The distribution of immunoreactive (ir) leucine-enkephalin (LE) and ir-methionine-enkephalin-Arg-Gly-Leu (ME-RGL) in 101 microdissected rat brain and spinal cord regions was determined using specific and sensitive radioimmunoassays. The highest concentrations of LE and ME-RGL were measured in globus pallidus (5190 and 4378.8 fmol/mg protein, respectively). Very high concentrations of LE and ME-RGL (greater than 750 fmol/mg protein) were found in the central amygdaloid nucleus, anterior hypothalamic nucleus, lateral preoptic area, nucleus of the solitary tract (medial and commissural parts), bed nucleus of stria terminalis, dorsomedial nucleus, parabrachial nuclei, periaqueductal gray and motor hypoglossal nucleus. Very high concentrations of ME-RGL were found in 14 additional brain regions including medial preoptic area, area postrema, nucleus ambiguus, periventricular nucleus, ventromedial nucleus, interpeduncular nucleus, paraventricular, arcuate and others. High concentrations of LE (between 500 and 750 fmol/mg protein) were found in 15 brain areas, among them the periventricular nucleus, medial preoptic area, suprachiasmatic nucleus, dorsal premamillary nucleus, ventromedial nucleus, arcuate nucleus, nucleus ambiguus, locus coeruleus, substantia nigra. High concentrations of ME-RGL were measured in 13 brain areas including the suprachiasmatic nucleus, lateral septal nucleus, raphe magnus, motor facial nucleus, lateral amygdaloid nucleus, sensory trigeminal nucleus, nucleus accumbens, caudate-putamen. Moderate concentrations of LE (between 250 and 500 fmol/mg) were found in 46 brain areas such as the lateral septal nucleus, nucleus accumbens, caudate-putamen, several amygdaloid nuclei, supraoptic nucleus, the perifornical nucleus, posterior hypothalamic nucleus, red nucleus. Moderate concentrations of ME-RGL were detected in 27 areas such as the median eminence, nuclei of the reticular formation, supraoptic nucleus, red nucleus and others.(ABSTRACT TRUNCATED AT 250 WORDS)