Golgi apparatus mediated polysaccharide secretion by outer root cap cells of Zea mays

Summary In the outer cap cells of roots of Zea mays, secretion is accompanied by hypertrophy of dictyosome cisternae with formation of large secretory vesicles. Vesicle contents are subsequently released from the protoplast by fusion of the vesicle membrane with the plasma membrane. The secreted material, a highly hydrated polysaccharide, was localized intracellularly by the periodic acid-Schiff reaction. Under appropriate conditions, the product moves outward through the cell wall after discharge from the protoplast, and appears as a droplet adhering to the root tip. Under conditions where the secretory product accumulates at the inner wall surfaces, no external droplet is formed.The secretory activity has an active phase that is sensitive to metabolic inhibitors and influenced by temperature (Q₁₀>2), and a passive phase that is independent of temperature, insensitive to metabolic inhibitors but sensitive to osmotic agents. The active phase is characterized by a temperature-independent periodicity (3 hours). Sucrose supplied to the growth medium increases the amount of polysaccharide secreted. Polysaccharide synthesis, segregation into vesicles, and discharge from the protoplast are assumed to require active metabolism; the step involving extrusion of polysaccharide through the cell wall region appears to be a passive process influenced by the degree of hydration of the polysaccharide and by cell turgor.Purdue University Agricultural Experiment Station Journal Paper No. 2967; Charles F. Kettering Research Laboratory Contribution No. 261.     

EXTRUSION OF NUCLEOLI FROM PRONUCLEI OF THE RAT

Electron microscope observations of osmium tetroxide-fixed rat eggs indicate that small nucleoli are extruded from pronuclei in a sharply demarcated time period after sperm penetration. Approximately 4½ hours after sperm penetration, fine fibrous material aggregated in distinct loci along the inner surface of the nuclear envelope and condensed into small, dense bodies. The term tertiary nucleolus or extrusion body is used to designate the forming bodies. The small tertiary nucleoli form distinct protrusions from the pronuclei during the following developmental period and finally bud off into the cytoplasm, carrying with them a small portion of the double nuclear envelope. The extrusion bodies can be observed only in the vicinity of the pronuclei and have not been seen near the cell membrane. The fate of the tertiary nucleoli is not known; apparently they transform or disappear after they have passed into the cytoplasm. Eleven hours after sperm penetration, tertiary nucleoli are not present near the nuclear membrane and the extrusion activity has apparently ceased. Large and small nucleoli react similarly to cytochemical reagents: they are Feulgen negative; they are positive to the Millon, Sakaguchi, brom-phenol blue, and PAS reactions. Azure B stain combined with nuclease extraction indicates the presence of small amounts of RNA in the nucleoli.     

THE FINE STRUCTURE OF BRUNNER''S GLANDS IN THE MOUSE

Examined with the electron microscope, the secretory cells of the submucosal glands of Brunner in the mouse present a curious combination of the fine-structural features of both serous and mucus-secreting cells. The cells have numerous mitochondria, abundant basal ergastoplasm, dense secretory granules that bear a superficial resemblance to pancreatic zymogen granules, and an unusually extensive Golgi apparatus. The prominence of the lamellar, vesicular, and vacuolar elements of the Golgi complex facilitates detailed observation of these components. More evident than in other glandular cells, aggregates of small vesicles appear to represent the transitional elements and are vehicles for transport of the product between the ergastoplasm and the Golgi complex. The numerous vesicular evaginations of smooth-surfaced regions on cisternae of the rough-surfaced endoplasmic reticulum and accumulations of innumerable vesicles of similar size in the area between the nearest profiles of the ergastoplasm and the Golgi complex support this contention. The cytological characteristics and physiologic properties of Brunner's glands in various species are discussed briefly. It is concluded that the submucosal glands of the mouse are excellent material for exploration of the ultrastructural correlates of both protein and carbohydrate secretion, and it is suggested that their secretion may have functions other than those generally attributed to them, namely, chemical and mechanical protection of the duodenal surface epithelium.     

RECURRENT CARCINOMA OF THE RECTUM—Surgical Treatment

Following operations on the rectum for carcinoma, approximately half of the patients have recurrence in the perineum, pelvis, abdomen or at the suture line of anastomosis. The prognosis is almost uniformly poor and although the problems of management are complicated, dealing with them may give the patient worthwhile physical, emotional and economic benefits. Surgical procedures used in the treatment of the common types of recurrence are discussed.     

Involvement of an Inducible Cytochrome P-450 in Precocene II Oxidation by Streptomyces Griseus

Streptomyces griseus oxidizes the insecticide precocene II to its cis- and trans-dihydrodiols and 3-chromenol after growth on an enriched medium containing soybean flour. Oxidation of precocene II is dependent on the level of cytochrome P-450 in this organism. Extracts of cells grown on media lacking soybean flour were devoid of cytochrome P-450 and could not oxidize precocene II. In an in vitro reconstituted system containing NADPH, spinach ferredoxin reductase, spinach ferredoxin and ammonium sulfate fractions enriched in cytochrome P-450, precocene II was oxidized to its dihydrodiols. An aerial mycelium-negative variant of S. griseus (AMY mutant), that was unable to elicit cytochrome P-450 when grown on soybean flour-enriched medium, failed to oxidize precocene II.     

Regulation of insulin synthesis in an insulin-producing cell line (RINm5F): Long-term experiments

Summary The insulin-producing cell line RINm5F, has been used in short-term experiments to evaluate insulin secretion. We sought to maintain the responsiveness of these cells to stimuli for up to 2 days. We examined the course of new insulin synthesis over this period by measuring at intervals immunoreactive insulin (IRI) in two parts: IRI in the medium (M) and IRI extracted from the cells (C). Control cells were incubated in RPMI 1640/2.8 mM glucose/10% fetal bovine serum/200 μg/ml bacitracin (to prevent insulin degradation). The addition of dibutyryl cAMP 10 mM to the experimental dishes significantly increased total (M+C) IRI at 48 hr to 37% above the insulin content of the control dishes (p<0.01). Theophylline 10 mM increased total (M+C) IRI by 24% over control (p<0.05) after 24 hrs. Glucose, glyceraldehyde, leucine, arginine, glucagon and tolbutamide, other stimulants of insulin production, had no effect. Under the experimental conditions reported here, including the use of bacitracin, IRI synthesis can be studied for up to 48 hr. Portions of this study have been published in abstract form for the 47th Annual Meeting of the American Diabetes Association, Indianapolis, Indiana, 1987. Supported in part by the American Diabetic Association, Maryland Affiliate.