Inhibition of focal adhesion kinase promotes adult olfactory stem cell self-renewal and neuroregeneration via ciliary neurotrophic factor
Location
Ballroom
Start Date
4-5-2018 8:00 AM
End Date
4-5-2018 12:00 PM
Poster Number
37
Name of Project's Faculty Sponsor
Dr Jia, Cuihong
Faculty Sponsor's Department
Deparment of Biomedical Sciences
Type
Poster: Competitive
Project's Category
Biomedical and Health Sciences
Abstract or Artist's Statement
The Olfactory Epithelium (OE) is a specialized epithelial tissue inside the nasal cavity that is involved in the smell sensation. The OE maintains neuroregeneration, i.e. producing new olfactory sensory neurons, throughout the adult life via neural stem cell self-renewal, proliferation, neuronal differentiation and maturation. The neural stem cell niche regulates stem cell self-renewal and proliferation, and consists of stem cells, blood vessels and multiple extracellular matrix proteins (ECMs). ECMs regulate stem cell adhesion, proliferation, differentiation and migration via integrins. One of the main mediators of intracellular integrin signaling is the Focal Adhesion Kinase (FAK). Our previous studies found that FAK inhibition increased cell proliferation in adult mouse olfactory epithelium (OE) via up-regulation of Ciliary Neurotrophic Factor (CNTF). Now we continue to test whether FAK inhibition increases neuroregeneration through CNTF in the adult mouse OE using BrdU-chase pulse method. Adult male and female C57BL/6, CNTF wildtype and CNTF knockout (lack the CNTF gene) mice were systemically injected with PBS or FAK inhibitor (FAK14) for 3 days. During these 3 days, BrdU was injected into mice 4 h following PBS or FAK on each day. BrdU acts as a thymidine analog and is incorporated into DNA during DNA syntheses. Using immunohistochemistry with anti-BrdU antibody, BrdU+ cells can be visualized in the tissue. The BrdU+ cells are the ones who are replicating during the time frame when BrdU was given. 20 days after last BrdU injection, we fixed the mice via cardiac perfusion. The whole heads of mice was decalcified with EDTA and then frozen cross head sections including OE were cut using cryostat and mounted onto slides. The OE sections were then stained with anti-BrdU antibody followed by FITC-conjugated secondary antibody. The BrdU+ cells in the OE were counted in three sections (both left and right sides) per mouse and normalized to linear length of OE basement membrane. The results of the experiment showed that FAK 14 significantly increased BrdU+ stem cells and olfactory sensory neurons in the OE of C57BL/6 and CNTF wildtype mice but not knockout mice, indicating that FAK inhibition promotes olfactory stem cell self-renewal and neuroregeneration via CNTF. Collectively, this data indicates that FAK normally inhibits OE neuroregeneration by inhibiting CNTF expression and identifies the OE is a good model to study neuroregenerative mechanisms in the CNS.
Inhibition of focal adhesion kinase promotes adult olfactory stem cell self-renewal and neuroregeneration via ciliary neurotrophic factor
Ballroom
The Olfactory Epithelium (OE) is a specialized epithelial tissue inside the nasal cavity that is involved in the smell sensation. The OE maintains neuroregeneration, i.e. producing new olfactory sensory neurons, throughout the adult life via neural stem cell self-renewal, proliferation, neuronal differentiation and maturation. The neural stem cell niche regulates stem cell self-renewal and proliferation, and consists of stem cells, blood vessels and multiple extracellular matrix proteins (ECMs). ECMs regulate stem cell adhesion, proliferation, differentiation and migration via integrins. One of the main mediators of intracellular integrin signaling is the Focal Adhesion Kinase (FAK). Our previous studies found that FAK inhibition increased cell proliferation in adult mouse olfactory epithelium (OE) via up-regulation of Ciliary Neurotrophic Factor (CNTF). Now we continue to test whether FAK inhibition increases neuroregeneration through CNTF in the adult mouse OE using BrdU-chase pulse method. Adult male and female C57BL/6, CNTF wildtype and CNTF knockout (lack the CNTF gene) mice were systemically injected with PBS or FAK inhibitor (FAK14) for 3 days. During these 3 days, BrdU was injected into mice 4 h following PBS or FAK on each day. BrdU acts as a thymidine analog and is incorporated into DNA during DNA syntheses. Using immunohistochemistry with anti-BrdU antibody, BrdU+ cells can be visualized in the tissue. The BrdU+ cells are the ones who are replicating during the time frame when BrdU was given. 20 days after last BrdU injection, we fixed the mice via cardiac perfusion. The whole heads of mice was decalcified with EDTA and then frozen cross head sections including OE were cut using cryostat and mounted onto slides. The OE sections were then stained with anti-BrdU antibody followed by FITC-conjugated secondary antibody. The BrdU+ cells in the OE were counted in three sections (both left and right sides) per mouse and normalized to linear length of OE basement membrane. The results of the experiment showed that FAK 14 significantly increased BrdU+ stem cells and olfactory sensory neurons in the OE of C57BL/6 and CNTF wildtype mice but not knockout mice, indicating that FAK inhibition promotes olfactory stem cell self-renewal and neuroregeneration via CNTF. Collectively, this data indicates that FAK normally inhibits OE neuroregeneration by inhibiting CNTF expression and identifies the OE is a good model to study neuroregenerative mechanisms in the CNS.