Distinctive subsets of glutamatergic neurons in cerebral cortex sequester the transition metal zinc within the synaptic vesicles of their axon terminals. In the present study we used histochemical localization of synaptic zinc to investigate normal postnatal development and experience-dependent plasticity of zinc-containing circuits in somatosensory barrel cortex of rats. First, we found that zinc-containing cortical circuits are dynamically reorganized between postnatal day (P) 0 and P28. Whereas most cortical laminae exhibited idiosyncratic increases in zinc histochemical staining with advancing age, lamina IV barrels were darkly reactive early in life and then lost much of their complement of synaptic zinc during postnatal weeks 2-4. Second, we established that sensory experience plays a major role in sculpting the zinc-containing innervation of cortical barrels. Trimming a particular facial whisker arrested the normal postnatal decline in synaptic zinc in the corresponding, deprived barrel. This resulted in more intense zinc staining in deprived barrels compared with adjacent, nondeprived barrels. Notably, the influence of experience on development of zinc circuits was most robust during a critical period extending from about P14, when an effect of whisker trimming first could be observed, through P28, after which time chronic deprivation no longer resulted in heightened levels of synaptic zinc in lamina IV. These findings indicate that sensory input can have a marked influence on development of cortical circuits, including those within lamina IV, throughout the first postnatal month.
Barrel hollows in the posteromedial barrel subfield of adult rat somatosensory cortex typically encompass two or three metabolically and structurally distinct regions, termed subbarrels. We used immunohistochemical staining for vesicular glutamate transporter 2 and the neuronal serotonin transporter, in conjunction with cytochrome oxidase (CO) histochemistry, to investigate the distribution of thalamocortical (TC) axon terminals in relation to subbarrel domains. We found, first, that CO-dark subbarrels are more intensely immunoreactive for thalamocortical terminals than the CO-light clefts that separate them. Second, during the first postnatal week, immunoreactivity for markers of TC terminals is relatively homogeneous throughout the barrel hollow; subbarrel patterns of distribution only become recognizable between P-8 and P-10. These observations extend previous findings that subbarrels denote barrel regions enriched in synaptic contacts. The data also indicate that allocation of TC terminals into subbarrel domains does not occur immediately upon thalamic axon ingrowth. Instead, refinement of TC arbors into subbarrels is a gradual process, the outcome of which is not manifest until the second week of postnatal life.
The transition metal zinc is an essential dietary constituent that is believed to serve an important intercellular signaling role at certain excitatory synapses in the central nervous system. In the present study, we used histochemical techniques to investigate the distribution of synaptic zinc during postnatal development of retinogeniculate projections in rats. From postnatal day (P) 1 until P-21, the pattern of zinc histochemical staining in the dorsal lateral geniculate nucleus (LGNd) precisely matched the distribution of axon terminals from the ipsilateral eye that were labeled by anterograde transport of horseradish peroxidase. Regions of the LGNd that contained only crossed axons were devoid of zinc staining. Abnormalities in the distribution of uncrossed retinogeniculate projections in albino versus pigmented rats were paralleled by identical variations in localization of synaptic zinc. Unilateral enucleation on P-10 was followed within 5 days by loss of zinc staining in the LGNd ipsilateral to the removed eye without affecting staining in the contralateral nucleus. Finally, the ability to detect zinc histochemically in the LGNd ceased at approximately P-24. These findings provide evidence that zinc is sequestered within synaptic boutons of a subpopulation of retinal ganglion cells whose axons terminate on the ipsilateral side of the brain. The duration of zinc staining overlaps with the major period of axonal remodeling in the LGNd, suggesting that synaptically released zinc may play a role in postnatal refinement of retinogeniculate projections.
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