Molecular and cellular physiology of aromatase in the brain and retina.
Due to exceptionally high brain aromatase activity, teleost fish are advantageous for studying neural aromatase regulation, localization, and physiology. To determine the molecular mechanism of enhanced expression, we have isolated, cloned and sequenced a 3 kb full-length aromatase cDNA from a goldfish (Carassius auratus) brain library using a human placental aromatase cDNA as probe. The deduced sequence of goldfish aromatase is 510 amino acids (predicted Mw, 58 kDa) with 69% overall sequence similarity, when compared to human placental aromatase, and higher homologies in presumptive functional domains. A major 3 kb mRNA species was abundant in brain and low or non-detectable in non-neural tissues, reflecting the order of enzyme activities. To determine the cellular basis of high enzyme activity in goldfish brain, a human placental aromatase antibody was used to immunolocalize labeled cells. This antibody immunoprecipitated a single 56 kDa in vitro translation product of goldfish brain poly(A+)RNA and revealed discrete clusters of intensely stained neurons, processes, and terminals concentrated in, but not limited to, reproductive brain centers. Close proximity of aromatase- and androgen receptor-positive neurons in certain regions provides anatomic evidence of a functional relationship between direct and indirect pathways of neural androgen action. Aromatase-positive neurons and fibers formed interconnected networks in novel loci (e.g. retina-->optic tract-->optic tectum), and catalytic activity was confirmed biochemically in these tissues, indicating that neuroestrogen may have a role in visual input and integration. Availability of goldfish-specific nucleotide and antibody probes will facilitate further studies using this model.
Callard
G V
GV
Department of Biology, Boston University, MA 02215.
Drygas
M
M
Gelinas
D
D
D000818:Animals; D001141:Aromatase / Q000235:genetics* / Q000378:metabolism*; D001921:Brain / Q000201:enzymology*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes; D005260:Female; D006054:Goldfish; D006801:Humans; D009474:Neurons / Q000201:enzymology*; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011944:Receptors, Androgen / Q000378:metabolism / Q000502:physiology; D012160:Retina / Q000201:enzymology*
D015342:DNA Probes; D011944:Receptors, Androgen; D004247:DNA; D001141:Aromatase