Porcine zonae pellucidae immunisation of tammar wallabies (Macropus eugenii): fertility and immune responses

Kitchener, Anne L.; Edds, L. M.; Molinia, F. C.; Kay, D. J.

doi:10.1071/rd01121

Cited by 28 publications

(13 citation statements)

References 37 publications

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“…Vaginal insemination 16-17 h before FSH/LH-induced ovulation resulted in a 40% fertilization rate in this species [5]. In FSH/LH-treated tammar wallabies, the fertilization rate was 45% and 70% after IUAI 4-6 h and 5-7 h, respectively, before the expected ovulation [2,3]. From these data, the optimal time of IUAI in both the brushtail possum and tammar wallaby appears to be less than 7 h before artificially induced ovulation.…”

Section: Introductionmentioning

confidence: 70%

“…In most other marsupials, however, ovulation is spontaneous and fertilization occurs long after mating [7]. After superovulation, fertilization success in the possum and tammar wallaby using AI is high [3]. However, more than 60% of embryos recovered from superovulated possums 4 days after IUAI had degenerated [2].…”

Section: Discussionmentioning

confidence: 94%

“…These egg coats appear critical during preimplantation embryo development and blastocyst expansion [11]. From the results of recent attempts at AI [2][3][4][5]32], it appears that superovulation may affect embryo viability in monovular marsupials, possibly by altering the structural integrity of the tertiary egg coats.…”

Section: Discussionmentioning

confidence: 98%

“…per female [2,3]. In natural cycles, because ovulation occurs 43-60 h p.p., birth can be used to time insemination of spermatozoa into different sites of the female reproductive tract relative to an expected ovulation and when spermatozoa are found in these regions after natural mating.…”

Section: Introductionmentioning

confidence: 99%

“…Artificial insemination (AI) has been used increasingly in wild species [1] but has only been attempted in three marsupials: the tammar wallaby (Macropus eugenii) [2,3], the brushtail possum (Trichosurus vulpecula) [2,4,5], and the koala (Phascolarctos cinereus) [6]. The only confirmed births after AI in marsupials have been in the koala, which appears to be an induced ovulator.…”

Section: Introductionmentioning

confidence: 99%

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Birth of Pouch Young after Artificial Insemination in the Tammar Wallaby (Macropus eugenii)1

Paris¹,

Taggart²,

Shaw³

et al. 2005

Biology of Reproduction

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Timing of artificial insemination (AI) in marsupials is critical because fertilization must occur before mucin coats the oocyte during passage through the oviduct. In this study, timing and the site of insemination were examined to develop AI in the tammar wallaby (Macropus eugenii). Birth and postpartum (p.p.) estrus was synchronized in 46 females. Epididymal spermatozoa (n=4) or semen collected by electroejaculation (n=42) were inseminated early (4-21 h p.p.) into the urogenital sinus (n=7), the anterior vaginal culs de sac (n=7), the uterus by transcervical catheter (n=5), or the uterus by injection (intrauterine artificial insemination, IUAI) (n=5). A further 16 females were inseminated late (19-48 h p.p.) by IUAI. All females were monitored for birth. A third group of six females was inseminated late (21-54 h p.p.) by IUAI and 0.4-6.6 h later, sperm had reached the oviduct in all animals. In total, an oocyte to which spermatozoa were attached was recovered and two young were born after IUAI using epididymal (n=1) or electroejaculated (n=2) spermatozoa, but no young resulted from insemination at other sites. Two females were successfully inseminated at 43 and 47 h p.p., later than most other animals, and the third was inseminated much earlier (18 h p.p.) but with highly motile spermatozoa. These young represent the first macropodids born by AI and the first marsupials conceived using epididymal spermatozoa.

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Section: Introductionmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Birth of Pouch Young after Artificial Insemination in the Tammar Wallaby (Macropus eugenii)1

Paris¹,

Taggart²,

Shaw³

et al. 2005

Biology of Reproduction

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Expression in Escherichia coli and immunological characterization of three zona pellucida proteins (ZP1, ZP2, and ZP3) from a marsupial, the brushtail possum (Trichosurus vulpecula)

Mate

Buist

Duckworth

2002

Molecular Reproduction Devel

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The brushtail possum (Trichosurus vulpecula) zona pellucida (ZP) is composed of three major glycoproteins, designated ZP1, ZP2, and ZP3 based on their size and homology with eutherian ZP proteins. These proteins are candidate antigens for the development of an immunocontraceptive vaccine to control the fertility of the brushtail possum in New Zealand, where it is an introduced pest. In order to further their immunological and functional characterization, recombinant possum ZP proteins were produced in Escherichia coli (E. coli) strain JM109, M15, SG13009, or BL21 codon plus. Each of the proteins produced possessed a N-terminal six histidine tag (His)(6) to facilitate purification and consisted of amino acid (aa) residues 18-471 of possum ZP1, aa residues 40-311 of ZP2 (ZP2-N), aa residues 305-634 of ZP2 (ZP2-C), and aa residues 23-342 of ZP3. Immunoblot using anti-RGS(His)(4) antibodies and polyclonal rabbit anti-porcine ZP antibodies detected major bands at 54 kDa for ZP1, 32 kDa for ZP2-N, 39 kDa for ZP2-C, and 40 kDa for ZP3. Immunization of male and female rabbits with ZP2-N, ZP2-C, and ZP3 purified on Ni-NTA resin under denaturing conditions generated antibodies reactive with recombinant ZP proteins on Western blot and with native ZP proteins in possum ovarian sections using immunofluorescence. Antibodies generated against ZP1 in the same way were reactive with recombinant ZP proteins on Western blot only. The recombinant possum ZP proteins and specific antibodies produced in this study give an indication of the antigenic relationship of the possum ZP proteins and are vital tools for future studies of sperm-ZP binding in marsupials and for the evaluation of ZP-based contraceptive vaccines in possums and other marsupials.

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Towards a ZP‐based contraceptive for marsupials: Comparative analysis and developmental expression of marsupial ZP genes

McCartney

Harris

Rodger

et al. 2007

Molecular Reproduction Devel

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Fertility control in the form of a zona pellucida (ZP)-based immunocontraceptive has shown potential as a humane form of control for overabundant marsupials including the brushtail possum and macropods. Further refinement and development of a ZP-based vaccine requires detailed knowledge of the protein structure and expression in order to ensure maximum efficacy and specificity. Sequencing and comparative analysis of the ZP3 protein from three marsupial orders in this study found a high overall level of conservation; within order Diprotodontia, the ZP3 protein is 86.9-98.9% identical. ZP3 identity falls to 56.6-57.2%, when the grey, short-tailed opossum (a Didelphimorphian) is compared to dasyurid and diprotodontan marsupials. This is similar to its amino acid identity with ZP3 from eutherian species (50.7-52.8%). Comparison of a 21 amino acid epitope in marsupial ZP3 that has shown contraceptive effects, reveals 95-100% identity between the four macropodid species, 81-86% amino acid identity between brushtail possum and the macropods and 67-71% identity between the diprotodontans and the fat-tailed dunnart (a dasyurid). This is comparable to the level of identity between related eutherian mammals. The expression pattern of three ZP genes during brushtail possum and tammar wallaby pouch young development was examined by RT-PCR. This analysis of ZP gene expression has confirmed that ZP mRNA transcription begins in the ovary during pouch young development by about 51 days of age. The presence of ZP transcripts at this stage in pouch young development suggests that marsupial ZP gene transcription begins before the onset of follicular development.

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Porcine zonae pellucidae immunisation of tammar wallabies (Macropus eugenii): fertility and immune responses

Cited by 28 publications

References 37 publications

Birth of Pouch Young after Artificial Insemination in the Tammar Wallaby (Macropus eugenii)1

Birth of Pouch Young after Artificial Insemination in the Tammar Wallaby (Macropus eugenii)1

Expression in Escherichia coli and immunological characterization of three zona pellucida proteins (ZP1, ZP2, and ZP3) from a marsupial, the brushtail possum (Trichosurus vulpecula)

Towards a ZP‐based contraceptive for marsupials: Comparative analysis and developmental expression of marsupial ZP genes

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