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The visual cortex is vulnerable to changes in visual input, especially during the critical period when numerous molecules drive the refinement of the circuitry. From a list of potential actors identified in a recent proteomics study, we selected 2 collapsin response mediator proteins (CRMP2/CRMP4) and 2 synaptic proteins, Dynamin I (Dyn I) and Synaptotagmin I (Syt I), for in-depth analysis of their developmental expression profile in cat visual cortex. CRMP2 and CRMP4 levels were high early in life and clearly declined toward adulthood. In contrast, Dyn I expression levels progressively augmented during maturation. Syt I showed low levels at eye opening and in adults, high levels around the peak of the critical period, and maximal levels at juvenile age. We further determined a role for each molecule in ocular dominance plasticity. CRMP2 and Syt I levels decreased in area 17 upon monocular deprivation, whereas CRMP4 and Dyn I levels remained unaffected. In contrast, binocular removal of pattern vision had no influence on CRMP2 and Syt I expression in kitten area 17. This study illustrates that not the loss of quality of vision through visual deprivation, but disruption of normal binocular visual experience is crucial to induce the observed molecular changes.

During early postnatal brain development, changes in visual input can lead to specific alteration of function and connectivity in mammalian visual cortex. In cat, this so-called critical period exhibits maximal sensory-driven adaptations around postnatal day 30 (P30), and ceases toward adulthood. We examined the molecular framework that directs age- and experience-dependent plasticity in cat visual cortex, by comparing protein expression profiles at eye opening (postnatal day 10 (P10), when experience-dependent plasticity starts), the peak of the critical period (P30), and in adulthood. Using 2-D DIGE, we performed comparisons of P10-P30 and P30-adult brain protein samples. Sixty protein spots showed statistically significant intensity changes in at least one comparison. Fifty-one spots were identified using quadrupole-TOF MS/MS or LC-MS/MS, containing 37 different proteins. The progressive increase or decrease in protein expression levels could be correlated to age-dependent postnatal brain development. Four spots containing transferrin, 14-3-3 alpha/beta and cypin, showed maximal protein expression levels at P30, thereby showing a positive correlation to critical period plasticity. Western analysis indeed revealed a clear effect of visual deprivation on cypin expression in cat visual cortex. Our results therefore demonstrate the power of 2-D DIGE as a tool toward understanding the molecular basis of nervous system development and plasticity.

We monitored the protein expression profiles of collapsin response mediator protein 2 and 4 (CRMP2 and CRMP4) throughout cat primary visual area 17 at different postnatal ages. Single immunocytochemical stainings revealed a clear effect of cortical maturation on the spatial and laminar distribution profile of CRMP2 and CRMP4. In kittens of postnatal day 10 (P10) and 30 (P30), CRMP2 and CRMP4 immunoreactivity was exclusively present in fibers running perpendicular to the cortical surface and crossing all cortical layers, but was never found in neuronal cell bodies. The immunoreactive fibers were embedded in an intensely and homogeneously stained neuropil. In contrast, mature visual cortex immunocytochemistry located CRMP2 and CRMP4 in the somatodendritic compartment of neurons with a clear CRMP-specific lamination pattern. Similar to kitten, neuropil staining was clearly observed but showed a decreasing gradient from layer I to VI in adult area 17. Detailed analysis of cellular morphology and size classified the CRMP2- and CRMP4-immunopositive cells in distinct neuronal populations. Double labeling of CRMP2 or CRMP4 with the typical interneuron marker parvalbumin (PV) showed many double-labeled cells immunoreactive for CRMP4 and PV, but not for CRMP2 and PV, corroborating the cell type-specific character of each CRMP. Our present results clearly illustrate that CRMP2 and CRMP4 may play an important role in visual cortex, possibly providing different classes of neurons with the potential to form a functionally meaningful network, not only during development, but also in adulthood, coincident with the belief that CRMPs are involved in neurite growth and guidance.