New Molecular Markers Involved in Regulation of Ovarian Granulosa Cell Morphogenesis, Development and Differentiation during Short-Term Primary In Vitro Culture—Transcriptomic and Histochemical Study Based on Ovaries and Individual Separated Follicles
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Collection of Porcine Ovaries and In Vitro Culture of Granulosa Cells
4.3. Histological Analysis
4.4. Microarray Expression Analysis and Statistics
4.5. Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR) Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene | FC 48 h/0 h | FC 96 h/0 h | FC 14 h4/0 h | p Value 48 h/0 h | p Value 96 h/0 h | p Value 144 h/0 h | Entrez ID |
---|---|---|---|---|---|---|---|
CXCL10 | 0.036689 | 0.027744 | 0.030979 | 3.47 × 106 | 1.07 × 106 | 1.53 × 106 | 3627 |
DAB1 | 0.112175 | 0.096043 | 0.092698 | 3.33 × 106 | 1.07 × 106 | 1.26 × 106 | 1600 |
DAPL1 | 0.03486 | 0.003772 | 0.004046 | 1.00 × 106 | 9.14 × 108 | 9.57 × 108 | 92196 |
IGF1 | 0.119841 | 0.180781 | 0.391369 | 9.96 × 105 | 0.000172 | 0.006284 | 3479 |
IHH | 0.049771 | 0.0538 | 0.047738 | 2.16 × 106 | 1.06 × 106 | 1.09 × 106 | 3549 |
ITM2A | 0.11303 | 0.043537 | 0.042942 | 7.74 × 106 | 7.05 × 107 | 8.20 × 107 | 9452 |
MCOLN3 | 0.114236 | 0.052489 | 0.068606 | 9.09 × 106 | 1.06 × 106 | 1.98 × 106 | 55283 |
NEBL | 0.037428 | 0.037303 | 0.034729 | 9.56 × 107 | 3.68 × 107 | 4.09 × 107 | 10529 |
SCUBE1 | 0.084451 | 0.063451 | 0.087118 | 4.12 × 106 | 1.09 × 106 | 2.53 × 106 | 80274 |
TGFBR3 | 0.065878 | 0.045288 | 0.04622 | 4.56 × 107 | 1.51 × 107 | 1.50 × 107 | 7049 |
CAV1 | 22.31557 | 10.4332 | 28.12126 | 4.28 × 107 | 4.59 × 107 | 1.50 × 107 | 857 |
FN1 | 35.37896 | 66.1651 | 68.74185 | 3.59 × 107 | 9.92 × 108 | 9.57 × 108 | 2335 |
IGFBP5 | 29.89599 | 20.97384 | 31.2743 | 4.42 × 107 | 3.09 × 107 | 2.28 × 107 | 3488 |
ITGA2 | 42.67896 | 41.0013 | 38.26346 | 3.52 × 107 | 1.21 × 107 | 1.26 × 107 | 3673 |
ITGB3 | 29.91255 | 39.85199 | 57.68212 | 3.52 × 107 | 1.21 × 107 | 9.57 × 108 | 3690 |
LAMB1 | 33.30078 | 60.54652 | 39.6371 | 3.52 × 107 | 9.14 × 108 | 9.57 × 108 | 3912 |
HPDPN | 18.98888 | 18.65575 | 22.39775 | 2.14 × 105 | 1.11 × 105 | 1.07 × 105 | 10630 |
POSTN | 95.22897 | 29.49005 | 88.94606 | 3.52 × 107 | 1.80 × 107 | 9.57 × 108 | 10631 |
TAGLN | 25.56666 | 53.77139 | 50.96687 | 3.52 × 107 | 9.14 × 108 | 9.57 × 108 | 6876 |
TMOD1 | 23.71613 | 5.946509 | 9.468941 | 1.00 × 106 | 6.63 × 106 | 2.71 × 106 | 7111 |
Gene | Gene ID | Primer Sequence (5′–3′) | Product Size (bp) |
---|---|---|---|
CXCL10 | 494019 | CTCCTGAAAGGCCCATCATA GCACATGGGATAGAGGAGGA | 193 |
DAB1 | 100037307 | TACGTTTGTGGGAAGGAAGG CTTCCTTCTTTTGGCTGGTG | 202 |
DAPL1 | 100157537 | CCTGCTCTGGAGAAGGTCAC GGGCCTAAGGAAAGTTTTGG | 151 |
IGF1 | 47523587 | TTCTACTTGGCCCTGTGCTT CTCCAGCCTCCTCAGATCAC | 222 |
IHH | 397174 | CTCCACTGCCCTCTCAGAAC AGCTCGCAGCTGTGTCACTA | 182 |
ITM2A | 595131 | TCTCGTAGGCCTTTCCTTCA AGGCAGGAAGTAGGGCTCTC | 163 |
MCOLN3 | 100625693 | TCCGAGTGCCTTTTCTCACT CGGATATAAACGTGCGGAGT | 238 |
NEBL | 100522395 | CAAACCCTTCAAGGCTACCA CTGAGAACACGCTTCCATCA | 177 |
SCUBE1 | 100524621 | AATCCAATGAAGCCAACAGC AGGGCCTTGATCAGCTTCTT | 160 |
TGFBR3 | 397512 | TTTGTTTTAGCTGGGGGTTG TGGCCACAGGGATTTTTAAG | 177 |
CAV1 | 404693 | TAGGTCAGCAGCCTCCCTAA CTGGTGAGAGGCAGGAAAAG | 243 |
FN1 | 397620 | TGAGCCTGAAGAGACCTGCT CAGCTCCAATGCAGGTACAG | 113 |
IGFBP5 | 397182 | TGGGGGTTTGTTTCTCTGAC TTCTGGCAGGTAGAGCAGGT | 181 |
ITGA2 | 397483 | CATGCCAGATCCCTTCATCT CGCTTAAGGCTTGGAAACTG | 153 |
ITGB3 | 397063 | GGCTTCAAAGACAGCCTCAC AGTCCTTTTCCGAGCACTCA | 175 |
LAMB1 | 396707 | CTTCACCACCTTGGACCACT AGCTGTGGCTCATAGCGAAT | 216 |
PDPN | 100738269 | AGCAGATGCTGTGTCCCTCT TATGGAACCTGGGCTGGTAG | 201 |
POSTN | 100152401 | ATTGACCGTGTCCTCACACA GCCACTTTGTCTCCCATGAT | 212 |
TAGLN | 397021 | TTAAAGGCCGCTGAGGACTA ATGACATGCTTTCCCTCCTG | 233 |
TMOD1 | 100316850 | AGCCCTAACGGAAGAAGAGC CCTTTGCTTGCTTTTCCAAG | 170 |
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Kulus, M.; Sujka-Kordowska, P.; Konwerska, A.; Celichowski, P.; Kranc, W.; Kulus, J.; Piotrowska-Kempisty, H.; Antosik, P.; Bukowska, D.; Iżycki, D.; et al. New Molecular Markers Involved in Regulation of Ovarian Granulosa Cell Morphogenesis, Development and Differentiation during Short-Term Primary In Vitro Culture—Transcriptomic and Histochemical Study Based on Ovaries and Individual Separated Follicles. Int. J. Mol. Sci. 2019, 20, 3966. https://doi.org/10.3390/ijms20163966
Kulus M, Sujka-Kordowska P, Konwerska A, Celichowski P, Kranc W, Kulus J, Piotrowska-Kempisty H, Antosik P, Bukowska D, Iżycki D, et al. New Molecular Markers Involved in Regulation of Ovarian Granulosa Cell Morphogenesis, Development and Differentiation during Short-Term Primary In Vitro Culture—Transcriptomic and Histochemical Study Based on Ovaries and Individual Separated Follicles. International Journal of Molecular Sciences. 2019; 20(16):3966. https://doi.org/10.3390/ijms20163966
Chicago/Turabian StyleKulus, Magdalena, Patrycja Sujka-Kordowska, Aneta Konwerska, Piotr Celichowski, Wiesława Kranc, Jakub Kulus, Hanna Piotrowska-Kempisty, Paweł Antosik, Dorota Bukowska, Dariusz Iżycki, and et al. 2019. "New Molecular Markers Involved in Regulation of Ovarian Granulosa Cell Morphogenesis, Development and Differentiation during Short-Term Primary In Vitro Culture—Transcriptomic and Histochemical Study Based on Ovaries and Individual Separated Follicles" International Journal of Molecular Sciences 20, no. 16: 3966. https://doi.org/10.3390/ijms20163966