Distribution of 5′-nucleotidase in the renal interstitium of the rat

Summary The hydrolysis of 5-AMP by 5-nucleotidase is the main source of adenosine. In various tissues adenosine is a local mediator adjusting the organ work to the available energy. In the kidney it regulates renal hemodynamics, glomerular filtration rate and renin release via specific receptors of the arteriolar walls. By immunocytochemistry we identified interstitial and tubular sites of 5-nucleotidase in the rat kidney. In the interstitium the enzyme was detected only in the cortical labyrinth, the compartment that comprises all arteriolar vessels besides other putative targets of adenosine. The 5-nucleotidase-positive cells of the interstitium were identified as fibroblasts. The fibroblasts are in close contact with the tubules as well as with the vessels. Thus, any 5-AMP released by the tubules into the interstitial space would be converted to adenosine in the direct vicinity of its assumed targets. Adenosine produced by tubular cells would hardly have access to its known targets, since 5-nucleotidase is restricted to the luminal cell surface. Pathological events affecting the fibroblasts might influence renal function by modifying the interstitial adenosine production.     

Primary structure of the major β-chain of rat haemoglobins

The amino acid sequence of the major β-chain, ^IIβ, from rat haemoglobins was established with an automated sequencer. Amino acid heterogeneities were found that appear to result from allelic variation at particular residues. We applied several new or unusual techniques in determining the sequence: (1) reaction of the polypeptide with dansylaziridine for detection of cysteine; (2) blockage of the N-terminal residue and the ε-amino group of lysine residues with 1-fluoro-2-nitro-4-trimethylammoniobenzene iodide and subsequent identification of the modified lysine phenylthiohydantoin by absorbance at 420nm; (3) identification of histidine phenylthiohydantoin by its blue fluorescence under long-wave u.v. light; (4) cleavage of the chain into two or three fragments and subsequent sequencing without purification [a detailed statement giving the major phenylthiohydantoins assigned at each step for each sequence run before their alignment in individual sequences has been deposited as Supplementary Publication SUP 50084 (10 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1978) 169, 5]; (5) separation of fragments produced by CNBr cleavage by cation-exchange chromatography; (6) peptide sequencing after attachment of the peptide to cytochrome c. The amino acid sequence was confirmed by amino acid compositions of the complete chain, of CNBr fragments 1 and 3, and of 11 purified tryptic peptides.     

Autoradiographic localization of α-bungarotoxin-binding sites in the carotid body of the rat

Summary Radioiodinated -bungarotoxin (-Bgt) was used to localize -Bgt-acetylcholine receptors in the carotid body of the rat. The gamma spectrometer analyses indicated a high uptake of [¹²⁵I] -Bgt in carotid bodies incubated in vitro (1.51 fmole per organ). Incorporation of the isotope was effectively blocked by pretreatment of carotid bodies with d-tubocurarine and unlabeled -Bgt, but not by atropine. Light microscopic autoradiography showed a heavy labeling of some parenchymal cells. Electron-microscopic autoradiography revealed that labeling was localized along the interface between parenchymal cells, especially where their cytoplasmic processes engage in complex interdigitations. The silver grain counts on electron-microscopic autoradiographs suggest that labelings are preferentially associated with the plasma membrane of certain Type I cells. It is suggested that these Type I cells in the rat's carotid body probably are provided with nicotinic acetylcholine receptors on their plasma membranes.This research was partly supported by a grant from the Edward G. Schleider Foundation of New Orleans.The authors extend appreciation to Drs. Akira Arimura, Dept. of Medicine, for supplying a part of the radioisotope and to James Fisher, Dept. of Pharmacology, Tulane Medical School, for use of the gamma spectrometer. Appreciation also is extended to Mrs. Lia Pedroza for technical assistance     

Effect of α-p-chlorophenoxyisobutyrate on the metabolism of isoprenoid compounds in the rat

1. Feeding of alpha-p-chlorophenoxyisobutyrate (CPIB) to rats increased ubiquinone concentration in the liver but not in other tissues. The increase was progressive with the time of feeding and related to the concentration of CPIB in the diet. 2. Incorporation of [1-(14)C]acetate, but not of [2-(14)C]mevalonate, into sterols in the liver in vivo or by liver slices in vitro was decreased on feeding the rats with CPIB. However, incorporation of mevalonate into ubiquinone increased. 3. CPIB, when added in low concentrations to liver slices, had no effect on isoprene synthesis from acetate; higher concentrations, however, were inhibitory. 4. No activation of ubiquinone synthesis from mevalonate was observed when CPIB was added to the liver slices synthesizing ubiquinone. 5. The increase in ubiquinone in CPIB-fed animals appears to be due to increased synthesis in the initial stages and to decreased catabolism in the later stages. 6. An inverse relationship was found between the concentration of ubiquinone in the liver and the serum sterol concentration in CPIB-fed rats.     

Effect of cortisone on the developmental pattern of the neutral and the acid β-galactosidases of the small intestine of the rat

1. The developmental pattern and effect of cortisone on acid beta-galactosidase and neutral beta-galactosidase were studied in postnatal rats by a recently proposed method for their independent determination. 2. After birth the acid beta-galactosidase activity increases in the ileum, whereas it decreases slightly in the jejunum. On day 16 after birth the activity in the ileum decreases and in 20-day-old rats activity in both parts of the intestine decreases to adult values. In suckling animals the activity in the ileum exceeds the jejunal activity severalfold and in adult animals the activity in the jejunum is slightly higher than that in the ileum. 3. Neutral beta-galactosidase activity is high after birth and decreases in both jejunum and ileum after day 20 after birth. In 12-20-day-old rats activity in both parts is essentially the same, but in adult animals jejunal activity exceeds ileal activity four-to five-fold. 4. Cortisone (0.5, 2.0 or 5.0mg/100g body wt. daily for 4 days) does not influence the activity of either enzyme in 60-day-old rats. Acid beta-galactosidase activity is decreased after cortisone treatment in 8-, 12-, 16-and 18-day-old rats, with sensitivity to cortisone increasing with the approach of weaning. No effect of cortisone on acid beta-galactosidase is seen in 8-day-old rats. Neutral beta-galactosidase activity is increased in the ileum of 8-, 12-, 16- and 18-day old rats, but only in the jejunum of 8-and 12-day-old rats.     

Effect of progesterone pretreatment on the uptake of estradiol-17β by the uterine epithelium of the rat

Summary Progesterone pretreatment of ovariectomised rats resulted in a moderate (44%) lowering of the level of nuclear estradiol receptors found in the uterine epithelium 2 h after a single injection of this estrogen.     

Effect of β-phenylethylamine on dopaminergic systems of the rat brain

The time course of changes in the dopamine concentration in dopaminergic neurons of the nigro-neostriatal and mesolimbic systems of the rat brain during 1 h after intraperitoneal injection of -phenylethylamine (100 mg/kg) was studied by quantitative fluorescence-histochemical analysis. The results showed that -phenylethylamine causes a marked fall in the dopamine level in neurons of dopaminergic systems of the brain. The dopamine level in the bodies of dopaminergic neurons changes more than in their axon terminals. The fall in the dopamine concentration in the dopaminergic systems of the brain during the first hour is irregular in character: in the terminals between 10 and 30 min and in the bodies between 30 and 45 min there is actually a temporary increase in the dopamine concentration. The rise in the dopamine concentration in the terminals coincides with a sharp fall in the dopamine level in the neuron bodies, and conversely, the fall in the dopamine concentration in the terminals after 30 min is accompanied by some increase in the dopamine concentration in the neuron bodies. The results suggest that the increase in motor activity described in the literature in animals after injection of -phenylethylamine is connected with its action on catecholaminergic, especially dopaminergic, brain systems.Institute of Biophysics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 11, No. 6, pp. 578–584, November–December, 1979.     

Strain-specific differences in the regulation of the hepatic cytoplasmic 17β-hydroxysteroid dehydrogenase activity of the rat

• 1.1. The effect of prepuberal gonadectomy of Sprague-Dawley/NIH/HAN rats on cytoplasmic 17β-hydroxysteroid dehydrogenase was examined on day 30, 45, 60, 75, 90 and 105 of life.
• 2.2. The activity in male rats was not significantly affected by gonadectomy, whereas the activity in females showed an age-dependent oestrogen dependency.
• 3.3. This age-dependent oestrogen dependency could also be demonstrated in 5α-dihydrotestosterone treated intact females.
• 4.4. Cytoplasmic 17β-hydroxysteroid dehydrogenase activity of female Chbb:THOM rats also showed an age-dependent oestrogen dependency, whereas the enzyme activity of male rats of this strain showed a distinct androgen dependency absent in Sprague-Dawley/NIH/HAN rats.
• 5.5. On the basis of previous investigations it is concluded that the androgen dependency of the enzyme activity of male Chbb:THOM rats has been bred into this strain in the period 1974–1977.

     

Immunochemical study of transforming growth factor-β in the kidney of the rat and chicken

Transforming growth factor- (TGF-) is a homodimeric polypeptide of 25 kDa, which regulates cell growth and differentiation and influences extracellular matrix metabolism. Using immunochemical techniques, we identified TGF- in the loops of Henle and the collecting and Bellini ducts of rat kidney and in the loops of Henle of chicken kidney. Furthermore, we detected two TGF--immunoreactive proteins on kidney blots of the rat of 12.5 and 47 kDa, and three on chicken kidney blots of 12.5, 34, and 47 kDa. We suggest that the precursor forms of rat and chicken TGF-2 or 3, chicken TGF-4, and the mature form of all of them are expressed in the collecting and Bellini ducts of rat kidney and the loops of-Henle of rat and chicken kidney.     

Involvement of binding lipoproteins in the absorption and transport of α-tocopherol in the rat

1. Specific lipoproteins binding alpha-tocopherol but not its known metabolites have been isolated and identified from cytosol of rat intestinal mucosa and from serum. 2. A timestudy of the appearance of the orally administered alpha-[(3)H]tocopherol with these lipoproteins indicates that very-low-density lipoprotein of serum acts as a carrier of the vitamin. 3. The involvement of the mucosal lipoprotein in the absorption of the vitamin from the intestine has been inferred from observations on the amounts of alpha-tocopherol in serum of orotic acid-fed rats where release of lipoproteins from the liver to serum is completely inhibited. A considerable decrease in the association of alpha-tocopherol with serum very-low-density lipoprotein under this condition is interpreted to mean that serum lipoproteins are limiting factors for the transport of the vitamin across the intestine and that this is possibly effected by exchange of alpha-tocopherol between serum very-low-density lipoprotein and mucosal lipoprotein.     

Electron microscopy of the paracervical (Frankenhäuser) ganglion of the adult rat

Summary The ultrastructure of the paracervical (Frankenhäuser) ganglion in the rat was studied after immersion or perfusion fixation with glutaraldehyde followed by post-osmification. This ganglion is located at the uterovaginal junction in the vicinity of arteria uterina and contains three neuronal cell types. (1) Principal neurons have a fine structure mainly similar to the ganglion cells of other autonomic ganglia. (2) Small granule-containing cells occur in clusters often close to fenestrated capillaries. They are divided into two subgroups according to the size of their cytoplasmic granules; those containing only small granulated vesicles of 800 to 1400 Å in diameter and those having also large granulated vesicles of 2000 to 3000 Å in diameter. (3) Vacuolated nerve cells are large cells that resemble the principal neurons in their cytoplasmic components, except that they contain one to ten vacuoles with corpuscles of different size and shape. The possible physiological significance of the small, granule-containing cells in the uterine function is discussed.     

Invivo β-adrenergic induction of the unmasking of the uncoupling protein in rat brown fat

1. In 28°C adapted rats (WA) both cold stress and norepinephrine (NE) led to a 4-fold increase of uncoupling protein dependent proton conductance which was abolished by propranolol (PRO).2. In 4-day warm re-exposed rats (after 10 days at 5°C) (WR) the same uncoupling by cold stress was observed but the NE effect was lower. Uncoupling by cold stress was not abolished by PRO.3. In WR rats, uncoupling was not due to the involvement of an α-adrenergic pathway.4. Both β-agonist isoproterenol and β-agonists BRL 35135A and ICI D7114 led to high levels of unmasking.5. Interscapular brown adipose tissue surgical denervation, which abolished cold stress unmasking both in WA and WR rats, indicates a mediation by direct sympathetic innervation.6. Depending on the thermal history of the rat, the possibility that unmasking by cold stress could be mediated by different types of β-receptors is discussed.     

Expression of K-Cl cotransporters in the α-cells of rat endocrine pancreas

The expression of K⁺-Cl⁻ cotransporters (KCC) was examined in pancreatic islet cells. mRNA for KCC1, KCC3a, KCC3b and KCC4 were identified by RT-PCR in islets isolated from rat pancreas. In immunocytochemical studies, an antibody specific for KCC1 and KCC4 revealed the expression of KCC protein in α-cells, but not pancreatic β-cells nor δ-cells. A second antibody which does not discriminate among KCC isoforms identified KCC expression in both α-cell and β-cells. Exposure of isolated α-cells to hypotonic solutions caused cell swelling was followed by a regulatory volume decrease (RVD). The RVD was blocked by 10 μM [dihydroindenyl-oxy] alkanoic acid (DIOA; a KCC inhibitor). DIOA was without effect on the RVD in β-cells. NEM (0.2 mM), a KCC activator, caused a significant decrease of α-cell volume, which was completely inhibited by DIOA. By contrast, NEM had no effects on β-cell volume. In conclusion, KCCs are expressed in pancreatic α-cells and β-cells. However, they make a significant contribution to volume homeostasis only in α-cells.