Localization and Regulation of the Growth Hormone Receptor and Growth Hormone‐Binding Protein in the Rat Growth Plate

Growth hormone (GH) has direct effects on the growth plate to stimulate longitudinal growth, but it is not clear which chondrocyte populations GH acts on. The dual effector theory suggests that GH would act primarily on the “stem cells.” However, staining with a GH receptor (GHR) antibody is found in all layers of the growth plate in rabbits and humans. We now have investigated the localization and regulation of GHR and the related GH binding protein (GHBP) in the rat growth plate using a sensitive immunohistochemical method involving tyramide signal amplification (TSA) and antibodies specific for GHR or GHBP. Both GHR and GHBP were shown in the germinal and proliferative chondrocytes, but most clearly in early maturing chondrocytes at the interface between proliferative and hypertrophic cells. Staining for GHR and GHBP was located in both the cytoplasm and the nucleus. Expression of GHR mRNA and GHBP mRNA in the growth plate was confirmed by reverse‐transcription polymerase chain reaction (RT‐PCR). Immunohistochemical staining for GHR and GHBP decreased with age; in 12‐week‐old normal rats, only the early maturing chondrocytes were stained. In GH‐deficient dwarf rats, staining seemed less than in normal rats, and in hypophysectomized (Hx) rats, staining for GHBP was clearly reduced. Treatment of Hx rats with thyroid hormones (T3 + T4), via subcutaneously (sc) implanted osmotic minipumps, induced little growth and induced a small layer of GHR‐positive and GHBP‐positive early maturing chondrocytes. Treatment with GH and thyroid hormones (TH) resulted in greater growth and a broader layer of GHR‐positive and GHBP‐positive cells, indistinguishable from normal rats. In contrast, dexamethasone treatment of normal rats inhibited their growth and reduced GHR and GHBP staining in the growth plate. These results show that GHR and GHBP in the growth plate are under hormonal control. The localization of GHR/GHBP suggests that in addition to actions on germinal and proliferative cells in young rats, GH also has effects on early maturing chondrocytes and may be involved in their differentiation to a fully hypertrophic chondrocyte.

[1]  P. Backeljauw,et al.  Therapy for 6.5-7.5 years with recombinant insulin-like growth factor I in children with growth hormone insensitivity syndrome: a clinical research center study. , 2001, The Journal of clinical endocrinology and metabolism.

[2]  J. Terwilliger,et al.  Roles of growth hormone and insulin-like growth factor 1 in mouse postnatal growth. , 2001, Developmental biology.

[3]  R. Baron,et al.  Bone homeostasis in growth hormone receptor-null mice is restored by IGF-I but independent of Stat5. , 2000, The Journal of clinical investigation.

[4]  E. Hunziker,et al.  Effect of Growth Hormone and Insulin-Like Growth Factor I (IGF-I) on the Expression of IGF-I Messenger Ribonucleic Acid and Peptide in Rat Tibial Growth Plate and Articular Chondrocytes in Vivo. , 2000, Endocrinology.

[5]  G. Werther,et al.  Growth hormone receptor abundance in tibial growth plates of uremic rats: GH/IGF-I treatment. , 2000, Kidney international.

[6]  Marcel Karperien,et al.  Expression of Indian Hedgehog, Parathyroid Hormone‐Related Protein, and Their Receptors in the Postnatal Growth Plate of the Rat: Evidence for a Locally Acting Growth Restraining Feedback Loop After Birth , 2000, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[7]  J. Compston,et al.  The Localization of the Functional Glucocorticoid Receptor α in Human Bone , 2000 .

[8]  J. Compston,et al.  The localization of the functional glucocorticoid receptor alpha in human bone. , 2000, The Journal of clinical endocrinology and metabolism.

[9]  D. Leroith,et al.  Normal growth and development in the absence of hepatic insulin-like growth factor I. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[10]  K. Sjögren,et al.  Liver-derived insulin-like growth factor I (IGF-I) is the principal source of IGF-I in blood but is not required for postnatal body growth in mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  R. Ross.  Truncated growth hormone receptor isoforms , 1999, Acta paediatrica (Oslo, Norway : 1992). Supplement.

[12]  J. Kopchick,et al.  Transgenic models of growth hormone action. , 1999, Annual review of nutrition.

[13]  C. Ohlsson,et al.  Dexamethasone Impairs Growth Hormone (gh)- Stimulated Growth by Suppression of Local Insulin-like Growth Factor (igf)-i Production and Expression of Gh- and Igf-i-receptor in Cultured Rat Chondrocytes* , 2022 .

[14]  C. Ohlsson,et al.  The CCAAT/enhancer-binding protein-alpha is expressed in the germinal layer of the growth plate: colocalisation with the growth hormone receptor. , 1997, The Journal of endocrinology.

[15]  Clifford J. Tabin,et al.  Regulation of Rate of Cartilage Differentiation by Indian Hedgehog and PTH-Related Protein , 1996, Science.

[16]  B. Lanske,et al.  PTH/PTHrP Receptor in Early Development and Indian Hedgehog--Regulated Bone Growth , 1996, Science.

[17]  I. Robinson,et al.  Single cell enzyme activity and proliferation in the growth plate: Effects of growth hormone , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[18]  C. Carter-Su,et al.  Dexamethasone-induced antagonism of growth hormone (GH) action by down-regulation of GH binding in 3T3-F442A fibroblasts. , 1995, Endocrinology.

[19]  C. Roberts,et al.  Localization of growth hormone receptor/binding protein messenger ribonucleic acid (mRNA) during rat fetal development: relationship to insulin-like growth factor-I mRNA. , 1995, Endocrinology.

[20]  S. Scully,et al.  Transforming growth factor‐β1 and fibroblast growth factors in rat growth plate , 1995, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  J. Wroblewski,et al.  Inhibitory effects of basic fibroblast growth factor on chondrocyte differentiation , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[22]  J. Wiegant,et al.  Ultra-sensitive FISH using peroxidase-mediated deposition of biotin- or fluorochrome tyramides. , 1995, Human molecular genetics.

[23]  H. Werner,et al.  Rat growth hormone receptor/growth hormone-binding protein mRNAs with divergent 5'-untranslated regions are expressed in a tissue-specific manner. , 1995, DNA and cell biology.

[24]  B. Bingham,et al.  One class of growth hormone (GH) receptor and binding protein messenger ribonucleic acid in rat liver, GHR1, is sexually dimorphic and regulated by GH. , 1995, Endocrinology.

[25]  I. Robinson,et al.  Steroid regulation of growth hormone (GH) receptor and GH-binding protein messenger ribonucleic acids in the rat. , 1995, Endocrinology.

[26]  C. M. Chen,et al.  Nuclear translocation and anchorage of the growth hormone receptor. , 1994, The Journal of biological chemistry.

[27]  Z. Hochberg,et al.  Differential effects of hypothyroidism on the cartilage and the osteogenic process in the mandibular condyle: recovery by growth hormone and thyroxine. , 1994, Endocrinology.

[28]  M. Lane,et al.  CCAAT/enhancer binding protein alpha is sufficient to initiate the 3T3-L1 adipocyte differentiation program. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Baron,et al.  Dexamethasone increases growth hormone receptor messenger ribonucleic acid levels in liver and growth plate. , 1994, Endocrinology.

[30]  M. Waters,et al.  Receptor-mediated nuclear translocation of growth hormone. , 1994, The Journal of biological chemistry.

[31]  M. Dattani,et al.  Inhibition of growth hormone bioactivity by recombinant human growth hormone-binding protein in the eluted stain assay system. , 1994, The Journal of endocrinology.

[32]  E. Hunziker,et al.  Differential effects of insulin-like growth factor I and growth hormone on developmental stages of rat growth plate chondrocytes in vivo. , 1994, The Journal of clinical investigation.

[33]  F. Talamantes,et al.  Expression of growth hormone-binding protein with a hydrophilic carboxyl terminus by the mouse placenta: studies in vivo and in vitro. , 1994, The Journal of endocrinology.

[34]  Icaf Robinson,et al.  Growth hormone (GH) receptors, GH binding protein and GH: an autoregulatory system? , 1993, Acta paediatrica (Oslo, Norway : 1992). Supplement.

[35]  C. Ohlsson,et al.  Endocrine regulation of longitudinal bone growth , 1993, Acta paediatrica (Oslo, Norway : 1992). Supplement.

[36]  T. Videman,et al.  Vital staining indicating cell migration towards the periphery in the growth plate. Studies of fibular heads in rabbits. , 1993, Acta orthopaedica Scandinavica.

[37]  I. Robinson,et al.  Growth hormone (GH)-binding protein in normal and GH-deficient dwarf rats. , 1992, The Journal of endocrinology.

[38]  C. Ohlsson,et al.  Growth hormone induces multiplication of the slowly cycling germinal cells of the rat tibial growth plate. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[39]  I. Robinson,et al.  Growth hormone (GH) binding protein and GH interactions in vivo in the guinea pig. , 1992, Endocrinology.

[40]  M. Waters,et al.  Cellular localization of the growth hormone binding protein in the rat. , 1992, Endocrinology.

[41]  B. Moats-Staats,et al.  Tissue-Specific Developmental Regulation of the Messenger Ribonucleic Acids Encoding the Growth Hormone Receptor and the Growth Hormone Binding Protein in Rat Fetal and Postnatal Tissues , 1992, Pediatric Research.

[42]  M. Waters,et al.  Prenatal expression of the growth hormone (GH) receptor/binding protein in the rat: a role for GH in embryonic and fetal development? , 1992, Development.

[43]  A. Herington,et al.  Tissue distribution, characterization, and regulation of messenger ribonucleic acid for growth hormone receptor and serum binding protein in the rat. , 1991, Endocrinology.

[44]  J. Bass,et al.  Passive immunization against insulin-like growth factor-I does not inhibit growth hormone-stimulated growth of dwarf rats. , 1991, Endocrinology.

[45]  H. Sadeghi,et al.  Identification of the origin of the growth hormone-binding protein in rat serum. , 1990, Molecular endocrinology.

[46]  B. Carlsson,et al.  Expression of the growth hormone-binding protein messenger RNA in the liver and extrahepatic tissues in the rat: Co-expression with the growth hormone receptor , 1990, Molecular and Cellular Endocrinology.

[47]  M. Waters,et al.  Regulation of growth hormone (GH) bioactivity by a recombinant human GH-binding protein. , 1990, Endocrinology.

[48]  R. Palmiter,et al.  Expression of insulin-like growth factor I stimulates normal somatic growth in growth hormone-deficient transgenic mice. , 1990, Endocrinology.

[49]  M. Iwamoto,et al.  Fibroblast growth factor is an inhibitor of chondrocyte terminal differentiation. , 1990, The Journal of biological chemistry.

[50]  J. Ketelslegers,et al.  Initial characterization and sexual dimorphism of serum growth hormone-binding protein in adult rats. , 1990, Endocrinology.

[51]  J. Logan,et al.  The growth hormone-binding protein in rat serum is an alternatively spliced form of the rat growth hormone receptor. , 1989, Genes & development.

[52]  R. Palmiter,et al.  Growth enhancement of transgenic mice expressing human insulin-like growth factor I. , 1988, Endocrinology.

[53]  B. Bloch,et al.  Growth hormone-deficient dwarfism in the rat: a new mutation. , 1988, The Journal of endocrinology.

[54]  M. Waters,et al.  The ontogeny of growth hormone receptors in the rabbit tibia. , 1988, Endocrinology.

[55]  William I. Wood,et al.  Growth hormone receptor and serum binding protein: purification, cloning and expression , 1987, Nature.

[56]  A. Lindahl,et al.  Mechanism of the stimulatory effect of growth hormone on longitudinal bone growth. , 1987, Endocrine reviews.

[57]  A. Lindahl,et al.  Effects of growth hormone and insulin-like growth factor-I on colony formation of rabbit epiphyseal chondrocytes at different stages of maturation. , 1987, Journal of Endocrinology.

[58]  A. Lindahl,et al.  Differential effects of growth hormone and insulin-like growth factor I on colony formation of epiphyseal chondrocytes in suspension culture in rats of different ages. , 1987, Endocrinology.

[59]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[60]  A. Dahlström,et al.  Regulation by growth hormone of number of chondrocytes containing IGF-I in rat growth plate. , 1986, Science.

[61]  M. Waters,et al.  Monoclonal antibodies to the rabbit liver growth hormone receptor: production and characterization. , 1984, Endocrinology.

[62]  J. Ketelslegers,et al.  Ontogeny of liver somatotropic and lactogenic binding sites in male and female rats. , 1983, Endocrinology.

[63]  O. Isaksson,et al.  Growth hormone stimulates the proliferation of cultured chondrocytes from rabbit ear and rat rib growth cartilage , 1983, Nature.

[64]  J. Jansson,et al.  Growth hormone stimulates longitudinal bone growth directly. , 1982, Science.

[65]  N. Kember.  Cell division in endochondral ossification. A study of cell proliferation in rat bones by the method of tritiated thymidine autoradiography. , 1960, The Journal of bone and joint surgery. British volume.