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Visualization and Objective Quantification of Autonomic Circuitry in Mouse Area Postrema Using a Genetic Method
Author(s) -
Gokozan Hamza,
Corcoran Sarah,
Czeisler Catherine,
Gygli Patrick,
Catacutan Fay,
Baig Faisal,
Otero Jose
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.613.10
Subject(s) - hindbrain , biology , neuroscience , fate mapping , recombinase , congenital central hypoventilation syndrome , neuroanatomy , brainstem , transgene , progenitor , area postrema , microbiology and biotechnology , progenitor cell , anatomy , central nervous system , genetics , gene , stem cell , hypoventilation , respiratory system , recombination
The transcription factor PHOX2B is a master regulator of autonomic nervous system function as well as the specification of noradrenergic neurons. Incomplete maturity and/or absence of noradrenergic neuron networks are a proposed mechanism of infant breathing disorders including apnea of prematurity, sudden infant death syndrome, and congenital central hypoventilation syndrome (CCHS). PHOX2B mutations result in the “chemosensation” loss characteristic of CCHS, resulting in poor respiratory drive secondary to hypercarbia (high pCO2). The objective of our work is to identify PHOX2B derived structures in the developing brain and generate new animal models capable of tracing the neuronal connections between neuronal nuclei during embryonic and postnatal development. We utilized a PHOX2B‐CRE X ROSA‐GFP mouse to trace the appearance of PHOX2B‐derived cells in nuclei of the hindbrain. In order to trace the connections between these nuclei other hindbrain nuclei, we utilized a transgenic mouse in which targeted neurons display traceable processes and synapses. This transgenic mouse reagent (Jackson Labs stock number 010590) shows CRE‐mediated expression of tdTomato in cell bodies and GFP expression at synapses. Using these conditional gene targeting approaches we were able to conclude that neurons derived from PHOX2B progenitor domains innervate certain hindbrain structures. By using our transgenic approach, we were able to (1) identify PHOX2B derived progenitors generate both neurons and glia, (2) show PHOX2B derived neuronal synapses in Ventral Medulla and Area Postrema. We also present an unbiased stereology method which can be used to determine the complexity of PHOX2B neuronal network.