Analysis of the heterogeneity within bovine pineal gland by immunohistochemistry and in situ hybridization

Abstract

In the present study, we demonstrate a cortical and medullary arrangement of parenchymal cells in the bovine pineal gland by using antibodies for neuron-specific enolase, synaptophysin, and hydroxyindole O-methyltransferase (HIOMT) as markers of pinealocytes, and glial fibrillary acidic protein (GFAP) as a marker of interstitial (glial) cells. Furthermore, by means of probes specific for HIOMT mRNA, we have examined possible differences in melatonin synthesis between the cortex and the medulla. Immunoreactive pinealocytes for each antigen investigated are more densely distributed in the cortex than in the medulla. In the cortex, GFAP-positive interstitial cells have large intenselystained somata endowed with several long, thin cytoplasmic processes, whereas in the medulla, they display smaller, less intensely labeled perikarya from which numerous fine short processes emerge. Golgi staining has confirmed these morphological differences between the interstitial cells in the cortex and those in the medulla. An analysis using confocal laser microscopy together with in situ hybridization for HIOMT mRNA has shown that the expression of mRNA transcripts in the cortex is more intense than that in the medulla. The expression of the HIOMT gene in a cluster of cells in the medial habenular nucleus is lower than that in pinealocytes of the pineal organ proper.