Thyrotropin‐releasing hormone (TRH) regulates the hypothalamic–pituitary–thyroid axis in mammals and also regulates prolactin secretion, directly or indirectly via tuberoinfundibular dopamine neurons. Although TRH is abundantly expressed in teleost brain and believed to mediate neuronal communication, empirical evidence is lacking. We analyzed pro‐TRH‐mRNA expression, mapped TRH‐immunoreactive elements in the brain and pituitary, and explored its role in regulation of hypophysiotropic dopamine (DA) neurons in the catfish, Clarias batrachus. Partial pro‐TRH transcript from C. batrachus transcriptome showed six TRH progenitors repeats. Quantitative real‐time polymerase chain reaction (qRT‐PCR) identified pro‐TRH transcript in a number of different brain regions and immunofluorescence showed TRH‐immunoreactive cells/fibers in the olfactory bulb, telencephalon, preoptic area (POA), hypothalamus, midbrain, hindbrain, and spinal cord. In the pituitary, TRH‐immunoreactive fibers were seen in the neurohypophysis, proximal pars distalis, and pars intermedia but not rostral pars distalis. In POA, distinct TRH‐immunoreactive cells/fibers were seen in nucleus preopticus periventricularis anterior (NPPa) that demonstrated a significant increase in TRH‐immunoreactivity when collected during preparatory and prespawning phases, reaching a peak in the spawning phase. Although tyrosine hydroxylase (TH)‐immunoreactive neurons in NPPa are hypophysiotropic, none of the TRH‐immunoreactive neurons in NPPa accumulated neuronal tracer DiI following implants into the pituitary. However, 87 ± 1.6% NPPa TH‐immunoreactive neurons were surrounded by TRH‐immunoreactive axons that were seen in close proximity to the somata. Superfused POA slices treated with TRH (0.5–2 μM) significantly reduced TH concentration in tissue homogenates and the percent TH‐immunoreactive area in the NPPa. We suggest that TRH in the brain of C. batrachus regulates a range of physiological functions but in particular, serves as a potential regulator of hypophysiotropic DA neurons and reproduction.