Edited by: Shawn Hayley, Carleton University, Canada
Reviewed by: Xin Wang, Stanford University and HHMI, USA; Zhihong Chen, Cleveland Clinic, USA
*Correspondence: Jocelien D. A. Olivier, Department of Behavioral Physiology, Center for Behaviour and Neurosciences, University of Groningen, Nijenborgh 7, 9747 AG, Groningen, Netherlands e-mail:
This article was submitted to the journal Frontiers in Cellular Neuroscience.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
The effects of antenatal depression and antidepressant treatment during pregnancy on both mother and child are vigorously studied, but the underlying biology for these effects is largely unknown. The placenta plays a crucial role in the growth and development of the fetus. We performed a gene expression study on the fetal side of the placenta to investigate gene expression patterns in mothers with antenatal depression and in mothers using antidepressant treatment during pregnancy. Placental samples from mothers with normal pregnancies, from mothers with antenatal depression, and from mothers using antidepressants were collected. We performed a pilot microarray study to investigate alterations in the gene expression and selected several genes from the microarray for biological validation with qPCR in a larger sample. In mothers with antenatal depression 108 genes were differentially expressed, whereas 109 genes were differentially expressed in those using antidepressants. Validation of the microarray revealed more robust gene expression differences in the seven genes picked for confirmation in antidepressant-treated women than in depressed women. Among the genes that were validated
Unfortunately pregnancy is not a lighthearted period for all women. About 10% of pregnant women in economically developed countries and up to 25% of pregnant women in poorer countries develop symptoms of depression, such as fatigue, trouble sleeping, sense of sadness or hopelessness, during pregnancy (O'Keane and Marsh,
The placenta plays a pivotal role in supporting fetal growth and development and is a crucial regulator of maternal-fetal interactions and fetal brain development (Hsiao and Patterson,
This study was carried out at the Department of Women's and Children's health, Uppsala University Hospital, as part of an ongoing longitudinal study on antenatal and postpartum depression: the Biology, Affect, Stress, Imaging and Cognition in Pregnancy and the Puerperium (BASIC) project. The BASIC project started in 2010 and aims to include a study population of 5000 pregnant women in the Uppsala County. Women attending the routine ultrasound examination (gestational week 16–17) at Uppsala University Hospital are approached for participation, enabling a population-based sampling. Upon informed consent, women fill out web-based questionnaires in gestational week 17 and 32 including questions on physical and socio-demographic characteristics, medical, psychiatric, gynecologic and obstetric history variables, lifestyle, medication parameters, and the Swedish version of the Edinburgh Postnatal Depression Scale (EPDS). Information concerning the maternal depression, SSRI use, delivery and neonatal outcome were retrieved from the medical records. Placental biopsies are collected at delivery.
For the specific aim of this sub-study, inclusion criteria were women of Caucasian origin, normal pregnancies and deliveries and healthy offspring (no diagnoses and no admittance to neonatal care). Exclusion criteria were smoking or alcohol use during pregnancy, any daily use of prescribed drugs during pregnancy, any other chronic condition or disease, gestational age <35 weeks, and maternal age <18 or >42 years. Women on antidepressants used their treatment during the entire pregnancy in clinically relevant doses (low-dose use was excluded). The study was approved by the Regional Ethics Committee, Uppsala, Sweden, and performed in accordance with relevant guidelines and regulations.
Women with pregnancies complicated by ongoing depression (
Age (years) | 31.4 ± 2.2 | 31.2 ± 2.4 | 29.2 ± 3.4 | 29.0 ± 3.0 |
Parity ( |
1 (0-2) | 0 (0-1) | 0 (0-2) | 0 (0-2) |
BMI (kg/m2) | 22.8 ± 3.0 | 23.5 ± 2.5 | 26.9 ± 5.8 | 24.5 ± 4.1 |
Birth weight (gram) | 3542 ± 461 | 3632 ± 342 | 3538 ± 225 | 3556 ± 275 |
Gender offspring (% boy) | 60 | 40 | 20 | 80 |
Gestational length | 276 ± 11 | 283 ± 5 | 275 ± 3 | 277 ± 5 |
EPDS score week 17 | 14.8 ± 7.3 |
2.6 ± 1.8 | 8.0 ± 3.2 | 3.4 ± 1.6 |
EPDS score week 32 | 18.8 ± 5.3 |
3.2 ± 1.6 | 6.3 ± 3.4 | 4.0 ± 1.2 |
The samples described above were extended to 24 women with pregnancies complicated by ongoing depression, 29 antidepressant-treated women and 31 women with normal pregnancies. The depressed group included the five microarray cases and 18 women with EPDS >12 in gestational week 17
Placental tissues (containing both maternal and fetal side) were obtained after delivery, rinsed carefully in sterile phosphate-buffered saline to wash off maternal and fetal blood, and frozen on dry ice within 60 min of delivery and stored at −70°C until further use. Each placenta was individually processed as a single biological replicate in the microarray and validation study.
A biopsy was taken with a 3 mm cube from the fetal side of the placenta. Total RNA was isolated using miRNeasy mini kit (Qiagen, Hilden, Germany). Tissue was lysed with QIAzol reagent (Qiagen) using a rotor-stator homogenizer (up to 33.000 rpm; Ingenieursbűro CAT M Zipper Gmbh, type x120, Staufen, Germany) and chloroform (Sigma Aldrich, St. Louis, MO, USA) was added for phase-separation. The rest of the procedure was performed as described in manufactures protocol.
A biopsy was taken from the fetal side of the placenta with a 3 mm cube. Total RNA was isolated using RNeasy mini (Qiagen, Hilden, Germany). Tissue was lysed with QIAzol reagent (Qiagen) using TissueLyser (20Hz, 2 × 5 min) with stainless steel beads (Qiagen) and chloroform was added for phase-separation. The rest of the procedure was performed as described in manufactures protocol.
For both studies RNA concentration was measured with ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, Delaware, USA) and RNA quality was evaluated using the Agilent 2100 Bioanalyzer system (Agilent Technologies Inc, Palo Alto, California, USA).
250 nanograms of total RNA from each sample were used to generate amplified and biotinylated sense-strand cDNA from the entire expressed genome according to the Ambion WT Expression Kit (P/N 4425209 Rev B 05/2009) and Affymetrix GeneChip® WT Terminal Labeling and Hybridization User Manual (P/N 702808 Rev. 4, Affymetrix Inc., Santa Clara, CA). GeneChip® ST Arrays (GeneChip® XXX Gene 1.0 ST Array) were hybridized for 16 h in a 45°C incubator, rotated at 60 rpm. According to the GeneChip® Expression Wash, Stain and Scan Manual (PN 702731 Rev 3, Affymetrix Inc., Santa Clara, CA) the arrays were then washed and stained using the Fluidics Station 450 and finally scanned using the GeneChip® Scanner 3000 7G.
The raw data was normalized in the free software Expression Console provided by Affymetrix (
cDNA was synthesized using SuperScriptIII reversed transcriptase (Invitrogen, Carlsbad, California, USA) according to manufacturer's protocol. Briefly, 250 ng of total RNA was used to reverse transcribe using the random primer to prepare 20 μl of cDNA.
The validity of the microarray results was tested via quantitative real-time PCR (qRT-PCR) employing the StepOne Plus qPCR machine (Applied Biosystems, Life Technologies, Carlsbad, California, USA). For validation we selected seven genes [
All samples were performed in triplicates and averaged for further calculations. Mean normalized expression (MNE) based on the ratio between
Clinical characteristics of women in the microarray study and the validation study were compared by means of One-Way ANOVA, followed by a Bonferroni
The demographic characteristics of the women in the microarray and validation studies are presented in Tables
Age (years) | 31.4 ± 3.9 | 31.1 ± 4.3 | 31.2 ± 4.1 |
Parity ( |
0 (0-3) | 1 (0-2) | 1 (0-3) |
BMI (kg/m2) | 26.0 ± 4.9 | 24.0 ± 6.2 | 27.2 ± 4.9 |
Systolic blood pressure in first trimester, mmHg | 119 ± 13 | 111 ± 12 | 118 ± 12 |
Diastolic blood pressure in first trimester, mmHg | 73 ± 9 | 69 ± 7 | 70 ± 7 |
Systolic blood pressure at last visit, mmHg | 125 ± 11 | 119 ± 11 | 125 ± 10 |
Diastolic blood pressure at first visit, mmHg | 79 ± 8 | 76 ± 6 | 77 ± 7 |
Lowest hemoglobin level during pregnancy, g/dl | 11.7 ± 0.8 | 11.4 ± 0.8 | 11.0 ± 0.9 |
Birth weight (gram) | 3577 ± 351 | 3546 ± 499 | 3589 ± 400 |
Gender offspring (% boy) | 65 | 54 | 45 |
Gestational length | 281 ± 7 | 275 ± 11 | 273 ± 8c |
EPDS score week 17 | 2.9 ± 1.8 | 15.0 ± 4.2 |
7.7 ± 4.9 |
EPDS score week 32 | 2.9 ± 1.8 | 15.9 ± 3.6a | 8.7 ± 5.3 |
At a Bonferroni-corrected significance threshold (
vault RNA 1-2 | 8108629 | 2.280673826 | |
placental growth factor | 7980233 | 0.695475147 | |
ribonuclease/angiogenin inhibitor 1 /// hypothetical protein FLJ23519 | 7945420 | 0.565671098 | |
inositol 1,3,4-triphosphate 5/6 kinase | 7980970 | 0.549075065 | |
− | 8175261 | 0.519417648 | |
RAD23 homolog A ( |
8026122 | 0.51907744 | |
acyloxyacyl hydrolase (neutrophil) | 8139160 | 0.50922813 | |
apolipoprotein C-I | 8029536 | 0.502399918 | |
killer cell immunoglobulin-like receptor, three domains, long cytoplasmic tail, 2 /// killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 2 /// killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 2 /// killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 1 /// killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 4 /// killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 3 /// similar to killer cell immunoglobulin-like receptor 3DL2 precursor (MHC class I NK cell receptor) (Natural killer-associated transcript 4) (NKAT-4) (p70 natural killer cell receptor clone CL-5) (CD158k antigen) /// killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 5 | 8031293 | −0.506084964 | |
sorting nexin 4 | 8090256 | −0.506251931 | |
NIMA (never in mitosis gene a)-related kinase 1 | 8103646 | −0.50748106 | |
Ral GEF with PH domain and SH3 binding motif 2 | 7907657 | −0.508099708 | |
ring finger protein 160 | 8069711 | −0.50811689 | |
− | − | 8104012 | −0.508994997 |
RAP1 interacting factor homolog (yeast) | 8045697 | −0.511151224 | |
low density lipoprotein-related protein 2 | 8056611 | −0.513852689 | |
polypyrimidine tract binding protein 2 | 7903188 | −0.517797539 | |
zinc finger, matrin type 1 | 8174119 | −0.519592242 | |
RUN and FYVE domain containing 2 | 7933999 | −0.524450233 | |
small nuclear RNA activating complex, polypeptide 1, 43kDa | 7974870 | −0.52596952 | |
ring finger protein 19A | 8152041 | −0.526515918 | |
superkiller viralicidic activity 2-like 2 ( |
8105353 | −0.530773126 | |
anaphase promoting complex subunit 4 | 8094408 | −0.533126207 | |
− | − | 7916667 | −0.533554256 |
NCK-associated protein 1 | 8057517 | −0.53822831 | |
diacylglycerol kinase, eta | 7968800 | −0.538998035 | |
membrane-associated ring finger (C3HC4) 7 | 8045919 | −0.550742909 | |
homer homolog 1 (Drosophila) | 8112841 | −0.551657356 | |
PHD finger protein 20-like 1 | 8148358 | −0.551986952 | |
YEATS domain containing 4 | 7957032 | −0.55366204 | |
cholinergic receptor, muscarinic 3 | 7910915 | −0.554183212 | |
phosphodiesterase 3B, cGMP-inhibited | 7938629 | −0.557038965 | |
breast cancer metastasis-suppressor 1-like | 7973948 | −0.55836529 | |
small nucleolar RNA, C/D box 30 | 7948900 | −0.560928764 | |
zinc finger protein 84 | 7960143 | −0.562597276 | |
meiotic nuclear divisions 1 homolog ( |
8097857 | −0.5650204 | |
adenylate cyclase 10 pseudogene | 8119423 | −0.565773386 | |
ubiquitin-like modifier activating enzyme 6 | 8100615 | −0.566611488 | |
NEDD4 binding protein 2-like 2 | 7970907 | −0.570303348 | |
zinc finger, RAN-binding domain containing 2 | 7916969 | −0.573773594 | |
eukaryotic translation initiation factor 5B | 8043861 | −0.574408988 | |
nucleosome assembly protein 1-like 1 | 7965048 | −0.581711065 | |
zinc finger protein 146 | 8028186 | −0.584609444 | |
bromodomain and WD repeat domain containing 3 | 8173766 | −0.592331241 | |
KIAA1109 | 8097148 | −0.602191356 | |
SUMO1/sentrin specific peptidase 7 | 8089203 | −0.603487152 | |
frizzled homolog 6 (Drosophila) | 8147766 | −0.605426406 | |
chromodomain helicase DNA binding protein 9 | 7995583 | −0.607612535 | |
KIAA1430 | 8103979 | −0.617503882 | |
ring finger protein 217 | 8121825 | −0.620970732 | |
protein-L-isoaspartate (D-aspartate) O-methyltransferase domain containing 1 | 8150714 | −0.635640315 | |
natural killer-tumor recognition sequence | 8079079 | −0.63772807 | |
kinesin family member 23 | 7984540 | −0.649229354 | |
PRP40 pre-mRNA processing factor 40 homolog A ( |
8055913 | −0.652162751 | |
− | − | 8098287 | −0.658215787 |
AT rich interactive domain 4A (RBP1-like) | 7974621 | −0.661691075 | |
LUC7-like 3 ( |
8008493 | −0.663829187 | |
Nipped-B homolog (Drosophila) | 8104944 | −0.6642255 | |
centromere protein E, 312kDa | 8102076 | −0.664227054 | |
biorientation of chromosomes in cell division 1-like | 8099410 | −0.665905703 | |
TATA box binding protein (TBP)-associated factor, RNA polymerase I, D, 41kDa | 7951008 | −0.668975157 | |
progesterone immunomodulatory binding factor 1 | 7969390 | −0.670731182 | |
SUMO1/sentrin specific peptidase 6 | 8120758 | −0.675080818 | |
structural maintenance of chromosomes 2 | 8156982 | −0.681747538 | |
DnaJ (Hsp40) homolog, subfamily C, member 10 | 8046759 | −0.690820266 | |
zinc finger protein 638 | 8042601 | −0.696757481 | |
syntaxin binding protein 3 | 7903541 | −0.698824428 | |
TTK protein kinase | 8120838 | −0.699258503 | |
SCY1-like 2 ( |
7957806 | −0.710364273 | |
erbb2 interacting protein | 8105681 | −0.71178458 | |
chromodomain helicase DNA binding protein 1 | 8113305 | −0.71533841 | |
membrane protein, palmitoylated 6 (MAGUK p55 subfamily member 6) | 8131927 | −0.715467135 | |
chromosome 1 open reading frame 27 | 7908330 | −0.715867564 | |
dynamin 1-like | 7954752 | −0.716232722 | |
centrosomal protein 152kDa | 7988537 | −0.717688618 | |
OTU domain containing 6B | 8147262 | −0.735794509 | |
alpha thalassemia/mental retardation syndrome X-linked (RAD54 homolog, |
8173673 | −0.740293061 | |
zinc finger protein 100 | 8035808 | −0.741413733 | |
kinesin family member 18A | 7947248 | −0.743358618 | |
DEK oncogene | 8124144 | −0.744034441 | |
− | − | 8083445 | −0.766235985 |
− | − | 8119580 | −0.77576534 |
serologically defined colon cancer antigen 1 | 7978866 | −0.776418378 | |
ankyrin repeat domain 36B | 8054064 | −0.777582301 | |
ankyrin repeat domain 26 | 7932637 | −0.779453671 | |
LysM, putative peptidoglycan-binding, domain containing 3 | 8113064 | −0.780972234 | |
CTAGE family, member 4 /// CTAGE family, member 6 /// CTAGE family member /// similar to CTAGE6 /// CTAGE family, member 4-like | 8136979 | −0.78257421 | |
succinate-CoA ligase, ADP-forming, beta subunit | 7971541 | −0.78881152 | |
structural maintenance of chromosomes 5 | 8155770 | −0.795143951 | |
polymerase (DNA directed) kappa | 8106303 | −0.814486061 | |
ERGIC and golgi 2 | 7962013 | −0.820896474 | |
RAD50 homolog ( |
8107942 | −0.827079801 | |
THO complex 2 | 8174893 | −0.836189906 | |
kinectin 1 (kinesin receptor) | 7974483 | −0.866627304 | |
jumonji domain containing 1C | 7933877 | −0.86686842 | |
ubiquitin specific peptidase 15 | 7956670 | −0.869012602 | |
protein phosphatase 1, regulatory (inhibitor) subunit 12A | 7965123 | −0.883024382 | |
formin binding protein 1-like | 7903092 | −0.886872231 | |
structural maintenance of chromosomes 6 | 8050443 | −0.915161746 | |
structural maintenance of chromosomes 4 | 8083709 | −0.920861102 | |
Rho-associated, coiled-coil containing protein kinase 2 | 8050302 | −0.924567683 | |
A kinase (PRKA) anchor protein (yotiao) 9 | 8134122 | −0.935693684 | |
zinc finger protein 252 | 8153935 | −0.939582046 | |
COP9 constitutive photomorphogenic homolog subunit 2 (Arabidopsis) | 7988605 | −0.975928968 | |
GRIP and coiled-coil domain containing 2 | 8044236 | −1.024026926 | |
CTAGE family, member 4 /// CTAGE family, member 6 /// CTAGE family member /// similar to CTAGE6 /// CTAGE family, member 4-like | 8129560 | −1.025580319 | |
Rho-associated, coiled-coil containing protein kinase 1 | 8022441 | −1.074633032 | |
FLJ45950 protein | 7952673 | −1.131057985 |
vault RNA 1-2 | 2.28 | 0.026 | |
Placenta growth factor | 0.70 | 0.013 | |
ribonuclease/angiogenin inhibitor 1 | 0.57 | 0.036 | |
inositol 1,3,4-triphosphate 5/6 kinase | 0.55 | 0.037 | |
microRNA 503 | 0.52 | 0.047 | |
RAD23 homolog A ( |
0.52 | 0.013 | |
apolipoprotein C-I | 0.50 | 0.037 | |
ubiquitin specific peptidase 15 | −0.87 | 0.026 | |
protein phosphatase 1. regulatory (inhibitor) subunit 12A | −0.88 | 0.021 | |
formin binding protein 1-like | −0.89 | 0.046 | |
structural maintenance of chromosomes 6 | −0.92 | 0.026 | |
structural maintenance of chromosomes 4 | −0.92 | 0.024 | |
Rho-associated. coiled-coil containing protein kinase 2 | −0.92 | 0.021 | |
A kinase (PRKA) anchor protein (yotiao) 9 | −0.94 | 0.028 | |
COP9 constitutive photomorphogenic homolog subunit 2 (Arabidopsis) | −0.98 | 0.008 | |
GRIP and coiled-coil domain containing 2 | −1.02 | 0.019 | |
Rho-associated. coiled-coil containing protein kinase 1 | −1.07 | 0.028 |
DNA replication, recombination, and repair, cellular assembly and organization, cell cycle | 27 | |
Cell cycle, cancer, connective tissue disorders | 18 | |
Cellular movement, hematological system development and function, immune cell T rafficking | 16 | |
Cardiovascular system development and function, organismal development, visual system development and function | 14 | |
Molecular transport, RNA trafficking, connective tissue disorders | 9 |
Actin nucleation by ARP-WASP complex | 0.003 | |
RhoA signaling | 0.029 | |
VEGF signaling | 0.011 | |
Protein kinase A signaling | 0.011 | |
1D-myo-inositol Hexakisphosphate Biosynthesis V (from Ins(1,3,4)P3) | 0.015 |
Similarly, we found 109 genes to be differentially expressed between the SSRI-treated women and their respective controls at a Bonferroni-corrected significance (
chromosome 12 open reading frame 39 | 7954398 | 1.274830514 | |
hypothetical FLJ34503 | 8121569 | 1.266115442 | |
RNA, U4 small nuclear 1 /// RNA, U4 small nuclear 1B | 7967030 | 0.911924177 | |
keratin associated protein 19-8 | 8069876 | 0.821169613 | |
olfactory receptor, family 2, subfamily A, member 7 /// olfactory receptor, family 2, subfamily A, member 4 /// similar to rho guanine nucleotide exchange factor 5 | 8143633 | 0.791137617 | |
keratin 81 | 7963353 | 0.778243623 | |
olfactory receptor, family 2, subfamily A, member 7 /// olfactory receptor, family 2, subfamily A, member 4 /// similar to rho guanine nucleotide exchange factor 5 | 8129558 | 0.774018568 | |
RNA, U4 small nuclear 2 | 7967028 | 0.707367007 | |
− | − | 7899484 | 0.693169774 |
serine incorporator 2 | 7899615 | 0.641601416 | |
apelin | 8175016 | 0.632933192 | |
angiopoietin-like 4 | 8025402 | 0.61857093 | |
tubulin, alpha 1c | 7955179 | 0.558532616 | |
− | − | 8139128 | 0.546472497 |
S100 calcium binding protein A3 | 7920278 | 0.543186629 | |
hypothetical protein LOC100127980 | 8036302 | 0.540856558 | |
trans-2,3-enoyl-CoA reductase | 8101622 | 0.539800494 | |
small Cajal body-specific RNA 10 | 7953383 | 0.537535155 | |
Ras-related associated with diabetes | 8001918 | 0.534226961 | |
ephrin-A5 | 8113433 | 0.533194259 | |
CDC42 effector protein (Rho GTPase binding) 1 | 8072817 | 0.529414661 | |
PCTAIRE protein kinase 1 | 8167103 | 0.528119471 | |
small nucleolar RNA, C/D box 116-16 | 7981980 | 0.525075622 | |
− | − | 8130181 | 0.511027698 |
− | − | 7953128 | 0.508366155 |
ORM1-like 3 ( |
8014916 | 0.507840044 | |
Rho guanine nucleotide exchange factor (GEF) 5 /// Rho guanine nucleotide exchange factor (GEF) 5-like /// similar to rho guanine nucleotide exchange factor 5 | 8136987 | 0.503949491 | |
mitogen-activated protein kinase kinase kinase kinase 5 | 7978997 | −0.503225433 | |
Fanconi anemia, complementation group L /// vaccinia related kinase 2 | 8052382 | −0.503353866 | |
AT hook containing transcription factor 1 | 7925622 | −0.503946371 | |
zinc finger protein 280D | 7989159 | −0.505508381 | |
sorting nexin 6 | 7978570 | −0.512758574 | |
COBW domain containing 3 /// COBW domain containing 5 /// COBW domain containing 6 /// similar to COBW domain containing 3 /// COBW domain containing 7 /// similar to COBW domain containing 1 /// COBW domain containing 2 | 8155422 | −0.513922351 | |
PHD finger protein 20-like 1 | 8148358 | −0.514646483 | |
Fas (TNF receptor superfamily, member 6) | 7929032 | −0.527079798 | |
methyltransferase like 14 | 8097066 | −0.531709394 | |
zinc finger protein 100 | 8035808 | −0.534310441 | |
RANBP2-like and GRIP domain containing 2 /// RANBP2-like and GRIP domain containing 5 /// RANBP2-like and GRIP domain containing 8 /// RANBP2-like and GRIP domain containing 3 /// RANBP2-like and GRIP domain containing 4 /// RANBP2-like and GRIP domain containing 6 /// RANBP2-like and GRIP domain containing 1 /// RAN binding protein 2 | 8044161 | −0.543369485 | |
soc-2 suppressor of clear homolog (C. elegans) | 7930470 | −0.550070146 | |
interleukin-1 receptor-associated kinase 1 binding protein 1 | 8120826 | −0.550763514 | |
vesicle-associated membrane protein 7 | 8171041 | −0.554437451 | |
vesicle-associated membrane protein 7 | 8176962 | −0.554437451 | |
zinc finger protein 791 | 8026007 | −0.55599566 | |
golgin B1, golgi integral membrane protein | 8089930 | −0.559994665 | |
RANBP2-like and GRIP domain containing 2 /// RANBP2-like and GRIP domain containing 5 /// RANBP2-like and GRIP domain containing 8 /// RANBP2-like and GRIP domain containing 3 /// RANBP2-like and GRIP domain containing 4 /// RANBP2-like and GRIP domain containing 6 /// RANBP2-like and GRIP domain containing 7 /// RANBP2-like and GRIP domain containing 1 /// RAN binding protein 2 | 8044304 | −0.562605545 | |
family with sequence similarity 133, member B /// family with sequence similarity 133, member B pseudogene /// similar to FAM133B protein | 8105504 | −0.56577534 | |
serine/threonine kinase 17b | 8057887 | −0.565889797 | |
− | − | 8054532 | −0.568088762 |
pericentriolar material 1 | 8144812 | −0.574364755 | |
polymerase (DNA directed) kappa | 8106303 | −0.576549694 | |
− | − | 8147650 | −0.578295577 |
chromosome 1 open reading frame 58 | 7909931 | −0.580015227 | |
RANBP2-like and GRIP domain containing 1 /// RANBP2-like and GRIP domain containing 2 /// RANBP2-like and GRIP domain containing 5 /// RANBP2-like and GRIP domain containing 8 /// RANBP2-like and GRIP domain containing 3 /// RANBP2-like and GRIP domain containing 4 /// RANBP2-like and GRIP domain containing 6 /// RANBP2-like and GRIP domain containing 7 /// RAN binding protein 2 | 8054414 | −0.582592072 | |
RANBP2-like and GRIP domain containing 2 /// RANBP2-like and GRIP domain containing 5 /// RANBP2-like and GRIP domain containing 8 /// RANBP2-like and GRIP domain containing 3 /// RANBP2-like and GRIP domain containing 4 /// RANBP2-like and GRIP domain containing 6 /// RANBP2-like and GRIP domain containing 7 /// RANBP2-like and GRIP domain containing 1 /// RAN binding protein 2 | 8054676 | −0.586951747 | |
RAN binding protein 2 | 8044263 | −0.589348972 | |
− | − | 8054557 | −0.592061813 |
meiosis-specific nuclear structural 1 | 7989146 | −0.595349364 | |
DnaJ (Hsp40) homolog, subfamily C, member 13 | 8082688 | −0.601103633 | |
B double prime 1, subunit of RNA polymerase III transcription initiation factor IIIB | 8106025 | −0.601882298 | |
natural killer-tumor recognition sequence | 8079079 | −0.602491778 | |
AT rich interactive domain 4A (RBP1-like) | 7974621 | −0.602522967 | |
COBW domain containing 3 /// COBW domain containing 5 /// COBW domain containing 6 /// COBW domain containing 7 /// similar to COBW domain containing 1 /// COBW domain containing 2 /// COBW domain containing 1 | 8161575 | −0.605839737 | |
B double prime 1, subunit of RNA polymerase III transcription initiation factor IIIB | 8177560 | −0.608777219 | |
− | − | 7942645 | −0.611731094 |
jumonji domain containing 1C | 7933877 | −0.612753894 | |
ATP-binding cassette, sub-family C (CFTR/MRP), member 9 | 7961710 | −0.613240551 | |
chromosome 15 open reading frame 5 | 7990636 | −0.631000244 | |
laminin, alpha 4 | 8128991 | −0.636829539 | |
family with sequence similarity 133, member B /// family with sequence similarity 133, member B pseudogene | 8055978 | −0.63846971 | |
PRP40 pre-mRNA processing factor 40 homolog A ( |
8055913 | −0.641694474 | |
ERGIC and golgi 2 | 7962013 | −0.65241833 | |
zinc finger protein 644 | 7917604 | −0.664132951 | |
coiled-coil domain containing 88A | 8052269 | −0.673093507 | |
LysM, putative peptidoglycan-binding, domain containing 3 | 8113064 | −0.675591155 | |
leucine rich repeat and coiled-coil domain containing 1 | 8147079 | −0.676201087 | |
zinc finger protein 146 | 8028186 | −0.679184287 | |
deoxynucleotidyltransferase, terminal, interacting protein 2 | 7917771 | −0.687693256 | |
− | − | 8167910 | −0.688561865 |
guanylate binding protein 3 /// interferon-induced guanylate-binding protein 1 pseudogene /// guanylate binding protein 1, interferon-inducible, 67kDa | 7917516 | −0.690433417 | |
centrosomal protein 290kDa | 7965264 | −0.694219538 | |
ATP-binding cassette, sub-family E (OABP), member 1 | 8097647 | −0.696244083 | |
multimerin 1 | 8096415 | −0.703832221 | |
coiled-coil domain containing 55 | 8006112 | −0.709824621 | |
− | − | 7989309 | −0.71000087 |
zinc finger protein 260 | 8036324 | −0.712626342 | |
kinectin 1 (kinesin receptor) | 7974483 | −0.712691264 | |
dynamin 1-like | 7954752 | −0.713859601 | |
fer (fps/fes related) tyrosine kinase | 8107208 | −0.716049908 | |
serologically defined colon cancer antigen 1 | 7978866 | −0.717656644 | |
zinc finger protein 254 | 8027368 | −0.723859521 | |
enoyl Coenzyme A hydratase domain containing 1 | 8129379 | −0.730176629 | |
zinc finger protein 518A | 7929562 | −0.730789996 | |
STE20-like kinase (yeast) | 7930276 | −0.731081254 | |
RAD50 homolog ( |
8107942 | −0.732319601 | |
Rho-associated, coiled-coil containing protein kinase 2 | 8050302 | −0.73784498 | |
centrosomal protein 170kDa | 7925525 | −0.756625495 | |
− | − | 8047401 | −0.771296325 |
ankyrin repeat domain 12 | 8020068 | −0.777177689 | |
zinc finger protein 721 /// ATP-binding cassette, sub-family A (ABC1), member 11 (pseudogene) | 8098758 | −0.777277839 | |
Rho-associated, coiled-coil containing protein kinase 1 | 8022441 | −0.804644966 | |
A kinase (PRKA) anchor protein (yotiao) 9 | 8134122 | −0.83694096 | |
synaptonemal complex protein 2 | 8067305 | −0.866613383 | |
early endosome antigen 1 | 7965436 | −0.883045052 | |
protein immuno-reactive with anti-PTH polyclonal antibodies /// ankyrin repeat domain 36B /// ankyrin repeat domain 36 /// hypothetical protein LOC100289777 /// hypothetical protein LOC100133923 /// hypothetical LOC645784 | 8053801 | −0.883294274 | |
COP9 constitutive photomorphogenic homolog subunit 2 (Arabidopsis) | 7988605 | −0.883531246 | |
GRIP and coiled-coil domain containing 2 | 8044236 | −0.919095326 | |
− | − | 8084878 | −0.91918252 |
nexilin (F actin binding protein) | 7902495 | −1.000773069 | |
− | − | 8098287 | −1.149966378 |
chromosome 12 open reading frame 39 | 1.28 | 0.009 | |
RNA, U4 small nuclear 1 | 0.91 | 0.026 | |
keratin 81 | 0.78 | 0.007 | |
RNA, U4 small nuclear 2 | 0.71 | 0.043 | |
serine incorporator 2 | 0.64 | 0.038 | |
apelin | 0.63 | 0.014 | |
angiopoietin-like 4 | 0.62 | 0.047 | |
tubulin, alpha 1c | 0.56 | 0.027 | |
S100 calcium binding protein A3 | 0.54 | 0.042 | |
trans-2,3-enoyl-CoA reductase | 0.54 | 0.048 | |
ankyrin repeat domain 12 | −0.78 | 0.042 | |
zinc finger protein 721 | −0.78 | 0.033 | |
Rho-associated, coiled-coil containing protein kinase 1 | −0.80 | 0.041 | |
A kinase (PRKA) anchor protein (yotiao) 9 | −0.84 | 0.017 | |
synaptonemal complex protein 2 | −0.87 | 0.042 | |
early endosome antigen 1 | −0.88 | 0.015 | |
ankyrin repeat domain 36B | −0.88 | 0.041 | |
COP9 constitutive photomorphogenic homolog subunit 2 (Arabidopsis) | −0.88 | 0.007 | |
GRIP and coiled-coil domain containing 2 | −0.92 | 0.032 | |
nexilin (F actin binding protein) | −1.00 | 0.044 |
Infectious disease, cellular assembly and organization, cellular function and maintenance | 13 | |
Cellular growth and proliferation, inflammatory response, lipid metabolism | 11 | |
Cell death and survival, inflammatory response, cellular movement | 9 | |
Cell death and survival, liver necrosis/cell death, hematological system development and function | 8 | |
Cardiovascular disease, skeletal and muscular disorders, cardiovascular system development and function | 2 |
Ephrin A signaling | 0.001 | |
RhoA signaling | 0.002 | |
PEDF signaling | 0.004 | |
Breast cancer regulation by Stathmin 1 | 0.012 | |
Signaling by Rho Family GTPases | 0.021 |
Of the 108 genes that were differentially expressed between the depressed and the control cases, and the 109 genes that were differentially expressed between the SSRI-treated and the control cases, only 20 genes were overlapping. These genes are displayed in Table
jumonji domain containing 1C | 7933877 | −0.612753894 | −0.86686842 | |
dynamin 1-like | 7954752 | −0.713859601 | −0.716232722 | |
ERGIC and golgi 2 | 7962013 | −0.65241833 | −0.820896474 | |
kinectin 1 (kinesin receptor) | 7974483 | −0.712691264 | −0.866627304 | |
AT rich interactive domain 4A (RBP1-like) | 7974621 | −0.602522967 | −0.661691075 | |
serologically defined colon cancer antigen 1 | 7978866 | −0.717656644 | −0.776418378 | |
COP9 constitutive photomorphogenic homolog subunit 2 (Arabidopsis) | 7988605 | −0.883531246 | −0.975928968 | |
Rho-associated, coiled-coil containing protein kinase 1 | 8022441 | −0.804644966 | −1.074633032 | |
zinc finger protein 146 | 8028186 | −0.679184287 | −0.584609444 | |
zinc finger protein 100 | 8035808 | −0.534310441 | −0.741413733 | |
GRIP and coiled-coil domain containing 2 | 8044236 | −0.919095326 | −1.024026926 | |
Rho-associated, coiled-coil containing protein kinase 2 | 8050302 | −0.73784498 | −0.924567683 | |
PRP40 pre-mRNA processing factor 40 homolog A ( |
8055913 | −0.641694474 | −0.652162751 | |
natural killer-tumor recognition sequence | 8079079 | −0.602491778 | −0.63772807 | |
− | − | 8098287 | −1.149966378 | −0.658215787 |
polymerase (DNA directed) kappa | 8106303 | −0.576549694 | −0.814486061 | |
RAD50 homolog ( |
8107942 | −0.732319601 | −0.827079801 | |
LysM, putative peptidoglycan-binding, domain containing 3 | 8113064 | −0.675591155 | −0.780972234 | |
A kinase (PRKA) anchor protein (yotiao) 9 | 8134122 | −0.83694096 | −0.935693684 | |
PHD finger protein 20-like 1 | 8148358 | −0.514646483 | −0.551986952 |
For validation of the microarray results we selected seven genes that were detected in top up- or down-regulated genes, in the pathway analysis or in the canonical pathway analysis in placental tissue of both depressed and antidepressant-treated women.
In the placenta of depressed women (Figure
We performed a gene expression study in the fetal placenta of depressed women and antidepressant-treated women, and compared them with the gene expression of the placentas from women with normal pregnancies. We found that antenatal depression and antidepressant exposure during pregnancy has an influence on the gene expression of the placenta. In the microarray 108 genes were differentially expressed in women with antenatal depression, while 109 genes were differentially expressed in antidepressant-treated women. Only 20 genes were overlapping between depressed women and women on antidepressant treatment. Among the genes we chose for validation, only 2 were validated with qPCR for depressed women. In the antidepressant-treated women, 6 genes were validated, indicating a more robust effect in alterations of these genes due to antidepressant treatment during pregnancy.
Antenatal depression is a relatively heterogeneous condition with different causes (primary or secondary to somatic disease) and differential degree of endocrine disturbances. Furthermore, women with antenatal depression may also differ between the pilot microarray and the validation study as to depression severity and duration of the depressive episode. These factors may have precluded the possibility to confirm the microarray findings, and it is a major limitation that not all women in the validation part of the study were diagnosed by a structured psychiatric interview. As depression
When the microarray was validated with a larger sample-size,
Interestingly, we also found that
Besides the role of the Rho kinase pathway in cardiovascular diseases, a role has also been proposed for the modulation of the placental vasculature. Although expression of
Another gene that was down-regulated in depressed women and to a lesser extent in antidepressant-treated women is the
Similarly,
The last gene that was validated was
Despite the strengths of our study, such as the longitudinal nature of the study and the information on the state of the mothers mood at multiple time points, some limitations need to be discussed. First, we investigated alterations in gene expression of the fetal side of the placenta due to antenatal depression and antidepressant treatment. The results of this study may give us an indication on altered pathways in the placenta. However, the placenta is a separate organ and is not part of the fetus itself, therefore findings need to be replicated in the developing fetus. In humans this is not easily feasible therefore experiments are ongoing in a rodent study. Second, antenatal depression is a relatively heterogeneous condition and outcome of diagnoses and treatment plans (before the women entered the study) were diagnosed by different doctors which may have biased the outcome. As a result dosages and types of medication may not have been appropriate for the diagnosed depression. Nevertheless, all women did undergo the EPDS screening providing comparable data between the groups concerning the mood state at different time points during (and after) pregnancy. Third, in the validation study we included different types of antidepressants, although they were mainly SSRIs, this may have influenced the outcome of the gene expression in the SSRI treatment group.
In conclusion, gene expression in the fetal placenta is altered by antenatal depression and SSRI treatment. As more placental genes alterations were validated in a larger subset of SSRI-treated women compared to those with antenatal depression we conclude that for these subset of genes, the effects of SSRI-intake during pregnancy are more robust. It remains to be established how these differentially affected genes influence the development of the child, and whether these differences are found in the fetus as well.
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
This work was supported by grants from the Swedish Research Council (Grant No. K2014-54X-20642-07-4), the Marianne and Marcus Wallenberg Foundation (2010:0031), KI fonder research (2013fobi37758), and the Swedish Society of Medicine (SLS-303881 & SLS-384001).
The Supplementary Material for this article can be found online at: