Animal Reproduction (AR)
Animal Reproduction (AR)
Original Article

Follicular blood flow, antrum growth and angiogenic mediators in mares from ovulation to deviation

Amal Mhmoud Abo El-Maaty, ElShymaa Ahmad Abdelnaby

Downloads: 1
Views: 818


This study assumed that vascular perfusion, antrum growth, leptin, nitric oxide (NO) and insulin like growth factor 1 (IGF-1) play an important role during selection and deviation of mares’ next dominant follicle. Five broad mares were subjected to daily rectal Doppler ultrasonographic examination and blood sampling for 2 successive estrous cycles (n = 10). Using electronic calipers, three diameters were taken to estimate area and volume of first (F1O) and second large follicles (F2O) on the ovulated ovary with first (F1C) and second large follicles (F2C) on contralateral ovary. Follicles’ area, circumference, antrum area, area of color- and power- Doppler were measured in pixels. Days after ovulation affected significantly (P < 0.0001) follicles blood flow, dimensions and measured hormones. On day 4 after ovulation, the follicle that had a mean diameter of 1.31 ± 0.06 and reached to 1.41 ± 0.06 cm on day 5, the lowest area/cm2 (1.38 ± 0.18), highest area/pixsel (10229 ± 366), antrum/pixel (7671 ± 357), highest volume (5.54 ± 0.09), lowest power blood flow area (2060.25 ± 8.52) and percent colored pixels of follicle without antrum (80.57 ± 0.72) was selected. Deviation started from day 9 and completed on day 10 where the dominant follicle attained the highest diameter, area, volume, area and antrum area in pixels, color blood flow red area, and percent of colored pixels of follicle without its antrum, leptin, IGF-1 and NO but the lowest power blood flow area and percent of total follicle colored pixels. Our assumption that follicle selection and deviation did not depend only on diameter but also on blood flow, antrum growth, leptin and IGF-1 was proved.


blood flow, deviation, follicle selection, mare, reproductive hormones.


Abo El-Maaty AM, Gabr FI. 2010. Relation between leptin and estradiol level in Egyptian lactating Arab mares during foaling heat. Anim Reprod Sci, 117:95-98.

Abo-El Maaty AM, El-Shahat KH. 2012. Hormonal and biochemical serum assay in relation to the estrous cycle and follicular growth in Arabian mare. Asian Pac J Reprod, 1:105-110.

Abo El-Maaty AM, Kotp MS, El-Tohamy, MM, El Natat WS, Mohamed AH. 2015. Role of leptin and insulin like growth factor-1 in breeding Egyptian horses along the year. J Med Biol Sci Res, 1:169-178.

Acosta TJ, Hayashi KG, Ohtani M, Miyamoto A. 2003. Local changes in blood flow within the preovulatory follicle wall and early corpus luteum in the cow. Reproduction, 125:759-767.

Acosta TJ, Beg MA, Ginther OJ. 2004a. Aberrant blood flow area and plasma gonadotropin concentrations during the development of dominantsized transitional anovulatory follicles in mares. Biol Reprod, 71:637-642.

Acosta TJ, Gastal EL, Gastal MO, Beg MA, Ginther OJ. 2004b. Differential blood flow changes between the future dominant and subordinate follicles precede diameter changes during follicle selection in mares. Biol Reprod, 71:502-507.

Acosta TJ, Hayashi KG, Matsui M, Miyamoto A. 2005. Changes in follicular vascularity during the first follicular wave in lactating cows. J Reprod Dev, 51:273- 280.

Agarwal SK, Vogel K, Weitsman SR, Magoffin DA. 1999. Leptin antagonizes the insulin-like growth factor-I augmentation of steroidogenesis in granulosa and theca cells of the human ovary. J Clin Endocrinol Metab, 84:1072-1076.

Almog B, Gold R, Tajima K, Dantes A, Salim K, Rubinstein M, Barkan D, Homburg R, Lessing JB, Nevo N, Gertler A, Amsterdam A. 2001. Leptin attenuates follicular apoptosis and accelerates the onset of puberty in immature rats. Mol Cell Endocrinol, 183:179-191.

Anteby EY, Hurwitz A, Korach O, Revel A, Simon A, Finci-Yeheskel Z, Mayer M, Laufer N. 1996. Human follicular nitric oxide pathway: relationship to follicular size, oestradiol concentration and ovarian blood flow. Hum Reprod, 11:1947-1951 Bächler M, Menshykau D, De Geyter Ch, Iber D. 2014. Species-specific differences in follicular antral sizes result from diffusion-based limitations on the thickness of the granulosa cell layer. Mol Hum Reprod, 20:208-221.

Barroso G, Barrionuevo M, Rao P, Graham L, Danforth D, Huey S, Abuhamad A, Oehninger S. 1999. Vascular endothelial growth factor, nitric oxide, and leptin follicular fluid levels correlate negatively with embryo quality in IVF patients. Fertil Steril, 72:1024-1026.

Beg MA, Bergfelt DR, Kot K, Wiltbank MC, Ginther OJ. 2001. Follicular-fluid factors and granulosa-cell gene expression associated with follicle deviation in cattle. Biol Reprod, 64:432-441.

Beg MA, Ginther OJ. 2006. Follicle selection in cattle and horses: role of intrafollicular factors. Reproduction, 132:365-377. Berisha B, Schams D, Rodler D, Pfaffl MW. 2016. Angiogenesis in the ovary - the most important regulatory event for follicle and corpus luteum development and function in cow - an overview.

Anat Histol Embryol, 45:124-130. Bollwein H, Weber F, Kolberg B, Stolla R. 2002. Uterine and ovarian blood flow during the estrous cycle inmares. Theriogenology, 57:2129-2138.

Bouloumie A, Drexler HC, Lafontan M, Busse R. 1998. Leptin, the product of Ob gene, promotes angiogenesis. Circ Res, 83:1059-1066.

Davidson TR, Chamberlain CS, Bridges TS, Spicer LJ. 2002. Effect of follicle size on in vitro production of steroids and insulin-like growth factor (IGF)-I, IGF-II, and the IGF-binding proteins by equine ovarian granulosa cells. Biol Reprod, 66:1640-1648.

Dayi A, Bediz CS, Musal B, Yilmaz O, Comlekci A, Celiloglu M, Cimrin D. 2005. Comparison of leptin levels in serum and follicular fluid during the oestrous cycle in cows. Acta Vet Hung, 53:457-467.

Donadeu FX, Ginther OJ. 2002. Changes in concentrations of follicular fluid factors during follicle selection in mares. Biol Reprod, 66:1111-1118.

Donadeu FX, Ginther OJ. 2004. Interrelationships of estradiol, inhibin, and gonadotropins during follicle deviation in pony mares. Theriogenology, 61:1395- 1405.

El-Sherry TM, Derar R, Bakry R. 2013. Changes in blood flow in ovine follicles and serum concentration of estradiol 17 beta (E2) and nitric oxide (NO) around the time of ovulation in Ossimi ewes. Anim Reprod Sci, 138:188-193.

Fahiminiya S, Gérard N. 2010. Follicular fluid in mammals. Gynecol Obstet Fertil, 38:402-404.

Feranil JB, Isobe N, Nakao T. 2004. Changes in the thecal vasculature during follicular atresia in the ovary of swamp buffalo. J Reprod Dev, 50:315-321.

Fortune JE, Sirois J, Turzillo AM, Lavoir M. 1991. Follicle selection in domestic ruminants. J Reprod Fertil Suppl, 43:187-198.

Fortune JE, Rivera GM, Evans AC, Turzillo AM. 2001. Differentiation of dominant versus subordinate follicles in cattle. Biol Reprod, 65:648-654.

Fraser HM. 2006. Regulation of the ovarian follicular vasculature. Reprod Biol Endocrinol, 4:18. doi: 10.1186/1477-7827-4-18.

Fruhbeck G. 2006. Intracellular signalling pathways activated by leptin. Biochem J, 393(pt. 1):7-20.

Gastal EL, Gastal MO, Bergfelt DR, Ginther OJ. 1997. Role of diameter differences among follicles in selection of a future dominant follicle in mares. Biol Reprod, 57:1320-1327.

Gastal EL, Donadeu FX, Gastal MO, Ginther OJ. 1999a, Echotextural changes in the follicular wall during follicle deviation in mares. Theriogenology, 52:803-814.

Gastal EL, Gastal MO, Wiltbank MC, Ginther OJ. 1999b. Follicle deviation and intrafollicular and systemic estradiol concentrations in mares. Biol Reprod, 61:31-39.

Gastal EL, Gastal MO, Beg MA, Ginther OJ. 2004. Interrelationships among follicles during the commongrowth phase of a follicular wave and capacity of individual follicles for dominance in mares. Reproduction, 128:417-422.

Gastal EL, Gastal MO, Ginther OJ. 2006. Relationships of changes in B-mode echotexture and color-Doppler signals in the wall of the preovulatory follicle to changes in systemic oestradiol concentrations and the effects of human chorionic gonadotrophin in mares. Reproduction, 131:699-709.

Gastal MO, Gastal EL, Beg MA, Ginther OJ. 2010. Short-term feed restriction decreases the systemic and intrafollicular concentrations of leptin and increases the vascularity of the preovulatory follicle in mares. Theriogenology, 73:1202-1209.

Ginther OJ. 1993. Major and minor follicular waves during the equine estrous cycle. J Equine Vet Sci, 13:18- 25.

Ginther OJ, Wiltbank MC, Fricke PM, Gibbons JR, Kot K. 1996. Selection of the dominant follicle in cattle. Biol Reprod, 55:1187-1194.

Ginther OJ. 2000. Selection of the dominant follicle in cattle and horses. Anim Reprod Sci, 60/61:61-79.

Ginther OJ, Beg MA, Bergfelt DR, Donadeu FX, Kot K. 2001. Follicle selection in monovular species. Biol Reprod, 65:638-647 Ginther OJ, Beg MA, Donadeu FX, Bergfelt DR. 2003. Mechanism of follicle deviation in monovular farm species. Anim Reprod Sci, 78:239-257.

Ginther OJ, Beg MA, Gastal MO, Gastal EL. 2004a. Follicle dynamics and selection in mares. Anim Reprod, 1:45-63.

Ginther OJ, Bergfelt DR, Beg MA, Meira C, Kot K. 2004b. In vivo effects of an intrafollicular injection of insulin-like growth factor 1 on the mechanism of follicle deviation in heifers and mares. Biol Reprod, 70:99-105.

Ginther OJ, Gastal EL, Gastal MO, Beg MA. 2004c. Critical role of insulin-like growth factor system in follicle selection and dominance in mares. Biol Reprod, 70:1374-1379.

Ginther OJ, Gastal EL, Gastal MO, Checura CM, Beg MA. 2004d. Dose-response study of intrafollicular injection of insulin-like growth factor-1 on follicularfluid factors and follicle dominance in mares. Biol Reprod, 70:1063-1069.

Ginther OJ, Beg MA, Gastal EL, Gastal MO, Baerwald AR, Pierson RA. 2005. Systemic concentrations of hormones during development of follicular waves in mares and women: a comparative study. Reproduction, 130:379-388.

Ginther OJ, Utt MD, Beg MA. 2007. Follicle deviation and diurnal variation in circulating hormone concentrations in mares. Anim Reprod Sci, 100:197-203.

Ginther OJ, Jacob JC, Gastal MO, Gastal EL, Beg MA. 2009. Development of one vs multiple ovulatory follicles and associated systemic hormone concentrations in mares. Reprod Domest Anim, 44:441-449.

Ginther OJ. 2012. The mare: a 1000-pound guinea pig for study of the ovulatory follicular wave in women. Theriogenology, 77:818-828.

Grazul-Bilska AT, Navanukraw C, Johnson ML, Vonnahme KA, Ford SP, Reynolds LP, Redmer DA. 2007. Vascularity and expression of angiogenic factors in bovine dominant follicles of the first follicular wave. J Anim Sci, 85:1914-1922.

Jablonka-Sharif A, Fricke PM, Grazul-Bilska AT, Reynolds LP, Redmer DA. 1994. Size, number, cellular proliferation and atresia of gonadotropininduced follicles in ewes. Biol Reprod, 51:531-540.

Jiang JY, Macchiarelli G, Tsang BK, Sato E. 2003. Capillary angiogenesisand degeneration in bovine ovarian antral follicles. Reproduction, 125:211-223.

Kerban A, Doré M, Sirois J. 1999. Characterization of cellular and vascular changes in equine follicles during hCG-induced ovulation. J Reprod Fertil, 117:115-123.

Kitawaki J, Kusuki I, Koshiba H, Tsukamoto K, Honj OH. 1999. Leptin directly stimulates aromatase activity in human luteinized granulosa cells. Mol Hum Reprod, 5:708-713.

Louis GW, Greenwald-Yarnell M, Phillips R, Coolen LM, Lehman MN, Myers MG Jr. 2011. Molecular mapping of the neural pathways linking leptin to the neuroendocrine reproductive axis. Endocrinology, 152:2302-2310.

Macchiarelli G, Jiang JY, Nottola SA, Sato E. 2006. Morphological patterns of angiogenesis in ovarian follicle capillary networks. A scanning electron microscopy study of corrosion cast. Microsc Res Tech, 69:459-468.

Marković, D. Pavlović M., Pavlović V. 2003. Seasonality, folliculogenesis and luteogenesis in mare ovaries. Facta Univ Ser Med Biol, 10:120-126.

Mazerbourg S, Bondy CA, Zhou J, Monget P. 2003. The insulin - like growth factor system: a key determinant role in the growth and selection of ovarian follicles? a comparative species study. Reprod Domest Anim, 38:247-258.

Mihm M, Austin EJ, Good TEM, Ireland JLH, Knight PG, Roche JF, Ireland JJ. 2000. Identification of potential intrafollicular factors involved in selection of dominant follicles in heifers. Biol Reprod, 63:811- 819.

Miura R, Haneda S, Lee HH, Miyamoto A, Shimizu T, Miyahara K, Miyake Y, Matsui M. 2014. Evidence that the dominant follicle of the first wave is more active than that of the second wave in terms of its growth rate, blood flow supply and steroidogenic capacity in cows. Anim Reprod Sci, 145:114-122.

Okuda Y, Okamura H, Kanzaki H, Takenaka A, Morimoto K, Nishimura T. 1982. [An ultrastructural study of capillary permeability of rabbit ovarian follicles during ovulation using carbon tracer (author's transl)]. Nihon Sanka Fujinka Gakkai Zasshi, 34:181- 186.

Pancarc SM, Güngör O, Atakişi O, Ciğremiş Y, Arı UÇ, Bollwein H. 2011. Changes in follicular blood flow and nitric oxide levels in follicular fluid during follicular deviation in cows. Anim Reprod Sci, 123:149- 156.

Redmer DA, Reynolds LP. 1996. Angiogenesis in the ovary. Rev Reprod, 1:182-192.

Rodgers RJ, Irving-Rodgers HF. 2010. Formation of the ovarian follicular antrum and follicular fluid. Biol Reprod, 82:1021-1029.

Roman EA, Ricci AG, Faletti AG. 2005. Leptin enhances ovulation and attenuates the effects produced by food restriction. Mol Cell Endocrinol, 242:33-41.

Rosselli M, Imthurn B, Macas E, Keller PJ, Dubey RK. 1994. Circulating nitrite/nitrate increase with follicular development: indirect evidence for estradiol mediated release. Biochem Biophys Res Commun, 202:1543-1552.

Rosselli M, Keller PJ, Dubey RK. 1998. Role of nitric oxide in the biology, physiology and pathophysiology of reproduction. Hum Reprod Update, 4:3-24.

Satué K, Montesinos P, Gardon JC. 2013. Influence of oestrogen and progesterone on circulating neutrophils and monocyte during ovulatory and luteal phase in healthy Spanish Purebred mares. In: Proceedings of XIX Congress of Societa Italiana Veterinari per Equini.

Cremona: SIVE. pp. 383-384. Shahiduzzaman AK, Beg MA, Palhao MP, Siddiqui MA, Shamsuddin M, Ginther OJ. 2010. Stimulation of the largest subordinate follicle by intrafollicular treatment with insulin-like growth factor 1 is associated with inhibition of the dominant follicle in heifers. Theriogenology, 74:194-201.

Shirazi A, Gharagozloo F, Ghasemzadeh-Nava H. 2004. Ultrasonic characteristics of preovulatory follicle and ovulation in Caspian mares. Anim Reprod Sci, 80:261-266.

Sierra-Honigmann MR, Nath AK, Murakami C, Garcia-Cadena G, Papapetropoulos A, Sessa WC, Madge LA, Schechner JS, Schwabb MB, Polverini PJ, Flores-Riveros JR. 1998. Biological action of leptin as an angiogenic factor. Science, ;281:1683-1686 Spicer LJ, Tucker KE, Henderson KA, Duby RT. 1991. Concentrations of insulin-like growth factor-I in follicular fluid and blood plasma of mares during early and late oestrus. Anim Reprod Sci, 25:57-65.

Uliani RC, Silva LA, Alvarenga MA. 2011. Mare’s Folliculogenesis: assessment of ovarian and perifollicular vascular perfusion by Doppler ultrasound. Acta Sci Vet, 39(suppl. 1):s113-s116.

Van Voorhis BJ, Dunn MS, Snyder GA Weiner CP. 1994. Nitric oxide: an autocrine regulator of human granulose-luteal cell steroidogenesis. Endocrinology, 135:1799-1806.

Wood KC, Cortese-Krott MM, Kovacic JC, Noguchi A, Liu VB, Wang X, Raghavachari N, Boehm M, Kato GJ, Kelm M, Gladwin MT. 2013. Circulating blood endothelial nitric oxide synthase contributes to the regulation of systemic blood pressure and nitrite homeostasis. Arterioscler Thromb Vasc Biol, 33:1861- 1871.

Wulff C, Wilson H, Wiegand SJ, Rudge JS, Fraser HM. 2002. Prevention of thecal angiogenesis, antral follicular growth, and ovulation in the primate by treatment with vascular endothelial growth factor Trap R1R2. Endocrinology, 143:2797-2807 Yu WH, Walczewska A, Karanth S, McCann SM. 1997. Nitric oxide mediates leptin-induced luteinizing hormone-releasing hormone (LHRH) and LHRH and leptin-induced LH release from the pituitary gland. Endocrinology,138:5055-5058.

Yuan W, Bao B, Garverick HA, Youngquist RS, Lucy MC. 1998. Follicular dominance in cattle is associated with divergent patterns of ovarian gene expression for insulin-like growth factor (IGF)-I, IGFII, and IGF binding protein-2 in dominant and subordinate follicles. Domest Anim Endocrinol, 15:55- 63.

Zachow RJ, Magoffin DA. 1997. Direct intraovarian effects of leptin: impairment of the synergistic action of insulin-like growth factor-I on follicle-stimulating hormone-dependent estradiol- 17 beta production by rat ovarian granulosa cells. Endocrinology, 138:847-850.

Zeleznik AJ, Schuler HM, Reichert LE. 1981. Gonadotropin-binding sites in the rhesus monkey ovary: role of the vasculature in the selective distribution of human chorionic gonadotropin to the preovulatory follicle. Endocrinology, 109:356-362.

5b73189c0e88259f178068a8 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections