Table 3.

To investigate the developmental expression profile of the novel transcripts, we used samples from the prefrontal cortex of nonpsychiatric controls encompassing almost the entire human lifespan, from the fetal 14th through 20th gestational weeks and from birth through old age (78 years)a

NameSequence
NKCC1a (1-27) forward primer5′-ACCAAGGATGTGGTAGTAAGTGTGG-3′
NKCC1a (1-27) reverse primer5′-GGATTCTTTTTTCAACAGTGGTTGA-3′
NKCC1a (1-27) probe6FAMTACACACAAAGTTGAGGAAGMGBNFQ
NKCC1b (1-27 (Δ21)) forward primer5′-AGGATGTGGTAGTAAGTGTGGAATATAGTAA-3′
NKCC1b (1-27 (Δ21)) reverse primer5′-TCAAGAAGCTTTTGGTCAGCTACAT-3′
NKCC1b (1-27 (Δ21)) probe6FAMTACACACAAAGAATCCAMGBNFQ
NKCC1 1-27 (21a) forward primer5′-GAATATAGTAAAAAGTCCGATTTAGATACTTCC-3′
NKCC1 1-27 (21a) reverse primer5′-CCTTCTCATAAAATTCATGTATGTTGACA-3′
NKCC1 1-27 (21a) probe6FAMTTACACACAAAGTCGTGCTGMGBNFQ
NKCC1 1-4a forward primer5′-TGACAACTATCACAGGATTGTCTACTTC-3′
NKCC1 1-4a reverse primer5′-ATTCTTTTCTTCTTGTTTTCAGGGATC-3′
NKCC1 1-4a probe6FAMAGAGGAGGAAGAGGACMGBNFQ
NKCC1 1-2a forward primer5′-CTGTGGTCACGTATACTGCAGAAAGTA-3′
NKCC1 1-2a reverse primer5′-AACTGATGAGAACGTTAGGAAGATTTTATA-3′
NKCC1 1-2a probe6FAMCAAGGGTGTATTAGTATGTATATMGBNFQ
  • aWe assessed mRNA expression levels of novel NKCC1 transcripts [isoforms 1–27 (21a), 1–4a, and 1–2a] as well as the known transcripts NKCC1a (1–27) and NKCC1b (1–27 (Δ21)) in the DLPFC by qRT-PCR (Table 3). We measured the expression of NKCC1a (1–27), using an assay spanning exon 20 and exon 21, expression of NKCC1b (1–27 (Δ21)), using an assay spanning exon 20 and exon 22, expression of 1–4a, using an assay spanning exon 4 and a unique exon 4a, and expression of NKCC1b 1–2a, using an assay spanning exon 2 and a unique exon 2a. We did not measure the isoform 1–27 (Δ5–26) by qPCR because of low abundance (Ct > 36) of mRNA expression in our preliminary study (data not shown). All data were normalized to a geometric mean of three “housekeeping genes.”