Active transport of glucose across the jejunal epithelium decreases with age in broiler chickens.

Intestinal glucose absorption varies with growth; however, the dynamics of these variations has not been yet fully elucidated in broiler chickens. The present study aimed to compare jejunal glucose uptake and maltose digestion in broilers of 2 different ages, i.e., 1- vs. 5 wk old. Oral D-maltose gavage, everted sac, and Ussing chamber experiments were carried out to investigate intestinal glucose absorption and mRNA expression of glucose-transport-related genes as well as jejunal maltase activity. Upon gavage, glucose concentrations peaked at 10 min post-administration in 1-wk-old chicks, while they peaked at 40 min in 5-wk-old chickens. Glucose concentrations at 10 min were significantly higher in the 1-wk-old chicks (P = 0.010). Using the everted sacs experimental setup, 5 intestinal regions i.e., duodenum, proximal jejunum, distal jejunum, proximal ileum, and distal ileum, were targeted to examine D-maltose digestion and glucose transport across the intestinal mucosa. In the distal and proximal ileum, glucose concentrations were found to be significantly higher in the serosal compartment of the 1-wk-old chicks upon incubation with D-maltose (25 mM) (P < 0.05), while in the mucosal compartment the levels were significantly higher in the 5-wk-old chickens (P < 0.05). An Ussing chamber setup was employed to measure glucose-induced short-circuit current (ΔIsc) in the mucosal epithelium of the jejunum. In response to the addition of D-maltose (10 mM) into the mucosal compartment, ΔIsc was significantly higher in the 1-wk-old chicks (P = 0.018). Furthermore, no variations in jejunal maltase activity were observed between the 2 age groups. While jejunal glucose absorption was lower in the 5-wk-old chickens, the mRNA expression levels of jejunal SGLT1, GLUT2, and Na+/K+-ATPase did not show any significant differences between the 2 age groups. Our results suggest that the active transport of glucose across the jejunal epithelium decreases upon growth in broiler chickens but is not accompanied by any variations in maltase activity or in the expression of glucose-absorption-related genes.

Mitochondrial deoxyribonucleic acid 3256C-T mutation in a Japanese family with noninsulin-dependent diabetes mellitus.

Accumulating reports indicate a relationship between mitochondrial DNA mutation and impaired glucose-induced insulin secretion leading to a subtype of noninsulin-dependent diabetes mellitus. DNA from a 45-yr-old Japanese woman with noninsulin-dependent diabetes mellitus and muscle atrophy was isolated and studied for mitochondrial DNA mutations. We identified a mitochondrial DNA C-T heteroplasmic mutation at nucleotide position 3256. The mutation was located in the transfer ribonucleic acidLeu in a region conserved in evolution. Eight other members of her family were examined for the mutation. Six of them had the same mutation together with noninsulin-dependent diabetes mellitus, and one teenage boy had the mutation and impaired glucose tolerance. The other family member who did not have this mutation had normal glucose tolerance. The enzyme activity of the mitochondrial oxidative phosphorylation pathway in the muscle of the proband was measured. The enzyme activity was decreased in the proband, especially in complex I. This mutation might be responsible for the abnormal glucose metabolism.

Prenatal stress and postnatal development of neonatal rats--sex-dependent effects on emotional behavior and learning ability of neonatal rats.

Maternal sound stress (800 Hz; 77 dB, every other minute for 15 min/day, from day 10 to 18 of gestation), combined with forced swimming stress (15 min/day), was found to cause potentiation of sound-induced loss of locomotor activity, referred to as emotional behavior, of male offspring, but not that of female offspring, at 4 weeks of age. Maternal stress also caused an increase in the total number of errors by male, but not female offspring in the water-maze test at 6 weeks of age. These effects of stress on emotional behavior and learning behavior were abolished when dams were pretreated with buspirone (30 min before the stress, from day 8 to 18 of gestation). Thus, prenatal stress might have sex-dependent effects on emotional behavior and learning ability of neonatal rats.

Oxidative damage to mitochondrial DNA and its relationship to diabetic complications.

Increased oxidative stress induced by hyperglycemia may contribute to the pathogenesis of diabetic complications. Oxidative stress is known to increase the conversion of deoxyguanosine (dG) to 8-hydroxydeoxyguanosine (8-OHdG) in DNA, which is linked to increased mitochondrial DNA (mtDNA) deletions. We investigated mtDNA deletions and 8-OHdG in the muscle DNA of non-insulin-dependent diabetes mellitus (NIDDM) patients. mtDNA deletion of 4977 bp (delta mtDNA4977) and the content of 8-OHdG in the muscle DNA of the NIDDM patients were much higher than those of the control subjects. There was a significant correlation between delta mtDNA4977 and the 8-OHdG content (P < 0.0001). Both delta mtDNA4977 and the 8-OHdG content were also correlated with the duration of diabetes. Delta mtDNA4977 and the 8-OHdG content in muscle DNA increased in proportion to the severity of diabetic nephropathy and retinopathy. This is the first report that an increase in delta mtDNA4977 and 8-OHdG is proportional to the severity of diabetic complications. Oxidative mtDNA damage is speculated to contribute to the pathogenesis of diabetic complications though a defect in mitochondrial oxidative phosphorylation or other mechanisms. 8-OHdG and delta mtDNA4977 are useful markers to evaluate oxidative mtDNA damage in the diabetic patients.

The process of otoconia formation in guinea pig utricular supporting cells.

In order to clarify the mechanism of otoconia formation, the pathway of calcium transport in the utricular supporting cells was investigated. Potassium pyroantimonate (PA) precipitation, which indicates the localization of calcium ions, was seen not only in otoconia but also in supporting cell cytoplasm. In the latter, deposits of PA were detected in the secretory granules and endoplasmic reticulum (ER). These deposits were not present in cells pretreated with ethylene glycol-O,O,-bis (2-aminoethyl)-N,N,N',N'-tetra-acetic acid (EGTA). Exposure of the supporting cells to streptomycin sulfate (SM) increased the number of lysosomes. These lysosomes contained many small deposits of PA. The remaining granules and ER in cytoplasm also contained small deposits of PA. The findings suggest that otoconia are formed by the vestibular supporting cells in which calcium ions might be transported via ER-secretory granule-lysosome-cytoplasmic protrusion.

Comparison and evolution of the lagena in various animal species.

The structure of the vestibular organs of the teleost fish (bluegill), newts (Japanese fire-belly newt), frogs (black-spotted pond frog), snapping turtles and birds (chicks) was morphologically compared, with particular attention to the lagena macula, and the differences between animal species with relation to evolution were considered. Teleost fish had no striola on the lagena macula. The striola of newts were short and restricted to the central area of the macula, but those of frogs, snapping turtles and chicks extended from the anterior to posterior edges of the macula. This indicates that the frog is more highly evolved than the newt. The length of the kinocilium of sensory hairs was equal to that of the longest stereocilium in teleost fish and newts, but the kinocilia of frogs, snapping turtles and chicks were longer than the longest stereocilium. This indicates that the function of the lagena of teleost fish and newts is for hearing whilst in the other animals they are for posture. The diameter of the sensory hair bundles is small in teleost fish and frogs, but large in newts and snapping turtles. This indicates that the sensitivity of the sensory cells of the lagena towards outer force is low in teleost fish and frogs, high in newts and snapping turtles and intermediate in birds. The lagena of snapping turtles protrudes from the basilar papilla into the vestibule but the lagena of chicks lies on the tip of the long projecting basilar papilla. From observation of the locations of lagenae it is natural to speculate that there must have been some species of animal now extinct that had the evolving location of the lagena prior to that of chicks. In future it will be very interesting and useful to identify this extinct animal using DNA techniques.

Effects of secretory stimulation on the Golgi apparatus and GERL of rat tracheal gland cells.

The morphological and cytochemical changes of the Golgi apparatus and GERL of rat tracheal serous cells were studied after in vivo secretory stimulation with pilocarpine. Discharge of PAS-positive secretory granules had started within 30 min and complete discharge was finished within 2 h after stimulation. Reaccumulation of secretory granules became evident at 6 h and the volume of granules was similar to that of unstimulated control cells at 8 h. At 6 h after pilocarpine administration, acid phosphatase (AcPase) activity was markedly increased in GERL and immature secretory granules as compared to uninjected controls and this phenomenon continued 12 h after stimulation. GERL appeared to increase in extent and numerous continuities with immature granules were observed. Thiamine pyrophosphatase (TPPase) activity, normally present in trans Golgi saccules was not altered during the secretory cycle. At 2 h after stimulation, tubular structures of GERL, which also react to AcPase, were seldom observed. At 6 h, these structures were more numerous than those of control cells. These results demonstrate that, in rat tracheal serous cells, pilocarpine induces alteration of only GERL, and reconstitution followed by resynthesis of secretory granules is performed without participation of the Golgi stack.

Three-dimensional observation of the Golgi apparatus in rat tracheal gland during the secretory cycle.

The purpose of the present work is three-dimensional clarification of the overall configuration of the Golgi apparatus in rat tracheal serous cells including the sequence of structural modification during the secretory cycle induced by pilocarpine. From the results of both transmission electron microscopy from the serial sections, and scanning electron microscopy using the osmium-DMSO-osmium method based on maceration in dilute osmium tetroxide, the overall configuration of the Golgi apparatus shows that the Golgi stacks are joined together to make an interconnected, single Golgi complex in a cell. Connections, not only between the TGN (trans Golgi network) and rough ER (endoplasmic reticulum), but also between rough ER and cis cisternae are observed. This finding offers alternative routes for movement of substances through the cellular endomembrane system. During the secretory cycle induced by pilocarpine injection, it is evidenced that the disappearance of the TGN is accompanied by rapid discharge of secretory granules and the reconstitution of the TGN is followed by reformation of secretory materials. The beginning of the reconstitution of the TGN is indicated by vesicular buds from the sacculotubular element. These results indicate that the TGN participates directly in membrane recycling and in formation of secretory granules.

Ultrastructural changes in aging rat tracheal gland.

Ultrastructural differences in the tracheal gland between young and aged rats were studied. Marked differences were observed in the serous cells between the young (8-17 week-old) and the aged (20-30 month-old) subjects. The secretory granules of the serous cells of the young subjects were homogeneously electron dense, whilst those of the aged subjects had a bizonal appearance. These bizonal granules contained abundant mucous material. The trans Golgi network of the aged rat serous cells was more developed and formed a fine meshwork in comparison with that of the young subjects. In the aged subjects, findings indicate alterations not only in the function of the Golgi apparatus including formation of secretory granules and addition of terminal sugars to the secretory protein but also the histochemical and ultrastructural properties of the secretory material. The changes in the Golgi apparatus are possibly responsible for depression of the tracheal host defense mechanism and might explain why various airway diseases increase with aging.