The action of various vitamin D3 metabolites on calcium and phosphorus metabolism in chick embryo calvariae

Summary Chick embryos from vitamin D-deficient hens given physiological doses of 1,25-dihydroxyvitamin D₃ or 24,25-dihydroxyvitamin D₃ or both become severely hypocalcemic, hyperphosphatemic and fail to hatch as compared to those derived from hens given 25-hydroxyvitamin D₃ or 24,25-difluoro-25-hydroxyvitamin D₃. Calvariae from the former contain less mineral and on incubation in vitro produce significantly lower calcium and higher phosphate concentration in the medium than do the calvariae derived from the embryos of hens supported on 25-hydroxyvitamin D₃ or 24,24-difluoro-25-hydroxyvitamin D₃.     

Vitamin D receptors in heart: Effects on atrial natiuretic factor

We report that receptors for vitamin D exist in distinct regions of the heart in female and male mice, predominantly in the right atrium where most of the cardial atrial natriuretic peptide (ANF) is produced. Tritiated 1,25-dihydroxyvitamin D₃ (1,25-D₃, vitamin D, soltriol) and ANF are colocalized in nuclei and cytoplasm respectively in identical cardiomyocytes. Changes of ANF tissue and blood levels under dietary deficiency and treatment with 1,25-D₃ suggest direct genomic actions of vitamin D on myoendocrine cells of the atrium for the regulation of ANF manufacture and secretion. These results were obtained by combining thaw-mount autoradiography with immunocytochemistry using tritiated 1,25-D₃ and an antibody against rat ANF. This antibody was also used in a radioimmunoassay to determine atrial natriuretic factor in plasma, atria and ventricles of normal or vitamin D-deficient mice.     

Acid-base homeostasis and the thyro-parathyroid glands]

Chronic metabolic acidosis entails hyperparathyroidism and osteopathy. In order to elucidate the role of the thyroparathyroids in this bone lesion production the effects of acidic diet for 7 weeks were studied in parathyroidectomized (PTX), thyroparathyroidectomized (TPTX) and shamoperated (Sh-O) growing rats. In all animals urinary excretion of calcium, phosphate, ammonium and titrable acidity was similarly increased. The rise in hydroxyproline excretion and urinary 85-sr (that was injected previous to acidic feeding) was more marked in PTX and TPTX rats. Moreover, in these animals the serum calcium level was increased, the blood pH was decreased. According to these data, an acidic diet intake that is not sufficient to elicit a fall in blood pH of normal young rats can induce severe acidosis in chronically parathyroidectomized or thyroparathyroidectomized animals; moreover the bone resorption appears more marked. It is concluded that parathyroids are involved in the extra-cellular fluid defense mechanism against acidosis by a no bone resorptive mechanism. We hypothesize that the parathyroids permit the necessary and adequate supply of bicarbonates by the bone to maintain blood pH homeostasis.     

Wirkungen von Gallensäure und Vitamin D auf die Kalziumresorption bei der Ratte

Summary Comparison of the effects of vitamin D and the bile salt Taurochenodeoxycholate on calcium absorption in young rats suggests that bile salt enhancement of calcium absorption occurs mainly in the ileum in normal animals but occurs in both duodenum and ileum in rachitic rats. Also suggested is that bile salts mainly assist the entry of calcium into mucosal cells from the gut lumen while vitamin D chiefly aids the egress of calcium from the cells into the extracellular fluid.     

Direct evidence for pH-dependent Fc receptors on proximal enterocytes of suckling rat gut

Summary By means of an erythrocyte-antibody rosette technique, Fc receptors, functional at pH 6.0 but not at 7.2, were shown to be present on enterocytes isolated from duodenum and jejunum (but absent from ileum) of 12–20-day-old suckling rats.     

Distribution of the 1,25 dihydroxy-vitamin D3 receptor in the bursa of Fabricius of chicken

The vitamin D₃ metabolite 1,25(OH)₂D₃ is probably involved in B lymphocyte ontogeny. We therefore determined the distribution of the 1,25(OH)₂D₃ receptor in the bursa of Fabricius and spleen cells of 7-day-old chicks, by immunohistochemistry using a monoclonal antibody against the chick intestinal cell 1,25(OH)₂D₃ receptor. The bursa cells of young (7-day-old) chicks contained large amounts of receptor while the spleen cells did not. The bursa cells of older (35-day-old) chicks contained fewer receptors, but the number of receptors in the spleen increased.     

Amino acid absorption in jejunum and ileum in vivo — a kinetic comparison of function on surface area and regional bases

Summary Maximum absorptive capacities (V_max) for the jejunum and ileum corrected for the presence of an unstirred layer of water have been calculated for glycine, valine and methioninein vivo in fowls per unit surface area and per region. V_max per cm² showed that ileal enterocytes had a greater absorbing capacity than jejunal for glycine and valine but not for methionine. V_max for glycine and valine, calculated for the whole jejunum and ileum, however, were not different but for methionine the jejunal value was 1.9 times greater than the ileal.MAM is supported by an A.R.C. research grant (No. A50/37).     

Participation of ACTH1–10 and ACTH4–10 on the melatonin modulation of benzodiazepine receptors in rat cerebral cortex

In this study, we examined the effect of intracerebroventricular (i.c.v) injection of melatonin and/or ACTH_1–10 and ACTH_4–10 on [³H]flunitrazepam binding sites in the cerebral cortex of hypophysectomized rats. Hypophysectomy increased the B_max (maximum number of binding sites) of benzodiazepine (BNZ) receptors for at least 7 days after surgery, without changing K_D (dissociation constant). The i.c.v. injection of melatonin to hypophysectomized rats significantly increased B_max, whereas the same doses of melatonin were ineffective in sham-operated animals. In both cases, K_D values were unchanged. The i.c.v injection of ACTH_1–10 to hypophysectomized animals significantly increased B_max, an effect that was enhanced by simultaneous i.c.v. injection of ACTH_1–10+melatonin, reaching higher values of B_max than the i.c.v. injection of these hormones individually. No significant changes in K_D values were found after ACTH_1–10 and/or melatonin administration. However, the i.c.v. injection of ACTH_4–10 to hypophysectomized rats did not change B_max, although it significantly increased K_D values, indicating a decrease in the BNZ binding affinity. Melatonin injection counteracted this effect of ACTH_4–10, returning K_D to the control value. Moreover, although the lower dose of i.c.v. melatonin used, 10 ng, was unable to modify B_max of BNZ binding in the ACTH_4–10-injected group, the higher dose, 20 ng, significantly increased B_max. The results suggest that these ACTH-derived peptides can modulate the effect of melatonin on brain benzodiazepine receptors.     

Bicarbonate and chloride transport across rat ileal basolateral membrane

The mechanisms of HCO ₃ ^– and Cl^– transport across basolateral membranes from rat ileum were investigated in isolated vesicles by means of uptake experiments. Neither Cl^–/HCO ₃ ^– exchanger nor Na⁺–(HCO ₃ ^– )_n cotransport seem to be present in ileal basolateral membranes. Moreover Cl^– uptake is unaffected bycis Na⁺ and/or K⁺ gradients, indicating the absence of Na⁺–Cl^–, K⁺–Cl^– and Na⁺–K⁺–2Cl^– symport activity. An electrically conductive pathway seems to be responsible for both HCO ₃ ^– and Cl^– fluxes. Evidence is also given for the presence of a Na⁺/H⁺ exchanger at the basolateral pole of ileal enterocytes.     

The role of maternal parathyroids and vitamin D3 metabolites in fetal growth and the deposition of glycogen reserves in the maternal and fetal liver in rats]

Thyro-parathyroidectomy of pregnant Rats at 12.5 days of gestation decreased maternal liver glycogen on 21.5 days of gestation and fetal weight as well as fetal liver glycogen stores. The graft of one parathyroid gland or the injection of 1 alpha-hydroxycholecalciferol in these thyro-parathyroidectomized mothers increased their liver glycogen stores at 21.5 days of gestation. These treatments also markedly increased both fetal weight and fetal liver glycogen stores. It was concluded that maternal 1,25-dihydorxycholecalciferol, which is synthesized under the control of parathyroid hormone secretion, controls fetal growth and liver glycogen stores. The mechanism of these effects (direct or indirect) requires further investigations.     

Role of bile in intestinal absorption of203Pb in rats

Summary The author studied absorption of²⁰³Pb after administering intraduodenally²⁰³Pb eliminated with bile. The results obtained were compared with absorption of203Pb administered into the duodenum as²⁰³PbCl₂ in rats forming 2 groups, one with bile ducts cannulated, the other intact. It was found that bile played an important role in absorption of Pb from the gastrointestinal tract. Absorption of Pb²⁰³ is significantly reduced if the bile is drained off by means of a canula.     

Epithelial-mesenchymal interaction in differentiation of duodenal epithelium of fetal rats in organ culture

Summary Epithelial-mesenchymal interaction in the differentiation of duodenal epithelium of fetal rats was investigated by recombination experiments in vitro. The proportion of goblet cells in duodenal epithelium was significantly greater on recombination of developing duodenal epithelium with mesenchyme of the glandular stomach than on recombination with that of the duodenum. Mesenchyme of the glandular stomach or forestomach was better than duodenal mesenchyme in supporting morphogenesis of duodenal epithelium. Treatment of tissues with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) did not affect these tissue interactions.This work was supported by grants-in-aid for cancer research from the Ministry of Education, Science and Culture, Japan.Acknowledgment. The authors wish to express their gratitude to Prof. T. Mizuno of the University of Tokyo for valuable suggestions.     

Direct evidence for pH-dependent Fc receptors on proximal enterocytes of suckling rat gut.

By means of an erythrocyte-antibody rosette technique, Fc receptors, functional at pH 6.0 but not at 7.2, were shown to be present on enterocytes isolated from duodenum and jejunum (but absent from ileum) of 12-20-day-old suckling rats.     

The effect of dietary lactose on the ileal absorption of sodium taurocholate in rats]

When Rats were fed a lactose containing diet, both the absorption rate of sodium taurocholate at the level of ileum and the contents of bile acids of the small intestine were increased. On the contrary, feeding of lactose did not modify the daily fecal excretion of bile acids. It therefore appears that dietary lactose increases the intestinal pool of bile acids by increasing their ileal absorption rate and that this effect of lactose is not subordinated to a modification of bile acid synthesis.     

Morphological appearance of fat in the epithelial cells of different portions of the intestines in mice

Summary Absorption of administered fat in the small intestine of mice as judged morphologically in semi-thin sections demonstrates a proximal to distal gradient, being greatest in the mid-jejunal area, but less in the duodenum and ileum. The criterion of the amount and size of fat droplets in intestinal cells, however, does not necessarily give a reliable indication of the efficiency of fat absorption in the different segments of intestine.     

Absorption of iron from gut into blood: Sex- and time-related studies in rats

Summary As judged from 2-h blood level curves, adult female rats absorbed more Fe^II per cm² of gross duodenal mucosa than adult male rats. By contrast, the 2-h blood level curves per cm² of mucosa of proximal jejunum did not differ significantly in male and female rats although in both sexes, iron was absorbed more efficiently from the duodenum. This research was supported by grant RO1 AM 12381 from the National Institute of arthritis, Diabetes and Digestive and Kidney Diseases, National Institutes of Health.     

The discovery of α-Klotho and FGF23 unveiled new insight into calcium and phosphate homeostasis

The traditional view of calcium homeostasis is that it is maintained by two essential reactions. First, changes in extracellular Ca²⁺ are sensed in several distinct cell types, stimulating the secretion of parathyroid hormone (PTH), 1,25(OH)₂ D and calcitonin in response to the body’s requirement. Second, these calcitropic hormones then act on the calcium-translocating cells of the kidney, bone, and intestine to restore calcium balance. Recent progress indicates that α-Klotho and fibroblast growth factor (FGF) 23 are key players that integrate the multi-step regulatory system of calcium homeostasis that rapidly adjusts the extracellular calcium concentration and continuously maintains its concentration within a narrow physiological range. α-Klotho and FGF23 are also found to be major players in the regulatory system of phosphate homeostasis. Here, the demonstration of the molecular functions of α-Klotho and FGF23 has recently given new insight into the field of calcium and phosphate homeostasis. Received 3 April 2008; received after revision 23 May 2008; accepted 5 June 2008     

Absorption of pancreatic lipase from the duodenum into lymphatics

Summary A significantly higher lipase activity was measured in the duodenal lymph samples of 15 dogs than in each of corresponding arterial blood plasma samples collected prior to, during and after maximal hormonal stimulation of pancreatic secretion. The result may be evaluated as a sign of pancreatic lipase absorption by the duodenum into lymphatics.     

Suppression of natural killer cell activity in mouse spleen lymphocytes by several dopamine receptor antagonists

The effects of dopaminergic receptor inhibitors such as thiothixine (D₁/D₂), fluphenazine (D₁/D₂), trifluoperazine (D₁/D₂), pimozide (D₂), flupenthixol (D₁/D₂), (+/–)-SKF 83566 (D₁), and spiperone (D₂) on splenic natural killer (NK) cell cytotoxic activities were assessed in vitro using mouse spleen lymphocytes or enriched NK cells. Both the activities of the splenic NK cell cytotoxicity and the effector-target cell conjugation were suppressed by thiothixine, fluphenazine, and trifluoperazine at concentrations from 2.64 to 14.78 M. In addition, the augmentation of the cytolytic activity of NK cells induced by interferon- or interleukin-2 was antagonized by pretreatment with these neuroleptic compounds. However, neither the splenic NK cell cytotoxicity nor the effector-target cell conjugation were affected by treatment with other neuroleptic compounds such as pimozide, flupenthixol, (+/–)-SKF 83566, and spiperone. Thus, it appears that neuroleptic compounds such as thiothixine, fluphenazine, and trifluoperazine may act through the mechanisms other than a dopaminergic pathway to affect the NK cell-target cell interaction.     

Effects of a bovine pineal peptidic fraction (E5) on plasma and pituitary levels of LH,FSH and prolactin

Summary Pituitary levels of prolactin were significantly diminished after 5 s.c. injections spaced 12 h apart of 5 g of a partially purified bovine pineal peptide fraction (E₅) in both intact and castrated rats. E₅ treatment did not significantly affect the castration-induced changes in plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH) or pituitary LH but did partially block the pituitary fall in FSH in castrated animals.Supported by NSF grant PCM 8003441 and NIH Center for Reproductive Biology (Bioassay Core) No.P30 HD 10202. Many thanks are due A. Moreno for his excellent technical assistance.