) cell line (100% LDH release). At a concentration of 600 ng ml −1 Tox B was shown to be cytotoxic to a mammalian GH(4)C(1) cell line (>30% LDH release), but not ichthyotoxic to zebrafish ( D. rerio) larvae up to a concentration of 250 ng ml −1. Although treatment with either algicidal copper or potassium permanganate caused significant lysis of K. micrum cells (>70%), toxic activity was released after treatment with copper and eliminated following treatment with potassium permanganate. This observation in cultures is consistent with observations made at HyRock Fish Farm where significantly higher mortality was observed following treatment of a K. micrum bloom with copper sulfate compared to treatment with potassium permanganate. This study represents the first direct evidence of the toxicity of K. micrum isolated from the Chesapeake Bay.

Recombinant bovine growth hormone treatment of tilapia: growth response, metabolic clearance, receptor binding and immunoglobulin production

Experiments were performed to examine the growth-promoting effects of recombinant bovine growth hormone (rbGH) in the euryhaline tilapia ( Oreochromis mossambicus). A radioreceptor assay using a crude membrane preparation of tilapia liver revealed that rbGH was about 100-fold less potent than native tilapia GH (tGH) in displacing 125I-labeled tGH. Bovine prolactin (bPRL) was equipotent to bovine GH indicating that the GH receptor of tilapia does not distinguish mammalian GH from mammalian PRL. When juvenile tilapia, weighing 1 g, were maintained at 28 °C and received intraperitoneal injection of rbGH at doses of 0.1, 1 or 10 μg/g weekly for 8 weeks, no significant effect on growth was observed. A second experiment examined weekly doses ranging from 1 to 50 μg/g for 16 weeks, using 1 g fish maintained at 23 °C. rbGH (50 μg/g) significantly increased growth after 14 and 16 weeks, although the growth rate was significantly less than those held at 28 °C. More pronounced growth-promoting effects were observed, however, when fish weighing 5 g and held at 29 °C were injected with rbGH at doses of 100 and 1000 μg/g once a week for 4 weeks. A single injection of a sustained-release formulation of rbGH (Posilac®, 100 and 1000 μg/g) also elicited growth-promoting effects in fish weighing 4 g and kept at 29 °C. Treatment with rbGH, Posilac® or bovine serum albumin (BSA) elicited significant increases in plasma levels of immunoglobulin (IgM) in a dose-dependent manner. By contrast, there was no change in plasma levels of lysozyme activity in rbGH- or Posilac®-injected fish compared with controls. An uptake and clearance study confirmed a slower decline in circulating levels of rbGH following Posilac® injection compared with rbGH in saline. There was no change in plasma concentration of tGH after rbGH treatment, indicating that GH secretion from the tilapia pituitary was unaffected by high plasma levels of rbGH. The relative refractoriness of juvenile tilapia to growth-promoting effects of rbGH compared with that of other species may be due to the specific nature of the tGH receptor in recognizing the homologous hormone.

Isolated GH deficiency with dominant inheritance: new mutations, new insights.

Autosomal dominantly inherited isolated GH deficiency is caused by mutations of GH-1 that alter the normal structure of GH. We studied 16 familial cases and 1 sporadic case with isolated GH deficiency type II from 1 Dutch and 4 German families by direct sequencing of PCR-amplified genomic DNA and ectopic transcript analysis of lymphocyte mRNA. In addition, the clinical data of the affected individuals were analyzed. Two previously reported mutations and 1 novel splice site mutation in intron III of GH-1 (+1G to C and +1G to A; new, +2T to C) were detected that cause exon 3 skipping. We also discovered a novel G6191 to T missense mutation in exon 4 of GH-1 that changes valine 110, which is highly conserved in mammalian and several nonmammalian GH, to phenylalanine. Splicing of the primary RNA transcript was not affected by this mutation, which is very likely to alter the normal GH structure at the protein level. The onset of growth failure was earlier, and the degree was more severe in affected children with GH-1 splice site mutations compared with those in children with the GH-1 missense mutation. In addition, the severity of the phenotype was variable, even within the same family. The age at diagnosis was between 0.8-9.6 yr (median, 5.1 yr); height at diagnosis was between -2.5 and -8.1 SD score (median, -4.0). Most of the children were lean at diagnosis, with a body mass index ranging from -1.7 to +3.3 SD score (median, -0.5). The 5 affected adults had final heights between -1.8 and -4.5 SD score (median, -3.6), centripetal obesity, and muscular hypotrophy. Before therapy, IGF-I and IGF-binding protein-3 serum levels of all affected children were severely diminished (<<5th percentiles for age). The maximum GH peak in a total of 25 stimulation tests was between 0.1-5.0 microg/liter (median, 0.9), indicating severe GH deficiency. The height of the adenohypophysis studied by magnetic resonance imaging was normal in 2 affected children and mildly decreased in 2 others. Substitution with GH resulted in good catch-up growth in all treated children. Children with severe GH and IGF-I deficiencies, but normal size of the adenohypophysis should be examined for GH-1 splice site and missense mutations. The observed discrepancy between the very homogeneous hormone data proving severe GH and IGF-I deficiencies and the high variability of growth failure even within the same family suggests that the onset and predominance of GH-dependent growth during infancy are individually different and modified by as yet unknown factors.

A beta-turn endocytic code is required for optimal internalization of the growth hormone receptor but not for alpha-adaptin association.

Intracellular trafficking of GH and its receptor, more particularly the chicken GH receptor (cGHR), was examined in COS-7 cells using biochemical and structural studies. Internalization of radioactive GH by the cGHR is reduced as compared with the rat GHR. On the contrary, activation of gene transcription through Janus kinase-2 was similar for both species. Secondary structures of the cytoplasmic domain of chicken and rat GHR were compared, since beta-turns were reported as internalization signals. The substitution of Pro335-Asp335, present in mammalian GH receptors, with Thr307-Gln308 in the cGHR leads to the loss of a beta-turn within a conserved cytoplasmic region. Mutational analysis indicated that the lower rate of internalization of cGHR, as compared with mammalian GHR, was due to this motif. Our data further show that alpha-adaptin, a subunit of adaptor protein AP-2, associates with the GHR upon hormone stimulation. The clathrin-coated pit pathway therefore seems to be involved in the endocytosis of cGHR, as AP-2 is known to intervene in the recruitment of receptors to these pits. Interaction with alpha-adaptin may occur through a common epitope of the chicken and mammalian GHR, since receptors from both species bind similar amounts of alpha-adaptin; alternatively, two different epitopes with similar affinity may be involved. Therefore, not alpha-adaptin but an uncharacterized factor, presumably interacting with the identified beta-turn endocytic code, is responsible for the difference in internalization kinetics. Finally, the present study illustrates that functional amino acid motifs of receptors can be derived from comparative studies.

Structure and functional analysis of a tilapia (Oreochromis mossambicus) growth hormone gene: activation and repression by pituitary transcription factor Pit-1.

A gene encoding the Tilapia mossambica (Oreochromis mossambicus) growth hormone (tiGH) was isolated and sequenced. The gene spans 5.6 kb, including 3.7 kb of 5' and 0.2 kb of 3' flanking sequences and a 1.7-kb transcription unit comprised of six exons and five introns. The gene and the 5' flanking region contain several potential binding sites for Pit-1, a key transcription activator of mammalian GH genes. One of these (-57/-42) is highly conserved in fish GH genes. It activates transcription in pituitary cells and binds Pit-1. Transfection of luciferase reporter plasmids containing either the -3602/+19 tiGH sequence or one of its 5' deletion mutants (-2863/, -1292/, and -463/+19) resulted in strong activity in Pit-1-producing rat pituitary GC cells. A dose-dependent activation of the tiGH promoter was achieved in nonpituitary fish EPC and monkey COS cells cotransfected with a rat Pit-1 expression vector, demonstrating the crucial role played by Pit-1 as an activator of the tiGH gene. Fusion of the tiGH promoter with the beta-galactosidase gene led to transient expression specifically in the nervous system of microinjected zebrafish embryos. The activity of the tiGH promoter in GC and EPC cells was strongly repressed by extending its 3' end from +19 to +40, a sequence in which a Pit-1-binding site was identified using gel retardation assays. Point mutations of the site that suppressed Pit-1 binding in vitro restored full tiGH promoter activity. Thus, a Pit-1-binding site located in the 5' untranslated region mediates Pit-1-dependent repression of the tiGH gene.

A dog growth hormone cDNA codes for a mature protein identical to pig growth hormone

Although methods for purification of dog ( Canis familiaris) growth hormone (cfGH) were described in the late Sixties, the cloning of its cDNA has not been achieved until now. In order to clone the cfGH cDNA, we capitalized on the high degree of nucleotide sequence conservation among mammalian G H genes to design a pair of consensus oligodeoxyri bonucleotide primers. With these, and starting with dog pituitary gland total RNA, we specifically amplified the cfGH cDNA using the reverse transcription-polymerase chain reaction. Its coding sequence (651 bp), as well as its 3' untranslated region (101 bp), resemble those of a typical mammalian GH cDNA. Interestingly, its encoded mature protei is identical to pig growth hormone (pGH).

Sequence of a cDNA encoding horse growth hormone

A cDNA encoding horse growth hormone (ecGH) was isolated and sequenced. The coding sequence resembles a typical mammalian GH pre-protein and contains a 3' untranslated region of 101 nucleotides carrying two contiguous polyadenylation signals.

Pit-1/GH factor-1 involvement in the gene expression of somatolactin.

A presumed pituitary hormone, somatolactin (SL), belonging to the GH/PRL family, is produced in the intermediate lobe of teleost pituitary. The pituitary-specific transcription factor, Pit-1, is indispensable not only for the expression of mammalian GH and PRL genes, but for the development of GH- and PRL-producing cells present in the anterior lobe of the pituitary as well. Thus, in this study, examination was made of the possible involvement of Pit-1 in the intermediate lobe-specific expression of the SL gene. Using antibodies against chum salmon Pit-1, the presence of the 40-kilodalton Pit-1 protein in anterior and neurointermediate lobes of rainbow trout pituitary was demonstrated. By immunohistochemical examination, Pit-1 protein was shown to be located in the nuclei of SL-producing cells in the intermediate lobe. In experiments of cotransfection into HeLa cells, rat Pit-1 enhanced the promoter activity of SL gene, and a 0.5-kilobase upstream region from the transcriptional start site was sufficient for this enhancement. It follows from these results that Pit-1 protein may possibly be involved in SL gene expression as well as the development of SL-producing cells.

Development and validation of a highly sensitive radioimmunoassay for chinook salmon ( Oncorhynchus tshawytscha) growth hormone

This study describes the development of a highly specific and very sensitive radioimmunoassay for salmonid growth hormone. Antiserum raised against chinook ( Oncorhynchus tshawytscha) GH2, which did not recognize 125I-sPRL and 125I-sGTH (at 1:1000 initial dilution), was able to inhibit growth when injected into rainbow trout ( Oncorhynchus mykiss). 125I-sGH2, used as tracer, was not recognized by anti-sGTH or by anti-sPRL. Mammalian GH and ACTH and salmonid GTH, TSH, and PRL did not cross-react in the sGH assay. The inhibition curves for pituitary extracts and plasma from salmonids were parallel to the salmon GH standard, whereas those from carp, tilapia, and catfish showed no significant cross reactivity. The RIA ED90 and ED50 values were 0.2 and 1.5 ng/ml, respectively. Using this RIA for measuring GH release by cultured pituitary cell we observed a strong inhibiting effect of SRIF (10 −6 M) and a stimulatory effect of hGRF (10 −6 M). This RIA allowed us also to detect daily fluctuations in the plasma GH concentration in cannulated rainbow trout.

Gene Structure Of Chum Salmon Somatolactin, A Presumed Pituitary Hormone Of The Growth Hormone/Prolactin Family

From a chum salmon genomic DNA library, clones coding for somatolactin (SL), a presumed pituitary hormone belonging to the GH/PRL family, were isolated and analyzed. The salmon SL gene was 16 kilobases (kb) in length, this being the largest among mammalian GH (approximately 2 kb) and PRL (approximately 10 kb) and fish GH (approximately 4 kb) genes. As in the case of mammalian GH/PRL genes, the salmon SL gene was comprised of five exons in contrast to rainbow trout and Atlantic salmon GH genes, which each have six exons. Upstream from the transcriptional initiation site were found one TATA box identical to those of mammalian PRL genes and four consensus sequences corresponding to the Pit-1/GHF-1 binding element which is essential to the expression of mammalian GH and/or PRL genes. The similarity in amino acid sequences and organization of the SL gene with mammalian GH/PRL genes indicates that the SL gene is quite likely produced from an ancestral gene common to GH/PRL genes by gene duplication. Based on these considerations, an evolutionary model for the GH/PRL/SL gene family was made and shows the common ancestral gene to originate not from duplication of a primordial gene but from shuffling of independent exons and a regulatory sequence.

Molecular Cloning of the Chicken Growth Hormone Receptor Complementary Deoxyribonucleic Acid: Mutation of the Gene in Sex-Linked Dwarf Chickens*

A novel complementary DNA (cDNA) encoding the chicken GH receptor was isolated from a chicken liver cDNA library, using polymerase chain reaction with primers derived from highly conserved sequences of the mammalian GH receptor. The nucleotide sequence predicts a mature protein of 592 amino acids and a 16 amino acid signal peptide that are partially homologous to the sequence reported for the rabbit (53%), rat (58%), and human (50%) GH receptors. Despite this low level of homology, a number of structural features of the GH receptor are conserved, including 7 cysteine residues in the extracellular domain and 5 in the intracellular region. Three transcripts of approximately 4.7, 4.0, and 1.0 kilobases are present on Northern blots of total RNA prepared from the livers of 35-week-old male chickens. Expression of the GH receptor was also detected in a wide range of tissues. The chicken GH receptor cDNA was then used as a probe in Southern and Northern blot analyses of DNA and RNA prepared from livers of sex-linked dwarf chickens, which have undetectable levels of hepatic GH-binding activity, in addition to other endocrine abnormalities. A restriction fragment length polymorphism was found in DNA, and an aberrantly-sized transcript was found in hepatic RNA of the dwarf chicken. These results indicate that a mutation in the GH receptor gene is responsible for the phenotype of the sex-linked dwarf chicken. This type of dwarfism resembles Laron-type dwarfism in humans, where a defect in the GH receptor gene has recently been identified. These receptor-deficient chickens should serve as a unique model system for studying the role of the GH receptor in growth and development.

Presence of specific growth hormone binding sites in rainbow trout (Oncorhynchus mykiss) tissues: Characterization of the hepatic receptor

The present work outlines the presence of specific binding for chinook salmon growth hormone (sGH) in different tissue preparations of rainbow trout. Optimal incubation conditions (pH, Tris, MgCl2) were determined. Specific binding was very sensitive to salt concentration during incubation. The specific binding reached a plateau after 15 and 25 hr of incubation at 12 and 4 degrees. At 20 degrees, specific and nonspecific binding were not stable. Specific binding dissociation was slower than association and was only partial. The binding was saturable (Bmax = 187 +/- 167 pmol), of high affinity (Ka = 2.4 +/- 0.8 10(9) M-1), and very specific for GH, properties which are in agreement with the characteristics of hormonal receptors. Sea bream and mammalian GH appeared 2- and 30-fold, respectively, less potent than cold sGH2 for displacing 125I-sGH2. Tissue preparations from ovary, testis, fat, skin, cartilage, gill, blood pellet, brain, spleen, kidney, and muscle showed significant saturable binding.

Evolutionary implications of two rainbow trout growth hormone genes

The primary structures of two rainbow trout growth hormone mRNAs (GH1 and GH2) have been deduced by direct sequencing of their respective cDNA clones and portions of the mRNA. Both GH1 and GH2 mRNA contain open reading frames comprised of 630 nucleotides and encode 210 amino acid residues of which 11 are variant. The translated regions of both mRNA are flanked by a short but rather conserved 5'-end, and a relatively long but highly diverged 3'-end. The differences at translated and 3'-untranslated regions suggest that the GH1 and GH2 mRNA originate from different loci. The GH1 and GH2 mRNA are likely transcribed from two distinct loci which were duplicated during tetraploidization of salmonid genome between 50 to 100 million years ago.The GH2 gene has been isolated and sequenced from a rainbow trout genomic library. This gene spans a region of approximately 4 kilobases. The trout GH gene is comprised of 6 exons and 5 introns, in contrast to 5 exons and 4 introns in mammals. The additional intron in the trout gene interrupts the translated regions that are analogous to the last exon of the mammalian counterpart. The alleged internally repeating sequences in mammalian GH, prolactin (Pr1) and placental lactogen (PL) are not observed in the predicted polypeptide sequence of trout GH. In addition, direct repeats that flank exons I, III and V of mammalian GH, Pr1 and PL genes are absent in trout gene. These findings indicate that the rainbow trout GH gene structure does not support the current hypothesis that internally repeated regions in GH, Pr1 and PL arose from a small primordial gene.

The complete nucleotide sequence of the growth-hormone gene from Atlantic salmon ( Salmo salar)

We report here the complete genomic nucleotide sequence for the Atlantic salmon growth-hormone gene ( asGH), including 600 bp of 5′ flanking sequences. The primary transcript (3651 nt) is significantly longer than that of the mammalian genes, mainly because of larger intron sizes, but also because the asGH gene contains an additional intron (intron 5). The coding regions of the asGH gene have been compared to the corresponding regions from rainbow trout (cDNA and genomic), coho salmon (cDNA) and chum salmon (cDNA). With the exception of the rainbow trout cDNA sequence, all results were in agreement with current classification of the four species. The results of a similar comparison with the mRNA leader and trailer regions were also consistent with current classification. Sequences upstream from the transcription start point have been compared to the corresponding regions from rainbow trout and mammalian GH gene ( maGH) upstream sequences. The results showed that the upstream sequences in the two fish species were very similar, while short stretches similar to conserved upstream sequences in the maGH genes were also found. Some of these conserved sequences are known to be involved in the specificity of expression of the mammalian genes.

Endocrine Factors Affecting Thyroid Economy of Teleost Fish

SYNOPSIS. This review examines the way in which the activity of the hypothalamo-pituitary-thyroid axis in teleost fishes is modified by pituitary, steroid and amine hormones. These factors may act at the level of the hypothalamus, the pituitary gland (thyrotrops), the thyroid gland, or the peripheral tissues, and affect thyroid hormone synthesis, release, metabolism, or degradation. With few exceptions, the studies are limited to only a few species, the results are often fragmentary and contradictory, are based on experiments in which pharmacological, rather than physiological, hormone ranges have been used, and there is too little information to establish a consistent pattern of response to endocrines other than those which are components of the hypothalamo-pituitary-thyroid axis itself. Most studies evaluating the effects of pituitary hormones on thyroid economy in fish have, of necessity, relied on mammalian preparations. Some of these ( e.g. , prolactin and the gonadotropins) elicit very different responses from those of the equivalent semipurified or purified piscine hormone, although mammalian and piscine TSH and GH respectively appear to elicit similar responses in the teleostean thyroid system. The elevation of plasma T4 levels in response to a challenge of exogenous TSH has been used in several studies as a way of evaluating the sensitivity of the thyroid to TSH; the response is modified by photoperiod, season, stage of development, estrogen and corticosteroids.

The primary structure of coho salmon growth hormone and its cDNA

Total RNA was extracted from coho salmon growth hormone (sGH) cell regions and used to synthesize double-stranded cDNA, which was inserted into a plasmid vector and used to transform Escherichia coli HB101. The total RNA was also separated according to size by electrophoresis on agarose gels and the fraction that directed the cell-free synthesis of protein in the size range of GHs of other species was isolated and used to screen the transformed colonies of E. coli. A clone containing the putative sGH cDNA was identified and its nucleotide sequence was determined. To verify that the cDNA was that of sGH, the GH cell region of coho pituitary glands was incubated in organ culture. The secreted GH was purified by HPLC and the sequence of its 42 amino-terminal amino acids was determined. Comparison of this sequence with the amino acid sequence derived from the cDNA showed that it encoded sGH. Medium containing the presumptive sGH as the only prominent protein was active in a GH radioreceptor assay that involved labeled bovine GH and pregnant mouse liver membranes: the sGH was approximately 10% as active as the bGH standard. RNA blotting analysis showed that sGH was the major species of RNA produced by the GH cell region of the salmon pituitary. The mRNA of sGH differed from those of human, rat, and bovine GH in that its 3′-untranslated region was unusually large (about 500 nucleotides) but the coding region showed significant homology with mammalian GHs and resembled them in having a strong (78%) preference for G and C in the third positions of the codons. The amino acid sequence of sGH showed 32–34% and 19–22% identical homology with mammalian GHs and prolactins, respectively. Several conserved regions between sGH and mammalian GH and PRL molecules were also revealed that could indicate conservation of structurally and/or functionally important domains. Hydropathy analysis disclosed that although sGH and the GH of a representative mammal (pig) had similar profiles in some regions, the sGH was overall more hydrophobic than the pig (p) GH. Similarities and differences were also noted in the predicted secondary structure of sGH and pGH.

Effect of mammalian growth hormone and prolactin on the growth of hypophysectomized chickens.

Body weight gain and shank-toe growth during a 26-day treatment period following hypophysectomy were 55 and 46%, respectively, of control values, but the body weight gain was unaffected and bone growth only slightly reduced when the hypophysectomized chickens were fed a low dose of corticosterone (5 ppm). Bovine growth hormone (0.5 mg GH/kg body wt/day for 18 days) enhanced body weight gain and shank-toe length increase (an estimate of bone growth) by 46 and 33%, respectively, compared to the growth of hypophysectomized chickens receiving only corticosterone. These same endpoints were increased approximately 24% after ovine growth hormone treatment in hypophysectomized chickens not receiving corticosterone. Body weight gain during 18 days of treatment with bovine prolactin (0.5 mg PRL/kg/day) was 27% greater than the value for corticosterone-treated hypophysectomized chickens, but bone growth was unaffected. The mammalian GH preparations increased heart weight of the hypophysectomized chickens (25-29%), but pectoralis muscle weight was unaffected. GH treatment enhanced thymal weights by 71% in corticosterone-treated hypophysectomized chickens, and by 93% in hypophysectomized animals not receiving corticosterone. GH had no significant effect on bursal weights, and PRL had no effect on either of these lymphoid organ weights in corticosterone-treated hypophysectomized chickens. GH increased liver and adipose tissue weights considerably more than the large increases that followed treatment of hypophysectomized chickens with corticosterone alone (69 and 126% greater, respectively), but had no effect on these endpoints in hypophysectomized chickens not receiving corticosterone. PRL also greatly increased liver and adipose tissue weights in corticosterone-treated hypophysectomized chickens (79 and 75%, respectively). These results provide evidence that mammalian GH enhances body weight gain, bone growth, and the growth of several organs in the hypophysectomized chicken. Mammalian PRL increased body weight gain, liver weight, and adipose tissue weight in corticosterone-treated hypophysectomized chickens, but did not influence bone growth or the weights of the heart, pectoralis, thymi, or bursa.

A heterologous radioimmunoassay for avian prolactin: application to the measurement of prolactin in the turkey.

A specific heterologous double-antibody radioimmunoassay has been developed to measure turkey prolactin (PRL) using a guinea pig anti-hPRL antiserum and 125I-labelled ovine PRL [125I]oPRL. Turkey pituitary prolactin and serum give parallel dose-response curves and no cross-reaction is seen with turkey growth hormone, LH or FSH, or mammalian LH, FSH, TSH, GH or placental lactogens. The RIA is accurate and precise and is sufficiently sensitive to measure PRL in all physiological situations investigated in the turkey. The RIA will measure PRL in several avian species including the chicken, duck, goose, pheasant, pheasant X chicken F1 hybrid, pigeon, quail and rock. Plasma PRL concentrations in laying and broody turkey hens were not significantly different (46.5 +/- 2.5 vs. 39.7 +/- 3.8 ng/ml) but both were significantly higher (P less than 0.001) than in non-laying turkey hens (4.6 +/- 0.7 ng/ml). Oestradiol injection into laying hens did not alter PRL levels while the same injection in non-laying hens caused a significant three-fold increase in plasma PRL levels.