In Situ Hybridization Analysis of ZPK Gene Expression During Murine Embryogenesis

ZPK is a recently described protein serine/threonine kinase that has been originally identified from a human teratocarcinoma cell line by the polymerase chain reaction and whose function in signal transduction has not yet been elucidated. To investigate the potential role of this protein kinase in developmental processes, we have analyzed the spatial and temporal patterns of expression of the ZPK gene in mouse embryos of different gestational ages. Northern blot analysis revealed a single mRNA species of about 3.5 KB from Day 11 of gestation onwards. In situ hybridization studies demonstrated strong expression of ZPK mRNA in brain and in a variety of embryonic organs that rely on epithelio-mesenchymal interactions for their development, including skin, intestine, pancreas, and kidney. In these tissues, the ZPK mRNA was localized primarily in areas composed of specific types of differentiating cells, and this expression appeared to be upregulated at a time concomitant with the onset of terminal differentiation. Taken together, these observations raise the possibility that the ZPK gene product is involved in the establishment and/or maintenance of a fully cytodifferentiated state in a variety of cell lineages.

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Neurogenesis and Increase in Differentiated Neural Cell Survival via Phosphorylation of Akt1 after Fluoxetine Treatment of Stem Cells

BioMed Research International ◽

10.1155/2013/582526 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 14

Author(s):

Anahita Rahmani ◽

Danial Kheradmand ◽

Peyman Keyhanvar ◽

Alireza Shoae-Hassani ◽

Amir Darbandi-Azar

Keyword(s):

Survival Rate ◽

Cell Survival ◽

Neural Cell ◽

Neural Cells ◽

Chain Reaction ◽

Serine Threonine Kinase ◽

Threonine Kinase ◽

Fluoxetine Treatment ◽

Polymerase Chain

Fluoxetine (FLX) is a selective serotonin reuptake inhibitor (SSRI). Its action is possibly through an increase in neural cell survival. The mechanism of improved survival rate of neurons by FLX may relate to the overexpression of some kinases such as Akt protein. Akt1 (a serine/threonine kinase) plays a key role in the modulation of cell proliferation and survival. Our study evaluated the effects of FLX on mesenchymal stem cell (MSC) fate and Akt1 phosphorylation levels in MSCs. Evaluation tests included reverse transcriptase polymerase chain reaction, western blot, and immunocytochemistry assays. Nestin, MAP-2, andβ-tubulin were detected after neurogenesis as neural markers. TenμM of FLX upregulated phosphorylation of Akt1 protein in induced hEnSC significantly. Also FLX did increase viability of these MSCs. Continuous FLX treatment after neurogenesis elevated the survival rate of differentiated neural cells probably by enhanced induction of Akt1 phosphorylation. This study addresses a novel role of FLX in neurogenesis and differentiated neural cell survival that may contribute to explaining the therapeutic action of fluoxetine in regenerative pharmacology.

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Cloning and widespread distribution of the rat rod-type cyclic nucleotide-gated cation channel

AJP Cell Physiology ◽

10.1152/ajpcell.1997.272.4.c1335 ◽

1997 ◽

Vol 272 (4) ◽

pp. C1335-C1344 ◽

Cited By ~ 57

Author(s):

C. Ding ◽

E. D. Potter ◽

W. Qiu ◽

S. L. Coon ◽

M. A. Levine ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

In Situ Hybridization ◽

Pineal Gland ◽

Northern Blot ◽

Blot Analysis ◽

Cyclic Nucleotide ◽

Northern Blot Analysis ◽

Chain Reaction ◽

Polymerase Chain

We used Northern blot analysis, ribonuclease protection assay (RPA), reverse transcriptase-polymerase chain reaction, and in situ hybridization to investigate the hypothesis that the CNG1 isoform of the cyclic nucleotide-gated nonselective cation channel may be widely distributed in tissues of the rat. A cDNA encoding the CNG1 isoform was isolated from rat eye and human retina, and partial sequences were isolated from rat pineal gland and human kidney. Northern blot analysis revealed a 3.1-kilobase (kb) CNG1 transcript in rat eye, pineal gland, pituitary, adrenal gland, and spleen, and a larger transcript of 3.5 kb was found in testis. RPA confirmed the identity of CNG1 mRNA in rat eye, lung, spleen, and brain. Polymerase chain reaction-based detection of the mRNA for CNG1 indicates that the channel is expressed in lower abundance in many other tissues, including thymus, skeletal muscle, heart, and parathyroid gland. The cellular distribution of CNG1 was further studied by in situ hybridization, which demonstrated expression of mRNA in lung, thymus, pineal gland, hippocampus, cerebellum, and cerebral cortex but not in heart or kidney.

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