Calbindin-D28k and calretinin immunoreactivity in the spinal cord of the lizard Gekko gecko: Colocalization with choline acetyltransferase and nitric oxide synthase

Abstract

The distribution of the calcium-binding proteins calbindin-D28k (CB) and calretinin (CR) was investigated in the spinal cord of the lizard Gekko gecko, by means of immunohistochemical techniques. Abundant cell bodies and fibers immunoreactive for either CB or CR were widely distributed throughout the spinal cord. Most neurons and fibers were labeled in the superficial dorsal horn, but numerous cells were also located in the intermediate gray and ventral horn. Distinct CB- and CR-containing cell populations were observed, although double immunohistochemistry revealed that 17–20% of the single-labeled cells for CB or CR in the dorsal horn contained both proteins. In addition, nitric oxide synthase was immunodetected in about 6% of the CB-positive neurons in the dorsal horn and in 10% in the ventral horn, whereas nitric oxide synthase was present in 9–13% of CR-positive cells in the dorsal horn and in 14% in the ventral horn. These doubly immunoreactive cells were restricted to areas IV, VII and VIII. Similar colocalization experiments revealed that 18–24% of the cholinergic cells in the ventral horn contained CB and 21–30% CR, with some variations throughout the length of the spinal cord. The pattern of distribution for CB and CR immunoreactivity in the spinal cord of the lizard, reported in the present study, is largely comparable to those reported for mammals, birds and anuran amphibians suggesting a high degree of conservation of the spinal systems modulated by these calcium-binding proteins.