Serotonin Genes
I am interested in 4 of the genes related to serotonin function in depression and food intake: Pet-1, Serotonin Transporter (5HTT), Tryptophan Hydroxylase 2 (TPH2), and 5HT2C Receptor (5HT2CR)
ISH probes
Pet-1
Reference:
Serotonin Transporter
Reference:
Tryptophan Hydroxylase 2
(see PCR primers below).
Reference: Kulikov AV, Osipova DV, Naumenko VS, Popova NK. Association between Tph2 gene polymorphism, brain tryptophan hydroxylase activity and aggressiveness in mouse strains. Genes Brain Behav. 2005 Nov;4(8):482-5. PMID 16268992
5HT2C Receptor
(see PCR primers below)
Was originally called the 5HT1C receptor when first discovered.
References:
Molineaux SM, Jessell TM, Axel R, Julius D. 5-HT1c receptor is a prominent serotonin receptor subtype in the central nervous system. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6793-7. PMID 2771958 Very nice in situ usinga 300 bp riboprobe. The sequence was taken from Julius et al 1988
Julius D, MacDermott AB, Axel R, Jessell TM. Molecular characterization of a functional cDNA encoding the serotonin 1c receptor. Science. 1988 Jul 29;241(4865):558-64. PMID: 339989
PCR primers
Tryptophan Hydroxylase 2 cDNA
for in situ hybridization
product size:
495 bp product corresponding to bases 319-813 of AY098915 (from Malek PMID 16115212).
502 bp product corresponding to bases 312-814 of NM173839 (based on our lookup)
Verified by sequencing on 2-12-09; RT-PCR product sequenced with for and rev primers matched bases 366-769 of AY098915.
- TPH2for base 312-334:
- 5’ -GAAGTTGGCGGGCTGGTGAGA–3’
- TPH2rev:bases 791-814
- 5’ -TCGGCAAGCATGAGTGGGGTAGA–3’
Reference:
Malek ZS, Dardente H, Pevet P, Raison S. Tissue-specific expression of tryptophan hydroxylase mRNAs in the rat midbrain: anatomical evidence and daily profiles. Eur J Neurosci. 2005 Aug;22(4):895-901. PMID 16115212
Tryptophan Hydroxylase 2 polymorphisms
linked to aggression. 5 min at 94°C then 40 cycles (30 s at 94°C, 30 s at 60°C, 30 s at 72°C)
Positive control, product size 523 bp
- ctrlTPH2for: 5’ -tttgacccaaagacgacctgcttgca –3’
- ctrlTPH2rev: 5’-tgcatgcttactagccaaccatgagaca-3’
- 1473C allele, product size 307bp
TPH2C1473: 5’-cagaatttcaatgctctgcgtgtggg-3’
- 1473G allele. product size 307 bp
TPH2g1473: 5’-cagaatttcaatgctctgcgtgtggc-3’
Mouse accession number: NM_173391 Rat accession Number: NM_173839
Reference: Zhang X, Beaulieu JM, Sotnikova TD, Gainetdinov RR, Caron MG. Tryptophan hydroxylase-2 controls brain serotonin synthesis. Science. 2004 Jul 9;305(5681):217. PMID 15247473
5HT2C Receptor cDNA
for in situ hybridization
product size: 207 bp, ascension number: M21410, bp 105-311
- 5HT2Cfor (bp 105-126): 5’ -CTATCCCTTACCTTCCTATTAC–3’
- 5HT2Crev (bp 311-290): 5’ -TACCCTACTCTAGCATAGAAAC–3’
References:
Heisler LK, Pronchuk N, Nonogaki K, Zhou L, Raber J, Tung L, Yeo GS, O'Rahilly S, Colmers WF, Elmquist JK, Tecott LH. Serotonin activates the hypothalamic-pituitary-adrenal axis via serotonin 2C receptor stimulation. J Neurosci. 2007 Jun 27;27(26):6956-64. PMID 17596444
Molineaux SM, Jessell TM, Axel R, Julius D.
5-HT1c receptor is a prominent serotonin receptor subtype in the central nervous system.
Proc Natl Acad Sci U S A. 1989 Sep;86(17):6793-7.
PMID 2771958
Julius,D., MacDermott,A.B., Axel,R. and Jessell,T.M. Molecular characterization of a functional cDNA encoding the serotonin 1c receptor, Science 241 (4865), 558-564 (1988) PMID 3399891
Antisera for IHC
NT-102 5HTrab rabbit serotonin antisera
PROTOS BIOTECH CORP (Archive link) PBTantibody Division 315 E. 65th Street, Suit 4K, New York, New York 10065 1-212-517-6801 1-212-772-1514 (fax) PBTantibody@verizon.net