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The purpose of the present study was to examine the regulation of three molecular variants of gonadotropin-releasing hormone (GnRH)-encoding mRNAs by testosterone in the male tilapia Oreochromis niloticus. Tilapias castrated for two weeks were injected intraperitoneally with sesame oil or 5 μg/g testosterone for 7 days. In situ hybridization histochemistry was performed using ³⁵S -labelled 30-mer antisense oligonucleotide probes complementary to exon two (bases 1–30) of salmon-, seabream-, and chicken II-GnRH. Computerized image analysis was performed to quantify GnRH mRNA expression in the terminal nerve ganglia (nucleus olfactoretinalis) and in individual cells of the preoptic area and the midbrain tegmentum. Testosterone treatment significantly elevated terminal nerve salmon-GnRH mRNA, reduced preoptic seabream-GnRH mRNA but had no effect on midbrain chicken II-GnRH mRNA levels …