The digestion and transmission of labelled immunoglobulin G by enterocytes of the proximal distal regions of the small intestine of young rats.

Density gradient centrifugation of samples prepared from proximal gut homogenates after intra-lumenal injection of 125I-labelled IgG, was used to prepare batches of IgG fragments according to sedimentation coefficients. 2. Ultrafiltration was employed to partition the radioactivity in the vascular compartments, viscera and carcasses of rats aged 14-15 days, 2 hr after the injection of standard doses of labelled IgG into the proximal and distal regions of the small intestine. 3. Radioactive samples prepared by these methods were re-introduced into young rats by intra-cardiac injection, and the rate at which they were removed from the vascular compartment was assessed. 4. Proximal enterocytes transmitted about 39% of the IgG which had been removed from the intestine in intact form. Most of this was retained in the vascular compartment; they degraded up to about 57% of the total removed into fragments less than 1000 mol. wt. and about 4% into intermediate sized fragments. 5. Distal enterocytes degraded almost 90% of the IgG processed into fragments less than 1000 mol. wt., about 8% as fragments greater than 100,000 mol. wt. 6. Fragments, of all sizes, were cleared rapidly from the circulation into the viscera and carcass. 7. The relevance of these results to protein transmission and digestion by the rat small intestine is discussed.     

Factors influencing the uptake of [125I]polyvinyl pyrrolidone by the intestine of the young rat

1. The uptake of orally administered [(125)I]polyvinyl pyrrolidone (PVP) of mean mol. wt. 160,000 (K. 60) by the small intestine of young rats was determined 4 hr after feeding by measurement of the radioactivity remaining in the wall after flushing out the intestinal contents.2. PVP uptake was proportional to the administered dose of PVP for doses of 1 mg and below. At higher doses, uptake was no longer in linear proportion, suggesting a degree of saturation of uptake.3. Litters were reared in one of three environmental temperatures: 13, 20 and 30 degrees C, and were tested to determine the age at which PVP uptake ceased (;closure'). The results were evaluated statistically (see Appendix).4. There was no significant difference between the time of closure in animals reared at 20 and 30 degrees C, but, relative to these groups, closure was significantly delayed, by approximately 48 hr, in animals reared at 13 degrees C.5. Once closure had begun, there were no significant differences in the rate of decline in PVP uptake with age between animals reared at different environmental temperatures.6. Separation of the young rats from their mother both before and after the test-feed of PVP caused a considerable reduction in PVP uptake. Provided that the body temperature of the young rats was maintained, the main factor contributing to the reduction in PVP uptake appeared to be the absence of milk, rather than separation from the mother.     

Structural changes in the small intestine associated with the uptake of polyvinyl pyrrolidone by the young ferret, rabbit, guinea-pig, cat and chicken

1. The entry of [(125)I]polyvinyl pyrrolidone (PVP) of mean mol. wt. 160,000 (K. 60) into the epithelial cells of the small intestine has been measured in new-born animals of five species.2. The distribution along the intestine of cells capable of taking up [(125)I]PVP and the decrease and eventual cessation of uptake (closure) with increasing age have been investigated, and have been related to changes in the histological appearance of the small intestine.3. The small intestine of the ferret took up PVP readily until 33-34 days after birth. From 34 to 37 days of age PVP uptake declined sharply and disappeared completely by 40-45 days.4. In the ferret, unlike other species studied, some PVP was taken up by the duodenum. This continued for the first 4 weeks after birth. Thereafter PVP uptake gradually became confined to the terminal ileum.5. In the guinea-pig, PVP uptake was limited to the first 48 hr after birth. During this period the site of uptake was progressively restricted to the terminal ileum.6. In the rabbit, PVP could be taken up in the distal two-thirds of the small intestine for at least 20 days after birth. A decline in uptake occurred between 20 and 22 days after birth in most animals.7. Wide individual variations were seen in the kitten, but PVP uptake was seen in some animals up to 14 days after birth.8. Newly hatched chicks and chicks tested 48 hr after hatching did not take up PVP.9. Histological examination of the small intestine with the light microscope demonstrated that in all species PVP uptake was associated with the presence of vacuoles in the epithelial cells of the villus.10. In the young guinea-pig, large PAS-positive granules were seen in the macrophages of the lamina propria. These appeared to migrate through the epithelium into the intestinal lumen. The significance of this finding and its relation to macromolecular uptake remain unclear.     

The transmission of -125-I-labelled immunoglobulin G by proximal and distal regions of the small intestine of 16-day-old rats.

1. Standard doses of -125-I-labelled rat IgG were injected into the intestinal lumen of rats aged 16 days, and their sera were sampled 2 and 3 hr later. High concentration quotients were obtained after injection into the proximal small intestime, whereas very little immunoglobulin was transmitted from doses injected into the terminal 20 cm of the small intestine. 2. The villi of the terminal 18--20 cm of the small intestine of 16-day-old rats, the region from which very little transmission of IgG occurred, were lined by tall columnar absorptive cells with very larg supra-nuclear vacuoles. The extent of the terminal intestine, in which this cell type predominated in the absorptive epithelium, varied with age. The importance of defining the precise location of the region of the intestine under examination is stressed. 3. The experimental results and the histological observations are discussed in relation to (a) the results which have been obtained using PVP, which is unsuitable as an indicator of immunoglobulin transport in the rat and (b) the histological composition of the absorptive epithelium and the maturation changes which affect the epithelium between 18 and 21 days.     

The absorption of polyvinyl pyrrolidone by the new-born pig intestine

1. The intestinal absorption of [(131)I]polyvinyl pyrrolidone of mean mol. wt. 160,000 (K. 60) and 40,000 (K. 30) after oral administration has been measured in unsuckled conscious pigs less than 20 hr old. Absorption was assessed by the measurement of the concentration of [(131)I]PVP in venous blood during the 6 hr after feeding and also by the distribution at the end of the experiment of [(131)I]PVP between homogenates of the alimentary tract and homogenates of the rest of the animal.2. The concentration of [(131)I]PVP in the peripheral blood after feeding was dependent upon the mol. wt. of the polymer, when comparable amounts had been absorbed from the intestine. PVP K. 60 attained higher blood concentrations than PVP K. 30 and the blood concentrations of PVP K. 60 were close to the values to be expected if all the material which had left the intestine had remained in the blood. The lower blood concentrations found when PVP K. 30 was fed were associated with the disappearance of labelled solute from the gut and were thus the consequence of the relatively rapid escape of labelled solute from the plasma after absorption had taken place.3. The ability of the intestine to absorb [(131)I]PVP K. 60 declined progressively after birth but did not terminate abruptly unless the animal was fed colostrum. In unsuckled animals the rate and extent of absorption at 3 hr was much greater than at 20 hr after birth, but some absorption was still present at least 65 hr after birth.4. The transfer of PVP K. 60 to the peripheral blood was dependent upon factors in sow colostrum, since significant absorption did not occur when PVP was fed in water or simple salt solutions.5. The factors which accelerated absorption were present in colostrum from the goat, cow and ewe as well as that from the sow; they remained in the whey, but, in contrast to the factors which accelerate absorption in the calf, were largely inactivated by boiling. Similarly, neither phosphate, lactate, pyruvate, nor lower volatile fatty acid salts, which were effective in the calf, accelerated absorption in the pig.6. The absorption of [(131)I]PVP K. 30 was found to be much less dependent upon the composition of the solvent than the absorption of [(131)I]PVP K. 60, although absorption was most rapid when PVP K. 30 was fed in colostrum.     

The use of [125I]polyvinyl pyrrolidone K. 60 in the quantitative assessment of the uptake of macromolecular substances by the intestine of the young rat

1. A method has been developed which allows the quantitative estimation of the uptake of labelled polyvinyl pyrrolidone (PVP) of mean mol. wt. 160,000 (K. 60) by the wall of the small intestine of young rats.2. Four hours after feeding a standard dose of [(125)I]PVP by stomach tube, the small intestine was thoroughly washed out, and the radioactivity of the intestinal wall measured. Under these conditions, the small intestine of animals less than 18 days old took up more than 50% of the radioactivity which had left the stomach. There was no increase in PVP uptake if the duration of absorption exceeded 4 hr. The PVP was taken up by the epithelial cells of the villus, and its intracellular localization has been demonstrated by fluorescence microscopy and can be related to vacuolation in the cells.3. In animals between 18 and 20 days old the uptake of PVP declined progressively, until, in animals more than 20 days old, less than 5% of the radioactivity was taken up by the intestinal wall.4. There is good agreement between the reported age of termination of antibody absorption in young rats and the age at which PVP uptake ceased in the present experiments. It is suggested that the loss of ability of the intestine to take up substances of high mol. wt. may be the factor which limits the duration of the period of antibody absorption in this species.     

Quantitative assessment of the transmission of labelled protein by the proximal and distal regions of the small intestine of young rats.

1. The plasma volume in rats aged 15-16 days was measured by dilution analysis using homologous, 125I-labelled immunoglobulin G. A mean plasma volume of 5-53 ml./100 g and a mean blood volume of 8-01 ml./100 g were obtained.2. After the injection of labelled immunoglobulin G into the heart, homogenates of various abdominal organs and of the carcass were prepared. Labelled immunoglobulin G left the vascular compartment at a rate of about 9-10%/hr over a 3 hr period. About 11% of the labelled immunoglobulin G was catabolized in 2 hr. 3. The data obtained from these studies was used to make quantitative estimates of the amount of intact immunoglobulin G transmitted from the proximal intestine and from the ileum after the intra-intestinal injection of 1000 mug of labelled immunoglobulin G.Homogenates of the experimental animals were prepared and it was estimated that over 40% of the labelled immunoglobulin G was transmitted as intact protein from the proximal intestine. The results suggest that no intact immunoglobulin G was transmitted from the ileum, but about 15% of the protein removed from the ileum was recovered in the whole body as degraded fragments precipitable with trichloroacetic acid. 4. These observations are discussed in the context of the transmission of antibodies, and their relevance to the receptor hypothesis is considered.     

Fractionation studies on the absorption of labelled immunoglobulin G by the gut of young rats.

1. Centrifugation in density gradients was used to study the fragments produced during intraluminal and intracellular digestion, after the injection of 125I-labelled immunoglobulin G (IgG) into different regions of the small intestine of 14 to 15-day-old (pre-closure) and 24-day-old (post-closure/ rats. 2. After injection into the proximal small intestine and into the ileum of pre-closure animals, the bulk of the radioactivity recorded for gut washes and gut homogenates was located at 4S-7S. The serum from animals which had received injections into the proximal small intestine had high radioactivity and one peak at 7S; the serum from animals which had received injections into the ileum had low radioactivity and no activity in the 7S region. 3. After injection into the proximal small intestine of post-closure animals, the bulk of the radioactivity recorded for gut wash samples was located at 3-5S--5S. Gut homogenates had peak activity at 2-5S--4S. Thus large molecular weight products can be absorbed by the proximal enterocytes of post-closure rats and degraded. The sera of these animals had low radioactivity. 4. After injection into the distal small intestine of post-closure animals, the bulk of the radioactivity recorded for gut wash and gut homogenate samples was located at 4S-7S and in this respect the radioactivity plots resembled those for (2) above. Serum radioactivity was low. 5. The effect of precipitation with trichloroacetic acid and incubation with specific antiserum upon the radioactivity of gut washes, gut homogenates and serum samples was recorded. 6. The relevance of these findings to studies on the transmission of protein by the rat small intestine is discussed.     

The absorption of 125I-labelled immunoglobulin G by different regions of the gut in young rats

1. (125)I-labelled homologous IgG was injected into different regions of the small intestine of rats aged 12, 16, 18, 20 and 22 days. At 12 days the proximal and middle regions of the intestine readily absorbed globulin and transmitted it to the circulation. The distal region of the intestine transmitted little to the circulation at all ages tested.2. The intestine loses its ability to transmit globulin to the circulation in a distal-proximal direction. At 16 and 18 days the ability of the middle region had declined significantly, and this decline continued so that little globulin was transmitted from this region at 20 and 22 days.3. The proximal intestine retained the ability to transmit globulin to the circulation in significant amounts up to 20 days.4. There is a close negative correlation between body weight and total radioactivity of the sera of rats which had received doses of labelled globulin into the proximal and middle regions of the intestine. There was no such correlation after injection into the distal intestine - suggesting a restricted throughput of radioactive material by the absorptive cells of this region.5. These results are discussed in the context of the termination of antibody absorption, and in relation to the results obtained using polyvinyl pyrrolidone.     

Sex-specific effects of corticosterone on hippocampally mediated learning in young rats

Glucocorticoids administered during development can have lasting consequences on learning performance and brain development. Whereas most studies administer glucocorticoids to the young rat during the so-called stress-hyporesponsiveness period (SHRP), we examined the effects of corticosterone pellets implanted at the conclusion of the SHRP on two forms of eye blink conditioning (EBC). Analysis of blood samples indicated that pellets implanted on Day 15 released the bulk of the corticosterone before Day 21. In tests of EBC beginning on Day 28, corticosterone-treated males, but not females, showed impaired performance in the hippocampally mediated "trace" version of the EBC paradigm. There were no effects of corticosterone on the "delay" version of the task. These results are consistent with earlier findings that the hippocampus is particularly sensitive to elevated glucocorticoid levels. Moreover, the findings suggest that glucocorticoids administered after the SHRP may produce subtle effects on learning performance akin to those that have been reported in children.     

Changes in metabolite transport by small intestine and kidney of young and old rats

We have studied the influence of the ageing phenomenon on metabolite absorption by the small intestine and the kidney of the rat, using isolated brush-border membrane vesicles prepared from 2 groups, one composed of 2 month, and the other of 24 month, animals. "Overshoot", which is typical of Na(+)-glucose cotransporter activity, disappeared in the duodenum and decreased in the kidney of the old rats. Short-circuiting of vesicles with valinomycin showed that, in the presence of K+ and valinomycin, "overshoot" decreased in both groups by about the same percentage. The Na(+)-dependent uptake of aspartate and phenylalanine showed contrasting pictures in the jejunum and kidney of the aged animals: aspartate transport decreased only in the kidney, while phenylalanine uptake was negatively affected in the jejunum. Na(+)-dependent citrate uptake, studied in renal brush-border membrane vesicles, was lower in the old rats. The Km values determined for Na(+)-dependent D-glucose and citrate uptake in the kidney did not meaningfully differ between the two groups. A continuous decrease in Na(+)-dependent D-glucose and citrate uptake in the rat kidney, during ageing, was demonstrated.     

The effects of cortisone acetate on stomach evacuation and the absorption of 125I-labelled globulins in young rats

1. After short exposure (3-5 hr) to I.P. cortisone acetate (5 mg), the reduced transmission of labelled globulin to the circulation in 14-day-old rats is due to the slow release of the oral dose from the stomach. The ability of the small intestine to absorb and transmit globulin to the circulation is comparable in control and experimental animals.2. About 26 hr after cortisone acetate treatment (5 mg), the greatly reduced absorption of labelled globulin from oral doses administered to rats aged 15 days is due to the combined effects of the slower release of the dose from the stomach and to changes which have occurred in the small intestine.3. About 50 hr after the administration of 5 mg cortisone acetate the effect on the rate of stomach evacuation is minimal in rats aged 16 days. When labelled globulin is introduced directly into the duodenum of these animals virtually no absorption occurs.4. The results obtained from the experiments in which labelled globulin was injected into the duodenum support the contention that the proximal half of the small intestine is an important site for macromolecular transport.     

The effects of cortisol and BUDR on cellular differentiation in the small intestine in suckling rats

In suckling rats absorptive cells in the distal two-thirds of the small intestine ingest colloids by pinocytosis. In the middle third of the intestine pinocytosis ceases 17 to 21 days after birth and can be halted prematurely by glucocorticoids. The adrenal cortex is relatively inactive in newborn rats and begins to secrete glucocorticoids two weeks after birth; this secretion has been suspected to cause the termination of pinocytosis. Glucocorticoids do not stop pinocytosis in absorptive cells already present on the villi but induce the renewing population of cells in the crypts to replace the cells on the villi with new ones that do not engage in pinocytosis. Therefore, glucocorticoids act by inducing a new direction of cellular differentiation. BUDR (5-bromodeoxyuridine) is reported to inhibit cellular differentiation reversibly in vitro. When injected into 8 to 9-day-old rats, simultaneously with cortisol, it inhibited the change in cellular differentiation induced by cortisol; that is, pinocytosis continued unabated. BUDR did not appear to be toxic and did not interfere with cellular replacement in the small intestine, as judged autoradio-graphically. Thus it appears that BUDR inhibits the initiation of cellular differentiation by cortisol but does not interfere with cellular proliferation or the pre-established program of cellular differentiation that leads to pinocytosis. The mechanism of action of BUDR is still unknown; it is discussed in light of the possibility that BUDR may after cellular differentiation without acting directly on the genome.     

Differential satiating effects of fats in the small intestine of obesity-resistant and obesity-prone rats.

The effects of duodenal infusions of fats on sham feeding was measured in two strains of rats that differ in their susceptibility to fat-induced obesity. Osborne-Mendel rats are prone to developing obesity on a high-fat diet and preferentially choose fats over carbohydrates in macronutrient selection paradigms. In contrast, S 5B/PL rats are resistant to developing obesity when eating a high-fat diet, and preferentially choose carbohydrates in macronutrient selection paradigms. To test the hypothesis that differences in the satiating potency of fats in the small intestine contributed to these differences between the two strains, we measured the effects of duodenal infusions of Intralipid and sodium linoleate on sham-feeding intakes. The results were consistent with the hypothesis. Duodenal infusions of either of these fats decreased intake significantly more in S5B/PL rats than in Osborne-Mendel rats. Both rat strains sham fed similar amounts when intestinally infused with 0.15 M NaCl. These results suggest that differences in responses to intestinal satiating mechanisms may contribute to the differences in susceptibility to fat-induced obesity in these rat strains.     

Effect of metabolic cage housing on immunoglobulin A and corticosterone excretion in faeces and urine of young male rats

Six 8-week-old Sprague-Dawley rats were studied for 9 days divided into three periods of 3 days each: before transferral to metabolism cages, during metabolic cage housing and after return to their home cages. Faeces were collected daily when the animals were housed in their home cages and every 6 h when the animals were housed in metabolic cages during which time urine was also collected every 6 h. The rate of weight gain was slightly reduced during the 3 days in metabolic cages and the animals produced significantly larger amounts of faeces when housed in metabolic cages than when housed in their home cages. The total faecal excretion of corticosterone (nanograms excreted per hour per kilogram body weight) and immunoglobulin A (IgA) (milligrams excreted per hour per kg body weight) quantified by enzyme-linked immunosorbent assays (ELISAs) exhibited a clear diurnal rhythm in the metabolic cage. Urinary excretions of corticosterone and IgA also followed a clear diurnal cycle. The mean daily amounts of corticosterone excreted were not significantly affected by cage change and by housing in metabolic cages. However, the excretion of faecal IgA was significantly reduced during the 3 days after the period in metabolic cages. Taken together the results indicate that metabolic cage housing is mildly stressful for young adult male rats.     

The influence of exogenous steroids on macromolecule uptake by the small intestine of the new-born rat

1. Plasma concentrations of cortisol and corticosterone measured by competitive protein binding in rats between 5 and 28 days after birth have been related to the intestinal uptake of [(125)I]polyvinyl pyrrolidone (PVP).2. Plasma cortisol concentration was consistently low throughout the period studied, but there was an increase in plasma corticosterone concentration at the time (18-21 days) when PVP uptake declined to zero (closure).3. Injection of a large dose of cortisone acetate 5 days after birth resulted in precocious closure; PVP uptake declined progressively to zero during the 6 days following the injection. Injection of this steroid at 12 days of age caused closure within 4 days.4. Precocious closure induced by cortisone acetate was closely comparable histologically with natural closure; the decline in PVP uptake was associated with the progressive displacement of vacuolated cells from the villi of the terminal intestine.5. Injection of corticosterone at either 5 or 12 days after birth also reduced PVP uptake. However, the reduction was transient and uptake returned to control levels some days after the injection.6. The temporary reduction in PVP uptake following corticosterone injection was not associated with any change in the histological appearance of the small intestine at the light microscope level.7. The injection of either cortisone acetate or corticosterone was followed by a period of impaired body growth and also a reduction of adrenal weight in animals injected at 12 days but not in animals injected at 5 days.