Date of Award
Doctor of Philosophy (PhD)
Chun Jiang - Committee Chair
Andrew Clancy - Committee Member
Deborah Baro - Committee Member
Zhi-Ren Liu - Committee Member
Patients with Rett Syndrome (RTT) show severe breathing disorders in addition to other neuropathological features, contributing to the high incidence of sudden unexplained death and abnormal brain development. However, the molecular and cellular mechanisms underlying the breathing disorders are still unknown. Recent studies indicate that the dysfunction of brainstem norepinephrine (NE) systems are closely associated with breathing disorders in RTT patients as well as its mice model, the Mecp2-null (Mecp2â”€/Y) mice. This as well as the fact the major group of NE-ergic neurons in the locus coeruleus (LC) is CO2 chemosensitive suggests that the breathing disorders in RTT may be related these LC neurons. To test this hypothesis, we took a multidisciplinary approach and systematically studied these neurons using molecular biology, in-vitro brain slices, acutely dissociated neurons, immunocytochemistry, and whole-body plethysmograph. To facilitate the electrophysiological studies, we developed a new strain of transgenic mice with GFP expression selectively in the LC neurons of both WT and Mecp2â”€/Y mice. Breathing activity of the Mecp2â”€/Y mice showed selective disruptions in responses to mild hypercapnia. The defect was alleviated with the NE uptake blocker desipramine, suggesting the involvement of NE in central CO2 chemosensitivity. In the LC region, the expressions of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) at both protein and mRNA levels reduced by ~50% in Mecp2â”€/Y mice. No evidence was found for selective deficiency in TH- or DBH-containing neurons in Mecp2â”€/Y mice, and no major loss of NE-ergic LC cells were found, indicating that the NE defect is likely to result from deficient expression of biosynthetic enzymes rather than a loss of neurons in the LC. Several intrinsic membrane properties were abnormal in Mecp2â”€/Y LC neurons in comparison to wild type cells, including stronger inward rectification, shorter time constant, extended action potential duration, smaller amplitude of medium afterhyperpolarization (AHP) and over-expression of fast AHP. These abnormalities seem to be associated with the altered K+ and Na+ currents. Most importantly, Mecp2â”€/Y LC neurons displayed defective CO2 chemosensitivity in agreement of in vivo CO2 response, likely due to excessive expression of the homomeric Kir4.1 channel. Thus, it seems that the global effect of MeCP2 on the A6 NE system contributes to the impaired systemic CO2 response as well as the breathing irregularities in Mecp2â”€/Y mice. Such an alteration allowed CO2 to be detected only when hypercapnia became severe, leading to periodical hyper- and hypoventilation. These findings not only provide a novel etiology for the breathing disturbances of Mecp2â”€/Y mice but also show direct evidence for the first time on a molecular mechanism for the central CO2 chemosensitivity.
Zhang, Xiaoli, "Locus Coeruleus Neurons in Autonomic Regulation of Breathing: Insight from a Mouse Model of Rett Syndrome." Dissertation, Georgia State University, 2010.