Effects of Matrix Proteins and Heparin-Binding Components in Fetal Bovine Serum upon the Proliferation of Ectoplacental Cone Cells in Mouse Blastocysts Cultured in Vitro1

Koi, Hideki; Tachi, Chikashi; Tojo, Hideaki; Kubota, Toshiro; Asô, Takeshi

doi:10.1095/biolreprod52.4.759

Cited by 6 publications

(5 citation statements)

References 55 publications

(66 reference statements)

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“…To investigate this, we isolated acinar cell clusters from control and b1 integrin-deficient pancreata and cultured them with or without FBS that contains rich ECM proteins. 19,20 b1 integrindeficient acinar cell clusters cultured with BSA showed significantly increased cell apoptosis when compared with the control cell clusters (*Po0.05; Figure 5c). However, the number of TUNEL-positive cells in b1 integrin-deficient acinar cell clusters was reduced and reached control levels when cultured with FBS medium (*Po0.05; Figure 5c).…”

Section: Significantly Reduced Amylase Expression Observed In B1 Intementioning

confidence: 95%

“…Acinar cell clusters were cultured in modified RPMI 1640 media 18 with either 1% BSA (Sigma) or 10% FBS (Invitrogen, Burlington, ON, Canada), reported to be enriched for ECM proteins, 19,20 for 24 h. Cell clusters from four experimental groups (b1KO-BSA, Ctrl-BSA, b1KO-FBS, and Ctrl-FBS) were harvested and processed for immunofluorescence with at least four mouse pancreata per experimental per group used.…”

Section: Acinar Cell Culture Experimentsmentioning

confidence: 99%

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β1 integrin–extracellular matrix interactions are essential for maintaining exocrine pancreas architecture and function

Riopel

Liu

et al. 2013

Laboratory Investigation

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Integrin receptors are responsible for integrating extracellular matrix signals inside the cell. The most prominent integrin receptor, b1 integrin, has a role in cell function, survival and differentiation. Recently, we demonstrated a profound in vivo role of b1 integrin expression in the pancreas on glucose homeostasis and islet function. Here, we extend these results by examining the role of b1 integrin in exocrine pancreatic structure and function. Adult C57Bl/6 mice hemizygous for a collagen type Ia2 (Col1a2) promoter-controlled tamoxifen-inducible Cre recombinase gene and homozygous for loxP-b1 integrin were injected with tamoxifen or corn oil to generate mice deleted or not for b1 integrin. Pancreata derived from these male mice were analyzed by quantitative reverse transcriptase-polymerase chain reaction, western blot and immunofluorescence. Our results showed that b1 integrin-deficient mice displayed a significant decrease in pancreas weight with a significant reduction of amylase, regenerating islet-derived protein II and carboxypeptidase-A expression (Po0.05-0.01). Compared with control pancreata, b1 integrin-deficient pancreata showed reduced mRNA expression of extracellular matrix (collagen type Ia2, fibronectin and laminin) genes (Po0.05), detached acini clusters and lost focal adhesion structure. Moreover, b1 integrin-deficient pancreatic acinar cells displayed decreased proliferation (Po0.05) and increased apoptosis (Po0.001). Apoptosis was reduced to that of controls when isolated exocrine clusters were cultured in media supplemented with extracellular matrix proteins. Taken together, these results implicate b1 integrin as an essential component for maintaining exocrine pancreatic structure and function. Integrin receptors have a significant role in cell-cell and cellextracellular matrix (ECM) contacts in many different tissue types. There are 18 a and 8 b receptors capable of forming 24 heterodimeric interactions; yet, half these interactions are made up by b1 integrin receptors. 1 This b1 integrin receptor group is largely responsible for attachment to the ECM. 2 Upon stimulation, b1 integrin has been shown to mediate cell motility, survival, proliferation and differentiation. [2][3][4][5] Pancreatic acini are enclosed round structures that produce digestive enzymes. 6 Basal lamina covers the acini at the basal surface, stimulating acini integrin receptors. 6 Pancreatic stellate cells (PSCs) are identified as periacinar fibroblast-like cells of the pancreas that express glial fibrillary acidic protein (GFAP) and produce ECM proteins in support of surrounding tissue. 7 PSCs are also, in part, responsible for the fibrosis observed in chronic pancreatitis. 8,9 Previous studies have shown that a3b1 integrin is essential for proper apical/ basolateral cell surface receptor organization and basement membrane formation in the submandibular gland. 10 As well, b1 integrin deficiency has shown to interfere with laminin-1 expression and basement membrane synthesis and assembly in embryoid bodies 11 and te...

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Section: Significantly Reduced Amylase Expression Observed In B1 Intementioning

confidence: 95%

Section: Acinar Cell Culture Experimentsmentioning

confidence: 99%

β1 integrin–extracellular matrix interactions are essential for maintaining exocrine pancreas architecture and function

Riopel

Liu

et al. 2013

Laboratory Investigation

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“…Attachment and Trophoblast Outgrowth Outgrowth of mouse blastocyst-derived trophoblast cells has been used as an in vitro model for implantation (Koi et al, 1995;Suenaga et al, 1996). About 70% of zona-free blastocysts attached and spread on ®bronectin-coated surface within 24 hr of culture in the absence and presence of SNP or L-NAME, except for a complete inhibition of blastocyst attachment and spread- ing at the concentration of 10 À 3 M SNP.…”

Section: Effects Of No and L-name On The Blastocystmentioning

confidence: 99%

Requirement of nitric oxide for murine oocyte maturation, embryo development, and trophoblast outgrowth in vitro

et al. 2001

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We investigated the extent to which NO participates in the developmental competence (oocyte maturation, fertilization and embryo development to blastocyst) using an in vitro culture system adding sodium nitroprusside (SNP), NO donor, and NOS inhibitor (N‐omega‐nitro‐L‐arginine methyl ester, L‐NAME). We also assessed the effects of NO/NOS system on blastocyst implantation using an in vitro trophoblast outgrowth assay. The treatment of low concentrations of SNP (10−7 M) significantly stimulated meiotic maturation to metaphase II stages in cumulus enclosed oocytes. In contrast, 10−3 and 10−5 M L‐NAME demonstrated a significant suppression in resumption of meiosis. This inhibition was reversed by the addition of SNP. No development beyond the four‐cell stage was observed by the addition of high concentration of SNP (10−3 M). Inhibition of embryo development, especially the conversion of morulae to blastocysts, was also observed in the treatment of lower doses of SNP (10−5 and 10−7 M). Similarly, inhibition of NO by NOS inhibitor resulted in the dose‐dependent inhibition of embryo development and hatching rates, but the concomitant addition of SNP with L‐NAME reversed the inhibitory effect by each SNP or L‐NAME treatment. Furthermore, low concentration of SNP (10−7 M) but not high concentration of SNP (10−3 M) significantly stimulated trophoblast outgrowth, whereas the addition of L‐NAME suppressed the spreading of blastocysts in a dose‐dependent manner. These results suggest that NO may have crucial roles in oocyte maturation and embryogenesis including the process of implantation. The observed differences in required amount of NO and the sensitivity to cytotoxicity of NO in each developmental stage embryos may also suggest that NO/NOS system is tightly regulated in developmental stage specific manner. Mol. Reprod. Dev. 58:262–268, 2001. © 2001 Wiley‐Liss, Inc.

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“…The method used for assessing effects on outgrowth was the well-characterized assay established by Armant et al [30] and used by other investigators [31][32][33][34]. These studies have shown that when intact mouse blastocysts are placed in culture on fibronectin, there is a significant outgrowth of trophoblast cells.…”

Section: Blastocyst Outgrowth Assaysmentioning

confidence: 99%

Transforming Growth Factor-β Stimulates Mouse Blastocyst Outgrowth through a Mechanism Involving Parathyroid Hormone-Related Protein1

Nowak¹,

Haimovici²,

Biggers³

et al. 1999

Biology of Reproduction

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The goals of this study were 1) to compare the effects of transforming growth factor-beta (TGF-beta) and parathyroid hormone-related protein (PTHrP) on mouse blastocyst attachment and outgrowth in vitro, 2) to determine whether TGF-beta acts through a mechanism involving PTHrP, 3) to examine effects of PTHrP on preimplantation mouse embryo development, and 4) to determine the pattern of expression of PTHrP protein in the uterus of the mouse during early gestation. In the first set of experiments, hatched blastocysts were placed in fibronectin-coated wells. Cultures were treated with PTHrP or TGF-beta1 and assessed at 24, 48, and 72 h for attachment and surface area of blastocyst outgrowth. Results showed that both PTHrP and TGF-beta1 increased blastocyst outgrowth significantly. A PTHrP-neutralizing antibody blocked the stimulatory effect of both PTHrP and TGF-beta1, suggesting that TGF-beta1 acts to increase endogenous production of PTHrP by the blastocyst. Immunoassay of conditioned medium from blastocysts treated with either TGF-beta1 or PTHrP 1-34 confirmed a 3- to 4-fold increase in levels of PTHrP 1-141. In the second series of experiments, pronuclear zygotes were cultured in various concentrations of PTHrP for 96 h. Blastocysts then were subjected to differential fluorescent staining of inner cell mass and trophectoderm cells. Treatment of mouse embryos with the various concentrations of PTHrP altered neither the number developing to the blastocyst stage nor the number of inner cell mass or trophectoderm cells in the resulting blastocysts. In the third experiment, pregnant mice were killed at Days 3, 4, 5, 6, and 7 of gestation, and uterine horns were processed for immunohistochemistry. Uterine sections were stained with antibodies to PTHrP, desmin, and laminin. On Days 3, 4, and 5, uterine luminal and glandular epithelial cells stained intensely for PTHrP, while stromal cells were negative. By Days 6 and 7, decidualized stromal cells stained positively for PTHrP, desmin, and laminin. These results support the hypothesis that TGF-beta and PTHrP play an important role in the process of implantation.

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Effects of Matrix Proteins and Heparin-Binding Components in Fetal Bovine Serum upon the Proliferation of Ectoplacental Cone Cells in Mouse Blastocysts Cultured in Vitro1

Cited by 6 publications

References 55 publications

β1 integrin–extracellular matrix interactions are essential for maintaining exocrine pancreas architecture and function

β1 integrin–extracellular matrix interactions are essential for maintaining exocrine pancreas architecture and function

Requirement of nitric oxide for murine oocyte maturation, embryo development, and trophoblast outgrowth in vitro

Transforming Growth Factor-β Stimulates Mouse Blastocyst Outgrowth through a Mechanism Involving Parathyroid Hormone-Related Protein1

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