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Biomedical subjects

D J Flint

Publications and source records attributed to D J Flint.

At least 91 records · Page 5Linked to original sources

Immunological approaches for manipulation of animal growth, body composition and fecundity.

The ability to manipulate certain physiological processes by using the immune system, so as to regulate endocrine secretions and/or actions is clearly possible. The dramatic effects of immunocastration and the ability to increase ovulation rates in sheep are probably the best examples. Other approaches along similar lines have produced equivocal results, the effects of immunization against somatostatin being the most notable case. Although anti-idiotypic antibody approaches to producing hormone mimics have also been shown to be attainable and, indeed, possibly involved in certain auto-immune dysfunctions of the endocrine system, to date no successful applications of this approach have been demonstrated in commercial livestock. The ability to enhance hormone action using antibodies is an extremely promising area. Its prospects probably hinge on the ability to synthesize suitable short peptides which will mimic epitopes on the hormone and so permit the development of active immunization techniques to produce polyclonal antibodies of restricted and enhancing specificity. It seems less likely that administration of hormones pre-complexed to monoclonal antibodies has any potential as a practical approach to manipulating animal productivity. All of these approaches involving active immunization suffer the same limitations: the highly variable response of individual animals and the general inability to regulate the duration of the response; a need to find suitable adjuvants to replace the almost universally used and commercially unacceptable Freund's adjuvant; and the problem of trying to generate what, in most cases, is an auto-immune response. A second group of approaches consists of attempts to use antibodies in their classical role, that is by targeting antigens or cells for destruction by the immune system. These include, for example, antibodies directed against adipose tissue or cytotoxic antibodies to specific hormones aimed at destruction of the hormone-secreting cells. Since these are passive immunization techniques, the antibodies can be assessed carefully in vitro and administered in appropriate doses. However, success in these applications is largely dependent on the inability of damaged tissues to regenerate, since retreatment is generally precluded because of the anti-immunoglobulin response induced in treated animals. Toleragenic forms of such antibodies or the use of appropriate immunosuppressants may ultimately remove this limitation. Perhaps the greatest current limitation to the use of all of these techniques in animal production systems, however, is public resistance to the use of such techniques.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Long-term reductions in GH, insulin-like growth factor-I and body weight gain in rats treated neonatally with antibodies to rat GH.

Treatment of neonatal rats on days 2-5 with antibodies against rat GH (rGH) markedly reduced body weight gain and serum concentrations of insulin-like growth factor-I for 6-8 weeks in both females and males, after which weight gain normalized without evidence of catch-up growth. There were no significant effects on serum prolactin, tri-iodothyronine or corticosterone. Testis and ovarian weights were reduced, although only in proportion to body size. In females, but not males, the treated rats, though lighter, had increased fat deposition in the parametrial depot. Pituitary weight was considerably reduced over 100 days later, as was the pituitary content of GH, but not prolactin. The response to GH-releasing factor of both male and female rats was also greatly reduced at this time. Taken together with the fact that these rGH antibodies can bind directly to somatotrophs, we propose that the long-term effects of the antibodies are induced by specific somatotroph destruction.

Animals↗

Production of anti-idiotypic antisera to rat GH antibodies capable of binding to GH receptors and increasing body weight gain in hypophysectomized rats.

Anti-idiotypic antibodies to rat GH antibodies were produced in both sheep and mice and shown to be capable of mimicking GH by inhibiting 125I-labelled ovine GH (oGH) binding to sheep liver membranes. The sheep anti-idiotypes were characterized further and shown to (1) inhibit 125I-labelled oGH binding to oGH antibodies, (2) inhibit 125I-labelled oGH binding to rat adipocytes and (3) be incapable of inhibiting the binding of either 125I-labelled ovine prolactin or 125I-labelled bovine insulin to sheep liver membranes. This indicated that the antibodies were not limited to certain species or tissues, but were hormone specific. Finally, these anti-idiotypic antibodies were also capable of stimulating an increase in body weight gain in hypophysectomized rats, suggesting that they may be functional as well as structural mimics of GH, although the increased body weight gain was not accompanied by any increase in circulating concentrations of insulin-like growth factor-I.

Animals↗

Hypoinsulinaemia in the lactating rat is caused by a decreased glycaemic stimulus to the pancreas.

An in-vitro perifusion system was devised in order to examine the secretory profiles of isolated islets of Langerhans, derived from different physiological states, when subjected to various stimuli relevant to lactation. Islets from pregnant rats secreted more insulin than did those from virgin animals; however, islets from lactating and virgin animals secreted similar amounts of insulin with all stimuli, including glucose, amino acids, cations and neurotransmitters. When virgin rats were pretreated for 5 days in vivo with GH or prolactin, insulin responses in vitro were unchanged. Cannulation of the hepatic portal vein and inferior vena cava in vivo revealed that both insulin and glucose concentrations were lower in the portal vein of the lactating rat compared with the virgin animal. It was therefore concluded that insulin concentrations are depressed during lactation as a consequence of the pancreas receiving a diminished glycaemic stimulus rather than because of any change in beta-cell sensitivity.

Animals↗

Cytotoxic antibodies to chicken adipocytes and their precursors: lack of tissue specificity.

1. Antisera raised in sheep against chicken adipocyte plasma membranes recognised adipocyte, liver and red blood cell membranes in enzyme immunoassays (EIA). 2. These antisera were cytotoxic when incubated with both adipocytes and their precursors grown in culture, and with red blood cells. 3. Adsorption of antisera with liver membranes completely abolished the response to liver and red blood cell membranes in the EIA. The response in adipocytes was reduced to only about 6% of the titre present in the unadsorbed sample. 4. Adsorption of antisera with liver membranes abolished the cytotoxic response to red blood cells and to adipocyte precursors, but some response to adipocytes still remained. 5. These results are in contrast to the rat for which adipocyte specific antisera were obtained which represented about 75% of the unadsorbed titre.

Adipose Tissue↗

Inhibition of neonatal rat growth and circulating concentrations of insulin-like growth factor-I using an antiserum to rat growth hormone.

Treatment of rats for 24 h on day 2, 10 or 20 of age with a specific antiserum to rGH (anti-(rGH], GH, bromocriptine (CB-154) or prolactin failed to influence body weight gain or serum concentrations of insulin-like growth factor-I (IGF-I). On day 28 of age, however, anti-(rGH) completely inhibited body weight gain and markedly reduced circulating IGF-I concentrations, effects which were completely prevented by exogenous ovine GH (oGH). When administered to control rats on day 28 oGH caused supranormal weight gain and serum IGF-I concentrations. These results suggested that GH does not play a significant role in growth or regulation of serum IGF-I until after day 20 of age. By contrast, when anti-(rGH) was given for 4 consecutive days beginning on day 2 of life, body weight gain was reduced within 48 h and remained so until at least 28 days of age. Tail length was also significantly reduced. The effect was due to inhibition of GH effects since serum GH concentrations were low and exogenous GH prevented the effect. Inhibition of growth during the first 14 days of life occurred in the absence of any effect on serum IGF-I although by 21 days of age serum IGF-I was considerably lower than in control rats. The prolonged reduction in growth after treatment has stopped appeared to be due to a cytotoxic effect on the pituitary gland since pituitary weight and GH but not prolactin content were significantly decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of extended lactation and diet on transferrin concentrations in rat milk.

Milk transferrin in the rat is immunochemically identical to serum transferrin. Its concentration in milk during normal lactation (10 pups for 21 d) varies biphasically, decreasing from a value of 1.5 mg/ml in colostrum to barely detectable values at d 4 and 8 of lactation, and thereafter increasing to reach values of 4 mg/ml at d 21. The effect of extended lactation on transferrin in milk was investigated in two experiments in which litters were replaced by 4-d-old litters at d 8 and 12 of lactation or at d 20 of lactation. Transferrin concentrations in milk in both experiments increased in a similar manner to reach values of 10 mg/ml at d 28 through d 36 of lactation. Serum transferrin and serum insulin and prolactin concentrations were not significantly altered in these experiments. Premature exposure of dams to older pups did not affect the pattern of milk transferrin concentrations. Milk transferrin concentrations were, however, modulated by altering the milk demand (changing litter sizes) and by restricting either the total food intake or the protein content of the diet. These restrictions led to lower transferrin concentrations.

Animals↗

Transferrin receptor activity in rat mammary epithelial cells.

The binding of 125I-transferrin to rat mammary cells isolated by collagenase and hyaluronidase digestion has been investigated. Surface binding was determined at 4 degrees C and total binding also at 4 degrees C but in the presence of 0.1% w/v saponin. KD values between 20 and 25 nM were obtained. Binding assays at 37 degrees C showed the internalisation of the receptor and the bound transferrin was occurring but also provided evidence for an impaired recycling of the receptors to the cell surface in the freshly isolated cells. No differences in total binding were observed in cells prepared at different stages of lactation with a mean value of 29 fmol transferrin bound/micrograms cellular DNA, equivalent to 180,000 receptors per cell.

Animals↗

Role of growth hormone in the adaptations of lipolysis in rat adipocytes during recovery from lactation.

Removal of the litter from lactating rats results in a fall in the rate of noradrenaline-stimulated lipolysis of adipocytes. This adaptation can be prevented by administration of growth hormone (somatotropin) to such rats and mimicked by injecting lactating rats with an antiserum to growth hormone, whereas lowering serum prolactin by injecting bromocriptine had no effect. The anti-lipolytic effect of adenosine is increased during lactation and is still increased by 2 days after litter removal. Injection of growth hormone into lactating rats decreased slightly the response to adenosine, whereas injection of growth hormone into rats after removal of their litters resulted in a much greater decrease in the response to adenosine, to that found in virgin and pregnant rats.

Adaptation, Physiological↗

Relationships between plasma hormone concentrations, udder development and the production of early mammary secretions in twin-bearing ewes on different planes of nutrition.

Ewes carrying twin fetuses were maintained during late pregnancy on a uniformly high plane (well fed), a uniformly low plane (underfed) or a low plane rising to a high plane (refed) of nutrition. The maternal plasma concentrations of hormones concerned with udder development, colostrum production and milk secretion were measured in samples obtained during the last 32 d before and the first 18 h after birth. Udder weights were derived from their linear dimensions, the yields of mammary secretions by hand milking during the first 18 h after birth and the transition from colostrum to milk was indicated by changes in the concentrations of lactose, lipid and immunoglobulin G. Underfeeding reduced prenatal udder development and colostrum accumulation and delayed the post-natal transition to milk secretion. Refeeding previously underfed ewes to a high plane during the last 5 d before birth had no effect on udder growth, but it did return the secretory functions of udder tissue to the levels observed in well-fed ewes. Nutritional effects on the plasma concentrations of insulin, placental lactogen, prolactin, cortisol and oestradiol-17 beta were small or absent. The relative changes in the plasma concentrations of growth hormone and insulin were such that the growth hormone: insulin ratio increased markedly during the last 11 d before birth in the underfed and refed ewes but not in the well-fed ewes. These differences in the growth hormone: insulin ratio were associated with greater increases in udder growth rate in the underfed and refed ewes than in the well-fed ewes. The plasma concentrations of progesterone decreased before birth in all ewes, but the decrease was delayed by underfeeding and refeeding rapidly overcame this effect. The slower progesterone withdrawal in the underfed ewes was associated with a delay in lactogenesis. It is concluded that the nutritionally induced changes in the growth hormone: insulin ratio and in the progesterone concentrations were the most likely endocrine determinants of the observed differences in the rates of mammogenesis and lactogenesis, respectively, but possible independent effects of substrate supply to the udder remain to be clarified.

Animals↗

Adipose tissue metabolism in sheep: response to season and its modulation by reproductive state.

Seasonal changes in subcutaneous adipose tissue metabolism and serum metabolite and hormone concentrations are described in virgin ewes fed a fixed amount of a cereal mixture plus hay ad libitum. Body weight, adipocyte mean cell volume, the rates of fatty acid and acylglycerol glycerol synthesis, and lipoprotein lipase activity increased from October to May and then decreased over the following five months. These changes are probably due to an increase in voluntary food intake leading to increased availability of acetate for fatty acid synthesis and also a probable rise in serum insulin concentration. Seasonal changes in adipose tissue metabolism in sheep are modulated by pregnancy and lactation, possibly mediated in part by changes in the serum insulin: growth hormone ratio. Although seasonal changes in adipose tissue metabolism are paralleled by changes in serum prolactin concentration, prolactin probably does not have a direct effect on adipose tissue metabolism.

Adipose Tissue↗

Plasma prolactin concentrations and cyclic activity in pony mares during parturition and early lactation.

Five pony mares were blood sampled during late pregnancy, foaling and early lactation. An homologous assay for horse prolactin was used to measure plasma prolactin concentrations in these samples. Regular estimates of cyclic activity were also made. Plasma prolactin concentrations rose markedly in the last week of pregnancy and remained high although variable in early lactation, before declining to basal levels by 1-2 months post partum. All mares showed a post-partum oestrus 7.0 +/- 0.9 days after parturition. One mare whose foal died shortly after birth showed a rapid decline in plasma prolactin values after death of the foal and an early oestrous period (4 days after parturition). The pattern of prolactin changes reported for the mare are in agreement with those reported for other mammalian species.

Animals↗

Inhibitory effects of bromocriptine on mammary development and function in lactating mice.

Five experiments were conducted. In the first, the plasma prolactin concentration of lactating mice was significantly reduced 3, 6 and 12 h after a single injection of 0.2 mg bromocriptine at 09.00 h, but recovered to normal levels at 24 h. A second injection at 17.00 h (i.e. total dose of 0.4 mg) completely prevented this recovery. Prolactin concentration was also reduced after the s.c. implantation of a solid pellet of bromocriptine. Once daily injections of bromocriptine for 7 days starting 1-day prepartum (early lactation group) or on day 5 of lactation (established lactation group) significantly reduced milk yield, assessed from the weight gain of the litter or estimated by a tritiated water dilution procedure. In early lactation the degree of inhibition was positively related to the size of the suckling litter (three pups, no inhibition; nine pups, 16.4% inhibition; 14 pups, 40.8% inhibition), but in established lactation an inhibition of 25-30% was observed, regardless of litter size. Twice-daily injections and s.c. implants of bromocriptine both reduced milk yield by a greater amount (approximately 45%), but in no case was secretion completely suppressed. The inhibitory effect of bromocriptine was prevented by the simultaneous administration of ovine prolactin (0.4 mg). Lactogenesis was apparently not affected by bromocriptine treatment. At the end of the early lactation treatment period, the mammary glands of bromocriptine-treated animals were significantly smaller and contained fewer cells than those of controls, but this was not the case in animals treated during established lactation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of an antiserum to rat growth hormone on lactation in the rat.

A highly specific antiserum to rat GH (anti-rGH) was used to assess the role of GH in lactation in the rat. When administered alone, anti-rGH had no effect on litter weight gain, whereas bromocriptine reduced serum prolactin concentrations and litter weight gain for up to 7 days when given on day 4 of lactation. When bromocriptine and anti-rGH were given in combination, however, litter weight gain declined even more dramatically so that pups were receiving virtually no milk 2-3 days after treatment. Daily litter exchange failed to prevent this effect. Concurrent injections of highly purified GH (prolactin contamination undetectable) prevented the dramatic decline in litter weight gain induced by combined bromocriptine and anti-rGH treatment, so that these litters grew as well as those receiving bromocriptine alone. Growth hormone did not act by influencing serum prolactin concentrations, which remained low during GH therapy. Direct effects of anti-rGH or GH on the pups (transferred through the milk) were ruled out since virtually identical results were obtained when milk yield was estimated during a 30-min suckling period after a 3-h separation of mother and pups. Lactation had virtually ceased 3 days after treatment with both bromocriptine and anti-rGH, but it could be reinitiated by a single injection of prolactin or GH, and subsequent recovery was virtually complete. The results of this study show that prolactin can maintain a full milk yield in the absence of GH, milk yield is reduced by approximately 50% in the absence of prolactin and milk yield is totally stopped in the absence of prolactin and GH.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Stimulatory and cytotoxic effects of an antiserum to adipocyte plasma membranes on adipose tissue metabolism in vitro and in vivo.

Antibodies to rat adipocyte plasma membranes raised in sheep had a dual effect in vitro; at low concentrations they mimicked the actions of insulin whilst higher concentrations inhibited glucose incorporation into lipid. The insulin-like effects of the antibody appeared to be due to direct activation of the glucose transport system since the antibodies did not bind to the insulin receptor, as judged by their inability to immunoprecipitate the receptor or to inhibit insulin binding, and antibodies were able to stimulate glucose transport in cells which had had their insulin receptors removed by trypsinization. The inhibitory effects of the antiserum were due to cytotoxicity since, in the presence of antiserum, adipocytes began to release large quantities of the intracellular enzyme, lactate dehydrogenase, and were ultimately lysed. This cytotoxic effect of the antiserum was complement-dependent since heat-inactivated antiserum or a crude immunoglobulin fraction of the serum possessed only stimulatory effects on lipid metabolism. When injected into rats for 4 days the antiserum produced gross abnormality of adipose tissue depots. Dissolution of adipocytes and massive infiltration by lymphocytes and polymorphs were evident. Preliminary observations suggest that such treatment results in long-term reduction of the number of adipocytes in internal fat depots.

Adipose Tissue↗