Distribution of histamine‐, 5‐hydroxytryptamine‐, and tyrosine hydroxylase‐immunoreactive neurons and nerve fibers in developing rat brain

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Abstract
Although the general patterns of the developing histaminergic system in the rat brain are known, no comparative studies between the development of the brain histaminergic system and the development of other neuroactive substances have yet been published. Interestingly, separate immunohistochemical studies on the development of the 5‐HT system and on the catecholaminergic system in the rat imply common features in the different aminergic systems. Therefore, the spatial distribution of histamine‐immunoreactive (HA‐ir) neurons and nerve fibers was compared to the distribution of 5‐hydroxytryptamine (5‐HT)‐, and tyrosine hydroxylase‐immunoreactive (TH‐ir) ones in the developing rat brain between embryonic days 12, (E12) and 20 (E20) by using a double‐immunostaining method. The high‐pressure liquid chromatography (HPLC) fluorometric method was used for determination of histamine concentration in different brain regions during the same period of development and synthetic oligonucleotide probes complementary to the rat histidine decarboxylase (HDC) to determine the origin of HA in the brain during the development with in situ hybridization. The immunohistoch6mical results revealed co‐localization of HA and 5‐HT within a subgroup of cells in the developing raphe nuclei between E14 and E18. From E18 onwards HA immunoreactivity started to gradually disappear from the rhombencephalon, and was totally abolished by E20, while 5‐HT‐ir cells continued to establish their adult positions. No significant colocalization of RA and TH immunoreactivities was detected. The biochemical results were in agreement with the immunohistochemical ones and confirmed that histamine detected in the early developing brain is authentic. A positive in situ hybridization signal for HDC was detected in a small area in the ventrolateral pons in the same areas as HA‐ and HDC‐ir cell bodies at E16, suggesting that at least some HA may be synthesized locally. These results confirm that HA is one of the first neurotransmitters to appear in the developing brain. In addition, the transient co‐localization of HA and 5‐HT immunoreactivities and the transient HDC expression at E16 within the developing pontine raphe nuclei may imply an interesting and a more general role for HA in modification of brain development.

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