The ATBF1 gene encodes transcription factors containing four homeodomains and multiple zinc finger motifs. However, the gene products have yet to be identified and the role remains unknown in vivo. In this study, we raised an antiserum for ATBF1 and found high levels of expression of ATBF1 in developing rat brain. Western and Northern blot analyses detected a 400 kDa protein and 12.5 kb mRNA in developing rat brain, respectively; both corresponding to ATBF1-A but not the B isoform. The protein was highly expressed in the midbrain and diencephalon and mRNA was highly expressed in the brainstem, mostly in embryo and neonatal brain. Immunohistochemistry identified postmitotic neurons in the brainstem as the major site of ATBF1 expression, and the expression levels varied depending on age of and location in the brain. Expression was transient and weak in the precursor cells at early neurogenesis. ATBF1 decreased postnatally, but remained in mature neurons, including those expressing DOPA decarboxylase (DDC). High levels of ATBF1 were expressed in precursor cells in accordance with neurogenesis and were continued to the mature neurons in specific areas such as the inferior colliculus. Expression was not significant from precursor cells to mature neurons in the cerebral cortex and hippocampus. ATBF1 and its Drosophila homolog, Zfh-2, are known to regulate cell differentiation and proliferation via the interaction with either of the basic helix-loop-helix transcription factors, c-myb, or the DDC gene. Together with these reported functions the expression features detected here suggest that ATBF1 may participate in the regulation of neuronal cell maturation or region-specific central nervous system differentiation.
The zinc finger-homeodomain (ZFH) transcription factors contain a zinc finger motif and a homeodomain that might regulate neural and mesenchymal cell differentiation. We have cloned the ZFH4 gene that encodes a protein with structures closely related to ATBF1. In order to study the expression pattern of ZFH4 in the developing rat brain, we raised an antibody against a glutathione-S-transferase (GST) fusion protein of ZFH4. Western blotting with this antibody identified a gene product of 390 kDa in the normal rat brain. Levels of the protein were high in the brainstem at embryonic and neonatal periods and in the midbrain and diencephalon in neonatal rat brain. In addition, the corresponding mRNA of 12.5 kb was detected by Northern blotting. An immunolocalization study showed that postmitotic neurons in the brainstem were the major site of ZFH4 expression, and the levels of expression varied depending on age and anatomical sites. Expression was transient and weak in precursor cells at early neurogenesis. Although ZFH4 levels decreased after birth, ZFH4 continued to be expressed in the mature neurons including DOPA decarboxylase-positive neurons. High levels of expression were also detected in non-neuronal cells of the subcommissural organ, but the expression was almost undetectable throughout precursor cells to mature neurons in the cerebral cortex and hippocampus. The spatial and temporal expression patterns closely resembled those of ATBF1, and we detected neurons that expressed ZFH4, ATBF1, or both. We postulate that ZFH4 participates in the regulation of neural cell maturation or of region-specific differentiation of the brain.
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