. 1998 Dec 1;18(23):9766–9779. doi: 10.1523/JNEUROSCI.18-23-09766.1998

Table 1.

RT-PCR gene-specific primers

Oligonucleotide Primers	Specificity^1-a	Sequence	Position^1-b	Predicted product size^1-c (bp)	Annealing temperature
PCP1	ProPACAP mRNA	5′-ATGCCTCTCTGGTTGTGATTC-3′	486–506	606/612	57°
PCP2	(rat neuronal proPACAP mRNA)	5′-CGCTATTCGGCGTCCTTTGTT-3′	1071–1091	606/612	57°

PCPX1	ProPACAP mRNA	5′-ATGACCATGTGTAGCGGAGCAAGGT-3′	573–597	430/433	61°
PCPX2	(5′ coding region; cross species)	5′-GGTAGCGGCTGTAGCTGTCTGTGAA-3′	978–1002	430/433	61°

PCPX5	ProPACAP mRNA	5′-AGGACGGAAACCCGCTGCAAGACTT-3′	703–727	399	62°
PCPX6	(3′ coding region; cross species)	5′-GCTACAAGTACGCTATTCGGCGTCCTTTGT-3′	1072–1101	399	62°

PACAPR1 PACAPR2	PAC1 receptor mRNA (third cytoplasmic loop splice variants; cross species)	5′-CTTGTACAGAAGCTGCAGTCCCCAGACATG-3′ 5′-CCGGTGCTTGAAGTCCATAGTGAAGTAACGG TTCACCTT-3′	1078–1107 1510–1548	303–471^1-d	60°

PACAPRX3 PACAPRX4	PAC1 receptor mRNA (amino terminal extracellular domain splice variants; cross species)	5′-TGCCCTGAGGTCTTCCGGATCTTCAAC-3′ 5′-AATGAACAGCCAGAAGTAGTTGGACAC-3′	316–342 799–825	447/510^1-d	58°

^F1-a

For cross-species primers, regions of high homology for proPACAP were selected using the rat (M63006), mouse (D14716), sheep (S83511), and human (S83513) cDNA sequences. Sequences used for identifying PAC1 receptor homologous regions included the rat (Z23272), mouse (D82935), human (D17516), and cow (D17290).

^F1-b

Positions are given as the nucleotide sequences for the rat cDNA with the following GenBank accession numbers: rat proPACAP transcript, M63006; rat PAC1 (PACAP₁)short HIP/HOP receptor transcript, Z23272.

^F1-c

The predicted amplified product sizes for each set of oligonucleotide proPACAP primer templates are based on the rat, mouse, sheep, and human cDNA sequences (see above). PACAP-selective receptor amplified product sizes are predicted from the cDNA sequences with the following GenBank accession numbers: rat PAC1short, Z23279; rat PAC1 short HIP/HOP1, Z23272; rat PAC1 shortHIP, Z23273; rat PAC1 shortHOP1,Z23274; rat PAC1 shortHOP2, Z23275; mouse PAC1shortHOP1, D82935; human PAC1 short, D17516; cow PAC1 shortHOP1, D17290.

^F1-d

The potential PAC1 receptor third cytoplasmic loop alternate splice variant product sizes are PAC1 (neither HIP nor HOP), 303 bp; PAC1HOP1, 387 bp; PAC1HIP, 387 bp; PAC1HOP2, 384 bp; PAC1HIPHOP1, 471 bp; PAC1HIPHOP2, 468 bp. The PAC1 receptor amino terminal extracellular domain very short transcript variant without exons 4 and 5, and short variant containing these alternatively spliced exons are predicted to produce 447 and 510 nucleotide amplified products, respectively (Chatterjee et al., 1997).