Supplementary Table 1: Sense and antisense sequences of oligonucleotide primers used in the quantitative real time polymerase chain reaction (qRT-PCR), the corresponding product size
and function of the selected genes. For microarrays confirmations, most genes were chosen based on their neuroplasticity-related function; those without neuroplasticity-related function were
randomly selected from the list of significantly altered genes to avoid bias in the microarrays confirmation. Moreover, genes with annotated function which expression was reversed by at least
three of the ADs were analyzed. For macrodissections’ accuracy determination, genes predominantly expressed in the DG comparing to the CA regions were chosen.
Gene
symbol
Gene name
Selected functions/features
Sense
Antisense
Product
size (bp)
Microarrays confirmation
HES1
Hairy and enhancer of split-1
Repressor of neuronal commitment in
stem cells (Nakamura et al, 2000).
TGTCAACACGACACCGGACAAAC
TGGAATGCCGGGAGCTATCTTT
161
GADD45B
Growth arrest and DNA-damageinducible, beta
Role in region-specific DNA
demethylation to control key aspects of
activity-dependent adult neurogenesis
(Ma et al, 2009).
GCTGCGACAATGACATTGAC
GTATGACAGTTCGTGACCAG
136
APOBEC1
Apolipoprotein B mRNA editing
enzyme, catalytic polypeptide 1
Involved in neuronal activity-induced
active DNA demethylation and
subsequent gene expression in the
dentate gyrus of the adult mouse brain in
vivo (Guo et al, 2011).
ACATAAGCTCCCGAGGAAGG
GGCTCAATTCTTCTCCTCAGAGT
102
NOS1
Nitric oxide synthase 1
Role in the manifestation of depressionlike behavior (Yazir et al, 2012)
GACAACGTTCCTGTGGTCCT
GAAGAGCTGGTCCTTTGTGC
276
AR
Androgen receptor
Role of androgens in synaptic plasticity
(Ooishi et al, 2012)
GGGTGACTTCTCTGCCTCTG
CCACAGATCAGGCAGGTCTT
210
PDYN
Prodynorphin
Involvement in neuroplasticity and
depression-like behavior (Schwarzer,
2009)
CCTGTCCTTGTGTTCCCTGT
AGAGGCAGTCAGGGTGAGAA
157
CYP2D1
Cytochrome P450 2D1
Role in response to centrally acting
drugs (Miksys and Tyndale, 2004)
GTGCAGCGCAGAGTCCAACAA
ATGGAAGCGGTGGGGCTTCT
270
EBAG9
Estrogen Receptor Binding Site
Associated, Antigen, 9
Role of estrogen in synaptic plasticity
(Ooishi et al, 2012)
TAAGCAGACAGACGTGGAGGA
TGCTACCATCTGGGACACCAA
218
SP1
Specificity protein 1
Role in neuronal plasticity by activation
of synapsin I gene (SYN1) (Paonessa et
al, 2013)
CTGCCACCATGAGCGACCAAGAT
CCCTGGGAGTTGTTGCTGTTCTC
259
DUSP1
Dual specificity protein
phosphatase 1
Reversed by at least three of the ADs.
Negative regulator of MAP kinase and
involvement in the development of
depressive-like behavior (Duric et al,
2010)
CTCCAAGGAGGATATGAAGCGTTTT
TGGTAAGCACTGCCCAGGTACA
203
MT4
Metallothionein 4
Reversed by at least three of the ADs.
CACGTGTATGTCTGGAGGGATCT
GCACTTGTCTGAGCCCCCTTTG
157
LRRC8E
Leucine rich repeat containing 8
family, member E
Reversed by at least three of the ADs.
CCCAGAGCAACAGGATGATCCCAG
TGCACCCGAACACCCCAATCA
144
PerfeCTa Universal PCR Primer (Quanta
MIR409-5P
microRNA 409, 5p
Reversed by all the ADs.
AGTGTCTCTCGACAGGCACAGGCT
MIR411-5P
microRNA 411, 5p
Reversed by at least three of the ADs.
AAAGAGTCTAAACAGGATGAGTCC
RNU6
RNA-U6 small nuclear 2
Reference gene for microRNAs
expression analysis
TGGGACAGGAAGTGATCTTGC
SIX1
Sine Oculis Homeobox Homolog
-
CAAGGAAAGGGAGAACACCGAAAA
CCGTTAGGCCCGGAAGAGAA
206
RSPO2
R-spondin 2
-
GGAGACGCAGTAAGCGAGCTAGT
TGGGGCTCGGTGTCCATAGT
188
MBD4
Methyl-CpG-binding domain
protein 4
-
CTCAATCGGACCTCAGGCAA
AGTTCTGACACGTCCCTCCA
110
B2M
Beta-2-microglobulin
Reference gene.
GTGCTTGCCATTCAGAAAACTCC
AGGTGGGTGGAACTGAGACA
136
DG-specific (Pleasure et al, 2000)
ACAACAGGAAGTGGAAACATGAC
ACTGAGGCACTCGTCTGTCC
102
Macrodissections’ accuracy confirmation
NEUROD1
Neurogenic differentiation 1
Biosciences*)
PerfeCTa Universal PCR Primer (Quanta
Biosciences*)
PerfeCTa Universal PCR Primer (Quanta
Biosciences*)
-
-
-
DSP
Desmoplakin
DG-specific (Lein et al, 2004)
CCAGCACCAGACAGTCACCAAAA
GCAGCCACGTCTCCTGTTTGT
116
PROX1
Prospero-related homeobox 1
DG-specific (Iwano et al, 2012)
CGGGAAGCGCAATGAAGGGCTAT
GATGGCTTGGCGCGCATACTT
211
* MicroRNAs expression analysis was assessed using the qScript™ microRNA Quantification System (Quanta Biosciences™, Gaithersburg, MD, USA).
References:
Duric V, Banasr M, Licznerski P, Schmidt HD, Stockmeier CA, Simen AA, et al (2010). A negative regulator of MAP kinase causes depressive behavior. Nat Med 16(11):
1328-1332.
Guo JU, Su Y, Zhong C, Ming GL, Song H (2011). Hydroxylation of 5-methylcytosine by TET1 promotes active DNA demethylation in the adult brain. Cell 145(3): 423434.
Iwano T, Masuda A, Kiyonari H, Enomoto H, Matsuzaki F (2012). Prox1 postmitotically defines dentate gyrus cells by specifying granule cell identity over CA3 pyramidal
cell fate in the hippocampus. Development 139(16): 3051-3062.
Lein ES, Zhao X, Gage FH (2004). Defining a molecular atlas of the hippocampus using DNA microarrays and high-throughput in situ hybridization. J Neurosci 24(15):
3879-3889.
Ma DK, Jang MH, Guo JU, Kitabatake Y, Chang ML, Pow-Anpongkul N, et al (2009). Neuronal activity-induced Gadd45b promotes epigenetic DNA demethylation and
adult neurogenesis. Science 323(5917): 1074-1077.
Miksys S, Tyndale RF (2004). The unique regulation of brain cytochrome P450 2 (CYP2) family enzymes by drugs and genetics. Drug Metab Rev 36(2): 313-333.
Nakamura Y, Sakakibara S, Miyata T, Ogawa M, Shimazaki T, Weiss S, et al (2000). The bHLH gene hes1 as a repressor of the neuronal commitment of CNS stem cells. J
Neurosci 20(1): 283-293.
Ooishi Y, Kawato S, Hojo Y, Hatanaka Y, Higo S, Murakami G, et al (2012). Modulation of synaptic plasticity in the hippocampus by hippocampus-derived estrogen and
androgen. J Steroid Biochem Mol Biol 131(1-2): 37-51.
Paonessa F, Latifi S, Scarongella H, Cesca F, Benfenati F (2013). Specificity protein 1 (Sp1)-dependent activation of the synapsin I gene (SYN1) is modulated by RE1silencing transcription factor (REST) and 5'-cytosine-phosphoguanine (CpG) methylation. J Biol Chem 288(5): 3227-3239.
Pleasure SJ, Collins AE, Lowenstein DH (2000). Unique expression patterns of cell fate molecules delineate sequential stages of dentate gyrus development. J Neurosci
20(16): 6095-6105.
Schwarzer C (2009). 30 years of dynorphins--new insights on their functions in neuropsychiatric diseases. Pharmacol Ther 123(3): 353-370.
Yazir Y, Utkan T, Aricioglu F (2012). Inhibition of neuronal nitric oxide synthase and soluble guanylate cyclase prevents depression-like behaviour in rats exposed to chronic
unpredictable mild stress. Basic Clin Pharmacol Toxicol 111(3): 154-160.