Developing Rat Pancreas Research Articles

Immunophenotypical analysis of pancreatic interstitial cells in the developing rat pancreas and myofibroblasts in the fibrotic pancreas in dogs and cats.

Pancreatic fibrosis develops as the results of the activity of myofibroblasts capable of producing collagens. The myofibroblasts derive from pancreatic interstitial cells, including pancreatic stellate cells (PSCs), which can express glial fibrillary acidic protein (GFAP). First, we investigated the expression patterns of vimentin, desmin, α-smooth muscle actin (α-SMA), Thy-1 and GFAP in the developing rat pancreas (in fetuses at 18 and 20 days, neonates from 1 to 21 days, and adults). Interstitial cells in the developing pancreas expressed vimentin, desmin, GFAP and Thy-1 at varying degrees; interestingly, the reactivity for desmin and vimentin was the highest in fetuses. GFAP expression was consistent between fetuses and neonates, and Thy-1 reactivity transiently increased after birth; however, α-SMA-positive interstitial cells were rarely seen. Next, we analyzed the immunophenotypical characteristics of myofibroblasts appearing in pancreatic fibrosis in dogs and cats. With increasing fibrotic grade, myofibroblasts showed increased expression of vimentin, desmin and α-SMA, in addition to increased GFAP expression. Collectively, pancreatic interstitial cells and myofibroblasts may have similar immunophenotypes, and myofibroblasts might originate partly from GFAP-expressing PSCs.

Detection of expressional changes induced by intrauterine growth restriction in the developing rat pancreas.

Intrauterine growth retardation (IUGR) is a disorder that can result in permanent changes in the physiology and metabolism of the newborn, which increased the risk of disease in adulthood. Evidence supports IUGR as a risk factor for the development of diabetes mellitus, which could reflect changes in pancreas developmental pathways. We sought to characterize the IUGR-induced alterations of the complex pathways of pancreas development in a rat model of IUGR. We analyzed the pancreases of Sprague Dawley rats after inducing IUGR by feeding a maternal low calorie diet from gestational day 1 until term. IUGR altered the pancreatic structure, islet areas, and islet quantities and resulted in abnormal morphological changes during pancreatic development, as determined by HE staining and light microscopy. We identified multiple differentially expressed genes in the pancreas by RT-PCR. The genes of the insulin/FoxO1/Pdx1/MafA signaling pathway were first expressed at embryonic day 14 (E14). The expressions of insulin and MafA increased as the fetus grew while the expressions of FoxO1 and Pdx1 decreased. Compared with the control rats, the expressions of FoxO1, Pdx1, and MafA were lower in the IUGR rats, whereas insulin levels showed no change. Microarray profiling, in combination with quantitative real-time PCR, uncovered a subset of microRNAs that changed in their degree of expression throughout pancreatic development. In conclusion, our data support the hypothesis that IUGR influences the development of the rat pancreas. We also identified new pathways that appear to be programmed by IUGR.

Expression and localization of paxillin in rat pancreas during development

To investigate the expression and localization of paxillin in rat pancreas during development. Pancreata from Sprague Dawley rat fetuses, embryos, young animals, and adult animals were used in this study. Expression levels of paxillin in pancreata of different development stages were detected by reverse transcription polymerase chain reaction and Western blotting. To identify the cell location of paxillin in the developing rat pancreas, immunohistochemistry and double-immunofluorescent staining were performed using antibodies for specific cell markers and paxillin, respectively. The highest paxillin mRNA level was detected at E15.5 (embryo day 15.5) following a decrease in the later developmental periods (P < 0.05 vs E18.5, P0 and adult, respectively), and a progressively increased paxillin protein expression through the transition from E15.5 to adult was detected. The paxillin positive staining was mainly localized in rat islets of Langerhans at each stage tested during pancreas development. The dynamic expression of paxillin in rat pancreas from different stages indicates that paxillin might be involved in some aspects of pancreatic development.

An appraisal of intermediate filament expression in adult and developing pancreas: vimentin is expressed in alpha cells of rat and mouse embryos.

Intermediate filaments are frequently used in studies of developmental biology as markers of cell differentiation. To assess whether they can be useful to identify differentiating pancreatic endocrine cells, we examined the pattern of expression of nestin, cytokeratin 20, and vimentin on acetone-fixed cryosections of rat adult and developing pancreas. We also studied vimentin expression in mouse embryonic pancreas at E19. Cytokeratin 20 was found in all pancreatic epithelial cell lineages during the entire development of the rat gland and in the adult animals. Under our experimental conditions, therefore, cytokeratin 20 is not an exclusive marker of rat duct cells. Nestin was detected exclusively in stromal cells either in the adult or developing rat pancreas. Vimentin was observed within cells located in the primitive ducts of rat pancreas starting from E12.5. Their number rapidly increased, reaching its highest level in newborn animals. Vimentin was also spotted in alpha cells starting from E12.5 but disappeared soon after birth, likely identifying immature or recently differentiated alpha cells. In addition, vimentin was observed in duct and alpha cells of mouse developing pancreas showing that its expression in such cells is not an event restricted to the rat. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.

Expression and localization ofWolfram syndrome 1gene in the developing rat pancreas

To investigate the expression and function of Wolfram syndrome 1 gene (WFS1) during the development of normal pancreas. Pancreas from Sprague-Dawley rat fetuses, embryos, young and adult animals were used in this study. Expression levels of WFS1 in pancreas of different development stages were detected by reverse transcription-polymerase chain reation (RT-PCR) and Western blotting. To identify the cell location of WFS1 in the developing rat pancreas, double-immunofluorescent staining was performed using antibodies to specific cell markers and WFS1, respectively. Compared to E15.5, the highest level of WFS1 mRNA was detected at E18.5, the level of WFS1 mRNA in E15.5 and P0 was less, and at a lowest at adult (P < 0.05 vs P0 and adult), respectively. Compare to E15.5, the highest level of WFS1 was at P14 and lowest at P21 (P < 0.05 vs P14 and P21), respectively. The WFS1 positive staining is expressed in the normal developing rat pancreas mainly in the islet beta-cells and mesenchyme at each stage tested. These results indicate that WFS1 may be involved in some aspects of pancreatic development and further research on WFS1 may provide new evidences to prove the interactions between mesenchyma and epithelia at the same time.

Expression and localization of Wolfram syndrome 1 gene inthe developing rat pancreas

Expression and localization of Wolfram syndrome 1 gene inthe developing rat pancreas

Expression and localization of mesothelin in developing rat pancreas

To define a genetic network that regulates development of the pancreas, we used high-density microarray (Affymetrix) to generate transcriptional profiles of rat pancreas from five biologically significant stages of development: embryonic day 12.5 (E12.5), E15.5, E18.5, postnatal day 0 (P0) and adult. Many genes were notably highly expressed in the later gestation when islet architecture and function are gradually forming. The expression and localization of mesothelin, one of these genes, was further examined. Reverse transcription-polymerase chain reaction and Western blot analysis revealed that mRNA and protein levels of mesothelin were high from later gestation to 2-3 weeks after birth, and with relatively low but detectable expression levels in adult rat pancreas. Immunolocalization indicated that mesothelin localized not only in islet beta-cells but also in the mesenchyme of developing rat pancreas. Transient mesothelin expression was concomitant with the development of islets architecture formation, remodeling and maturation. These findings indicate that mesothelin is dynamically expressed in the developing rat pancreas and that mesothelin might be involved in some developmental events during development of rat pancreas.

Early Expression of Adenosine 5′-Triphosphate-Gated P2X7 Receptors in the Developing Rat Pancreas

Extracellular adenosine 5'-triphosphate modulates the functions of the adult pancreas via 2 nucleotide receptor families, the P2X and P2Y receptors. Expression of the P2X7 receptor has been demonstrated in islet cells of the pancreas, particularly the mature alpha cells that secrete glucagon. In the streptozotocin-induced diabetic model, a loss of insulin-secreting cells was accompanied by an increase in alpha cells that expressed the P2X7 receptor. In the present study, we have examined the expression of P2X7 receptors in the developing pancreas from embryonic days 10 (E10) to E18. We detected P2X7 receptor-immunoreactive cells in pancreatic islet cells as early as E11' before glucagon expression. Subsequently, P2X7 receptors were expressed in glucagon-secreting cells at E12, and complete colocalization was observed at E14. Occasional colocalization of P2X7 receptors and insulin was observed in scattered cells at E12 and E14, but not at E18, when the glucagon- and insulin-secreting cells were almost completely segregated. It was found that P2X7 receptors were expressed early in a subpopulation of glucagon- and insulin-immunopositive cells in developing islets and subsequently became restricted to glucagon-expressing cells as development proceeded. The possible functional significance of these changes is discussed.

Cocaine- and amphetamine-regulated transcript (CART) is expressed in several islet cell types during rat development.

Cocaine- and amphetamine-regulated transcript (CART) is an anorexigenic peptide widely expressed in the central and peripheral, including the enteric, nervous systems. CART is also expressed in pituitary endocrine cells, adrenomedullary cells, islet somatostatin cells, and in rat antral gastrin cells. We used immunocytochemistry (IHC) and in situ hybridization (ISH) to study CART expression in developing rat pancreas. We also examined co-expression of CART and islet hormones and developmental markers and the effect of CART on proliferation using clonal insulin cells (INS-1 832/13). A major portion of each of the islet cell types, except the ghrelin cells, expressed CART during a period before and around birth. Two weeks postnatally, CART expression was restricted to somatostatin cells. Pre- and early postnatally, many of the CART-expressing cells co-expressed cytokeratin 20 (CK20), a marker of duct cells and islet precursor cells, the trophic hormone gastrin, and a smaller subpopulation also harbored the proliferation marker Ki67. CART was also expressed in pancreatic nerve fibers, both sensory and autonomic, and in ganglion nerve cell bodies. Although highly expressed in the developing islets, CART did not affect proliferation of INS-1 cells. We have demonstrated that CART is expressed in several islet cell types during rat development but is restricted to somatostatin cells and neurons in the adult rat.

Carboxyl-Methylation of Rab3D in the Rat Pancreatic Acinar Tumor Cell Line AR42J

Rab3D is a small GTPase implicated in regulated exocytosis, and is a marker of secretory granules in exocrine cells. We have previously shown that rab3D undergoes reversible carboxyl-methylation in adult rat pancreatic acinar cells, and that carboxyl-methylation of rab3D is developmentally regulated concomitantly with the maturation of the regulated secretory apparatus in rat pancreas. We also observed that dexamethasone treatment of the rat pancreatic acinar tumor cell line, AR42J, led to a significant increase in the size of the unmethylated pool of a rab3-like protein. The current study was designed to further characterize this rab3-like protein. Here we show that AR42J cells express rab3D, and that the protein focuses on 2D gels as two spots with pI values of 4.9 and 5.0. Treatment of AR42J cells with N-acetyl-S-geranylgeranyl-l-cysteine, an inhibitor of carboxyl-methylation, led to a decrease in the basic form of rab3D and a proportional increase in the acidic form. In contrast, N-acetyl-S-farnesyl-l-cysteine, which inhibits carboxyl-methylation of farnesylated proteins, had no effect. Lovastatin, an inhibitor of geranylgeranylation, also induced an accumulation of the acidic form of rab3D. Taken together, these data indicate that rab3D can undergo reversible carboxyl-methylation in AR42J cells by a geranylgeranyl-specific methyltransferase. The 2D gel and immunoblotting analyses indicated that dexamethasone treatment of AR42J cells led to an increase in the proportion of the unmethylated form of rab3D concurrent to inducing a regulated secretory pathway, similar to the rab3D profile change in developing rat pancreas. Our data, along with previous studies done on developing rat pancreas, indicate that the tumor cell line AR42J represents a good model system for studying the regulated secretory pathway, and that carboxyl-methylation of rab3D may play a role in the acquisition of stimulus–secretion coupling.

Cellular distribution and ontogeny of insulin-like growth factors (IGFs) and IGF binding protein messenger RNAs and peptides in developing rat pancreas.

To determine the role of insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) in the development of the pancreas, and specifically of the islets of Langerhans, we have examined the cellular distribution and developmental changes in the expression of IGFs and IGFBPs in the pancreas of the fetal and neonatal rat between 19.5 days of gestation and postnatal day 28. This represents a period of substantial growth and restructuring of the beta cell component in islets of this species. IGF-I, IGF-II, and IGFBPs-1 to -6 mRNAs were localized by in situ hybridization, and peptides by immunohistochemistry, in histological sections. IGF-II mRNA was highly expressed in islet cells and some ductal epithelial cells in late fetal and early neonatal life, but was barely detectable by postnatal day 28. IGF-II peptide showed a similar distribution. IGF-I mRNA was barely detected in the fetus or neonate and was localized predominantly in the ductal and acinar tissues after postnatal day 7. IGF-I immunoreactivity was associated with some islet cells in the fetus and neonate, suggesting an endocrine rather than a paracrine source. We performed co-localization studies to assess whether the distribution of IGFs within the pancreas might be due to a sequestration by locally produced IGFBPs. The presence of mRNAs for both IGFBPs-1 and -2 was minimal in the pancreas prior to postnatal day 7, although subsequently IGFBP-1 mRNA was seen in islet cells, while IGFBP-2 mRNA was localized in both islets and acinar tissues. In contrast, both IGFBPs-1 and -2 immunoreactivities were identified in islets from late fetal life, suggesting a circulatory source for these IGFBPs during early pancreatic development. IGFBPs-3 to -5 mRNAs and immunoreactivities were identified within islet cells throughout fetal and neonatal life, with IGFBPs-3 and -5 being mainly associated with the alpha cell-rich islet mantle. The results show a compartmentalization of IGFs within pancreatic tissue, reflecting both paracrine and endocrine sources. The localization and action of IGFs in pancreas likely involves sequestration and distribution by endogenous as well as circulating IGFBPs.

Impact of Protein Deprivation on Protein and DNA Synthesis in the Developing Rat Pancreas

The metabolic effects of protein malnutrition on growth and development of the exocrine pancreas are largely unknown. The aim of the present study was to evaluate the effects of protein malnutrition on pancreatic protein and DNA synthesis during postnatal development. Rat dams and their offspring were fed a protein-deficient diet (6% casein) or a control diet (25% casein) during gestation, lactation and after weaning. Pancreatic protein and DNA synthesis were measured in vitro at postnatal ages 1, 3, 10, 23, 36 and 60 days, by assessing [³H]leucine and [³H]thymidine incorporation in freshly isolated acini. Different patterns of protein synthesis were seen in the two groups. At birth, pancreatic protein synthesis was low in both control and malnourished animals. At day 3, protein synthesis in the control acini increased 10-fold while synthesis in acini of the malnourished animal group was only 50% of age-matched control values. No differences in protein synthesis were detected between the control and malnourished groups between 10 and 36 days of age. At 60 days (adulthood), acinar protein synthesis declined in the control-fed rats, but a significant increase was observed in the malnourished animals (p < 0.0005). At birth, DNA synthesis was high in the acini from both control and malnourished animals. The low-protein diet induced a slight reduction in DNA synthesis at day 3, without altering the general pattern during later stages of development. In conclusion, protein deprivation has variable effects on pancreatic protein and DNA synthesis at different stages of postnatal development. Furthermore, the mechanisms of control within acini appear to be intrinsically regulated.

Rat endocrine pancreatic development in relation to two homeobox gene products (Pdx-1 and Nkx 6.1).

We studied the distribution of the homeodomain proteins Pdx-1 and Nkx 6.1 in the developing rat pancreas. During early development, nuclear staining for both Pdx-1 and Nkx 6.1 occurred in most epithelial cells of the pancreatic anlage. Subsequently, Nkx 6.1 became more beta-cell-restricted, and Pdx-1 also occurred in other islet cell types and in the duodenal epithelium. During early pancreatic development, cells co-storing insulin and glucagon were regularly detected. The vast majority of these did not possess nuclear staining for either Pdx-1 or Nkx 6.1. Subsequently, cells storing insulin only appeared. Such cells displayed strongly Pdx-1- and Nkx 6.1-positive nuclei. Therefore, Nkx 6.1, like Pdx-1, may be an important factor in pancreatic development and in mature insulin cell function.

Immunocytochemical localization of the NPY/PYY Y1 receptor in the developing pancreas.

Neuropeptide Y, peptide YY, and pancreatic polypeptide are structurally related peptides that are considered to play a role in the regulation of pancreatic secretion and blood flow. Several receptor subtypes for these peptides have been identified, and the Y1, Y2, Y4/PP1, Y5, and Y5/PP2/Y2b receptors are cloned. We have prepared polyclonal peptide antibodies that recognize the Y1 receptor and now report on its localization in the adult and developing rat pancreas. In the adult pancreas, Y1 receptors were detected both in some centroacinar and intralobular duct cells and in endothelial cells. In the developing pancreas (E12.5-E16.5), Y1 receptor immunoreactivity was observed in numerous nonendocrine epithelial cells. These cells occurred in the immediate vicinity of peptide YY-positive endocrine cells. At E16.5, a fraction of these Y1 receptor-containing cells co-stored amylase. One day later, Y1 receptor immunoreactivity became restricted to pancreatic duct-like cells that occurred in close proximity to peptide YY cells. In fetal rats, intense Y1 receptor staining was also observed in endothelial cells. These observations, together with the finding of early pancreatic peptide YY expression, suggest that peptide YY produced by fetal endocrine cells may exert an action on exocrine cells, duct cells and endothelial cells during development.

Expression of ornithine decarboxylase in pancreatic development.

We examined the relationship between ornithine decarboxylase (ODC) and growth and differentiation in the developing rat exocrine pancreas. The ODC activity profile showed 2 distinct stages of increases with the first occurred at 14-16 days of age, and a second at 21-23 days of age. Growth parameters evaluated as gains in tissue mass, protein and DNA content in the pancreas indicated a low growth rate soon after birth with a transition to a much more rapid growth rate around the age of 20-21 days, a time corresponded to the second rise in ODC activity. Differentiation parameters evaluated as the accumulation of trypsinogen, amylase and lipase showed different temporal changes. While the rate of accumulation of all three enzymes was relatively low following birth, a rapid rate of accumulation of trypsinogen and amylase started around 15-16 days, a time corresponding to the first rise in ODC activity. Lipase, however, did not show an increase in its accumulation until around age 20 days. These results indicate that a rise in ODC activity is closely associated with growth and differentiation in the developing rat pancreas. To further examine this issue, the steady state levels of ODC mRNA in developing rats were evaluated by Northern blots probed with an ODC cDNA. The developmental profile of ODC mRNA showed a broad peak with a pronounced shoulder occurring at 10 days of age. A higher peak was reached around 20 days of age, then dropped precipitously to a very low level at the age of 24 days. This temporal changes in the level of ODC mRNA show good relationship to the changes in ODC activity suggesting that the control of ODC expression occurs at least in part at the pre-translational level.

Beta cells arise from glucose transporter type 2 (Glut2)-expressing epithelial cells of the developing rat pancreas.

We observe Glut2 protein in day 11 (E11) rat embryos in an endodermal domain containing the pancreatic primordium. Glut2 expression continues as the endodermal epithelium evaginates into the surrounding mesenchyme to form the pancreatic buds. Cells of the dorsal and ventral pancreatic buds maintain Glut2 expression as the epithelium grows and branches to form ducts. As acini form at the ends of the ducts, acinar cells cease Glut2 expression. Insulin protein is first detected in small clusters at the interface between pancreatic epithelium and mesenchyme. These clusters disperse into the interstitial tissue between E13 and E17. At E17 a distinct, larger population of insulin-expressing cells arises in the Glut2-expressing ductal network. Insulin- and Glut2-coexpressing cells then appear to segregate into large aggregates to form the beta cells of the islets of Langerhans. These observations support the hypothesis that two biologically distinct populations of insulin-expressing cells arise during pancreas formation.

Autologous regulation of pancreatic glucocorticoid receptors in suckling rats.

The exact mechanism of glucocorticoid action in pancreatic ontogeny is not fully understood. To investigate whether glucocorticoid receptors are subjected to autologous regulation during the early stage of life, the effects of glucocorticoid administration and adrenalectomy (Adx) were examined. Rat pups adrenalectomized on day 10 were given either dexamethasone (DX, 20 micrograms/100 g body weight) or vehicle daily for 3 days before being killed at 15 days of age. Adx increased and DX replacement decreased glucocorticoid receptor binding in the developing rat pancreas. Scatchard analysis showed that Adx led to an increase in receptor number without changing its affinity (Bmax, from 0.385 +/- 0.073 to 0.553 +/- 0.086 pmol/mg protein; Kd, from 2.2 +/- 0.5 x 10(-8) M to 2.7 +/- 0.6 x 10(-8) M), whereas DX down-regulated its own receptor by decreasing the receptor number and lowering the affinity (Bmax, from 0.553 +/- 0.086 to 0.140 +/- 0.025 pmol/mg protein; Kd, from 2.7 +/- 0.6 x 10(-8) M to 8.8 +/- 2.5 x 10(-8) M). The degree of down-regulation was dependent on the dose of DX. Maximal down-regulation (80%) was found at 20 micrograms/100 g body weight, whereas a dose of 1 microgram/100 g body weight caused only 40% down-regulation. Down-regulation was also ligand specific. DX was the most effective, whereas even at 100 times the DX dose, corticosterone had only 70% of the effect of DX. DX-induced down-regulation occurred rapidly (within hours), whereas up-regulation in Adx occurred relatively slowly (days), suggesting these two phenomena involve different mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)

Pharmacological and biochemical characterization of cholecystokinin/gastrin receptors in developing rat pancreas

Cholecystokinin/gastrin receptors in the pancreas of newborn (3-day-old) rats are of type A, as in control mature rats, revealed by pharmacological analysis of specific 125I-Bolton-Hunter-reagent-labelled [Thr34,Ahx37]cholecystokinin(31-39) (Ahx, aminohexanoic acid) binding. Also, by 1 day post-partum, pancreatic cholecystokinin receptors were shown to be coupled to guanine-nucleotide-binding regulatory (G) proteins. Scatchard analysis of 125I-Bolton-Hunter-reagent-labelled [Thr34,Ahx37]cholecystokinin(31-39) binding to pancreatic membranes from rats at different times after birth showed a slight increase in the binding capacity of cholecystokinin receptors between days 3 and 14 and a sixfold increase in 21-day-old rats, with no change in receptor affinity during development. SDS/PAGE analysis of pancreatic membranes affinity labelled with the photoactivable ligand 125I-[2-(p-azidosalicylamido)-1,3'-dithiopropionate]-labelled [Thr34,Ahx37]cholecystokinin-(31-39) identified cholecystokinin receptors of 100-135 kDa in 3-day-old rats, 96-130 kDa in 7-day-old rats, 90-125 kDa in 10-day-old rats and 85-100 kDa in 14-day-old and 21-day-old rats, as found in control adult rats. Endo-beta-N-acetylglucosaminidase F treatment yielded a core protein of 42 kDa in all developmental stages. These findings are consistent with an age-related postnatal expression of distinct glycoforms of pancreatic cholecystokinin receptors. Furthermore, it was observed that the period 2-3 weeks after birth, characterized by stabilization of the mass of the cholecystokinin receptor, precedes the dramatic increase in the receptor number.

Age and enzyme specificity of the response of developing rat pancreas to the trophic effects of bombesin.

The immediate postnatal period is a time of rapid pancreatic growth and development. Gastrointestinal regulatory peptides have been shown to exert trophic effects on the pancreas, and one such peptide, bombesin, has been shown to exert trophic effects on the pancreas of both the suckling and adult rat. Our previous studies had suggested that the sensitivity of the rat pancreas to bombesin might be changing during the suckling period. We therefore conducted experiments to determine if age-related changes in the responsiveness of rat pancreas to chronic administration of exogenous bombesin occur, and to characterize these changes. Beginning on day 3 ("suckling") or 24 ("weanling") postnatally, groups of rats were injected subcutaneously with several doses of bombesin tetradecapeptide every 12 h for 7 days. Bombesin injection produced a dose-dependent increase in pancreatic protein content and trypsin activity in both suckling and weanling groups, although the degree of increase was greater in the weanling group than in the sucklings. Significant increases in pancreatic contents of DNA and amylase activity were observed only in the weanling group. We have thus demonstrated for the first time that the pancreas of the suckling rat exhibits a diminished response to chronic parenteral administration of bombesin in terms of changes in protein and DNA contents, as well as enzyme composition, compared to the pancreas of weanlings. The elucidation of the physiologic basis for these differences may provide specific information regarding the mechanism of bombesin-stimulated pancreatic growth, as well as general information concerning the control of pancreatic growth during development.

Ontogenetic expression of peptidyl-glycine α-amidating monooxygenase mRNA in the rat pancreas

Qualitative and quantitative expression of m.RNA coding for Peptidyl-Glycine alpha-Amidating Monooxygenase (PAM) in the developing rat pancreas was investigated by Northern and dot blot hybridization, with a bovine PAM c.DNA probe (0.7 kb fragment). A specific hybridization signal was evidenced for a 3.7 kb m.RNA species. Measurement of PAM m. RNA rate during the rat pancreas ontogenesis revealed a biphasic profile which appeared corelated with that of gastrin and TRH m.RNA respectively. On the other hand, streptozotocin-treatment resulted in a 50% decrease of PAM m.RNA levels.