Muscle afferent activity modulates bioassayable growth hormone in human plasma

Immunoassayable and bioassayable growth hormone responses to vibration-induced activation of muscle spindle afferents of the soleus (Sol) or tibialis anterior (TA) muscles were studied in 10 men. Subjects were supine while a 10-min vibration stimulus (100 Hz; 1.5-mm amplitude) was applied to the muscle, with each of the muscles tested on separate days. Blood samples were collected before, during, immediately after, and after 5 and 10 min of vibration. Plasma growth hormone concentrations were determined by radioimmunoassay (IGH) for all sampling periods and by bioassay (BGH; measurement of tibial epiphysial cartilage growth in hypophysectomized rats) for samples obtained before and immediately after vibration. Plasma IGH concentrations were similar at all time points during the Sol or TA experiments. After 10 min of muscle vibration, mean plasma BGH was elevated 94% [1,216 ± 148 (SD) to 2,362 ± 487 μg/l; P = 0.0001] for TA and decreased 22% (1,358 ± 155 to 1,058 ± 311 μg/l; P = 0.09) for Sol. These data demonstrate that activation of TA muscle spindle afferents increases circulating BGH but not IGH. The absence of a similar vibration-induced BGH response for the Sol indicates a differential regulation of BGH release by these two predominantly slow muscles, perhaps related to their respective flexor and extensor functions. These data indicate that a muscle afferent-pituitary axis modulates the release of BGH, but not IGH, from the pituitary in humans and that this axis is muscle specific, similar to that observed in rats.

Mammalian cartilage synthesizes both proteoglycan and non-proteoglycan forms of type IX collagen

Bovine epiphysial cartilage synthesizes both proteoglycan (PG) and non-PG forms of type IX collagen in a ratio of approx. 2:1. The PG form with its attached glycosaminoglycan on the alpha 2(IX) chain is the major form in the medium, whereas both forms are found in the tissue. The results are discussed with regard to cartilage matrix organization.

Inhibitory effect of octreotide on growth hormone-induced IGF-I generation and organ growth in hypophysectomized rats

We investigated the effect of a somatostatin analogue octreotide (SMS) on the stimulatory effect of recombinant human growth hormone (hGH) on insulin-like growth factor I (IGF-I) generation and growth in hypophysectomized rats. Two weeks after hypophysectomy, treatment was given for 11 days with either saline, SMS (100 micrograms/day), hGH (140 micrograms/day), or hGH plus SMS. Compared with saline-injected controls, hGH stimulated body weight gain [1.1 +/- 0.7 vs. 40.9 +/- 0.8 (SE) g, P less than 0.001] and width of epiphysial cartilage (138.0 +/- 4.5 vs. 356 +/- 3.8 microns, P less than 0.001). Combined treatment with hGH and SMS significantly reduced both body weight gain (29.1 +/- 2.5 g, P less than 0.001) and width of epiphysial cartilage (315.3 +/- 5.8 microns, P less than 0.001) compared with the effects of hGH alone. During 11 days of hGH treatment, serum IGF-I increased from 22 +/- 5 to 1,288 +/- 92 micrograms/l (P less than 0.001) but increased only 40% (513 +/- 71 vs. 1,288 +/- 92 micrograms/l, P less than 0.001) when SMS was given in combination with hGH. In gastrocnemius muscle, heart, and lung, but not in liver, kidney, and brain, SMS suppressed organ weight increase when given both with and without hGH substitution. Thymus and gastrointestinal tract weight were significantly reduced in the group receiving SMS alone and tended to be reduced in the hGH-substituted group given SMS as well. Tissue IGF-I was increased in liver, lung, kidney, and heart with hGH treatment (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Hormone receptors in the epiphysial cartilage

ABSTRACT In order to assess which hormones may exert direct effects on skeletal growth at the epiphysial growth plate, the specific binding of hormones to the epiphysial cartilage of growing dogs and rabbits was studied. Membrane fractions obtained by centrifugation of homogenates prepared from dog and rabbit growth plate cartilage at 600, 15 000 and 105 000 g showed significant specific binding of serum insulin-like activity and insulin. Binding of growth hormone and prolactin by the three membrane fractions was negligible. Saturable binding sites for triiodothyronine could be demonstrated in nuclei from the dog growth plate. Nuclear binding showed an apparent Kd of 11 ±3·6 nmol/l and a maximum binding capacity of 4·1 ± 1·6 pmol/mg DNA, a level comparable to dog liver. Using a viable chondrocyte suspension prepared from dog epiphysial cartilage, specific steroid binding in the cells could be demonstrated for [3H]dexamethasone but not 17α-methyltrienolone, oestradiol-17β or 1α,25-di-hydroxycholecalciferol. Scatchard analysis of dexamethasone binding showed high affinity binding sites having a Kd of 1·2 ± 0·35 nmol/l and a capacity of 1700 sites/cell, and a low affinity binding with a Kd of 109 ± 57 nmol/l and a capacity of 24 000 sites/cell. Steroid competition for the specific binding showed the following sequence of affinity: dexamethasone > corticosterone > 11-deoxycortisol > testosterone > oestradiol-17β. Although all of the hormones examined except prolactin have well-established physiological effects on skeletal growth, our present results suggest that some of the hormonal effects observed in intact animals are secondary and do not involve receptor–hormone interaction in cartilage as such. J. Endocr. (1984) 103, 125–131

Adrenal/parathyroid regulation of DNA, collagen and protein synthesis in rat epiphysial cartilage and bone

ABSTRACT Young, growing rats which had been chronically (2 weeks) adrenalectomized or parathyroidectomized were used to define the roles of the adrenal and parathyroid glands on the maintenance of normal circadian rhythms of DNA, collagen and non-collagen protein synthesis in the skeleton. The animals were conditioned to food being available ad libitum and to 12 h light: 12 h darkness (lights on from 08.00 to 20.00 h). The pace of DNA, collagen and non-collagen protein synthesis in different regions of the tibia (tibial growth cartilage, metaphysial bone and diaphysial bone) was measured by the in-vivo incorporation of tritiated thymidine (1 h) and radioactive proline (48 h). In intact rats there were no regional differences in the phasing of the circadian profiles; peak DNA and non-collagen protein synthesis occurred at the onset of the dark period while peak collagen synthesis occurred during the middle of the period of light. Adrenalectomy selectively abolished the regional DNA synthesis rhythms without altering the phases of the serum Ca and phosphorus (P) rhythms, which peak at mid-day and at the onset of darkness respectively. Parathyroidectomy abolished the regional rhythms for collagen and non-collagen protein synthesis and serum Ca rhythms, without altering the phase of the serum P and corticosterone rhythms. Dietary Ca-lactate supplements, which raised serum Ca levels towards normal in parathyroidectomized rats, were able to correct serum corticosterone values but did not normalize bone collagen and non-collagen protein synthesis values. These data indicate that the adrenal rhythm governs the proliferative activities of bone and cartilage cells, and that parathyroid hormone is essential to maintain normal collagen and non-collagen protein synthesis rhythms. J. Endocr. (1984) 103, 49–57

Radioimmunoassay of the 148-kilodalton cartilage protein. Distribution of the protein among bovine tissues

A sensitive and specific radioimmunoassay for the 148 kDa cartilage protein (previously referred to as ‘cartilage matrix protein’) was developed. The protein is insoluble in conventional buffers and was therefore dissolved in sodium dodecyl sulphate. Excess sodium dodecyl sulphate was bound in mixed micelles with Triton X-100 before assay. Antibodies raised against the 148 kDa cartilage protein did not react with other cartilage macromolecules. Neither did they react with guanidinium chloride extracts of a number of non-cartilaginous tissues. The protein, then, is unique for cartilage. It was demonstrated in tracheal, nasal-septum, xiphisternal, auricular and epiphysial cartilage. Surprisingly, the protein was detected neither in extracts of articular cartilage nor in extracts of the anulus fibrosus or the nucleus pulposus of the intervertebral disc.

Active extrusion of Ca2+ from epiphysial chondrocytes of normal and rachitic chickens

Chondrocytes isolated from the epiphysial cartilage of chickens were exposed to either the ionophore A23187 or KCN, in the presence of 0.4 mM-extracellular Ca2+. This treatment elicits a prompt release of cell Ca2+, which can be measured as net cation efflux by a highly sensitive Ca2+-selective electrode system. Pre-exposure of chondrocytes to the metabolic inhibitors 2-deoxy-D-glucose or oligomycin causes a parallel decrease in both cell ATP and ionophore-mediated Ca2+ extrusion. Conversely, substitution of extracellular Na+ with choline produces only a very small decrease in the rate of Ca2+ efflux. This indicates that the ionophore A23187 and cyanide induce the mobilization of intracellular Ca2+, which is then extruded from the cell mainly by an ATP-driven pump system. Chondrocytes isolated from the cartilage of rachitic chickens also exhibit the capacity for extruding Ca2+ by the same mechanism, with an efficiency comparable with that of their normal counterpart.