Transcriptome Sequencing in the Preoptic Region of Rat Dams Reveals a Role of Androgen Receptor in the Control of Maternal Behavior
Abstract
:1. Introduction
2. Results
2.1. Pup-Deprived Mothers Showed Heavily Impaired Pup-Directed Behavior
2.2. Preoptic Transcriptome of Lactating and Pup-Deprived Mothers
2.3. Differentially Expressed Genes in the MPOA of Lactating and Pup-Deprived Mothers
2.4. Validation of RNA-Seq
2.5. Distribution of Differentially Expressed Genes in the Preoptic Area
2.6. Decreased Androgen Receptor (AR) Protein Levels in MPOA of Lactating Mothers
2.7. Intracerebroventricular (icv.) Administration of Androgen-Receptor Antagonist Flutamide Enhanced Pup-Directed Behavior
3. Discussion
3.1. Gene Expressional Changes in Mothers
3.2. DEGS in the MPOA of Lactating and Pup-Deprived Mothers
3.2.1. Ndufs5-NADH Dehydrogenase (Ubiquinone) Fe-S Protein 5
3.2.2. Nwd1-NACHT and WD Repeat Domain Containing Protein 1
3.2.3. Rbm3-RNA-Binding Motif Protein-3
3.3. Distribution of DEGS with the Preoptic Area
3.4. Androgen Receptor (AR) as a Possible Modulator of Central Maternal Adaptation
4. Materials and Methods
4.1. Animals
4.2. Analysis of Maternal Behavior
4.3. Microdissection of Brain Tissue Samples
4.4. RNA-Sequencing
4.4.1. Library Construction and Quality Check
4.4.2. Alignment and Quantification
4.5. Quantitiative RT-PCR
4.6. Production of Hybridization Probes for Ndufs5, Nwd1, and Rbm3
4.7. In Situ Hybridization Histochemistry
4.8. Androgen Receptor (AR) Immunohistochemistry
4.9. Western Blot Analysis
4.10. Implantation of Intracerebroventricular Cannulae
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ac | anterior commissure |
ACTH | adrenocorticotropin |
CP | caudatus putamen |
Cx | cerebral cortex |
DAB | 3,3-diaminobenzidine |
f | fornix |
FITC | fluorescein isothiocyanate |
GAPDH | glyceraldehyde-3-phosphate-dehydrogenase |
GnRH | gonadotropin releasing hormone |
LH | gonadotropin releasing hormone |
LS | lateral septal nucleus |
LV | lateral ventricle |
MPA | medial preoptic area |
MPN | medial preoptic nucleus |
och | optic chiasm |
PB | phosphate buffer |
PVN | paraventricular hypothalamic nucleus |
vBNST | ventral subdivision of the bed nucleus of the stria terminalis |
3V | third ventricle |
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Total Raw Reads (Mb) | Clean Reads (Mb) | Clean Reads Ratio (%) | Total Reads Mapping Ratio (%) | Uniquely Reads Mapping Ratio (%) | Total Gene Mapping Ratio (%) | |
---|---|---|---|---|---|---|
D1 | 57.27 | 52.8 | 92.19 | 75.12 | 61.22 | 57.25 |
D2 | 58.08 | 54.54 | 93.9 | 74.14 | 60.16 | 58.19 |
D3 | 51.25 | 48.8 | 95.23 | 94.28 | 76.39 | 70.65 |
D4 | 60.63 | 57.1 | 94.17 | 93.51 | 76.47 | 72.67 |
D5 | 62.55 | 59.42 | 95 | 74.3 | 60.26 | 58.78 |
D6 | 61.46 | 58.47 | 95.14 | 74.43 | 60.17 | 57.58 |
M1 | 102.26 | 96.28 | 94.15 | 94.69 | 77.19 | 75.08 |
M2 | 90.68 | 87.18 | 96.14 | 94.52 | 76.76 | 74.15 |
M3 | 92.07 | 87.73 | 95.28 | 93.71 | 76.05 | 72.68 |
M4 | 120.24 | 114.73 | 95.42 | 94.58 | 76.41 | 75.51 |
M5 | 100.04 | 96.01 | 95.97 | 94.58 | 76.04 | 75.19 |
M6 | 60.36 | 57.6 | 95.42 | 73.27 | 59.06 | 56.96 |
Gene ID | Unigen ID | Gene Symbol | Protein | Expression in Lactating Mothers | Expression in Pup-Deprived Mothers | Log2 (Lactating/Pup-Deprived) | PPEE | Function |
---|---|---|---|---|---|---|---|---|
103694869 | Rn.1166. | LOC103694869 | Isochorismatase domain-containing protein 1 | 254.936 | 0,000 | 8.612 | 0 | Metabolic process |
100910978 | Rn.106785. | Ecm2 | Ecm2 (Extracellular matrix protein 2) | 35.792 | 0.46 | 6.281 | 0.00102 | Organization of extracellular matrix, collagen binding |
100363268 | Rn.156413. | Ndufs5 | NADH dehydrogenase (Ubiquinone) Fe-S protein 5 | 155.64 | 5.81 | 4.744 | 0.0186 | Accessory subunit of the mitochondrial membrane respiratory chain |
100910540 | - | LOC100910540 | - | 525.556 | 135.613 | 1.954 | 0.000000966 | - |
100364350 | - | Nwd1 | NACHT And WD Repeat Domain Containing 1 | 2644.637 | 1919.119 | 0.463 | 0.0048 | Control of androgen receptor (AR) protein steady-state levels. |
114488 | Rn.18057 | Rbm3 | RNA-binding protein 3 | 1530.588 | 2190.024 | -0.517 | 0.00834 | Regulation of translation, cold-inducible mRNA binding protein |
554353 | Rn.163444 | Gpr34 | G Protein-Coupled Receptor 34 | 235.54 | 364.333 | -0.629 | 0.0384 | G-protein coupled signal transduction |
102553657 | - | LOC102553657 | - | 20.299 | 42.477 | -1.065 | 0.0105 | - |
304361 | Rn.198943. | Mepce | Mepce protein (Methylphosphate Capping Enzyme) | 243.175 | 631.98 | -1.378 | 0.00006 | RNA-binding methyltransferase |
681224 | - | LOC681224 | - | 13.044 | 105.041 | -3.01 | 0.0281 | - |
24994 | Rn.222446 | RT1-S2 | RT1 Class 1 S2 | 19.498 | 182.707 | -3.228 | 0.0002 | Involved in the presentation of foreign antigens to the immune system. |
103691822 | - | LOC103691822 | - | 0.178 | 31.412 | -7.463 | 0.00000003 | - |
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Lékó, A.H.; Kumari, R.; Dóra, F.; Keller, D.; Udvari, E.B.; Csikós, V.; Renner, É.; Dobolyi, A. Transcriptome Sequencing in the Preoptic Region of Rat Dams Reveals a Role of Androgen Receptor in the Control of Maternal Behavior. Int. J. Mol. Sci. 2021, 22, 1517. https://doi.org/10.3390/ijms22041517
Lékó AH, Kumari R, Dóra F, Keller D, Udvari EB, Csikós V, Renner É, Dobolyi A. Transcriptome Sequencing in the Preoptic Region of Rat Dams Reveals a Role of Androgen Receptor in the Control of Maternal Behavior. International Journal of Molecular Sciences. 2021; 22(4):1517. https://doi.org/10.3390/ijms22041517
Chicago/Turabian StyleLékó, András H., Rashmi Kumari, Fanni Dóra, Dávid Keller, Edina B. Udvari, Vivien Csikós, Éva Renner, and Arpád Dobolyi. 2021. "Transcriptome Sequencing in the Preoptic Region of Rat Dams Reveals a Role of Androgen Receptor in the Control of Maternal Behavior" International Journal of Molecular Sciences 22, no. 4: 1517. https://doi.org/10.3390/ijms22041517