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Immunocytochemical techniques identify Na+-coupled HCO3– transporters (NCBTs) in chemosensitive neurons of the medullary raphé

Coley, Austin A.
http://rave.ohiolink.edu/etdc/view?acc_num=case1322943857

Abstract Details

2011, Master of Sciences, Case Western Reserve University, Physiology and Biophysics.
The medullary raphé (MR) of the medulla oblongata contains chemosensitive (serotonin/5-Hydroxytryptamine [5HT], GABA) neurons that are extremely sensitive to CO2. Serotonergic neurons respond to increases in arterial [CO2] by exhibiting a major increase in firing rate, whereas GABAergic neurons respond with a decrease in firing rate, both contributing to an increase in pulmonary ventilation. The electroneutral Na+/HCO3- cotransporters (nNCBTs) respond to intracellular acid loads (such as that produced by an increase of CO2) by actively moving HCO3- into the cell, and thereby tending to restore the pHi. I hypothesized that the nNCBTs are less abundant in serotonergic and GABAergic neurons than in non-chemosensitive neurons. A reduction in nNCBT activity would be expected to inhibit the restoration of pHi and thereby sustained the fall in pHi, maintaining a high firing rate in 5HT neurons and a low firing rate in GABAergic neurons. I developed an immunocytochemical protocol to identify cell types by using primary antibodies directed against MAP2, a neuronal marker; TPH, a 5HT neuronal marker; GAD2, a GABA neuronal marker; and GFAP, an astrocytic marker. I colocalized the cell specific antibodies with polyclonal antibodies directed against nNCBTs: NBCn1, NBCn2, and NDCBE. I used image analysis techniques as a quantitative approach for determining if a cell is positive or negative for an antibody. Contrary to my hypothesis, however, I found that the fraction of neurons that were positive for nNCBTs were not substantially different for serotonergic neurons vs GABAergic neurons vs neurons that were negative for either TPH or GAD2.
Walter Boron, MD, PhD (Advisor)
Corey Smith, PhD (Committee Chair)
Ulrich Hopfer, MD, PhD (Committee Member)
David Katz, PhD (Committee Member)
146 p.
Biophysics; Neurosciences; Physiology
nNCBTs; NBCn1; NBCn2; NDCBE; Serotonergic neurons; GABAergic neurons; pHi; Immunocytochemistry; Image processing

Recommended Citations

Citations

  • Coley, A. A. (2011). Immunocytochemical techniques identify Na+-coupled HCO3– transporters (NCBTs) in chemosensitive neurons of the medullary raphé [Master's thesis, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1322943857

    APA Style (7th edition)

  • Coley, Austin. Immunocytochemical techniques identify Na+-coupled HCO3– transporters (NCBTs) in chemosensitive neurons of the medullary raphé. 2011. Case Western Reserve University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1322943857.

    MLA Style (8th edition)

  • Coley, Austin. "Immunocytochemical techniques identify Na+-coupled HCO3– transporters (NCBTs) in chemosensitive neurons of the medullary raphé." Master's thesis, Case Western Reserve University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=case1322943857

    Chicago Manual of Style (17th edition)

case1322943857
543
© 2011, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.