Transcriptome comparative analysis of ovarian follicles reveals the key genes and signaling pathways implicated in hen egg production

Abstract Background Ovarian follicle development plays an important role in determination of poultry egg production. The follicles at the various developmental stages possess their own distinct molecular genetic characteristics and have different biological roles in chicken ovary development and function. In the each stage, several genes of follicle-specific expression and biological pathways are involved in the vary-sized follicular development and physiological events. Identification of the pivotal genes and signaling pathways that control the follicular development is helpful for understanding their exact regulatory functions and molecular mechanisms underlying egg-laying traits of laying hens. Results The comparative mRNA transcriptomic analysis of ovarian follicles at three key developmental stages including slow growing white follicles (GWF), small yellow follicles (SYF) of recruitment into the hierarchy, and differentiated large yellow follicles (LYF), was accomplished in the layers with lower and higher egg production. Totally, 137, 447, and 229 of up-regulated differentially expressed genes (DEGs), and 99, 97, and 157 of down-regulated DEGs in the GWF, SYF and LYF follicles, including VIPR1, VIPR2, ADRB2, and HSD17B1 were identified, respectively. Moreover, NDUFAB1 and GABRA1 genes, two most promising candidates potentially associated with egg-laying performance were screened out from the 13 co-expressed DEGs in the GWF, SYF and LYF samples. We further investigated the biological effects of NDUFAB1 and GABRA1 on ovarian follicular development and found that NDUFAB1 promotes follicle development by stimulating granulosa cell (GC) proliferation and decreasing cell apoptosis, increases the expression of CCND1 and BCL-2 but attenuates the expression of caspase-3, and facilitates steroidogenesis by enhancing the expression of STAR and CYP11A1. In contrast, GABRA1 inhibits GC proliferation and stimulates cell apoptosis, decreases the expression of CCND1, BCL-2, STAR, and CYP11A1 but elevates the expression of caspase-3. Furthermore, the three crucial signaling pathways such as PPAR signaling pathway, cAMP signaling pathway and neuroactive ligand-receptor interaction were significantly enriched, which may play essential roles in ovarian follicle growth, differentiation, follicle selection, and maturation. Conclusions The current study provided new molecular data for insight into the regulatory mechanism underlying ovarian follicle development associated with egg production in chicken.

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Transcriptome Analysis of Ovarian Follicles Reveals Potential Pivotal Genes Associated With Increased and Decreased Rates of Chicken Egg Production

Frontiers in Genetics ◽

10.3389/fgene.2021.622751 ◽

2021 ◽

Vol 12 ◽

Author(s):

Xiaoxia Chen ◽

Xue Sun ◽

Ignatius Musenge Chimbaka ◽

Ning Qin ◽

Xiaoxing Xu ◽

...

Keyword(s):

Transcriptome Analysis ◽

Egg Production ◽

Ovarian Follicle ◽

Ovarian Follicles ◽

Laying Hen ◽

Egg Laying ◽

Differentially Expressed ◽

Molecular Evidence ◽

Follicle Development ◽

Ovarian Follicle Development

Egg production is an important economic trait in the commercial poultry industry. Ovarian follicle development plays a pivotal role in regulation of laying hen performance and reproductive physiology. However, the key genes and signaling pathways involved in the various-stages of laying hen follicular development remain poorly understood. In this study, transcriptomes of ovarian follicles at three developmental stages, the large white follicle (LWF), small yellow follicle (SYF), and large yellow follicle (LYF), were comparatively analyzed in hens with high (HR) and low (LR) egg-laying rates by RNA-sequencing. Eighteen cDNA libraries were constructed and a total of 236, 544, and 386 unigenes were significantly differentially expressed in the LWF, SYF, and LYF follicles of HR and LR hens, respectively. Among them, 47 co-transcribed differentially expressed genes (DEGs) in LWF and SYF, 68 co-expressed DEGs in SYF and LYF, and 54 co-expressed DEGs in LWF and LYF were mined. Thirteen co-expressed DEGs were found in LWF, SYF, and LYF follicles. Eighteen candidate genes, including P2RX1, CAB39L, BLK, CSMD3, GPR65, ADRB2, CSMD1, PLPP4, ATF3, PRLL, STMN3, RORB, PIK3R1, PERP1, ACSBG1, MRTO4, CDKN1A, and EDA2R were identified to be potentially related to egg production. Furthermore, Kyoto Encyclopedia of Genes and Genomes analysis indicated neuroactive ligand-receptor interaction, cell adhesion molecules, peroxisome proliferator-activated receptor pathway, and cAMP signaling pathway might elicit an important role in formation of egg-laying traits by influencing ovarian follicle development. This study represents the first transcriptome analysis of various-sized follicles between HR and LR hens. These results provide useful molecular evidence for elucidating the genetic mechanism underlying ovarian follicle development associated with egg production in chicken.

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The Signaling Pathways Involved in Ovarian Follicle Development

Frontiers in Physiology ◽

10.3389/fphys.2021.730196 ◽

2021 ◽

Vol 12 ◽

Author(s):

Liyuan Li ◽

Xiaojin Shi ◽

Yun Shi ◽

Zhao Wang

Keyword(s):

Signaling Pathways ◽

Signaling Pathway ◽

Ovarian Follicle ◽

Follicular Development ◽

Follicle Cells ◽

Follicle Development ◽

Akt Signaling Pathway ◽

Ovarian Follicle Development ◽

High Possibility

The follicle is the functional unit of the ovary, which is composed of three types of cells: oocytes, granulosa cells, and theca cells. Ovarian follicle development and the subsequent ovulation process are coordinated by highly complex interplay between endocrine, paracrine, and autocrine signals, which coordinate steroidogenesis and gametogenesis. Follicle development is regulated mainly by three organs, the hypothalamus, anterior pituitary, and gonad, which make up the hypothalamic-pituitary-gonadal axis. Steroid hormones and their receptors play pivotal roles in follicle development and participate in a series of classical signaling pathways. In this review, we summarize and compare the role of classical signaling pathways, such as the WNT, insulin, Notch, and Hedgehog pathways, in ovarian follicle development and the underlying regulatory mechanism. We have also found that these four signaling pathways all interact with FOXO3, a transcription factor that is widely known to be under control of the PI3K/AKT signaling pathway and has been implicated as a major signaling pathway in the regulation of dormancy and initial follicular activation in the ovary. Although some of these interactions with FOXO3 have not been verified in ovarian follicle cells, there is a high possibility that FOXO3 plays a core role in follicular development and is regulated by classical signaling pathways. In this review, we present these signaling pathways from a comprehensive perspective to obtain a better understanding of the follicular development process.

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Proteomic Analysis Identifies Potential Markers for Chicken Primary Follicle Development

Animals ◽

10.3390/ani11041108 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1108

Author(s):

Armughan Ahmed Wadood ◽

Jingyuan Wang ◽

Liping Pu ◽

Qaisar Shahzad ◽

Muhammad Waqas ◽

...

Keyword(s):

Proteomic Analysis ◽

Egg Production ◽

Potential Role ◽

Protein Localization ◽

Follicular Development ◽

Egg Laying ◽

Follicle Development ◽

Phosphate Binding ◽

Primary Follicle ◽

Mannose Metabolism

Follicles’ development in chicken imparts a major impact on egg production. To enhance the egg-laying efficiency, comprehensive knowledge of different phases of follicular development is a prerequisite. Therefore, we used the tandem mass tag (TMT) based proteomic approach to find the genes involved in the primary follicular development of chicken. The primary follicles were divided into two groups—small primary follicles (81–150 μm) and developed primary follicles (300–500 μm). Differential expression analysis (fold change > 1.2, p-value < 0.05) revealed a total of 70 differentially expressed proteins (DEPs), of which 38 were upregulated and 32 were downregulated. Gene ontology (GO) enrichment analysis disclosed that DEPs were intricate with cellular protein localization, the establishment of protein localization, and nucleoside phosphate-binding activities. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway indicated the involvement of DEPs in different metabolic pathways such as glycolysis, pyruvate metabolism, galactose metabolism, and fructose and mannose metabolism. The current proteomic analysis suggested suitable markers such as Anxa2, Pdia3, and Capzb, which may serve as a potential role for primary follicle development. The present study provides the first insight into the proteome dynamics of primary follicle development and would play a potential role for further studies in chicken to improve egg productivity.

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224 OVARIAN FOLLICLE DEVELOPMENT DURING SEASONAL TRANSITION IN WAPITI

Reproduction Fertility and Development ◽

10.1071/rdv18n2ab224 ◽

2006 ◽

Vol 18 (2) ◽

pp. 220

Author(s):

R. McCorkell ◽

M. Woodbury ◽

G. Adams

Keyword(s):

Breeding Season ◽

Ovarian Follicle ◽

Follicular Development ◽

Maximum Diameter ◽

Follicle Development ◽

Estrous Cycles ◽

Ovarian Follicle Development ◽

Array Transducer ◽

October 17 ◽

Ovulatory Follicles

Wapiti are seasonally polyestrous. The transition into and out of the breeding season is marked by resumption of ovulation in autumn and cessation of ovulation in winter. Onset of ovulatory cyclicity is distinct and associated with aggressive breeding behavior of stags in rut. Cessation of ovulation at the end of the breeding season is not distinguished by behavioral patterns. The objective of the present study was to characterize follicular and luteal dynamics in wapiti during the transitional periods into and out of the breeding seasons. Transition from anestrus to estrus was monitored in 15 hinds, aged 2 to 14 years, over two successive seasons (11 in year 1, with 5 hinds from year 1 used again in year 2 along with 4 new hinds; n = 20 observations). Transition from estrus to anestrus was monitored in 11 hinds over 1 season (n = 11 observations). Hinds were maintained on a farm near Saskatoon, Saskatchewan (52°07′N, 106°38′W). The ovaries were examined daily during September through October by transrectral ultrasonography using a B-mode ultrasound machine and a 7.5 MHz linear array transducer for transition to estrus, and December through April for transition to anestrus. The first ovulation was recorded on September 15 and all hinds had ovulated for the first time by October 7. In 17 of 20 observations, the duration of the first interovulatory interval (IOI) was 9.3 ± 0.4 days (mean ± SEM). With one exception, these IOIs were characterized by one wave of follicular development. The remaining three IOIs ranged from 16 to 23 days and consisted of two or three waves of follicle development. The second ovulation occurred by October 15 in hinds with a short IOI and by October 17 in all remaining hinds. The mean dates of first and last ovulation were September 25 and February 7, respectively, an interval of 135 days. The median date of the last ovulation was February 15 and the range was from December 3 to March 22. Duration of the last IOI of the season (21.2 ± 0.6 days) was similar to the notional 21-day cycle for wapiti, but longer (P < 0.05) than the duration of the first IOI (10.9 ± 1.0 days). Maximum diameters of the first 2 ovulatory follicles were similar (11.3 ± 0.4 vs. 11.3 ± 0.2 mm), but were larger (P < 0.05) than the last 2 ovulatory follicles of the breeding season (10.3 ± 0.3 vs. 10.1 ± 0.4 mm). Maximum diameter of the corpus luteum (CL) tended (P = 0.06) to be smaller for the short IOI than for longer IOI of the first and last cycles. Diameter of the last CL of the season was not different from that of the previous CL (12.8 ± 0.6 vs. 12.5 ± 0.6 mm); however, it was detected for a longer period (22.3 ± 1.2 vs. 19.3 ± 0.7 days; P < 0.05). Estrous cycles during transition into the breeding season have been described as being irregular and those out of the breeding season as increasingly long. In the present study, the transition periods were characterized by regular events. Transition to regular estrous cycles was preceded by one short (9 days) IOI. The last IOI of the breeding season was the same as that reported during the rut. Transition to anestrus occurred most commonly in February and was marked by a failure of the dominant follicle to ovulate after luteal regression.

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The Increased Expression of Connexin and VEGF in Mouse Ovarian Tissue Vitrification by Follicle Stimulating Hormone

BioMed Research International ◽

10.1155/2015/397264 ◽

2015 ◽

Vol 2015 ◽

pp. 1-13 ◽

Cited By ~ 6

Author(s):

Yanzhou Yang ◽

Jie Chen ◽

Hao Wu ◽

Xiuying Pei ◽

Qing Chang ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Caspase 3 ◽

Ischemia Reperfusion ◽

Ovarian Tissue ◽

Ovarian Follicle ◽

Follicle Stimulating Hormone ◽

Vegf Receptor ◽

Follicle Development ◽

Ovarian Follicle Development

Ovarian follicular damages were caused by cryoinjury during the process of ovarian vitrification and ischemia/reperfusion during the process of ovarian transplantation. And appropriate FSH plays an important role in antiapoptosis during ovarian follicle development. Therefore, in this study, 0.3 IU/mL FSH was administered into medium during mouse ovarian cryopreservation by vitrification to ascertain the function of FSH on ovarian vitrification and avascular transplantation. The results suggested that the expressions of Cx37, Cx43, apoptotic molecular caspase-3, and angiogenesis molecular VEGF were confirmed using immunohistochemistry, western blotting, and real-time PCR, and the results suggested that the treatment with FSH remarkably increased the number of morphologically normal follicles in vitrified/warmed ovaries by upregulating the expression of Cx37, Cx43, VEGF, and VEGF receptor 2, but downregulating the expression of caspase-3. In addition, the vitrified/warmed ovaries were transplanted, and the related fertility was analyzed, and the results suggested that the fertility, neoangiogenesis, and follicle reserve were remarkably increased in the FSH administrated group. Taken together, administration of 0.3 IU/mL FSH during ovarian cryopreservation by vitrification can maintain ovarian survival during ovarian vitrification and increases the blood supply with avascular transplantation via upregulation of Cx43, Cx37, and VEGF/VEGFR2, as well as through its antiapoptotic effects.

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Orphan nuclear receptors in angiogenesis and follicular development

Reproduction ◽

10.1530/rep-21-0118 ◽

2021 ◽

Author(s):

Adrian Guzmán ◽

Camilla H.k. Hughes ◽

Bruce D. Murphy

Keyword(s):

Nuclear Receptors ◽

Molecular Mechanisms ◽

Reproductive Tract ◽

Ovarian Follicle ◽

Follicular Development ◽

Antral Follicle ◽

Follicle Development ◽

Orphan Nuclear Receptors ◽

Ovarian Follicle Development ◽

Wide Range

Orphan nuclear receptors (ONRs) are a subset of the nuclear receptor family that lack known endogenous ligands. Among 48 nuclear receptors identified in humans, 25 are classified as ONRs. They function as transcription factors and control expression of a wide range of genes to regulate metabolism, fertility, immunity, angiogenesis, and many other functions. Angiogenic factors are essential during ovarian follicle development, including follicle growth and ovulation,. Correct development of blood vessels contributes to preantral and antral follicular development, selection of the dominant follicle or follicles, follicular atresia, and ovulation. Although progress has been made in understanding the molecular mechanisms that regulate follicular angiogenesis, the role of ONRs as regulators is not clear. Based on their functions in other tissues, the ONRs NR1D1 (REV-ERBß), NR2C2 (TR4), NR2F2 (COUP-TF-II) and NR3B1, 2, and 3 (ERRα, ERRß and ERRγ) may modulate angiogenesis during antral follicle development. We hypothesize that this is achieved by effects on the expression and function of VEGFA, ANGPT1, THBS1, and soluble VEGFR1. Further, angiogenesis during ovulation is expected to be influenced by ONRs. NR5A2 (LRH-1), which is required for ovulation, regulates angiogenic genes in the ovary, including VEGFA and the upstream regulator of angiogenesis, PGE2. These angiogenic molecules may also be regulated by NR5A1 (SF-1). Evidence from outside the reproductive tract suggests that NR2F2 and NR4A1(NUR77) promote VEGFC and PGF respectively and NR4As (ΝUR77, NOR1) seem to be necessary for the angiogenic effects of VEGFA and PGE2. Together, the data suggest that ONRs are important regulators of follicular angiogenesis.

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Leptin receptor signaling inhibits ovarian follicle development and egg laying in chicken hens

Reproductive Biology and Endocrinology ◽

10.1186/1477-7827-12-25 ◽

2014 ◽

Vol 12 (1) ◽

pp. 25 ◽

Cited By ~ 17

Author(s):

Ming M Lei ◽

Si Q Wu ◽

Xiao W Li ◽

Cong L Wang ◽

Zhe Chen ◽

...

Keyword(s):

Leptin Receptor ◽

Ovarian Follicle ◽

Egg Laying ◽

Follicle Development ◽

Receptor Signaling ◽

Ovarian Follicle Development

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Differential expression of mRNAs encoding the putative inhibin co-receptor (betaglycan) and activin type-I and type-II receptors in preovulatory and prehierarchical follicles of the laying hen ovary

Journal of Endocrinology ◽

10.1677/joe.1.06525 ◽

2006 ◽

Vol 188 (2) ◽

pp. 241-249 ◽

Cited By ~ 8

Author(s):

T M Lovell ◽

P G Knight ◽

R T Gladwell

Keyword(s):

Mrna Expression ◽

Transforming Growth Factor ◽

Ovarian Follicle ◽

Follicular Development ◽

Transforming Growth Factor Β ◽

Receptor Expression ◽

Type I ◽

Follicle Development ◽

Ovarian Follicle Development ◽

Real Time Quantitative Pcr

Ovarian follicle development is primarily regulated by an interplay between the pituitary gonadotrophins, LH and FSH, and ovary-derived steroids. Increasing evidence implicates regulatory roles of transforming growth factor-β (TGFβ) superfamily members, including inhibins and activins. The aim of this study was to identify the expression of mRNAs encoding key receptors of the inhibin/activin system in ovarian follicles ranging from 4 mm in diameter to the dominant F1 follicle (~40 mm). Ovaries were collected (n=16) from mid-sequence hens maintained on a long-day photoschedule (16 h of light:8 h of darkness). All follicles removed were dissected into individual granulosa and thecal layers. RNA was extracted and cDNA synthesized. Real-time quantitative PCR was used to quantify the expression of mRNA encoding betaglycan, activin receptor (ActR) subtypes (type-I, -IIA and -IIB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH); receptor expression data were normalized to GAPDH expression. Detectable levels of ActRI, -IIA and -IIB and the inhibin co-receptor (betaglycan) expression were found in all granulosa and thecal layers analysed. Granulosa ActRI mRNA peaked (P < 0.05) in 8–9.9 mm follicles, whereas ActRIIA rose significantly from 6–7.9 mm to 8–9.9 mm, before falling to F3/2; levels then rose sharply (3-fold) to F1 levels. Granulosa betaglycan mRNA expression rose 3-fold from 4–5.9 mm to 8–9.9 mm, before falling 4-fold to F3/2; levels then rose sharply (4-fold) to F1 levels. ActRIIB levels did not vary significantly during follicular development. Thecal ActRI mRNA expression was similar from 4–7.9 mm then decreased significantly to a nadir at the F4 position, before increasing 2-fold to the F1 (P < 0.05). Although thecal ActRIIB and -IIA expression did not vary significantly from 4 mm to F3, ActRIIB expression increased significantly (2-fold) from F3 to F1 and ActRIIA increased 2-fold from F2 to F1 (P < 0.05). Thecal betaglycan fell to a nadir at F6 after follicle selection; levels then increased significantly to F2, before falling ~50% in the F1. In all follicles studied expression of betaglycan and ActRI (granulosa: r=0.65, P < 0.001, n=144/group; theca: r=0.49, P < 0.001, n=144/group) was well correlated. No significant correlations were identified between betaglycan and ActRIIA or -IIB. Considering all follicles analysed, granulosa mRNA expression of betaglycan, ActRI, ActRIIA and ActRIIB were all significantly lower than in corresponding thecal tissue (betaglycan, 11.4-fold; ActRIIB, 5.1-fold; ActRI, 3.8-fold; ActRIIA, 2.8-fold). The co-localization of type-I and -II activin receptors and betaglycan on granulosa and thecal cells are consistent with a local auto/paracrine role of inhibins and activins in modulating ovarian follicle development, selection and progression in the domestic fowl.

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Involvement of adipokines, AMPK, PI3K and the PPAR signaling pathways in ovarian follicle development and cancer

The International Journal of Developmental Biology ◽

10.1387/ijdb.120134jd ◽

2012 ◽

Vol 56 (10-11-12) ◽

pp. 959-967 ◽

Cited By ~ 51

Author(s):

Jolle Dupont ◽

Maxime Reverchon ◽

Lucie Cloix ◽

Pascal Froment ◽

Christelle Ramé

Keyword(s):

Signaling Pathways ◽

Ovarian Follicle ◽

Follicle Development ◽

Ovarian Follicle Development ◽

Ppar Signaling

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Notch Signaling Is Involved in Ovarian Follicle Development by Regulating Granulosa Cell Proliferation

Endocrinology ◽

10.1210/en.2010-1182 ◽

2011 ◽

Vol 152 (6) ◽

pp. 2437-2447 ◽

Cited By ~ 60

Author(s):

Chun-Ping Zhang ◽

Jun-Ling Yang ◽

Jun Zhang ◽

Lei Li ◽

Lin Huang ◽

...

Keyword(s):

Cell Proliferation ◽

Notch Signaling ◽

Granulosa Cell ◽

Granulosa Cells ◽

Ovarian Follicle ◽

Butyl Ester ◽

Follicular Development ◽

Follicle Development ◽

Ovarian Follicle Development

Notch signaling is an evolutionarily conserved pathway, which regulates cell proliferation, differentiation, and apoptosis. It has been reported that the members of Notch signaling are expressed in mammalian ovaries, but the exact functions of this pathway in follicle development is still unclear. In this study, primary follicles were cultured in vitro and treated with Notch signaling inhibitors, L-658,458 and N-[N-(3,5-Difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT). We found that the cultured follicles completely stopped developing after L-658,458 and DAPT treatment, most of the granulosa cells were detached, and the oocytes were also degenerated with condensed cytoplasma. Further studies demonstrated that the proliferation of granulosa cells was dependent on the Notch signaling. L-658,458 and DAPT treatment inhibited proliferation of in vitro cultured primary granulosa cells and decreased the expression of c-Myc. Lentivirus mediated overexpression of Notch intracellular domain 2, and c-Myc could promote the proliferation of granulosa cells and rescue the growth inhibition induced by L-658,458 and DAPT. In conclusion, Notch signaling is involved in follicular development by regulating granulosa cell proliferation.

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