Supplementary Material for: Aggregates of cAMP-Dependent Kinase Isoforms Characterize Different Areas in the Developing Central Nervous System of the Chicken, Gallus gallus

2017-03-24T06:21:40Z (GMT) by Mucignat-Caretta C. Caretta A.
The intracellular second messenger adenosine 3′,5′-cyclic monophosphate (cAMP) acts mainly through cAMP-dependent protein kinases (PKA). In mammals and reptiles, the PKA regulatory isoforms (RI and RII) are differentially distributed among the various brain areas and cell types, according to the age of the animal. Since PKA distribution may be an additional marker for homologous areas, PKA regulatory subunit types RI and RII were examined in the chicken brain, a species not yet investigated. Chicken brains were examined from prehatching to adult age, by means of immunohistochemistry and biochemical characterization. Most PKA regulatory subunits were segregated in discrete non-soluble clusters that contained either RI or RII. While RII aggregates were present also in non-neuronal cells, RI aggregates were detected only in neurons of some brain areas that are mainly related to the telencephalon. They appeared later than RII aggregates; their presence and location varied during development. RI aggregates were detected first in the olfactory bulb, around embryonic day 14; within 3 days they appeared in the hyperpallium and nidopallium, where the most intense labeling was observed in the perihatching period. Fainter RI aggregates persisted up to 3 years in the olfactory bulb and nidopallium caudale. Less intense RI aggregates were present for a shorter time, from 2 weeks to 3 months, in the septal nuclei, thalamic medial nuclei, periventricular hypothalamus, optic tectum periventricular area, brainstem reticular formation and spinal cord substantia gelatinosa. RI aggregates were not detected in many brain areas including the arcopallium, striatum and cranial nerve nuclei. RII distribution showed less variation during development. From embryonic day 12, some insoluble RII aggregates were detected in the brain; however, only minor modifications were observed in positive structures once they started to harbor insoluble RII aggregates. The present results suggest that the distribution of PKA aggregates may assist in characterizing phylogenetically homologous structures of the vertebrate central nervous system and may also unravel biochemical differences among areas considered homologous.