Morphological and biochemical studies of B cells of fetal rat endocrine pancreas in organ culture. Evidence for (pro) insulin biosynthesis.
Fetal rat pancreases explanted on the 18th day of gestation and maintained in organ culture for 1-10 days were utilized for this series of studies. Ultrastructurally, at the time of explantation, the majority of fetal B cells was sparsely granulated and characterized by numerous free ribosomes and undeveloped rough endoplasmic reticulum (RER) and Golgi complexes. During the culture period, extensive development of the RER and Golgi complexes preceded an increasing accumulation of beta-granules. This later increase in the number of beta-granules and in the concentration of immunoreactive insulin was paralleled by a reduction of RER and Golgi complex activity. High resolution radioautographic studies of pulse-chase experiment over a 1 hr period demonstrated the shift of silver grains from the elements of the RER, through the Golgi region, and finally to the beta-granules. Incubation with (14)C-labeled leucine demonstrated the incorporation of radioactivity into molecules possessing the immunological and electrophoretic properties of insulin. These studies indicate that de novo synthesis of (pro)insulin occurs also during culture of fetal rat pancreas explanted relatively late in gestation.
D000818:Animals; D001345:Autoradiography; D002247:Carbon Isotopes; D002478:Cells, Cultured; D003470:Culture Media; D046508:Culture Techniques; D003594:Cytoplasmic Granules; D004721:Endoplasmic Reticulum; D005260:Female; D005333:Fetus; D005865:Gestational Age; D006056:Golgi Apparatus; D006168:Guinea Pigs; D006651:Histocytochemistry; D007106:Immune Sera; D007328:Insulin / Q000032:analysis / Q000302:isolation & purification; D007330:Insulin Antibodies / Q000032:analysis; D007930:Leucine / Q000378:metabolism; D008854:Microscopy, Electron; D009924:Organ Culture Techniques; D010179:Pancreas / Q000032:analysis / Q000166:cytology / Q000378:metabolism*; D011384:Proinsulin / Q000096:biosynthesis*; D011506:Proteins / Q000032:analysis; D011817:Rabbits; D051381:Rats; D012270:Ribosomes; D013552:Swine; D013997:Time Factors
D002247:Carbon Isotopes; D003470:Culture Media; D007106:Immune Sera; D007328:Insulin; D007330:Insulin Antibodies; D011506:Proteins; D011384:Proinsulin; D007930:Leucine