Postnatal development of substance-P immunoreactivity in the rat superior colliculus

Immunocytochemical techniques have been used to examine the distribution of substance-P (SP)-labeled neurons in the superior colliculus of rats from birth to adulthood. At birth, there are almost no SP-immunopositive neurons in the tectum. A small number of SP neurons appear over the next several days. However, the vast majority of SP neurons appear between P9 and P10, and by P12 have attained adult-like numbers and distribution. Neurons are confined to the superficial layers of the colliculus, specifically the upper two-thirds of the stratum griseum superficiale (SGS). There is no indication of a differential developmental sequence along rostrocaudal or mediolateral axes. Neuronal types can be distinguished as early as P6 and include horizontal, vertical, and multipolar cells.Substance-P-immunoreactive axons and boutons are also present in the superior colliculus at birth, and are for the most part confined to the deep layers. Boutons are generally of the en-passant type. The density of labeled axons and boutons increases progressively, and by P10 there is an almost adult-like lamination and patchiness. In the adult, labeled axons and boutons are most dense in the stratum opticum and stratum griseum intermedium. Bridges of dorsoventrally oriented labeled axons span the relatively label-poor stratum album intermedium. SP label in the stratum griseum profundum is dense and patchy, and there is also dense label in the stratum album profundum bordering the periaqueductal grey.The role of substance-P-labeled neurons in the superior colliculus is still a matter of speculation. The findings of this study indicate that SP neurons may play a role in intrinsic collicular circuitry.

Immunocytochemistry. A Review of Methodology for Electron Microscopic Applicaiton

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.