TABLE 1.List of the primers used to detect the mRNA expression of 24 genes in the two populations of layer V pyramidal cells by RT-PCR single cell
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Gene Name | Sense and Antisense Primer Sequences (Initial Position) | Primer Size, pb | Amplicon Size, pb | GenBank Reference |
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Kv1.2 | GAG AAC CTC AGC TCC TGC CT (501) | 20 | 590 | M30440 |
CTT CCT TGA TGT AGC CTT CAT (1523) | 21 | |||
Kv4.2 | GTT ACC GTT TCG TGC GCA GT (751) | 20 | 450 | AF107780 |
TAG CGC AAT GAC CAA GAC TC (1200) | 20 | |||
Kv4.3M/L | GAA GCT CCA ATG CCT ACC TG (1307) | 20 | 421/478 | AF107781 |
CAA ATT AAG GCT GGA GCG AC (1728/1785) | 20 | |||
α1A | CAC TCA AGC TGG TGT CTG GA (467) | 20 | 333 | NM_007578 |
GGA AAA CAG TGA GCA CAG CA (799) | 20 | |||
α1B | ACA TTC GTG GTC TCT CCA CC (4375) | 20 | 306 | NM_007579 |
GAG GCG AAG GAAGCT TAG GT (4680) | 20 | |||
α1C | CCA GCC CAG AAA AGA AAC AG (3530) | 20 | 271 | NM_009781 |
ACT GCC TTT TCC TTA AGG TGC A (3780) | 22 | |||
α1D | ATT GCC AGA AAA GAA AGC CTA GA (2450) | 23 | 321 | NM_028981 |
AAT GAG CTT GTG GCA ACC CAC (2770) | 21 | |||
α1E | ATG GAG ACT CGG ACC AGA GC (509) | 20 | 248 | NM_009782 |
GGT GGC CAG GAT CAT GTA CTC (757) | 21 | |||
α1G | GAT GGT ACC TCA CAC TGA GG (5916) | 20 | 207 | NM_009783 |
GGT TGG GAG TGA ACA GAC AA (6122) | 20 | |||
α1H | AGG ACG CAG CAG AGT TTG AT (3753) | 20 | 287 | NM_021415 |
GTT GGA GAC GCT GAG AAA GG (4029) | 20 | |||
α1I | ATG CTG GTG ATC CTG CTG AAC (79) | 21 | 300 | AF393329hu |
GCA CGC GGT TGA TGG CTT TGA G (400) | 22 | |||
HCN1 | AGG TTA ATC AGA TAC ATA CAC C (1008) | 22 | 231 | AJ225123 |
GAG TGC GTA GGA ATA TTG TTT T (1238) | 22 | |||
HCN2 | CGG CTC ATC CGA TAT ATC CA (978) | 20 | 230 | AJ225122 |
AGC GCG AAC GAG TAG AGC TC (1207) | 20 | |||
mGluR3 | CAA GAA AAC ATC CCA CTG CTC A (767) | 22 | 364 | NM_181850 |
TAG GAC TTG CGG ATG TTG GAG C (1131) | 22 | |||
mGluR5a/b | GCC TTC GTG CCT ATC TAC TTT G (2623) | 22 | 377/473 | NM_017012 |
TTG GGG TTC TCC TTC TTG TTG A (2999) | 22 | Rat | ||
Neurotrypsin | GAG AGG CCA CAG AAA ACA GC (1947) | 20 | 337 | Y13192 |
AAG GTA CAA AAG CGG GGA CT (2283) | 20 | |||
I-1 | CGA CAA CAG CCC ACG GAA GAT C (13) | 22 | 486 | AF281676 |
GAA TCC AGT GGT AGC ATG TG (498) | 20 | |||
PN-1 | CTT TCC TGT TTT CCA TCC GA (1272) | 20 | 254 | X70296 |
GCC CTT TCT TCT GAC ACA GC (1525) | 20 | |||
CCK | CGC ACT GCT AGC GCG ATA CA (244) | 20 | 217 | NM_031161 |
TTT CCT CAT TCC ACC TCC TC (460) | 20 | |||
COX4 | ATG TTG GCT TCC AGA GCG CTG A (56) | 22 | 507 | M37829 |
CTT CTT CCA CTC ATT CTT GTC ATA G (562) | 25 | |||
OTX1 | GTT CGC AAA GAC TCG CTA CC (1659) | 18 | 242 | AF424700 |
AAG CAG AGC TAG ATA CGGT CG (3211) | 20 | |||
Bcl6 | TTC TTC CAG TTG CAG GCT TT (1210) | 20 | 775 | U41465 |
TGC AGG AAG TTC ATC AAG GC (461) | 20 | |||
Calbindin-28K | GAC GGA AGT GGT TAC CTG GA (212) | 20 | 416 | NM_009788 |
AGT TCC AGC TTT CCG TCA TTA (627) | 21 | |||
Ten-m3 | ACC AAC GGG TGT CTG AAA AG (4868) | 20 | 216 | AB025412 |
GTC CAC CGT GAT AGC CTT GT (5083) | 20 |
Detailed in this table are the abbreviated names of genes, the sequences of the sense and reverse primers given from the 5'
and 3' and their initial position (number in parentheses), the size of the primer, the size of the PCR amplified product,
and the accession number in GenBank. The amplified genes correspond to the potassium channel subunits Kv1.2, Kv4.2, and Kv4.3;
the calcium channel subunits α1A, α1B, α1C, α1D, α1E, α1G, α1H, and α1I; the hyperpolarization-activated channel subunits
HCN1 and HCN2; the glutamate metabotropic receptor subunits mGluR3 and
mGluR5; the neurotrypsin (neuro.); the inhibitor of phosphatase 1 (1I); the protease nexin 1 (PN1); cholecystokinin (CCK);
polypeptidic complex IV of the cytochrome C oxidase (Cox-4); orthodenticle homolog 1 (Otx-1); transcriptional repressor Bcl
6, calcium-binding protein calbindin 28K; and a substrate of the tyrosine kinase II (Ten-m3).