The ontogeny and distribution of gastrin- and serotonin-immunoreactive (IR) cell in the proventriculus of chicks (Gallus gallus domestica, n = 60) in different growth periods was examined immunohistochemically using antisera specific to gastrin and serotonin. Gastrin and serotonin-IR cells were detected in chick proventriculus. Gastrin-IR cells were first evident after 12 days of incubation in lamina epithelialis and compound glands, while serotonin-IR cells were observed only in compound glands at that same time. Gastrin-IR and serotonin-IR cells increased in frequency on incubation day 14 and 16, respectively. Towards the end of incubation, gastrin- and serotonin-IR cell numbers decreased. In adult chicken, both IR cells were present but not lower numbers. The observations demonstrate the presence of gastrin- and serotonin-IR cells in the proventriculus of developing chicks in temporally changing frequencies.
Animals ; Chick Embryo/*metabolism ; Endocrine Cells/cytology/metabolism ; Gastrins/*metabolism ; Gene Expression Regulation, Developmental/physiology ; Proventriculus/*embryology/*metabolism ; Serotonin/*metabolism

OBJECTIVETo construct a method to culture pancreatic progenitor cells in hanging drop and on floating filter,and to examine if pancreatic progenitor cells can differentiate into mature endocrine cells with this method.

METHODSMurine embryos at day 12.5 were isolated and digested into single cells,which were then cultured in hanging drop for 24h and formed spheres.Spheres were cultured on the filter for 6 days,which floated in the dish containing medium.During culture,the expressions of pancreas duodenum homeobox-1(PDX-1)and neurogenin3(Ngn3)were determined.The expressions of endocrine and exocrine markers,insulin,glucagon,and carboxypeptidase(CPA)were determined on day 7 by immunohistochemistry.Insulin secretion of spheres stimulated by glucose was detected by ELISA.The changes of pancreatic marker expressions during culture were monitored by real-time polymerase chain reaction(PCR).

RESULTSOne day after the culture,there were still a large amount of PDX-1 positive cells in pancreatic spheres,and these cells proliferated.On day 3,high expression of Ngn3 was detected,and the Ngn3-positive cells did not proliferate.On day 7,The expressions of endocrine and exocrine markers in the differentiated pancreatic progenitor cells were detected,which were consistent with that in vivo.Insulin was secreted by spheres upon the stimulation of glucose.

CONCLUSIONIn hanging drop and on floating filter,pancreatic progenitor cells can differentiate into mature endocrine cells.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Endocrine Cells ; cytology ; Homeodomain Proteins ; metabolism ; Insulin ; metabolism ; Mice ; Pancreas ; cytology ; Stem Cells ; cytology ; Trans-Activators ; metabolism

Here we report the detection and distribution of synaptophysin (SPY), non-neuronal enolase (NNE), glial fibrillary acidic protein (GFAP), vimentin (VIM), neuropeptide Y (NPY), and vasoactive intestinal peptide (VIP) expression in the goat forestomach during prenatal development. A total of 140 embryos and fetuses were examined to evaluate protein expression from the first stage of prenatal life until birth. In all cases, SPY immunoreactivity was detected at 53 days gestation in the lamina propria-submucosa, tunica muscularis, serosa, and myenteric plexuses. Immunoreactivity to NNE was observed at 64 days gestation in the same locations as well as the epithelial layer. Glial cells were found at 64 days as indicated by signals corresponding to GFAP and VIM at 39 days. Positive staining for NPY and VIP was observed at 113, 75, and 95 days in the rumen, reticulum, and omasum, respectively, in the lamina propria-submucosa, tunica muscularis, and myenteric plexuses of each of these gastric compartments. These findings indicate possible preparation of the fetal goat forestomach for postnatal function. Compared to other ruminant species, neuroendocrine cells, glial cells and peptidergic innervations markers were detected earlier compared to sheep but at around the same stage as in deer.
Animals ; Biological Markers/metabolism ; Embryo, Mammalian ; Endocrine Cells/*metabolism ; Fetus/metabolism ; Gene Expression Regulation, Developmental ; Goats/*embryology/genetics ; Immunohistochemistry ; Neuroendocrine Cells/*metabolism ; Neuroglia/*metabolism ; Proteins/genetics ; Rumen/*embryology/metabolism

OBJECTIVETo establish a new culture method to induce the differentiation of embryonic pancreatic cells into mature endocrine cells.

METHODSMouse embryos at day 12.5 were used and embryonic pancreata were isolated. The isolated embryonic pancreata were cultured on the filter for 7 days, which floated in the dish containing medium. During culture, the expression of pancreas duodenum homeobox-1 (PDX-1), a pancreatic stem cell marker, was examined at day 1. The expression of neurogenin 3 (Ngn3), a pancreatic progenitor cell marker, was examined at day 3. The expressions of endocrine and exocrine markers, insulin, glucagon, and carboxypeptidase (CPA) were examined at day 7 by immunohistochemistry. The kinetics of pancreatic marker expression during culture was assayed by real-time PCR.

RESULTSMany pancreatic stem cells still existed in embryonic pancreata cultured for 1 day; meanwhile, these pancreatic stem cells proliferated in high rate. A large amount of pancreatic progenitor cells were found in embryonic pancreata cultured for 3 days.Pancreatic stem/progenitor cells differentiated into mature endocrine and exocrine cells in embryonic pancreata after having been cultured for 7 days. Furthermore, the expression pattern of pancreatic marker is consistent with that in vivo.

CONCLUSIONWe successfully established a new culture method, with which embryonic pancreatic cells can efficiently differentiate into mature endocrine cell.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Endocrine Cells ; cytology ; Female ; Homeodomain Proteins ; metabolism ; Male ; Mice ; Nerve Tissue Proteins ; metabolism ; Pancreas ; cytology ; Trans-Activators ; metabolism

BACKGROUND AND OBJECTIVES: Insulin is a peptide hormone that regulates the metabolism of carbohydrates and fats by promoting the absorption of glucose from the blood to skeletal muscles. Insulin has been reported to be closely related to cardiovascular, respiratory, and endocrine disease. However, the effect of insulin on production of major mucins in human airway epithelial cells has not been reported. Therefore, this study investigated the relationship between high levels of insulin and mucin in human airway epithelial cells. MATERIALS AND METHODS: This study analyzed the effect of high level of insulin on MUC4, MUC5AC, and MUC5B expression using reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay in human airway epithelial cells. RESULTS: In human NCI-H292 airway epithelial cells, high level of insulin significant increased MUC4, MUC5AC, and MUC5B mRNA expression and glycoprotein production. In the primary cultures of normal nasal epithelial cells, high level of insulin also increased MUC4, MUC5AC, and MUC5B expression. CONCLUSION: These results suggest that insulin plays a role in control of mucus hypersecretion in human airway epithelial cells.
Absorption ; Carbohydrates ; Endocrine System Diseases ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells* ; Fats ; Glucose ; Glycoproteins ; Humans ; Insulin ; Metabolism ; Mucins ; Mucus ; Muscle, Skeletal ; RNA, Messenger

BACKGROUNDMenin is a ubiquitously expressed protein encoded by the multiple endocrine neoplasia type 1 (MEN1) gene. Besides its importance in endocrine organs, menin has been shown to interact with the mixed lineage leukemia (MLL) protein, a histone H3 lysine 4 methyltransferase, and plays a critical role in hematopoiesis and leukemogenesis. Previous studies have shown that menin promotes transforming growth factor beta (TGF-β) signaling in endocrine cells. However, little is known regarding the impact of TGF-β pathway on menin in hematopoietic system. Here, with leukemia cell lines generated from conditional MEN1 or TGF-β receptor (TβRII) knockout mouse models, we investigated the possible cross-talk of these two pathways in leukemia cells.

METHODSMEN1 or TβRII conditional knockout mice were bred and the bone marrow cells were transduced with retroviruses expressing oncogeneic MLL-AF9 (a mixed lineage leukemia fusion protein) to generate two leukemia cell lines. Cell proliferation assays were performed to investigate the effect of TGF-β treatment on MLL-AF9 transformed leukemia cells with/without MEN1 or TβRII excision. Menin protein was detected with Western blotting and mRNA levels of cell proliferation-related genes Cyclin A(2) and Cyclin E(2) were examined with real-time RT-PCR for each treated sample. In vivo effect of TGF-β signal on menin expression was also investigated in mouse liver tissue after TβRII excision.

RESULTSTGF-β not only inhibited the proliferation of wild type MLL-AF9 transformed mouse bone marrow cells, but also up-regulated menin expression in these cells. Moreover, TGF-β failed to further inhibit the proliferation of Men1-null cells as compared to Men1-expressing control cells. Furthermore, excision of TβRII, a vital component in TGF-β signaling pathway, down-regulated menin expression in MLL-AF9 transformed mouse bone marrow cells. In vivo data also confirmed that menin expression was decreased in liver samples of conditional TβRII knockout mice after TβRII excision.

CONCLUSIONThese results provided the first piece of evidence of cross-talk between menin and TGF-β signaling pathways in regulating proliferation of leukemia cells, suggesting that manipulating the cross-talk of the two pathways may lead to a novel therapy for leukemia.

Animals ; Blotting, Western ; Cells, Cultured ; Humans ; Leukemia ; metabolism ; Mice ; Mice, Knockout ; Multiple Endocrine Neoplasia Type 1 ; genetics ; metabolism ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, Transforming Growth Factor beta ; genetics ; metabolism ; Transforming Growth Factor beta ; genetics ; metabolism