ABSTRACT

This study was designed to test the hypothesis that oestrogens inhibit growth by reducing hepatic somatomedin generation. We have attempted to deliver oestrogen preferentially to the liver by transplanting ovarian tissue into the spleen. Four groups of rats were compared: intact, ovariectomized, ovariectomized with successful ovarian transplants, and animals where due to adhesions between the transplant and body wall and/or viscera oestrogens reached the general circulation. Plasma levels of LH and FSH, uterine weights, ovarian weights and vaginal smears supported this classification of animals. Ovariectomy increased body weight, body length, bioassayable serum somatomedin levels and ³⁵SO₄ ²⁻ uptake into costal cartilage in vivo compared with intact rats and animals with adhesions. Preferentially exposing the liver to oestrogen did not suppress the increased growth, serum somatomedin activity or uptake of ³⁵SO₄ ²⁻ in vivo observed in ovariectomized animals. The results suggest that the presence of oestrogen in the general circulation is associated with growth suppression and lowered somatomedin bioactivity while the presence of oestrogen in the hepatic portal circulation has little effect on body growth. We conclude that oestrogen does not appear to inhibit growth in the rat by influencing the release of somatomedin from the liver. It also seems that serum somatomedin concentration may not reflect liver somatomedin generation but overall production throughout the body.

J. Endocr. (1984) 103, 43–47

ABSTRACT

The 'Little' mouse is characterized by a body growth rate 60% of normal due to a defect in the synthesis and storage of GH in the anterior pituitary gland. We have now investigated the effects of GH releasing factor (GRF) in these mice and in normal animals. The pituitary GH content in Little mice was only 4% of that in normal C57: +/+ mice, and was not affected by twice daily i.p. injections of human (h) GRF1-29NH₂ (0·2−2 μg) for 14 days. This treatment also had no effect on body growth. In anaesthetized normal mice, single i.v. injections of 0·1 or 2 μg hGRF1-29NH₂ released large amounts of GH into the plasma, whereas this peptide was ineffective in Little mice, whether or not they had been pretreated with GRF. Therefore, although pituitaries of Little mice contain significant amounts of GH, this pool is not releasable by GRF. This suggests that the dwarfism in Little mice may be partly due to a pituitary defect in GRF receptors or their stimulus-secretion coupling, rather than a deficiency in hypothalamic GRF.

J. Endocr. (1985) 106, 1–5

ABSTRACT

The GH responses to single i.v. injections of GH-releasing factor (GRF) in conscious male rats are highly variable. Although normal male rats show a pulsatile secretory pattern of GH with pulses occurring at intervals of 3–3·5 h, the peaks occur at different times in individual animals. We have compared the GH responses of young conscious male and female rats to multiple i.v. injections of 1 μg human (h) GRF1-29NH₂. The peak GH responses occurred 3–5 min after hGRF1-29NH₂ injection and were lower in female than in male rats. Both males and females responded uniformly to hGRF1-29NH₂ injections given 180 min apart and the GH responses became entrained with no endogenous GH pulsing. Female rats produced consistent GH peaks in response to hGRF1-29NH₂ injections at 90-min intervals, whereas male rats responded only to alternate injections, so that GH peaks occurred only every 180 min despite giving GRF every 90 min. When the frequency of hGRF1-29NH₂ administration was increased to once every 40 min female rats again responded consistently to each injection. Male rats responded intermittently, being able to respond to two injections 40 min apart, after which they became refractory to hGRF1-29NH₂. This cycle of varying sensitivity to GRF in male rats probably underlies their 3-hourly endogenous GH secretory rhythm. Female rats can respond uniformly to repeated GRF injections, consistent with their more continuous pattern of endogenous GH secretion. Introducing a pulse of 10 μg rat GH into a series of hGRF1-29NH₂ injections did not induce refractoriness to hGRF1-29NH₂, suggesting that GH does not itself desensitize the pituitary to GRF. Whether the different patterns of GH secretion in males and females result from different patterns of GRF and/or somatostatin secretion remains to be determined.

J. Endocr. (1985) 106, 281–289

This paper presents an investigation into the effects of prolonged oestrogen treatment (20 days) on basal growth and on growth stimulated by GH in hypopituitary dwarf mice. Body ³⁵SO₄ ²⁻ weight and tail length were measured during the treatment period and uptake of S04 into costal cartilage in vivo at the end of the treatment period. This study confirmed that treatment with human GH promotes a dose-dependent increase in body weight, tail length and uptake of ³⁵SO₄ ²⁻ in vivo; there was a highly significant correlation between these responses. Treatment with oestrogen alone had no significant effect on any of the parameters measured. All groups receiving combined oestrogen and human GH treatment showed a significant increase in body weight and tail length compared with animals receiving the same dose of oestrogen alone. However, the increase in body weight and tail length was significantly less in animals given the highest dose of oestrogen plus human GH than that observed in animals treated with the same dose of human GH alone. Treatment with oestrogen had no significant effect on the uptake of ³⁵SO₄ ²⁻ stimulated by human GH. Possible mechanisms for the growth-inhibiting effects of oestrogens are discussed.

ABSTRACT

A system is described for the automatic collection of small samples of blood from conscious rats. Rats bearing chronic indwelling i.v. catheters were connected via swivels to a multichannel peristaltic pump, solenoid valves and a fraction collector. A microcomputer controlled the operations involved in the removal of blood and its deposition into a fraction collector for subsequent direct radioimmunoassay for GH. Blood samples of 10–20 μl could be collected, into a total volume of 100 μl heparinized saline, from up to eight rats simultaneously every few minutes for many hours. This collection method avoided major blood loss and did not require transfusions of donor blood to maintain blood volume. Using a doublelumen cannula it was possible to inject or infuse into the animals while sampling blood. The system was used to investigate in detail the secretion of GH in conscious male rats. The 3-hourly endogenous secretory rhythm of GH was maintained for up to 44 h with episodes of GH secretion being multicomponent. Endogenous secretion was suppressed by constant i.v. infusions of somatostatin, with repetitive sampling showing in detail a rapid rebound secretion of GH after terminating the somatostatin infusions. Four injections of a fragment of GH-releasing factor, given at 3-hourly intervals, produced entrained GH responses, but the subsequent recovery of endogenous GH pulsing was delayed for up to 12 h. This method for the automatic microsampling of blood in small animals gives a very detailed description of the blood levels of hormones secreted in a highly episodic fashion, and could be widely applicable to other endocrine studies.

J. Endocr. (1986) 111, 27–35

ABSTRACT

An automatic method for repetitive microsampling of blood from conscious animals was used to obtain detailed GH secretory profiles from normal female rats, which were compared with those in males and ovariectomized females. Female rats showed a highly variable GH secretory pattern, with sustained periods of low, almost continuous secretion, followed by very rapid bursts of high amplitude and short duration, occurring mostly at night. There was no clear relationship between the pattern of GH secretion and the phase of the oestrous cycle in rats continuously sampled over a 5-day period. In ovariectomized rats, the day:night difference was maintained, though the nocturnal GH surges were larger and of longer duration than in intact females. Male rats produced multicomponent GH bursts which continued unchanged throughout the day and night. This study shows for the first time that female rats switch to a rapid, highly pulsatile pattern of GH release at night, which can only be resolved by rapid blood sampling over extended periods in conscious undisturbed animals.

J. Endocr. (1987) 114, 399–407

ABSTRACT

Growth hormone inhibits its own secretion in animals and man but the mechanism for this inhibition is unclear: both stimulation of somatostatin release and inhibition of GH-releasing factor (GRF) release have been implicated. We have now studied the GRF responsiveness of conscious male and female rats under conditions of GH feedback induced by constant infusion of exogenous human GH (hGH). Intravenous infusions of hGH (60 μg/h) were maintained for 3 to 6 h whilst serial injections of GRF(1–29)NH₂ (0·2–1 μg) were given at 45-min intervals. The GH responses were studied by assaying blood samples withdrawn at frequent intervals using an automatic blood sampling system. We have confirmed that male and female rats differ in their ability to respond to a series of GRF injections; female rats produced consistent GH responses for up to 13 consecutive GRF injections, whereas male rats showed a 3-hourly pattern of intermittent responsiveness. In female rats, multiple injections of GRF continued to elicit uniform GH responses during hGH infusions, whereas hGH infusions in male rats disturbed their intermittent pattern of responsiveness to GRF, and their regular 3-hourly cycle of refractoriness was prolonged. We suggest that this sex difference in GH feedback may be due to GH altering the pattern of endogenous somatostatin release differentially in male and female rats. Such a mechanism of GH autofeedback could be involved in the physiological control of the sexually differentiated pattern of GH secretion in the rat.

Journal of Endocrinology (1990) 126, 27–35

ABSTRACT

The in-vivo biological activity of recombinant methionyl insulin-like growth factor I (met-IGF-I) was demonstrated in hypophysectomized rats by following blood glucose after an i.v. bolus injection of met-IGF-I; a dose-dependent decrease in blood sugar was seen. Membrane transport was studied using the non-metabolizable amino acid α-aminoisobutyric acid; stimulation was obtained with the highest dose used (90 μg/rat). To test the original somatomedin hypothesis, growth studies were performed in hypophysectomized rats. Two or three doses of met-IGF-I were given with three different administration regimes (i.v. or s.c. infusion, or s.c. injections twice daily) for 6 or 8 days. Little growth-promoting activity was observed, with a significant effect on body weight gain obtained only when met-IGF-I was given continuously at the highest dose used (180 μg/day). No effect was seen on the in-vivo uptake of radioactive sulphate into cartilage. Epiphyseal cartilage width increased slightly at the highest dose of met-IGF-I, but only when the hormone was given by infusion. When 180 μg met-IGF-I/day were given by injections, a significant effect on longitudinal bone growth was obtained (90 μm above control). The levels of IGF in the serum were not measurably increased after s.c. administration of met-IGF-I, whereas after i.v. infusion, significantly raised levels were obtained at the higher dose rates (3·0 ± 0·3 and 2·8 ± 0·1 units/ml). Growth hormone was much more effective than met-IGF-I even at 50-fold lower doses.

Priming the animals with 10 mu. bovine GH/day followed by combined infusions of GH and met-IGF-I did not reveal any potentiating effects of met-IGF-I in the presence of GH. We conclude that met-IGF-I is a relatively poor growth-promoting agent when given systemically, and that somatomedins are more likely to act as local growth factors rather than as circulating mediators of the growth-promoting effects of GH.

J. Endocr. (1987) 112, 123–132

ABSTRACT

Young hypophysectomized rats were maintained with chronic indwelling i.v. cannulae attached via swivels to a multichannel pumping system programmed to deliver GH in a continuous or pulsatile pattern for several days. Continuous i.v. infusions of human GH for 5 days produced dose-dependent increases in body weight and tail length, without increasing food intake. A comparison of GH infusions by the s.c. or i.v. route showed that the direct i.v. route was threefold more effective. Pulsatile i.v. infusions of human or bovine GH at two doses (12 or 36 mu./day, eight pulses/day, 5-min duration, every 3 h) produced greater increases in body weight than continuous i.v. infusions of GH at the same daily dose. Continuous infusions of bovine GH produced a lower growth rate in the second of two consecutive 5-day treatment periods, whereas the responses to pulsatile GH did not diminish with time. Both body weight gain and long-bone growth were affected by the frequency of GH pulses; nine pulses per day were more effective than three pulses per day which in turn produced larger growth responses than one pulse per day. Keeping GH pulse frequency constant and varying pulse duration (4, 16 or 64 min) did not affect growth rates. In conclusion, long-term pulsatile i.v. infusions of GH mimic the endogenous secretory pattern, and are most effective when given at the physiologically appropriate pulse frequency.

J. Endocr. (1985) 104, 53–61

ABSTRACT

The effects of streptozotocin-induced diabetes on weight gain, bone growth and GH secretion have been studied in conscious chronically cannulated male rats. In addition to the classic diabetic symptoms (hyperphagia, polydipsia, polyuria, glycosuria and hyperglycaemia), the slow body weight gain (0·95 ± 0·5 compared with 2·63 ± 0·5 g/day in non-diabetic controls) was associated with a reduction in bone growth (from 162 ± 9 to 48 ± 4 μm/day) and a reduced pituitary GH content (from 1·5 ± 0·2 to 0·6 ± 0·06 mg/gland). Serial blood sampling during the day or overnight showed that the normal male episodic GH secretory pattern was obliterated in the diabetic animals. The constant osmotic stimulation of hyperglycaemia and high fluid turnover was reflected in a significant reduction in pituitary oxytocin and arginine vasopressin (AVP) stores. Intravenous insulin infusions (67–1340 pmol/h for 4 or 7 days) caused a large initial weight gain (> 20 g in 2 days) followed by a slower increase, and stimulated tibial bone growth (to 100 ± 16 and 126 ± 8 μm/day after 4 or 7 days respectively). Insulin infusion for 7 days also increased pituitary GH content (to 1 ± 0·15 mg/gland), and the normal episodic GH secretory pattern returned. Intravenous infusions of insulin which reduced, but did not completely normalize, blood glucose levels, allowed the resumption of growth and pulsatile GH secretion. Continuous infusion of recombinant human insulin-like growth factor-I (hIGF-I) at 1110 pmol/h for 54 h also caused a large initial rise in body weight in diabetic rats (17·1 ± 1·6 compared with 7·5 ± 2·8 g in saline-infused controls) due primarily to increased fluid retention. This effect of hIGF-I occurred without any significant changes in pituitary GH, AVP, oxytocin, blood glucose or bone growth over this short-term infusion, nor was there any obvious effect on spontaneous GH secretion, monitored over the entire infusion period. We conclude that the diabetic rat is not a good model to study growth stimulation by short-term insulin or IGF-I treatments because the insulin-like effects of these peptides obscure their specific growth-promoting activities in this model.

Journal of Endocrinology (1989) 122, 661–670