Level and distribution of microtubule associated protein-2 (MAP2) as an index of dendritic structural dynamics
G. Di Stefano, T. Casoli, P. Fattoretti, M. Balietti, Y. Grossi, B. Giorgetti, C. Bertoni-Freddari
Neurobiology of Aging Laboratory, INRCA Research Department, Via Birarelli 8, 60121 Ancona Italy
In the fully differentiated adult central nervous system (CNS) neuronal
processes need to maintain a fine balance between stability and
plasticity. In addition to being very plastic to adapt to the changing
environmental stimulation, neuronal wiring diagrams must be also
sufficiently stable to accomplish specific functional tasks on which
they are tuned. In this dynamic status, the neuronal cytoskeleton and
its components (actin filaments, neurofilaments and microtubules
together with the respective associated proteins) are reported to play
a major and critical role. MAP2 is selectively located at the
somatodendritic compartment of neurones where it stabilizes polymerized
tubulin and participates in the regulation of both microtubule spacing
and actin filaments crosslinking, thus contributing significantly to
the stabilization of dendritic processes. By quantitative
immunohistochemical analysis, MAP2 levels and distribution were
measured in the hippocampus and olfactory bulb of rats of different
ages to seek age-related changes in dendritic structural dynamics and
to test the reliability of MAP2 quantitative immunohistochemistry as a
laboratory procedure to estimate the actual CNS plastic condition.
Immunocytochemical localization of MAP2 was performed by the
avidin-biotin peroxidase complex method. Optical density of MAP2
immunoreactivity (OD), the ratio between the MAP2 stained area/total
test area (area fraction: AF), the total length of MAP2 labeled
profiles (TL), the ratio between the perimeter/area of the
immunostained profiles (pleomorphism index: PI) were the parameters
measured in 15 fields/animal yielding a total area of about 300 mm2. In
the dentate gyrus molecular layer and CA1 stratum radiatum as well as
in the granular layer of the olfactory bulb of old rats OD and AF
values were significantly lower than in young and adult rats, on the
contrary the PI value that was significantly higher. TL was
significantly deceased in the olfactory bulb of old rats, while in both
the hippocampal areas showed a not significant reduction. Taken
together, the present findings show that dendritic structural dynamics
are significantly impaired in aging. Considering that both the
hippocampus and the olfactory bulb are reported to be very plastic
zones of the CNS, i.e. they are capable of consistent rearrangements
during the individual's lifespan, these data lend further support to
the many converging results on the higher vulnerability to aging of the
CNS areas featuring higher plasticity. Although the present findings
should be compared with data from studies conducted in CNS zones
reported to be less plastic and less sensitive to age-related changes
before proposing a tenable conclusion, the marked changes (e.g. the
several fold decrease of OD and AF values) of all the parameters
measured in the present investigation support that MAP2
immunohistochemistry may serve as a reliable method to test the actual
plastic status of discrete CNS zones in different experimental
conditions.
Key words:
MAP2, Dendritic structural dynamics, CNS plasticity, Hippocampus, Olfactory bulb
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