The blood supply of the cat's visual cortex and its postnatal development.

We examined the blood supply of the cat's visual cortex using alkaline phosphatase histochemistry to demonstrate the capillary endothelial cells. In the adult, layer 4 is marked by a band that is of obviously greater density, extends throughout areas 17 and 18, and ends abruptly at the 18/19 border. We quantified blood vessel density in area 17, observing a 23% greater density in layer 4 than in supragranular and infragranular layers. This difference reflects a laminar difference in metabolic rate. In three animals studied using the metabolic marker 2-deoxyglucose, layer 4 was 25% denser than the other layers. The band of greater density in layer 4 is not present in newborn kittens, but becomes apparent at about 5 weeks of age. Early in development, the endothelial cells form filopodia as the capillaries grow and branch. The density of blood vessels decreases slightly during the first week of postnatal life, but increases between 1 and 6 weeks of age, so that by 6 weeks, the blood supply of the visual cortex resembles that seen in the adult. This pattern resembles that of cortical metabolism seen with 2-deoxyglucose [J. Cereb. Blood Flow Metab. 11 (1991) 35], but the increase in vascular density precedes that in glucose metabolism.

Intracellular injection in fixed slices: obtaining complete dendritic arbors of large cells.

Intracellular injection of Lucifer yellow into fixed brain slices is widely used to demonstrate dendritic morphology. A major limitation of this technique is that large dendritic arbors are usually truncated at the cut surfaces. Here we describe modifications that allowed us to obtain complete dendritic arbors of large spiny stellate cells. Lucifer Yellow cadaverine biotin-X (LY-X) was injected into individual neurons within 300-1000 microm thick aldehyde-fixed slices of kitten visual cortex. Subsequently, the LY-X was histochemically reacted using standard ABC methods to obtain a permanent record of the injected cells. Dendrites, studded with a variety of dendritic spines, were darkly labeled and well defined against virtually no background. Somatic spines, dendritic varicosities and growth cones were common in the younger animals. Computer-assisted reconstructions demonstrated that, in older animals, the dendritic arbors of cells injected in 300 microm slices were truncated, whereas the arbors of cells injected deep within thick slices were complete. The modifications described here remove the most critical limitation of intracellular injection in slices, allowing quantitative analysis of even large dendritic arbors.