An intrabursal injection procedure for the in vivo study of ovulation in hamsters

Martin, Gary G.; Talbot, Prudence; Pendergrass, Paula B.

doi:10.1002/jez.1402160315

Cited by 22 publications

(13 citation statements)

References 15 publications

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“…One hour before hCG administration, mice (n = 209) received intrabursal (i.b.) injections, per ovary, of 2 pIl PBS containing 1, 10, 50, 100, 200, or 500 ng rhTGF[ 1 and 3 mg/ml BSA (fraction V) used as carrier [32,49]. TGF3 1 was administered either into both ovaries, in which case results were compared to vehicle-treated controls (group I), or into one ovary, in which case the contralateral ovary served as the vehicletreated control (group II).…”

Section: Tgf/31 Administrationmentioning

confidence: 99%

Ovarian Intrabursal Administration of Transforming Growth Factor β1 Inhibits Follicle Rupture in Gonadotropin-Primed Mice1

Juneja

Chegini

Williams

et al. 1996

Biology of Reproduction

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We hypothesized that over-expression and/or activation of latent transforming growth factor betas (TGF betas) by various ovarian cell types may lead to disturbances in ovulation and fertilization. To test this hypothesis, active TGF beta ranging from 1 to 500 ng was administered intrabursally into the ovaries of gonadotropin-primed mice, and the rates of ovum recovery and fertilization were determined. Furthermore, the presence and cellular distribution of endogenous TGF betas and TGF beta type I and type II receptors were determined immunohistochemically in the ovarian tissues of TGF beta 1-treated and untreated groups. The total number of ova recovered per ovary from ovaries treated as pairs or treated singly with TGF beta 1 at 1 to 10 ng/ovary was similar to that from controls, whereas the number recovered from ovaries treated as pairs or singly with 50 or 100 ng of TGF beta 1 per ovary was significantly lower than the number from respective controls (p < 0.05, 0.001). The number of ova recovered per ovary from ovaries treated as pairs or singly with TGF beta 1 at 200 or 500 ng/ovary was similar to the number of ova obtained from ovaries treated with TGF beta 1 at 100 ng/ovary. The rate of in vitro fertilization was low in ova recovered from ovaries treated with 50, 100, 200, and 500 ng/ovary of TGF beta 1, compared to that in ova from untreated ovaries. Histologically, the TGF beta 1-treated ovaries contained large numbers of unruptured follicles, whereas untreated ovaries contained large numbers of corpora lutea. Immunohistochemically, the endogenous TGF beta 1 and TGF beta 2 was localized in theca, granulosa, and luteal cells, without a substantial difference in intensity or distribution, in both TGF beta 1-treated ovaries and in controls. Theca cells were the primary site of immunoreactive TGF beta protein. TGF beta type I and type II receptors were also present in these cells, and their relative immunoreactive intensity was considerably reduced, particularly in granulosa cells in TGF beta 1-treated ovaries compared to controls. The results support our hypothesis and suggest that TGF betas play an important regulatory role in follicular development, oocyte maturation, and the ovulatory process.

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Section: Tgf/31 Administrationmentioning

confidence: 99%

Ovarian Intrabursal Administration of Transforming Growth Factor β1 Inhibits Follicle Rupture in Gonadotropin-Primed Mice1

Juneja

Chegini

Williams

et al. 1996

Biology of Reproduction

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“…injection of 0.3-0.4 ml of Nembutal (50 mg/ml solution of sodium pentobarbital; Abbott, N. Chicago, IL) delivered gradually. Surgical procedures and injection into the right oviduct were performed using a sterile technique as described previously [46]. A total of 125 or 250 g of IL2 IgG, 250 g of preimmune IgG, or 250 g of the IgG fraction of an antibody against the zona pellucida glycoprotein ZP1 (obtained by courtesy of Dr. Jurrien Dean at National Institutes of Health, Bethesda, MD) in 50 l of DPBS was injected into the oviduct via the infundibulum using a 1-ml tuberculin syringe fitted with a 30-gauge/1.27-cm needle.…”

Section: In Vivo Functional Studiesmentioning

confidence: 99%

Relationship Between p62 and p56, Two Proteins of the Mammalian Cortical Granule Envelope, and Hyalin, the Major Component of the Echinoderm Hyaline Layer, in Hamsters1

Hoodbhoy

Carroll

Talbot

2000

Biology of Reproduction

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Mammalian cortical granules contain two polypeptides (p62 and p56) that are incorporated into the cortical granule envelope after fertilization and function in cleavage of the zygote and the preimplantation blastomeres. Since the echinoderm hyaline layer and mammalian cortical granule envelope are analogous, and since the hyaline layer protein, hyalin, functions in early echinoderm embryogenesis, this study was done to determine whether p62 and p56 and/or other components of the mammalian cortical granule envelope are related to hyalin. A polyclonal antibody (IL2) against purified S. purpuratus hyalin was shown by confocal scanning laser microscopy to bind to hamster cortical granules and to the cortical granule envelope of fertilized hamster oocytes and preimplantation embryos up to the blastocyst stage. In immunoblots, IL2 bound only to 62- and 56-kDa cortical granule proteins that were incorporated into the cortical granule envelope after fertilization. IL2 binding antigens appeared to be resynthesized by preimplantation embryos starting at the 2-cell stage of development. In vivo treatment of 2-cell-stage hamster embryos with IL2 inhibited blastomere cleavage, but treatment of morulae did not inhibit blastocyst implantation. These results support the idea that the mammalian cortical granule envelope proteins, p62/p56, share a common antigenic epitope(s) with echinoderm hyalin, and that p62/p56, like hyalin, play a role in early embryogenesis.

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“…Forty microliters of tyrphostin dissolved in PBS or vehicle were injected into the bursal cavity of the ovary with a 30 gauge needle attached to a Hamilton syringe (Hamilton Co., Reno, NV, USA). The ovary was gently replaced in the EUROPEAN JOURNAL OF ENDOCRINOLOGY (1998) 138 peritoneal cavity, and the abdominal wall was closed (20). The left ovary received no treatment and served as a control.…”

Section: Intrabursal Injection Technique and Oocyte Retrievalmentioning

confidence: 99%

The role of tyrosine kinase in gonadotropin-induced ovulation in the rat ovary

Shimamoto¹,

Yamoto²,

Nakano³

1998

European Journal of Endocrinology

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Objectives: Our purpose was to elucidate the involvement of the tyrosine kinase pathway in gonadotropin-induced ovulation in the rat ovary. Study design: We investigated the effect of a tyrosine kinase inhibitor, tyrphostin, on the rat ovulatory process in vivo and in vitro. Methods: In cultured rat granulosa cells, the effect of tyrphostin on LH-, dibutyryl cyclic AMP ((Bu) 2 cAMP)-or forskolin-stimulated tissue type plasminogen activator (tPA) activities was examined by using a fibrin autography technique. In an in vivo system, tyrphostin was injected into the bursal cavity of the ovary in pregnant mare serum gonadotropin-treated rats, just before human chorionic gonadotropin administration. After 24 h, the number of oocytes in the oviduct was counted and the tyrphostin-treated ovaries were examined histologically. Results: Tyrphostin inhibited LH-stimulated tPA activity but did not affect (Bu) 2 cAMP-or forskolinstimulated ones. In an in vivo study, tyrphostin suppressed oocyte release dose-dependently. Histological observations revealed that tyrphostin-treated ovaries contained many large unruptured follicles and a few corpora lutea. Conclusion: This study suggests that the suppressive effect of tyrphostin on ovulation may be partly due to tPA activity inhibition in the granulosa cells via the suppression of tyrosine kinase activity. Additionally, tyrosine kinase phosphorylation may be involved in gonadotropin-activated signaling systems in the rat ovulatory process.

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An intrabursal injection procedure for the in vivo study of ovulation in hamsters

Cited by 22 publications

References 15 publications

Ovarian Intrabursal Administration of Transforming Growth Factor β1 Inhibits Follicle Rupture in Gonadotropin-Primed Mice1

Ovarian Intrabursal Administration of Transforming Growth Factor β1 Inhibits Follicle Rupture in Gonadotropin-Primed Mice1

Relationship Between p62 and p56, Two Proteins of the Mammalian Cortical Granule Envelope, and Hyalin, the Major Component of the Echinoderm Hyaline Layer, in Hamsters1

The role of tyrosine kinase in gonadotropin-induced ovulation in the rat ovary

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