Role of Zip Kinase in the Development of Early Embryonic Cells

Poster Number

34

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Douglas Weiser

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Reversible phosphorylation of the Type II Myosin Light Chain 2 (MLC2) is a crucial mechanism for regulating the Type II Myosin and the actin cytoskeleton. Zipper Interacting Protein Kinase (ZipK) is a major regulator of Myosin Light Chain 2 through phosphorylation of the MLC2 complex. This mechanism is imperative for proper myosin and actin cytoskeleton contraction and other cellular processes that contribute to morphogenetic cell movements through development, smooth muscle contraction, and tumor cell invasion. ZipK and ROCK are regulators of Myosin Phosphatase, which is responsible for dephosphorylating MLC2. This occurs when ZipK/ROCK phosphorylates Mypt1 (Myosin Phosphatase Target Subunit 1) at the conserved threonines 696 and 850, which will inhibit Myosin Phosphatase activity. Therefore, when ZipK is inactivated, cells exhibit irregularities such as disorganization of the cell membrane in addition to improper functioning of actin and myosin. This could lead to defects in mitosis, cell movement, and initiate the process of apoptosis. To analyze the morphogenetic defects in Zebrafish lacking Zip Kinase, in-situ hybridization probes are needed to mark specific regions of the embryo. The probes that are currently in the process of being built are hgg1 (prechordal plate), shh(midline), pax 2.1 (midbrain-hindbrain boundary), dlx3 (neural plate), and papc (presomatic mesoderm). To create the probes, restriction digests were carried using appropriate enzymes for anti-sense and sense probes. Phenol chloroform extraction was carried out to isolate the RNA so that a transcription reaction could be performed. We hope to continue our research by carrying out staging of the fish, permeabilization, and hybridization of the embryo. By viewing morphant embryos next to Wild type embryos, we will be able to determine the morphological difference using probes specifics to a region of the embryo

Location

DeRosa University Center, Ballroom

Start Date

30-4-2016 1:30 AM

End Date

30-4-2016 3:30 PM

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Apr 30th, 1:30 AM Apr 30th, 3:30 PM

Role of Zip Kinase in the Development of Early Embryonic Cells

DeRosa University Center, Ballroom

Reversible phosphorylation of the Type II Myosin Light Chain 2 (MLC2) is a crucial mechanism for regulating the Type II Myosin and the actin cytoskeleton. Zipper Interacting Protein Kinase (ZipK) is a major regulator of Myosin Light Chain 2 through phosphorylation of the MLC2 complex. This mechanism is imperative for proper myosin and actin cytoskeleton contraction and other cellular processes that contribute to morphogenetic cell movements through development, smooth muscle contraction, and tumor cell invasion. ZipK and ROCK are regulators of Myosin Phosphatase, which is responsible for dephosphorylating MLC2. This occurs when ZipK/ROCK phosphorylates Mypt1 (Myosin Phosphatase Target Subunit 1) at the conserved threonines 696 and 850, which will inhibit Myosin Phosphatase activity. Therefore, when ZipK is inactivated, cells exhibit irregularities such as disorganization of the cell membrane in addition to improper functioning of actin and myosin. This could lead to defects in mitosis, cell movement, and initiate the process of apoptosis. To analyze the morphogenetic defects in Zebrafish lacking Zip Kinase, in-situ hybridization probes are needed to mark specific regions of the embryo. The probes that are currently in the process of being built are hgg1 (prechordal plate), shh(midline), pax 2.1 (midbrain-hindbrain boundary), dlx3 (neural plate), and papc (presomatic mesoderm). To create the probes, restriction digests were carried using appropriate enzymes for anti-sense and sense probes. Phenol chloroform extraction was carried out to isolate the RNA so that a transcription reaction could be performed. We hope to continue our research by carrying out staging of the fish, permeabilization, and hybridization of the embryo. By viewing morphant embryos next to Wild type embryos, we will be able to determine the morphological difference using probes specifics to a region of the embryo