Biomaterial Database

Quantitative evaluation of the in vitro biosynthesis of gastric mucus glycoproteins. Standardization of the methodology. 1979

C Decaens, and D Waldron-Edward, and G Godeau, and J Labat-Robert

For the evaluation of a normal or pathological function of gastric mucosa, a reproducible method for the estimation of the biosynthesis of mucus glycoproteins appears to be necessary. Rat gastric mucosal scrapings incorporate in vitro several labeled compounds such as 14C-U-D-glucose, 35SO4, 14C-I-L-fucose and L-G-3H-proline in glycoproteins similar to those synthetized in vivo by native mucosa. The aim of our present work is to describe in detail the procedure we use and to show its reproducibility in 9 separate experiments. Secreted glycoproteins (fraction II) and intracellular glycoproteins (fraction III) obtained during a 4 hours in vitro incubation of rat gastric mucosal scrapings at 37 degrees C in standardized conditions were separately studied. The protein and hexose contents, total incorporated radioactivity and specific radioactivity (cpm/mg protein) were determined in these two fractions. The protein content of fraction II from 2 rat gastric mucosal scrapings incubated in 5 ml was 2 mg +/- 0.4 SEM and its hexose was 38.7 mg per 100 mg protein +/- 3.9. Fraction III contained 6.3 mg protein +/- 1.3 and 15.7 mg hexose per 100 mg protein +/- 3.5. About 27% of the total radioactivity incorporated (from 14C-glucose) was in fraction II (+/- 2.6 SEM) and 73% (+/- 2.6 SEM) in the cell bound fraction III. The distribution of total radioactivity was quite similar to that of total proteins : about 25% proteins were in fraction II and 75% in fraction III with a SEM of about 5 to 10% of the average value. If the biosynthetic activity is expressed as specific radioactivity, i.e. the ratio of incorporated radioactivity to mg proteins, the average for both fraction II and fraction III was about 10,000 cpm/mg protein and the standard error of the mean values are +/- 5.2% of the mean for in vitro secreted mucus glycoproteins (fraction II) and 9.1% in the case of intracellular glycoproteins (fraction III). These values can be considered as a satisfactory index of reproducibility of the method of mucosal scrapings if performed as described above.

UI	MeSH Term	Description	Entries
D008297	Male		Males
D008722	Methods	A series of steps taken in order to conduct research.	Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D009093	Mucus	The viscous secretion of mucous membranes. It contains mucin, white blood cells, water, inorganic salts, and exfoliated cells.
D002250	Carbon Radioisotopes	Unstable isotopes of carbon that decay or disintegrate emitting radiation. C atoms with atomic weights 10, 11, and 14-16 are radioactive carbon isotopes.	Radioisotopes, Carbon
D005753	Gastric Mucosa	Lining of the STOMACH, consisting of an inner EPITHELIUM, a middle LAMINA PROPRIA, and an outer MUSCULARIS MUCOSAE. The surface cells produce MUCUS that protects the stomach from attack by digestive acid and enzymes. When the epithelium invaginates into the LAMINA PROPRIA at various region of the stomach (CARDIA; GASTRIC FUNDUS; and PYLORUS), different tubular gastric glands are formed. These glands consist of cells that secrete mucus, enzymes, HYDROCHLORIC ACID, or hormones.	Cardiac Glands,Gastric Glands,Pyloric Glands,Cardiac Gland,Gastric Gland,Gastric Mucosas,Gland, Cardiac,Gland, Gastric,Gland, Pyloric,Glands, Cardiac,Glands, Gastric,Glands, Pyloric,Mucosa, Gastric,Mucosas, Gastric,Pyloric Gland
D005947	Glucose	A primary source of energy for living organisms. It is naturally occurring and is found in fruits and other parts of plants in its free state. It is used therapeutically in fluid and nutrient replacement.	Dextrose,Anhydrous Dextrose,D-Glucose,Glucose Monohydrate,Glucose, (DL)-Isomer,Glucose, (alpha-D)-Isomer,Glucose, (beta-D)-Isomer,D Glucose,Dextrose, Anhydrous,Monohydrate, Glucose
D006023	Glycoproteins	Conjugated protein-carbohydrate compounds including MUCINS; mucoid, and AMYLOID glycoproteins.	C-Glycosylated Proteins,Glycosylated Protein,Glycosylated Proteins,N-Glycosylated Proteins,O-Glycosylated Proteins,Glycoprotein,Neoglycoproteins,Protein, Glycosylated,Proteins, C-Glycosylated,Proteins, Glycosylated,Proteins, N-Glycosylated,Proteins, O-Glycosylated
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D051381	Rats	The common name for the genus Rattus.	Rattus,Rats, Laboratory,Rats, Norway,Rattus norvegicus,Laboratory Rat,Laboratory Rats,Norway Rat,Norway Rats,Rat,Rat, Laboratory,Rat, Norway,norvegicus, Rattus
D066298	In Vitro Techniques	Methods to study reactions or processes taking place in an artificial environment outside the living organism.	In Vitro Test,In Vitro Testing,In Vitro Tests,In Vitro as Topic,In Vitro,In Vitro Technique,In Vitro Testings,Technique, In Vitro,Techniques, In Vitro,Test, In Vitro,Testing, In Vitro,Testings, In Vitro,Tests, In Vitro,Vitro Testing, In