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VOLUME 6, NUMBER 2
ABSTRACTS
Human fertility protein PUMILIO2 interacts in vitro with testis mRNA encoding Cdc42 effector 3 (CEP3)
Anna Spik2, Slawomir Oczkowski3, Agata Olszak2, Piotr Formanowicz3,4, Jacek Blazewicz3,4, Jadwiga Jaruzelska 1,2
2Institute of Human Genetics, Polish Academy of Science, Poznan, Poland;
3Institute of Computing Science, Poznan University of Technology, Poland;
4Institute of Bioorganic Chemistry Polish Academy of Science, Poznan, Poland
Summary
PUMILIO protein regulates translation of specific mRNAs in morphogenesis and in development
of the germ-line of model organisms such as flies and worms. Given that a human homologue
(PUMILIO2) was recently identified in the germ-line stem cells, the question was raised
whether it regulates translation of fertility mRNAs similarly to Drosophila Pumilio. Here,
we describe a candidate mRNA encoding Cdc42 effector protein 3 (CEP3), however, a function
for this protein in reproduction has previously not been reported. We detected three CEP3
transcripts in the testis tissue including one which was highly expressed and testis specific
by northern blotting. We found that CEP mRNA contains GUUGU (A) and AUUGUA (B) motifs (ABB)
within the 3' untranslated region (3'UTR), which are also present in mRNA targets of Pumilio
in Drosophila. Interaction of PUMILIO2 with the fragment of CEP3 transcript containing the
ABB array was tested by mobility shift assay and we found that PUMILIO2 binds the 3'
untranslated region of the CEP3 mRNA. These results support the hypothesis that CEP3 mRNA
may be a target of PUMILIO2 protein in the human male gonad and be under translational
control mediated by specific nucleotide motifs within the 3'UTR.
Reproductive Biology 2006 6 (2): 103-113
1Institute of Human Genetics Polish Academy of Sciences, ul. Strzeszynska 32, 60-479 Poznan, Poland;
e-mail: jaruzjad@man.poznan.pl
Effects of oxytocin alone and in combination with selected hypothalamic hormones on ACTH, β-endorphin, LH and PRL secretion by anterior pituitary cells of cyclic pigs
Genowefa Kotwica1, Jaroslaw Staszkiewicz, Mariusz T. Skowronski, Gabriela Siawrys, Iwona Bogacka, Anita Franczak, Beata Kurowicka, Bartlomiej Krazinski, Stanislaw Okrasa
Department of Animal Physiology, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland
Summary
Oxytocin (OT) is involved in the stimulation of secretion of anterior pituitary hormones in
females during the periovulatory and periparturient periods. In the present study we examined
the role of OT in control of ACTH, β-endorphin, LH and PRL secretion in vitro from dispersed
anterior pituitary cells collected from gilts during the luteal (Days 10-12; n=6) and follicular
(Days 18-20; n=5) phases of the estrous cycle. Isolated anterior pituitary cells (1x106/ml)
were transferred into 24-well plates, separately for each animal, and were pre-incubated for
three days at 37°C in atmosphere of 5% CO2 and 95% air. The cells which attached to the dishes
were incubated (3.5 h, 37°C) in McCoy's medium in the absence (control) or in the presence
of the following factors: CRH alone (10-10, 10-9, 10-8, 10-7M), OT alone
(10-8, 10-7, 10-6M),LVP alone (10-7M), OT (10-7M) plus CRH
(10-9M) and LVP (10-7M) plus CRH (10-9M) for
studying ACTH and β-endorphin secretion; OT alone (10-8, 10-7, 10-6M), GnRH alone (100 ng/ml),
CRH alone (10-9M), OT (10-7M) plus GnRH (100 ng/ml) and OT (10-7M) plus CRH (10-9M)
for studying LH and PRL secretion. Concentrations of the studied hormones in media were
analyzed by RIA. Oxytocin alone increased ACTH (at doses 10-7, 10-6M), β-endorphin (at
dose 10-8M), LH (at dose 10-8M) and PRL (at doses 10-7, 10-6M) secretion by pituitary
cells isolated only from luteal-phase gilts. None of the studied hormone concentrations
in the medium was increased in response to OT when pituitary cells of follicular-phase
gilts were examined. Oxytocin in combination with CRH exerted an additive effect on β-endorphin
secretion during the luteal phase. Summarizing, in the present study the stimulatory
effect of oxytocin on ACTH, β-endorphin, LH and PRL secretion by pituitary cells isolated
from gilts during the luteal phase was demonstrated. However, the cells collected from
follicular-phase gilts appeared to be unresponsive to OT. Moreover, interaction between OT and
CRH in affecting β-endorphin secretion was shown. These results suggest that OT may be
transiently involved in the modulation of anterior pituitary hormone secretion in cyclic pigs.
Reproductive Biology 2006 6 (2): 115-131
1Corresponding author: Department of Animal Physiology, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn-Kortowo, ul. Oczapowskiego 1A, Poland; e-mail:
e-mail: gkotwica@uwm.edu.pl
Age, reproduction and fecundity of the spined loach Cobitis taenia L. (Pisces, Cobitidae) from Lake Klawoj (Poland)
Dorota Juchno1, Alicja Boron
Department of Zoology, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland
Summary
This is the first study concerning the features of the reproduction process of the karyologically
identified spined loach C. taenia (2n=48). The histology of 71 ovaries, and gonadosomatic index
(GSI) of karyologically identified spined loach Cobitis taenia L. from Lake Klawoj (Northern Poland)
were examined. The absolute and relative fecundity of 25 females was estimated by gravimetric method.
The age of fish was determined according to the annual increments of otholits. The spawning of C. taenia
from Lake Klawoj took place from May to July, at a water temperature exceeding 18.5°C. The GSI values
at the beginning of the reproduction period ranged from 7 to 19%. The average absolute fecundity of
females was 2078 eggs, with the number ranging from 869 to 3371 eggs. High individual variability
in the gonad histology and the GSI values during the reproductive period was observed. Such variability
could be the result of beginning the reproduction process in the fish at various times and, probably,
due to the various numbers of batches laid and various numbers of eggs per batch.
Reproductive Biology 2006 6 (2): 133-148
1Corresponding author: Department of Zoology, University of Warmia and Mazury, ul. Oczapowskiego 5, 10-957 Olsztyn, Poland;
e-mail: juchno@uwm.edu.pl
Retrograde transfer of testosterone to the porcine ovary in follicular and luteal phase of the estrous cycle in vivo
Barbara Wasowska1, Stanislawa Stefanczyk-Krzymowska
Division of Reproductive Endocrinology and Pathophysiology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland
Summary
This study was designed to determine the efficiency and rate of testosterone (T) retrograde
transfer during the follicular and luteal phase of the estrous cycle in gilts (n=27).
The efficiency and the rate of the retrograde transfer of T from the ovarian effluent
into blood supplying the ovary were determined for the first time under in vivo conditions.
Ovarian arterial blood concentration of T was higher than that in systemic blood during both,
the follicular phase (p<0.01) and luteal phase (p<0.0001). The efficiency of the retrograde
transfer of T into ovarian arterial blood was not dependent on concentration of testosterone
in the ovarian venous blood. However, the efficiency and the rate of the retrograde
transfer of T differed between phases of the estrous cycle. The presented results suggest that
local retrograde transfer enriched the porcine ovary with an additional amount of T which may
affect the ovarian secretory function.
Reproductive Biology 2006 6 (2): 149-159
1Corresponding author: Division of Reproductive Endocrinology and Pathophysiology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-747 Olsztyn, ul. Tuwima 10, Poland;
e-mail: bwasow@pan.olsztyn.pl
Oogenesis in pig ovaries during the prenatal period: ultrastructure and morphometry
Zofia Bielanska-Osuchowska1
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Department of Histology and Embryology, Warsaw Agricultural University, Warsaw, Poland
Summary
Oogenesis in fetal pig ovaries comprises the successive changes from the primordial germ
cells to the dictyotene oocytes in primordial ovarian follicles. In this study the
observations were carried out with an electron microscope and stereological analysis
was performed. At the ultrastructural level there are no differences between the primordial
germ cells and oogonia, but oogonia are connected with the intercellular bridges.
The onset of the dictyotene phase was accompanied by the changes in the cytoplasm of oocytes.
Near the nucleus, the yolk nucleus is formed containing numerous Golgi bodies,
endoplasmic reticulum (ER), mitochondria and granules. ER proliferates in contact with
the external leaflet of the nuclear envelope forming the narrow ER cisterns. Between
the nuclear envelope and ER cisterns, the vesicles with grey content are visible. The
proliferating ER forms numerous concentric cisterns around the nucleus. Next, the most
external cisterns fragment, detach, and then form the cup-like structures. These
structures separate the distinct areas of cytoplasm-compartments, which contain mitochondria,
ribosomes and lipid droplets. The cells of cortical sex cords of the ovary, which
encloses the oocyte, form the follicles. The volume of oocytes in forming follicle
increases due to the increase in the number of the cell inclusions: lipid droplets,
vacuoles and yolk globules. In the oocytes of primordial ovarian follicles, the
compartments are transformed into the yolk globules, which are encountered by a sheath
of ER cisterns and the grey vesicles; they contain the mitochondria, lipid droplets and
light vacuoles. The role of the compartments and yolk globules as metabolic units is
discussed in comparison with similar structures of the mature eggs of pigs and other
mammal species.
Reproductive Biology 2006 6 (2): 161-193
1Corresponding author: Department of Histology and Embryology, Warsaw Agricultural University, ul. Nowoursynowska 159, 02-776 Warszawa, Poland;