scholarly journals Two alkaline phosphatase genes are expressed during early development in the mouse embryo

Development ◽  
1990 ◽  
Vol 110 (2) ◽  
pp. 555-564 ◽  
Author(s):  
A.C. Hahnel ◽  
D.A. Rappolee ◽  
J.L. Millan ◽  
T. Manes ◽  
C.A. Ziomek ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Germ Cells ◽  
Inner Cell Mass ◽  
Primordial Germ Cells ◽  
Cell Mass ◽  
Mouse Embryos ◽  
Preimplantation Development ◽  
Chain Reaction ◽  
Polymerase Chain

Alkaline phosphatase (AP) activity is stage specific in mouse embryos and may be associated with compaction and separation of trophectoderm from inner cell mass in preimplantation development. We previously sequenced a cDNA and two mouse AP genes that could contribute to the AP activity in embryos. Oligonucleotide primers were constructed from the three sequences and used in the reverse transcription-polymerase chain reaction technique to establish that two of the three AP isozymes are transcribed during preimplantation development. The predominant transcript (E-AP) is from a gene highly homologous to the human tissue-specific APs, but different from the mouse intestinal AP. Tissue non-specific (TN) AP also is transcribed, but there is approximately 10 times less TN-AP than E-AP transcript. The TN-AP isozyme is the predominant transcript of 7 to 14 day embryos and primordial germ cells. A switch in predominance from E-AP to TN-AP must occur during early postimplantation development. This study establishes a framework for experiments to determine the functions of the two isozymes during preimplantation development.

Nuclear transplantation of male primordial germ cells in the mouse

Development ◽  
1989 ◽  
Vol 107 (2) ◽  
pp. 407-411 ◽  
Author(s):  
Y. Tsunoda ◽  
T. Tokunaga ◽  
H. Imai ◽  
T. Uchida
Keyword(s):  
Germ Cells ◽  
Inner Cell Mass ◽  
Primordial Germ Cells ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Nuclear Transplantation ◽  
Embryonic Genome ◽  
Donor Nucleus ◽  
Implantation Sites ◽  
Genome Activation

We examined the developmental ability of enucleated eggs receiving embryonic nuclei and male primordial germ cells (PGCs) in the mouse. Reconstituted eggs developed into the blastocyst stage only when an earlier 2-cell nucleus was transplanted (36%) but very rarely if the donor nucleus was derived from a later 2-cell, 8-cell, or inner cell mass of a blastocyst (0–3%). 54–100%, 11–67%, 6–43% and 6–20% of enucleated eggs receiving male PGCs developed to 2-cell, 4-cell, 8-cell and blastocyst stage, respectively, in culture. The overall success rate when taking into account the total number of attempts at introducing germ cells was actually 0–6%. Live fetuses were not obtained after transfer of reconstituted eggs to recipients, although implantation sites were observed. The developmental ability of reconstituted eggs in relation to embryonic genome activation and genomic imprinting is discussed.

scholarly journals Transient benign hyperphosphatasemia due to COVID-19: the first case report

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Tugba Erat ◽  
Müge Atar ◽  
Tugba Kontbay
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Case Report ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Benign Condition ◽  
Case Presentation ◽  
Normal Ranges ◽  
First Case ◽  
Polymerase Chain

AbstractObjectivesCoronavirus disease (COVID-19) rapidly spread worldwide in a few months and was declared as a worldwide pandemic by WHO in March 2020. Transient benign hyperphosphatasemia (THI) is a benign condition associated with marked elevation of alkaline phosphatase (ALP) without any other kidney, bone, and liver pathologies.Case presentationHerein, we report a previously healthy 16-month-old female patient who developed a secondary transient benign hyperphosphatasemia associated with SARS-CoV-2. Patient whole family’s SARS-CoV-2 real-time reverse transcription-polymerase chain reaction (RT-PCR) results were positive. Since THI is a diagnosis of exclusion, other reasons that may cause ALP elevation should be ruled out. ALP activity decreased and turned to normal ranges within the following month. THI has been reported to be in association with various conditions. Its relationship with many viruses has been reported previously.ConclusionsIf ALP elevation is detected in patients with COVID 19 due to the increasing number of infections, THI should be considered if there is no other accompanying pathology.

The Two mRNAS Encoding Rat Intestinal Alkaline Phosphatase Represent Two Distinct Nucleotide Sequences

1992 ◽  
Vol 38 (12) ◽  
pp. 2506-2509 ◽  
Author(s):  
M J Engle ◽  
D H Alpers
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Duodenal Mucosa ◽  
Differential Regulation ◽  
Untranslated Regions ◽  
Terminal Sequence ◽  
Intestinal Alkaline Phosphatase ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Fat Feeding

Abstract Rat duodenal mucosa contains two mRNAs (2.7 and 3.0 kb) encoding intestinal alkaline phosphatase (IAP), but the mechanism for their production has not been clear. By means of the polymerase chain reaction (PCR), we isolated a fragment that identifies a second rat IAP (rIAP-II), differing from the rIAP-I sequence in the coding and 3' untranslated regions and encoding a COOH-terminal sequence predicted to be hydrophilic. By means of probes unique to each sequence, rIAP-I identified the 2.7-kb mRNA, and rIAP-II the 3.0-kb mRNA. The different structures and differential regulation of the mRNAs after fat feeding demonstrate the presence of two rat IAP transcripts.

scholarly journals Use of a HpaII-polymerase chain reaction assay to study DNA methylation in the Pgk-1 CpG island of mouse embryos at the time of X-chromosome inactivation.

1990 ◽  
Vol 10 (9) ◽  
pp. 4987-4989 ◽  
Author(s):  
J Singer-Sam ◽  
M Grant ◽  
J M LeBon ◽  
K Okuyama ◽  
V Chapman ◽  
...  
Keyword(s):  
Dna Methylation ◽  
X Chromosome ◽  
Inner Cell Mass ◽  
Cpg Island ◽  
Cell Mass ◽  
X Chromosome Inactivation ◽  
Mouse Embryos ◽  
Chromosome Inactivation ◽  
Individual Mouse ◽  
Polymerase Chain

A HpaII-PCR assay was used to study DNA methylation in individual mouse embryos. It was found that HpaII site H-7 in the CpG island of the X-chromosome-linked Pgk-1 gene is less than or equal to 10% methylated in oocytes and male embryos but becomes 40% methylated in female embryos at 6.5 days; about the time of X-chromosome inactivation of the inner cell mass.

scholarly journals Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells

Medicina ◽  
2021 ◽  
Vol 57 (11) ◽  
pp. 1271
Author(s):  
Hyun-Jin Lee ◽  
Young-Min Song ◽  
Seunghoon Baek ◽  
Yoon-Hee Park ◽  
Jun-Beom Park
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Vitamin D ◽  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Cellular Viability ◽  
Chain Reaction ◽  
Cell Spheroids ◽  
Polymerase Chain

Background and Objectives: Vitamin D is a bone modulator widely used in regenerative medicine. This study aimed to analyze the effects of vitamin D on the osteogenic differentiation and mineralization of human mesenchymal stem cells. Materials and Methods: Spheroids were fabricated using human bone marrow-derived stem cells, and were cultured in the presence of vitamin D at concentrations of 0, 0.1, 1, 10, and 100 nM. Stem cell spheroids were fabricated and the morphological evaluation was conducted on days 1, 3, 7 and 14. Determination of qualitative cellular viability was performed with Live/Dead Kit assay on days 1 and 7. Quantitative cellular viability was evaluated with Cell Counting Kit-8 on days 1, 3, 7, and 14. To analyze the osteogenic differentiation of cell spheroids, alkaline phosphatase activity assays were performed with commercially available kit on days 7 and 14. Real-time polymerase chain reaction was used to determine the expression levels of RUNX2, BSP, OCN, and COL1A1 on days 7 and 14. Results: The stem cells produced well-formed spheroids, and addition of vitamin D did not result in any noticeable changes in the shape. The addition of vitamin D did not significantly change the diameter of the spheroids at 0, 0.1, 1, 10, or 100 nM concentrations. Quantitative cell viability results from days 1, 3, 7 and 14 showed no significant difference between groups (p > 0.05). There was significantly higher alkaline phosphatase activity in the 0.1 nM group when compared with the control group on day 14 (p < 0.05). Real-time polymerase chain reaction results demonstrated that the mRNA expression levels of RUNX2, OCN, and COL1A1 were significantly increased when vitamin D was added to the culture. Conclusions: Based on these findings, we concluded that vitamin D could be applied to the increased osteogenicity of stem cell spheroids.

scholarly journals Transmission of maternal blood cells to the fetus during pregnancy: detection in mouse neonatal spleen by immunofluorescence flow cytometry and polymerase chain reaction

Blood ◽  
1994 ◽  
Vol 83 (4) ◽  
pp. 926-930
Author(s):  
M Shimamura ◽  
S Ohta ◽  
R Suzuki ◽  
K Yamazaki
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Blood Cells ◽  
Maternal Blood ◽  
Mouse Embryos ◽  
Chain Reaction ◽  
Maternal Cell ◽  
Mhc Antigens ◽  
Histocompatibility Complex ◽  
Polymerase Chain

Mouse embryos were transferred to allogeneic pseudopregnant foster mothers and the cells of the resultant neonates were analyzed for the expression of maternal major histocompatibility complex (MHC) antigens to elucidate maternal cell transmission to the fetus during pregnancy. Expression of maternal-type MHC antigens was detected in the spleen but not in the liver or thymus of the neonates in analyses by immunofluorescence flow cytometry and polymerase chain reaction. These findings provide evidence, on the molecular basis, that maternal blood cells are transmitted to the fetus through the placenta.

Alkaline phosphatase activity in the preimplantation mouse embryo

Development ◽  
1977 ◽  
Vol 40 (1) ◽  
pp. 83-89
Author(s):  
Lincoln V. Johnson ◽  
Patricia G. Calarco ◽  
Margaret L. Siebert
Keyword(s):  
Alkaline Phosphatase ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Mouse Embryos ◽  
Frozen Sections ◽  
Mouse Development ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse ◽  
Morula Stage

Alkaline phosphatase (AP) activity has been assayed in frozen sections of preimplantation mouse embryos by an azo-dye cytochemical method. The results indicate that during preimplantation mouse development AP activity is first expressed between the 8- and 16-cell stages and develops in all cells by the late morula stage. During blastocyst formation AP activity is lost or greatly reduced in trophoblast cells while activity is maintained in the inner cell mass.

Expression of Forssman antigen in the post-implantation mouse embryo

Development ◽  
1981 ◽  
Vol 61 (1) ◽  
pp. 117-131
Author(s):  
M. G. Stinnakre ◽  
M. J. Evans ◽  
K. R. Willison ◽  
P. L. Stern
Keyword(s):  
Sertoli Cells ◽  
Inner Cell Mass ◽  
Primordial Germ Cells ◽  
Cell Mass ◽  
Mouse Embryos ◽  
Related Distribution ◽  
Forssman Antigen ◽  
Post Implantation ◽  
Embryonic Ectoderm ◽  
Positive Population

The expression of Forssman antigen on the surface of cells of post-implantation mouse embryos between 5 and 7½ days old and of cells of the gonads from 10½ days has been followed using the monoclonal antiserum M1/22.25. In the early post-implantation embryo a lineage-related distribution is found. The inner cell mass of the blastocyst was previously shown to be Forssman antigen positive and its derivative tissues the epiblast of the 5-day embryo and the primary embryonic endoderm are also positive. The endoderm cells remain positive both over the embryonic and extraembryonic portions of the embryo but the epiblast becomes Forssman antigen negative as it differentiates into embryonic ectoderm. The extraembryonic ectoderm which is derived from the Forssman negative trophectoderm remains negative throughout. The primordial germ cells are Forssman antigen positive from their first appearance in the germinal ridge until day 14 when they become negative but after that time it is other cells not related by direct lineage which become Forssman antigen positive. These are tentatively identified as Sertoli cells precursors as it is the Sertoli cells which are the antigen-positive population in the adult testis.

scholarly journals Transmission of maternal blood cells to the fetus during pregnancy: detection in mouse neonatal spleen by immunofluorescence flow cytometry and polymerase chain reaction

Blood ◽  
1994 ◽  
Vol 83 (4) ◽  
pp. 926-930 ◽  
Author(s):  
M Shimamura ◽  
S Ohta ◽  
R Suzuki ◽  
K Yamazaki
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Blood Cells ◽  
Maternal Blood ◽  
Mouse Embryos ◽  
Chain Reaction ◽  
Maternal Cell ◽  
Mhc Antigens ◽  
Histocompatibility Complex ◽  
Polymerase Chain

Abstract Mouse embryos were transferred to allogeneic pseudopregnant foster mothers and the cells of the resultant neonates were analyzed for the expression of maternal major histocompatibility complex (MHC) antigens to elucidate maternal cell transmission to the fetus during pregnancy. Expression of maternal-type MHC antigens was detected in the spleen but not in the liver or thymus of the neonates in analyses by immunofluorescence flow cytometry and polymerase chain reaction. These findings provide evidence, on the molecular basis, that maternal blood cells are transmitted to the fetus through the placenta.

Osteogenic Activity of MG63 Cells on Hydroxyapatite/Collagen Nanocompsosite under Pressure/perfusion Culture

2007 ◽  
Vol 330-332 ◽  
pp. 317-320 ◽  
Author(s):  
Teruaki Yoshida ◽  
Masanori Kikuchi ◽  
Yoshihisa Koyama ◽  
Kazuo Takakuda
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Perfusion Culture ◽  
Collagen Sponge ◽  
Perfusion Technique ◽  
Chain Reaction ◽  
Osteogenic Activity ◽  
Self Organized ◽  
Mg63 Cells ◽  
Polymerase Chain

The purpose of this study is to analyze osteogenic activity of human osteoblastic MG63 cells on the bone-like self-organized hydroxyapatite/collagen (HAp/Col) nanocomposite sponge cultured by a pressure/perfusion technique using collagen sponge as a control. Histological analyses, alkaline phosphatase (AlkP) protein analysis and real-time polymerase chain reaction (PCR) analyses for AlkP and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to evaluate the osteogenic activity of MG63. The MG63 cells were attached well and showed good proliferation on the HAp/Col sponge as well as in the control. The MG63 cells on HAp/Col sponge demonstrated higher osteogenic activity than those on the control. The results suggested that the HAp/Col sponge is expected to be a good scaffold for bone tissue engineering.

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