Tillet Y, Krieger M, Thibault J
Laboratoire de Neuroendocrinologie Sexuelle, INRA, Nouzilly, France.
J Neurochem. 1997 May;68(5):2161-9. doi: 10.1046/j.1471-4159.1997.68052161.x.
Tyrosine hydroxylase (TH) cDNA has been characterized in rodents and primates, but only a few studies have been developed in ungulates, except in cows. Because sheep is a species used for many physiological studies, it was of interest to clone TH cDNA in this species. Ovine TH cDNA was purified from a library of sheep adrenal glands. The entire cDNA was 1,721 bp long. It presented a higher percentage of similarity with bovine TH cDNA (93%) than with rodent cDNAs (75%). The deduced amino acid sequence was 490 amino acids long and had 96% similarity with the bovine amino acid sequence. The entire cDNA and different fragments obtained with endonuclease restriction enzymes were cloned in plasmid pUC 18 and were labeled with 35S-dATP to detect TH mRNA by in situ hybridization. Strong labelings were observed on adrenal medulla and on noradrenergic and dopaminergic neurons in the sheep but also in the cow and pig. This labeling matched completely TH immunohistochemical staining obtained on the same sections with anti-TH antibodies. Ovine TH cDNA is a useful tool to study the variations of TH mRNA levels in sheep catecholaminergic neurons.
酪氨酸羟化酶（TH）cDNA已在啮齿动物和灵长类动物中得到表征，但除了牛之外，在有蹄类动物中开展的研究较少。由于绵羊是用于许多生理学研究的物种，因此克隆该物种的TH cDNA很有意义。绵羊TH cDNA是从绵羊肾上腺文库中纯化得到的。整个cDNA长1721 bp。与牛TH cDNA的相似性百分比（93%）高于与啮齿动物cDNA的相似性百分比（75%）。推导的氨基酸序列长490个氨基酸，与牛氨基酸序列的相似性为96%。用核酸内切限制酶获得的整个cDNA和不同片段被克隆到质粒pUC 18中，并用35S-dATP进行标记，通过原位杂交检测TH mRNA。在绵羊的肾上腺髓质以及去甲肾上腺素能和多巴胺能神经元上观察到强标记，在牛和猪中也观察到了。这种标记与在同一切片上用抗TH抗体获得的TH免疫组织化学染色完全匹配。绵羊TH cDNA是研究绵羊儿茶酚胺能神经元中TH mRNA水平变化的有用工具。
