Department of Histology and Embryology, Pomeranian Medical University in Szczecin, Poland

SUMMARY
Spermatogenesis is a complex series of cellular changes leading to the formation of haploid male gametes (spermatozoa) and includes mitotic, meiotic and post-meiotic phases. Spermatogonial stem cells (SSCs) are essential for the continuous lifelong production of spermatozoa. Spermatogenesis is initiated when SSC is triggered to undergo mitosis that gives rise to progenitors, which further differentiate into spermatogonia. In this review, we describe the origin of SSCs and other spermatogonia populations and summarize the knowledge concerning their markers.

Reproductive Biology 2012 12 1: 5-23.

¹ Corresponding author: Department of Histology and Embryology, Pomeranian Medical University in Szczecin, 72 Powstańców Wielkopolskich Str., 70-111 Szczecin, Poland; e-mail: ak1712@pum.edu.pl

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Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Suwon, Republic of Korea

SUMMARY
The objective of this study was to examine the effect of different sera and serum-like substances on the preimplantation development of porcine parthenogenetic embryos. Chemically activated (calcium ionophore A23187+cytochalasin B) pig oocytes were pre-cultured for five days. On day 5, the parthenogenetic embryos were treated with porcine follicular fluid (PFF), fetal bovine serum (FBS), horse serum (HS) or porcine serum albumin (PSA), and were cultured two more days. Horse serum was found to be the most effective protein source in enhancing parthenote development judging by blastocyst formation and hatching. Next, three different concentrations of HS (10, 20 and 30%) were used to determine the optima HS concentration needed to improve the development of porcine parthenogenetic embryos. All HS concentrations increased the blastocyst cell number and decreased the incidence of blastocyst apoptotic cells with 20% being the most effective. In conclusion, horse serum enhanced parthenogenetic embryo development and the quality of porcine parthenogenetic embryos.

Reproductive Biology 2012 12 1: 25-39.

¹ Corresponding author: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Suwon, 441-706, Republic of Korea; e-mail: vetjack@korea.kr

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² Federal University of Minas Gerais, Belo Horizonte,

³ Embrapa Dairy Cattle Research Center, Juiz de Fora,

⁴ Federal University of Juiz de Fora, Juiz de Fora, Brazil

SUMMARY
Follicular atresia is a key event in the selection of the ovulatory follicles and occurs during all developmental stages. The aims of the study were to evaluate the follicular population as well as the rates of follicular recruitment and atresia in different strains of mice. Ovaries were obtained from four strains of mice: G1/ Swiss, G2/ F1 Swiss×C57BL/6, G3/ inbred strain C57BL/6, and G4/ F1 C57BL/6×Swiss. All mice used in the study were 60 days old. Ovaries collected from the mice were fixed and processed for histological analysis. The G2 ovaries were also used to examine immunolocalization of active caspase-3. The pimordial follicle population was smaller in G3 mice than in G1, G2 and G4 groups (7 565±1 845 vs. 17 180±3 159, 14 785±3 319 and 13 325±2 685, respectively; p<0.05). The rate of follicular recruitment in G3, however, was higher than in the other groups (29.2% vs. 18.2%, 17.3% and 13.0% in G1, G2 and G4, respectively; p<0.05), resulting in a similar (p>0.05) number of antral follicles among groups. The small follicular pool in G3 mice was also associated with a lower rate of follicular atresia (11.4% vs. 17.2%, 16.7% and 13.6% for G3, G1, G2 and G4, respectively; p<0.05). The number of follicles stained with active caspase-3 was higher (p<0.05) during the final stage of preantral folliculogenesis than in other stages of follicular development suggesting that apoptosis in mice occurs earlier in comparison to large animals. Thus, it was concluded that differences in follicle reservoir among mice strains are compensated by an increased rate of follicular recruitment and a decreased rate of follicular atresia; and atresia occurs in mice mainly at the end of the preantral stage of folliculogenesis.

Reproductive Biology 2012 12 1: 41-55.

¹ Corresponding author: Embrapa Dairy Cattle Research Center, R. Eugenio do Nascimento 610, Juiz de Fora, MG, 36038-330 Brazil; e-mail: jhmviana@cnpgl.embrapa.br

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² Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences;

³ Department of Biochemistry and Molecular Biology, Poznan University of Medical Sciences;

⁴ Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland

SUMMARY
Protamine 1 (PRM1) and 2 (PRM2) are major nuclear proteins in spermatozoa known to bind to chromatin during early spermatogenesis. The aim of this study was to evaluate the impact of protamine transcripts and proteins on human spermatozoa and their fertilization ability as well as the development of preimplantation embryos. Oocytes selected from 92 patients were fertilized in vitro (IVF) using their partners’ sperm after evaluating its concentration, motility and morphology. Reverse transcription and real-time quantitative PCR (RQ-PCR) were applied to determine protamine mRNA level, while sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were used to quantify the protamine protein concentration. Next, protamine mRNA and protein levels were correlated with sperm concentration, motility and morphology as well as with the fertilization ability of mature spermatozoa and embryos of different quality obtained from the IVF program. A significant correlation has been found between protamine transcripts/proteins and: sperm concentration, progressive sperm motility, sperm fertilization ability as well as embryo quality. Protamine transcripts and proteins contribute to the quality of spermatozoa, successful fertilization and may have a significant influence on the development of preimplantation embryos.

Reproductive Biology 2012 12 1: 57-72.

¹ Corresponding author: Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences, 33 Polna Str., 60-535 Poznan, Poland; e-mail: magdzik_24@o2.pl

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SUMMARY
In this paper we report for the first time the results of histology of the golden loach (S. baltica) gonads (25 females and 8 males) and absolute fecundity of females from the Bug River during the reproductive season. The golden loach has an asynchronous ovary and spawns in batches. The absolute fecundity of the golden loach ranged from 1507 to 7220 eggs (3050±1377). We hypothesize that the golden loach spawns twice a year.

Reproductive Biology 2012 12 1: 73-79.

¹ Corresponding author: Department of Zoology University of Warmia and Mazury, Olsztyn, Oczapowskiego 5, Poland; e-mail: juchno@uwm.edu.pl

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² Department of Animal Physiology, University of Warmia and Mazury, Olsztyn;

³ Department of Public Health and Epidemiology, University of Warmia and Mazury, Olsztyn;

⁴ Department of Oncology, MSWiA Hospital in Olsztyn;

⁵ Department of Human Physiology, University of Warmia and Mazury, Olsztyn, Poland

SUMMARY
The aim of this study was to investigate the expression of aquaporins (AQPs) in uterine tissues of premenopausal women. We demonstrated for the first time the expression of AQP1, AQP5 and AQP9 in uterine leiomyomata and in the adjacent normal endometrium and myometrium. The expression of AQP1 and 5 was higher in leiomyomata than in unaffected uteri. AQP9 was expressed only in the unaffected endometrium. It is possible that AQP1 and AQP5 contribute to the formation of leiomyomata in premenopausal women.

Reproductive Biology 2012 12 1: 81-89.

¹ Corresponding author: University of Warmia and Mazury in Olsztyn, Department of Animal Physiology, Oczapowskiego 1A, 10-718 Olsztyn, Poland; e-mail: skowron@uwm.edu.pl

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ART Audits Australasia, Northbridge, New South Wales, Australia

SUMMARY
Accreditation of laboratories who perform diagnostic semen analysis in Australia and New Zealand is a requirement of the healthcare system. Within the accreditation process laboratories are required to set ISO standards within their policies and procedures. In order to achieve their aims, laboratories need to be able to measure a number of defined semen parameters both accurately and repetitively, especially around the lower limit of the reference intervals. The methods documented in the WHO-manual are used almost universal as the laboratory standard. Some laboratories incorporate minor method variations into their procedures. As part of the ISO requirements all variations require validation using internally approved processes that are documented and that incorporate appropriate statistical analysis and comparison of results. Validation is an ongoing process and regular review is essential. Evidence of the validation must be available for review by external auditors during accreditation. Where any validated variant method returns results that are significantly different to any method within the WHO-manual, the laboratory needs to develop its own, in-house reference interval for that method.

Reproductive Biology 2011 Suppl. 3: 5-15.

¹ Address for correspondence: PO Box 729, Joondalup, WA 6919, Australia; e-mail: ipik8058@bigpond.net.au

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School of Veterinary Studies, Murdoch University, Murdoch, Australia

SUMMARY
This article traces the history of the work on mammalian embryos carried out by the author, his students and related scientists. It traces the work from the initial experiments at the King Ranch Laboratories, University of Pennsylvania in 1966, the set up of an embryo culture laboratory at the Department of Veterinary Physiology, University of Sydney and the work within that laboratory from 1967 to 1974. This is followed by an account of the author’s subsequent work at Murdoch University from 1975 till his retirement in 1996. The significant role of some of the author’s graduate students in human IVF is also documented.

Reproductive Biology 2011 Suppl. 3: 17-29.

¹ Address for correspondence: Professor Emeritus, School of Veterinary and Biomedical Sciences, Murdoch University, 90 South Street, Murdoch, Western Australia 6150, Australia; e-mail: ray.wales@bigpond.com

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PIVET Medical Centre, Perth, Western Australia, and Cairns Fertility Centre, Cairns, Queensland, Australia

SUMMARY
This invited presentation is intended to cover clinical developments in the evolution of assisted reproductive technology (ART), a process which was attempted during the 1940’s and 50’s and culminated in the first fruition in 1978. The first in vitro fertilisation (IVF) child ensued following the partnership by a scientist with a focussed ambition (Nobel laureate Robert Edwards) joining with the gynaecologist who introduced laparoscopy to Britain in the late 60’s (Patrick Steptoe). My journey commenced in 1976 as a clinician who became immersed in the embryological and endocrinological science, whence most progress in ART emanates, and continued into a medical directorship position from which this personal view is documented. Several clinical advances have been important developments in the understanding and management of sub-fertile patients. However evolution of the various laboratory sciences has been the major key essential to meeting both the immediate as well as the long-term needs for human reproduction. The future requires a much better understanding and control over gametogenesis and a laboratory process which much more closely duplicates intrinsic reproductive physiology, avoiding gamete and embryo exposure to the atmosphere.

Reproductive Biology 2011 Suppl. 3:31-42.

¹ Address for correspondence: Pivet Medical Centre, 166-168 Cambridge Street, Leederville WA 6007, Australia; e-mail: jlyovich@pivet.com.au

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² School of Animal Biology (M085), Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, Australia;

³ UWA Institute of Agriculture (M082), The University of Western Australia, Crawley, Australia

SUMMARY
For the emu, where monogamous mating is normal, artificial insemination (AI) promises much faster genetic improvement and a considerable reduction in production costs by reducing the number of male birds needed for mating. Semen collection is now a routine procedure so the next step is to develop successful protocols for sperm storage. In this paper, we briefly overview our recent progress on the development of protocols for liquid storage and cryopreservation of emu spermatozoa. We have shown that emu semen can be stored at 10°C for up to 48 h with a minimal loss of viability, and that cryopreservation with dimethylacetamide (DMA) as a cryoprotectant is feasible because we have observed no adverse effects of this cryoprotectant on the emu sperm membrane integrity, morphology and motility. We now need to establish the predictability of the various tests in vivo, but the proportions of live normal and motile sperm with good egg membrane penetration potential suggest that acceptable numbers of competent sperm are preserved and that this will be sufficient for AI.

Reproductive Biology 2011 Suppl. 3: 43-49.

¹ Corresponding author: School of Animal Biology M085, Faculty of Natural & Agricultural Sciences, The University of Western Australia, Crawley, Australia. e-mail: irek.malecki@uwa.edu.au

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