Ontological study of calbindin‐D28k‐like and parvalbumin‐like immunoreactivities in rat spinal cord and dorsal root ganglia

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Abstract
The calcium ion plays an important role in some critical developmental events in the nervous system, such as neurulation and neurite elongation. Therefore, as the intracellular calcium‐binding proteins calbindin‐D28k (CaB) and parvalbumin (PV) may be expressed in these developmental events. Accordingly, the ontological expression of CaB and PV was examined immunocytochemically in the spinal cord and dorsal root ganglia (DRG) of the rat, in order to evaluate the relationship between CaB and PV expression, and other important developmental events. During the ontogenesis of the spinal cord, the CaB‐like immunoreactivity was mainly observed in the cell somata. The immunoreactive cells in the ventral horn of the cervical and thoracic, lumbar, and sacral segments first appeared at embryonic day (E)‐12, E‐13, and E‐14, respectively. However, these cells were not detected in the intermediate gray matter of the same segments at E‐14, E‐15, and E‐16, respectively, and in the dorsal horn at E‐14‐E‐15, E‐16, and E‐17, respectively. The peak of immunoreactive cells, both as to number and intensity, occurred in the perinatal period. However, from postnatal day (P)‐14 on, the number and intensity of the positive cells decreased, the adult levels being reached at P‐35. The PV‐like immunoreactivity was mainly detected in the fibers and punctata during the ontogenesis of the spinal cord. The immunoreactive fibers first appeared on the surface of the dorsal horn in the cervical and thoracic segments at E‐14, then entered the dorsal horn at E‐15, and reached the intermediate gray matter and ventral horn at E‐16. The first appearance of these fibers in the same areas of the lumbar and sacral segments occurred 1 day later than in the cervical and thoracic segments. During the perinatal period, the maximum content of PV‐like immunoreactive fibers, together with many punctata, was seen in the gray matter. However, between P‐14 and P‐17, most of them lost immunoreactivity rapidly, with the exception of the medial region of the intermediate gray matter, where the PV‐immunoreactive punctata remained up to the adult stage. In DRG neurons, both CaB and PV was expressed, but in different neurons. Neurons labeled with anti‐CaB and anti‐PV sera were first detected at E‐16 and E‐14, respectively. These neurons were large or medium‐sized in the prenatal period. Small neurons were also labeled with these sera after birth. Positive neurons of different sizes in DRG increased in number in the postnatal period. Up to P‐35, the expression of CaB and PV in DRG reached the adult levels. The appearance of and developmental changes in CaB‐like and PV‐like immunoreactivities in the spinal cord and DRG may be maintained during the neurogenesis of the immunoreactive regions, suggesting the possible developmental roles of CaB and PV.