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Association of cumulus–oocyte complexes with the intrafollicular levels of a blood proten in Bubalus bubalis

Published online by Cambridge University Press:  26 September 2008

R. Talevi ,
P. Bergamo ,
G. Sansone ,
A. Savarese ,
P. Venditti  and
P. Abrescia
R. Talevi*
Affiliation:
Istituto Internazionale di Genetica e Biofisica, Naples, Italy.
P. Bergamo
Affiliation:
Istituto Internazionale di Genetica e Biofisica, Naples, Italy.
G. Sansone
Affiliation:
Istituto Internazionale di Genetica e Biofisica, Naples, Italy.
A. Savarese
Affiliation:
Istituto Internazionale di Genetica e Biofisica, Naples, Italy.
P. Venditti
Affiliation:
Istituto Internazionale di Genetica e Biofisica, Naples, Italy.
P. Abrescia
Affiliation:
Istituto Internazionale di Genetica e Biofisica, Naples, Italy.
*
R. Talevi, Dipartimento di Biologia Evolutiva e Comparata, Via Mezzocannone 8, I-80134 Naples, Italy.
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Summary

The protein pattern of the follicular fluid (FF) and the ultrastructure of the inner cumulus–oocyte complex (COC) has been analysed in single antral follicles (n = 146) of buffalo B. Bubalis ovaries. The protein population of FF was fractionated by SDS-PAGE; the resulting pattern was Coomassie stained and processed for densitometry. Comparartive analysis of sera and autologous FFs showed a marked difference in the level (measured as the percentage of total proteins) of one 21 kDa polypeptide band, called ‘L’. Concentration of L, which was mainly higher in the serum (2.05 ± 1.5%) than in the surrounding FF(0.98 ± 0.94%), fluctuated widely in fluids from the in the same ovary. On gel filtration of FF and SDS-PAGE of ther factions collected, the L polypeptide was found and eluted together with a 36 kDa polypeptide, called ‘H’, with an exclusion volume lower than that of albumin. The levels of both polypeptides in the eluted fractions were measured by gel densitometry and the same ratio H/L was found (2:1). These data suggest that H and L are subunits of a complex high-molecular-weight protein. The presence of L levels in male sera comparable to those detected in females indicates that this putative protein does not originate in the ovary but is transported from the blood. Moreover, a correlation between the increase in the percentage of Lf (calculated as %L in FF/%L in serum) and atresia was observed. COCs (n = 86) obtained during the collection of the single FF samples were processed for transmission electron microscopy. The ultrastructure of each COC was compared with the SDS-PAGE data of the associated FF. Healthy COCs were found to be related to very low levels of Lf (between 0 and 14% of those measured in serum). COCs with an early atretic ultrastructure undetectable at the dissection microscope, were associated with FFs having Lf levels between 24% and 60%; advanced atresia was associated with Lf values up to 70%. Finally, the acrosome reaction of buffalo precapacitated spermatozoa in vitro was monitored by adding one volume of FF with high (FF +; Lf = 80%) or undetectable (FF - ) values of Lf to the sperm suspension. It was found that 96.2% of spermatozoa treated with FF - showed a normal acrosome reaction, without any change in their motility pattern; spermatozoa treated with FF +, in contrast, exhibited poor motility, and only 66.4% of them underwent the acrosome reaction. The ionophore A23187, in a control experiment, resulted in lower percentages of acrosome reactions (38%). In conclusion, our results suggest that enhanced influx of L from serum into the follicle antrum causes an increase in the polypeptide level in FF. This might be due to a loss of selectivity in the blood-follicle barrier of early atretic follicles. The use of the L polypeptide as a biochemical marker for identifying the earlier stage of follicular atresia is proposed.

Keywords

Cumulus–oocyte complexFollicular fluidproteinUltrastructure
Type
Article
Information
Zygote , Volume 2 , Issue 2 , May 1994 , pp. 167 - 178
Copyright
Copyright © Cambridge University Press 1994

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References

Aitken, J. (1991). Do sperm find eggs attractive? Nature 351, 19–20.CrossRefGoogle ScholarPubMed
Andersen, M.M., Krøll, J., Byskov, A.G. & Faber, M. (1976). Protein composition of individual bovine follicles. J. Reprod. Fertil. 48, 109–18CrossRefGoogle ScholarPubMed
Bohnet, H.G. & Baukloh, V. (1985). Prolactin concentration in follicular fluid following ovarian hyperstimulation for in vitro fertilization. Hormone Res. 22, 189–95.CrossRefGoogle ScholarPubMed
Centola, G.M. (1982). Light microscopic observation of alterations in staining of the zona pellucida of porcine follicular oocytes: possible early indication of atresia. Gamete Res. 6, 293–304.CrossRefGoogle Scholar
Cross, N.L., Morales, P., Overstreet, J.W. & Hanson, F.W. (1986). Two simple methods for detecting acrosomereacted human sperm. Gamete Res. 15, 213–26.CrossRefGoogle Scholar
de Loos, F., van Vliet, C., van Maurik, P.Krip, T.A.M. (1989). Morphology of immature bovine oocytes. Gamete Res. 24, 197–204.CrossRefGoogle ScholarPubMed
de Loos, F., van Maurik, P., van Beneden, T.Kruip, T.A.M. (1992). Structural aspects of bovine oocyte maturation in vitro. Mol. Reprod. Dev. 31, 208–14.CrossRefGoogle ScholarPubMed
De Sutter, P., Dhont, M., Vanluchene, E.& Vandekerckhove, D. (1991). Correlation between follicular fluid steroid analysis and maturity and cytogenetic analysis of human oocytes that remained unfertilized after in vitro fertilization. Fertil. Steril. 55, 958–63.CrossRefGoogle ScholarPubMed
Franchimont, P., Hazee-Hagelstein, B.S., Hazount, A., Frydman, R., Schatz, B. & Demerle, F. (1989). Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. I. Sex steroids. Fertil. Steril. 52, 1006–11.CrossRefGoogle ScholarPubMed
Gondos, B. (1982). Ultrastructure of follicular atresia in the rat. Gamete Res. 5, 199–206.CrossRefGoogle Scholar
Goto, k. & Iritani, A. (1992). Oocyte and maturation and fertilization. Animal Reprod. Sci. 28, 407–13.CrossRefGoogle Scholar
Gougen, A. & Testart, J. (1986). Germinal vesicle breakdown in oocytes of human atretic follicles during the menstrual cycle. J. Reprod. Fertil. 78, 389–401.CrossRefGoogle Scholar
Greenwald, G.S. & Terranova, P.F. (1988). Follicular selection and its control. In: The Physiology of Reproduction, ed. Knobil, E.Neil, J., pp. 387–445. New York: Raven Press.Google Scholar
Grimes, R.W., Matton, P.Ireland, J.J. (1987). A comparison of histological and non-histological indices of atresia and follicular function. Biol. Reprod. 37, 82–8.CrossRefGoogle ScholarPubMed
Gupta, S.K., Gilbert, A.B. & Walker, M.A. (1988). Histological study of follicular atresia in the ovary of the domestic hen (Gallus domesticus). J. Reprod. Fertil. 82, 291–25.Google ScholarPubMed
Hartshorne, G.M. (1989). Preovulatory follicle fluid: relationship to ovarian stimulation protocol, fertilization and sperm penetration in vitro. Fertil. Steril. 48, 430–4.Google Scholar
Henderson, K.M., McNeilly, S.A. & Swanston, A.I. (1982). Gonadotrophin and steroid concentrations in bovine follicular fluid and their relationship to follicle size. J. Reprod. Fert. 65, 467–73.CrossRefGoogle ScholarPubMed
Hyttel, P., Westergaard, L. & Byskov, A.G. (1986). Ultrastructure of human cumulus-oocyte complexes from healthy and atretic follicles. Hum. Reprod. 1, 153–7.CrossRefGoogle ScholarPubMed
Kobayashi, T., Oda, T., Yoshimura, Y., Takehara, Y., Natori, M. & Nozawa, S. (1991). Androstenedione and progesterone concentrations in preovulatory follicular fluid correlate with successful fertilization and cleavage of human oocytes in vitro. Fertil. Steril. 56, 301–5.CrossRefGoogle ScholarPubMed
Kruip, T. & Dieleman, S.J. (1982). Macroscopic classification of bovine follicles and its validation by micromorphological and steroid biochemical procedure. Reprod. Nutr. Dev. 22, 465–73.CrossRefGoogle Scholar
Kruip, T., Cran, D.G., van Beneden, X. & Dieleman, S.J. (1983). Structural changes in bovine oocytes. Gamete Res. 8, 29–47.CrossRefGoogle Scholar
Kulkarni, B.A. (1988). Follicular fluid proteins of the Indian buffalo. In: Proceedings of the Second World Buffalo Congress, ed. Lokeshwar, R.R. & Kumar, A.T., vol.3, pp. 233–8.Google Scholar
Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–5.CrossRefGoogle ScholarPubMed
Lee, S.L., Kuo, Y.M., Kao, C.C., Hong, C.Y. & Wei, Y.H. (1992). Purification of a sperm motility stimulator from porcine follicular fluid. Comp. Biochem. Physiol. B 101, 591–4.Google ScholarPubMed
Lipner, H. (1988). Mechanism of mammalian ovulation. In: The Physiology of Reproduction, ed. Knobil, E.Neill, J. et al. vol.1, pp. 447–88. New York: Raven Press.Google Scholar
Lobo, R.A., diZerega, G.S. & Marrs, R.P. (1985). Follicular fluid steroid levels in dysmature and mature follicles from spontaneous and hyperstimulated cycles in normal and anovulatory women. J. Clin. Endocrinol. Metab. 49, 851–60.Google Scholar
Martin, T.L., Fogwell, R.L. & Ireland, J.J. (1991). Concentrations of inhibins and steroids in follicular fluid during development of dominant follicles in heifers. Biol. Reprod. 44, 693–700.CrossRefGoogle ScholarPubMed
Mbizvo, M.T., Burkman, L.J. & Alexander, N.J. (1990). Human follicular fluid stimulates hyperactivation motility in human sperm. Fertil. Steril. 54, 708–12.CrossRefGoogle ScholarPubMed
Mendoza, C. & Tesarik, L. (1990). Effect of follicular fluid on sperm movement characteristics. Fertil. Steril. 54, 1135–9.CrossRefGoogle ScholarPubMed
Miller, K.F., Xie, S. & Pop, W.F. (1991). Immunoreactive inhibin in follicular fluid is related to meiotic stage of the oocyte during final maturation of the porcine follicle. Mol. Reprod. Dev. 28, 35–9.CrossRefGoogle ScholarPubMed
Mukhopadhyay, A.K., Holstein, K., Szkudlinski, M., Brunswing-Spickenheier, B. & Leidenberger, F.A. (1991). The relationship between prorenin levels in follicular fluid and follicular atresia in bovine ovaries. Endocrinology 129, 2367–75.CrossRefGoogle ScholarPubMed
O'Farrell, P.H. (1975). High resolution two-dimensional electrophoresis of protein. J. Biol. Chem. 250, 4007–21.CrossRefGoogle Scholar
Pavlok, A., Lucas-Hahn, A. & Niemann, H. (1992). Fertilization and developmental competence of bovine oocytes derived from different categories of antral follicles. Mol. Reprod. Dev. 31, 63–7.CrossRefGoogle ScholarPubMed
Peluso, J.J., England-Charlesworth, C., Bolender, D.L. & Steger, R.W. (1980). Ultrastructural alterations associated with the initiation of follicular atresia. Cell Tissue Res. 211, 105–15.CrossRefGoogle ScholarPubMed
Phillips, D.M. & Dekel, N. (1991). Maturation of the rat cumulus–oocyte complex: structure and function. Mol. Reprod. Dev. 28, 297–306.CrossRefGoogle ScholarPubMed
Ralt, D., Goldenberg, M., Fetterolf, P., Thompson, D., Dor, J., Mashiach, S., Garbers, D.L. & Eisenbach, M. (1991). Sperm attraction to a follicular factor(s) correlates with human fertilizability. Proc. Natl. Acad. Sci. USA 88, 2840–4.CrossRefGoogle ScholarPubMed
Sato, E., Miyamoto, H. & Koide, S.S. (1990). Glycosaminoglycans in porcine follicular fluid promoting viability of oocytes in culture. Mol. Reprod. Dev. 26, 391–7.CrossRefGoogle ScholarPubMed
Shalgi, R., Kraicer, P., Rimon, A., Pinto, M. & Soferman, N. (1973). Proteins of human follicular fluid: the blood–follicle barrier. Fertil. Steril. 24, 429–34.CrossRefGoogle ScholarPubMed
Sitieri, J.E., Gottleb, W. & Meizel, S. (1988). Partial characterization of a fraction from human follicular fluid that initiates the human sperm acrosome reaction in vitro. Gamete Res. 20, 25–42.CrossRefGoogle Scholar
Suarez, S.S., Wolf, P.D. & Meizel, A. (1986). Induction of the acrosome reaction in human spermatozoa by a fraction of human tollicular fluid. Gamete Res. 14, 107–21.CrossRefGoogle Scholar
Talevi, R., Bergamo, P., Venditti, P., Sansone, G. & Abrescia, P. (1992). II fluido follicolare induce il rilascio di rivestimento e la reazione acrosomiale in umani. Atti del X congresso Nazionale ABCD Bari 57.Google Scholar
Totey, S.M., Singh, G., Taneja, M. & Talvar, C.P. (1991). In vitro maturation and fertilization of follicular oocytes from buffalo. Theriogenology 35, 284.CrossRefGoogle Scholar
Tsafriri, A. & Braw, R.H. (1984). Experimental approaches atresia in mammals. Oxford Rev. Reprod. Biol. 6, 226–65.Google ScholarPubMed
Vanluchene, E., Hinting, A., Dhont, M., de Sutter, P., Maele, G. & Vandekerckhove, D. (1991). Follicular fluid steroid levels in relation to oocyte maturity and in vitro fertilization. J. Steroid Biochem. Mol. Biol. 38, 83–7.CrossRefGoogle ScholarPubMed
Watson, E.D., Sertich, P.L. & Zanecosky, H.G. (1991). Detection of chemotactic factors in preovulatory follicular fluid from mares. Am. J. Vet. Res. 52, 1412–15.CrossRefGoogle ScholarPubMed
Westergaard, L. (1985). Follicular atresia in relation to oocyte morphology in non-pregnant and pregnant women. J.Reprod. Fertil. 74, 113–18.CrossRefGoogle ScholarPubMed
Wise, T. (1987). Biochemical analysis of bovine follicular fluid: albumin, total protein, lysosomal enzymes, ions, steroids and ascorbic acid content in relation to follicular size, rank, atresia classification and day of estrous cycle. J.Anim.Sci. 64, 1153–69.CrossRefGoogle ScholarPubMed
Yanagimachi, R. (1969). In vitro capacitation of hamster spermatozoa by follicular fluid. J. Reprod. Fertil. 19, 275–6.CrossRefGoogle Scholar