Expression of estrogen receptor alpha exon-deleted mRNA variants in the human and non-human primate frontal cortex.
Although estrogen receptor alpha (ERalpha) mRNA has been detected in the primate frontal cortex, the types of ERalpha transcripts expressed, including exon-deleted variants (Delta), have not been determined in the monkey or human frontal cortex. Because the types of ERalpha mRNA expressed in brain could define neuronal responses to estrogens, we examined the transcript pool of ERalpha mRNAs expressed in normal adult and developing human and macaque frontal cortex. We reverse transcribed total RNA from the postmortem frontal cortex of 29 normal adult humans, 12 rhesus macaques, and 19 people ranging from infants to adults and employed two rounds of nested polymerase chain reaction (PCR) to generate ERalpha products spanning the coding domain. In a third nested PCR, we used primers specific for novel sequences of exon-exon junctions created when whole exons are missing. By sequencing PCR products, we detected 60 instances of 12 distinct DeltaERalpha mRNAs in adult humans and 94 instances of 13 distinct DeltaERalpha mRNAs in monkeys in differing patterns from one individual to another. In adult humans, 83% of individuals expressed at least 1 DeltaERalpha mRNA variant, and 100% of the monkeys expressed at least 1 DeltaERalpha mRNA variant. The single Delta2, Delta5, and Delta7 variants were frequently expressed in both human and monkey frontal cortex, Delta3 variants were rare in both species, and Delta6 variants were more frequently expressed in monkeys. In both species, we detected double, triple and quadruple Deltas, but these were less common than single Deltas. The pattern of human variant expression did not appear to change dramatically as a function of age. These findings imply the potential to produce different ERalpha proteins in frontal cortex, possibly with altered structure and function which may have physiological relevance for gene transcription by virtue of altered functional interactions with each other, other steroid hormone receptors, and genomic DNA.
Perlman
W R
WR
Clinical Brain Disorders Branch, National Institute of Mental Health, National Institutes of Health, United States Department of Health and Human Services, Bethesda, MD 20892-1385, USA. perlmanw@intra.nimh.nih.gov
Matsumoto
M
M
Beltaifa
S
S
Hyde
T M
TM
Saunders
R C
RC
Webster
M J
MJ
Rubinow
D R
DR
Kleinman
J E
JE
Weickert
C S
CS
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000818:Animals; D015152:Blotting, Northern; D002648:Child; D002675:Child, Preschool; D002896:Chromosomes, Human, Pair 6; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D005091:Exons / Q000235:genetics*; D005260:Female; D005625:Frontal Lobe / Q000378:metabolism*; D017353:Gene Deletion; D015870:Gene Expression / Q000502:physiology; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006801:Humans; D007223:Infant; D008253:Macaca mulatta; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012737:Sex Factors
D047628:Estrogen Receptor alpha; D012333:RNA, Messenger