13 research outputs found
Placental Origin of Prostaglandin F 2 α
In the present study, the question was addressed whether the feline placenta can synthesize prostaglandin F2 α (PGF2 α ). The PGFS protein was elevated, particularly at 2.5-3 weeks of pregnancy compared to 7-8 (P < 0.05) and 8.5-9 weeks (P < 0.001). Transcripts for PGFS were significantly upregulated at 2.5-3 weeks of pregnancy and then gradually declined towards the end of gestation (P < 0.001). Transcripts for PTGS2 were only upregulated in placentas from queens close to term (P < 0.001) compared with earlier phases. Staining of PTGS2 showed distinct positive signals in placentas obtained during the last week before labor, particularly in the strongly invading trophoblast surrounding blood vessels, and also in decidual cells. Shortly after implantation, signals for PGFS were localized in the trophoblast cells. Near term, PGFS staining was seen mainly in decidual cells. Both placental PGF2 α and plasma PGFM were elevated towards the end of pregnancy (P < 0.001) compared with earlier weeks of pregnancy. The content of PGF2 α in extracted placenta mirrored the PGFM level in plasma of pregnant females. During late gestation there is a significant increase in PGFM levels in maternal blood and of PGF2 α levels in placental tissue concomitant with an upregulation of placental PTGS2
The contribution of steroids and prostaglandins to the lifespan of corpora lutea in domestic cats and lynxes
Iberische und Eurasische Luchse zeigen einen saisonalen Monoöstrus. Nach der Ovulation findet man frisch gebildete (freshCL) und physiologisch persistierende Gelbkörper (corpora lutea, perCL). Funktionelle perCL verhindern eine Ovulation außerhalb der Zuchtsaison durch konstant erhöhte Progesteron-(P4)-Plasmawerte. Hauskatzen zeigen einen saisonalen Polyöstrus. Nach der Ovulation werden CL gebildet, deren Lebensspanne in Abhängigkeit von einer Trächtigkeit durch unterschiedliche P4-Plasmaprofile charakterisiert ist. Ziel der Dissertation war es, die Synthese und Rezeption von Steroiden und Prostaglandinen (PG) in CL von Feliden zu untersuchen, um potentiell luteotrophe und luteolytische Faktoren zu identifizieren. Während der Gelbkörperphase trächtiger und nicht-trächtiger Katzen weisen CL gleicher Histomorphologie, unabhängig vom Vorhandensein einer Trächtigkeit, ähnliche steroidogene Kapazitäten auf. Die Abnahme der CL-Funktion spiegelt sich im graduellen Verlust der Steroidbiogenese wider. Bei Luchsen ist die steroidogene Kapazität der perCL im Proöstrus herabgesetzt, aber im Metöstrus wieder verstärkt. Die steroidogene Kapazität ist demnach mit verschiedenen CL-Stadien und dem Reproduktionszyklus assoziiert. Die Synthese und Rezeption von PGE2 erfolgen bei Katze und Luchs unabhängig vom CL-Stadium und dem Reproduktionszyklus. Hohe Werte an luteotrophem PGE2 in perCL könnten für die funktionelle und strukturelle CL-Persistenz beim Luchs verantwortlich sein. Der feline CL ist zur Bindung von luteolytischem PGF2alpha fähig, jedoch ist die Kapazität zur Synthese begrenzt. Feliden weisen keine PGF2alpha-assoziierte luteale Regression in Abwesenheit einer Trächtigkeit auf. Allerdings wurden Höchstwerte an PGF2alpha in der Plazenta, wie auch im Plasma (PGFM), im letzten Trächtigkeitstrimester der Katze gemessen. Folglich ist die feline Plazenta zur Synthese von luteolytischem PGF2alpha fähig, welches die CL-Regression und Geburt am Ende der Trächtigkeit ermöglicht.Iberian and Eurasian lynxes exhibit a seasonal monooestrus. After ovulation, freshly formed (freshCL) coexist with physiologically persistent luteal bodies (corpora lutea, perCL). Functional perCL prevent ovulation outside the breeding season through constantly elevated plasma progesterone (P4) levels. Domestic cats show a seasonal polyoestrus. After ovulation, CL are built with lifespans being characterised by different plasma P4 profiles dependent on pregnancy. The aim of the dissertation was to characterise the synthesis and reception of steroids and prostaglandins (PGs) in CL of felids to identify potential luteotrophic and luteolytic factors. During the luteal lifespan of pregnant and non-pregnant cats, CL of equal histomorphology exhibit similar steroidogenic capacities, irrespectively of an ongoing pregnancy. The functional demise of CL mirrors the gradual loss of steroid biogenesis. In lynxes, the steroidogenic capacity of perCL is limited at prooestrus, but is enhanced again during metoestrus. The steroidogenic capacity is thus associated with different CL stages and the reproductive cycle. The synthesis and reception of PGE2 in cat and lynx is independent on the CL stage and reproductive cycle. High levels of luteotrophic PGE2 in perCL might be responsible for the functional and structural CL persistence in lynxes. The feline CL is capable of binding luteolytic PGF2alpha; however, the capacity to synthesise PGF2alpha is limited. Felids show no PGF2alpha-associated luteal regression in the absence of pregnancy. Interestingly, peak levels of PGF2alpha in the placenta, as well as in plasma (PGFM), were measured during the last trimester of pregnancy in the cat. Therefore, the feline placenta is capable of synthesising luteolytic PGF2alpha, which enables CL regression and parturition at the end of pregnancy
Placental Origin of Prostaglandin F2α in the Domestic Cat
In the present study, the question was addressed whether the feline placenta can synthesize prostaglandin F2α (PGF2α). The PGFS protein was elevated, particularly at 2.5–3 weeks of pregnancy compared to 7-8 (P<0.05) and 8.5–9 weeks (P<0.001). Transcripts for PGFS were significantly upregulated at 2.5–3 weeks of pregnancy and then gradually declined towards the end of gestation (P<0.001). Transcripts for PTGS2 were only upregulated in placentas from queens close to term (P<0.001) compared with earlier phases. Staining of PTGS2 showed distinct positive signals in placentas obtained during the last week before labor, particularly in the strongly invading trophoblast surrounding blood vessels, and also in decidual cells. Shortly after implantation, signals for PGFS were localized in the trophoblast cells. Near term, PGFS staining was seen mainly in decidual cells. Both placental PGF2α and plasma PGFM were elevated towards the end of pregnancy (P<0.001) compared with earlier weeks of pregnancy. The content of PGF2α in extracted placenta mirrored the PGFM level in plasma of pregnant females. During late gestation there is a significant increase in PGFM levels in maternal blood and of PGF2α levels in placental tissue concomitant with an upregulation of placental PTGS2
Apoptosis-Related Factors in the Luteal Phase of the Domestic Cat and Their Involvement in the Persistence of Corpora Lutea in Lynx.
The corpus luteum (CL) is a transient gland formed in the ovary after ovulation and is the major source of progesterone. In the Iberian and Eurasian lynx, CL physiologically persist after parturition and retain their capacity to produce progesterone, thus suppressing the ovarian activity. This unique reproductive characteristic has a big impact on the success of assisted reproduction techniques in the endangered Iberian lynx. The mechanisms behind CL persistence are not yet understood and require extensive studies on potential luteotropic and luteolytic factors in felids. Because the apoptosis system has been shown to be involved in structural regression of CL in many species, we aimed to investigate the capacity of perCL to undergo apoptosis. In addition, we performed initial studies on the apoptosis system in the luteal phase of the domestic cat. No previous research on this system has been made in this species. Our factors of interest included agents of the intrinsic apoptosis pathway, i.e., pro-survival B-cell CLL/lymphoma 2 (BCL2) and pro-apoptotic BCL2-associated X protein (BAX), the executioner caspase-3 (CASP3), as well as of the extrinsic pathway, i.e., pro-apoptotic receptor FAS, and tumor necrosis factor (TNF) and its receptors (pro-apoptotic TNFRSF1A and pro-survival TNFRSF1B). We analyzed the relative mRNA levels of these factors, as well as protein localization of CASP3 and TNF during stages of pregnancy and the non-pregnant luteal phase in CL of domestic cats. The same factors were investigated in freshly ovulated CL (frCL) and perCL of Iberian and Eurasian lynx, which were histologically analyzed. All factors were present in the CL tissue of both domestic cat and lynx throughout all analyzed stages. The presence of pro-apoptotic factors BAX, CASP3, FAS and TNFRSF1A in perCL of the Eurasian and Iberian lynx might indicate the potential sensitivity of perCL to apoptotic signals. The expression of pro-survival factors BCL2 and TNFRSF1B was significantly higher in perCL compared to frCL of studied Iberian lynx, suggesting the potential involvement of these factors in the structural integrity of perCL. In both Iberian lynx and pregnant and non-pregnant domestic cats, the expression of TNFRSF1A was significantly higher in forming CL compared to other stages, suggesting the conserved involvement of this factor in the tissue reorganization during formation of the feline CL. The mRNA levels of CASP3 and TNFRSF1B were highest during regression stages of domestic cat CL. The current study provides initial results on the possible involvement of the apoptosis system in the structure and function of the feline CL and in its physiological persistence
Synthesis and reception of prostaglandins in corpora lutea of domestic cat and lynx
Felids show different reproductive strategies related to the luteal phase. Domestic cats exhibit a seasonal polyoestrus and ovulation is followed by formation of corpora lutea (CL). Pregnant and non-pregnant cycles are reflected by diverging plasma progesterone (P4) profiles. Eurasian and Iberian lynxes show a seasonal monooestrus, in which physiologically persistent CL (perCL) support constantly elevated plasma P4 levels. Prostaglandins (PGs) represent key regulators of reproduction, and we aimed to characterise PG synthesis in feline CL to identify their contribution to the luteal lifespan. We assessed mRNA and protein expression of PG synthases (PTGS2/COX2, PTGES, PGFS/AKR1C3) and PG receptors (PTGER2, PTGER4, PTGFR), and intra-luteal levels of PGE2 and PGF2α Therefore, CL of pregnant (pre-implantation, post-implantation, regression stages) and non-pregnant (formation, development/maintenance, early regression, late regression stages) domestic cats, and prooestrous Eurasian (perCL, pre-mating) and metoestrous Iberian (perCL, freshCL, post-mating) lynxes were investigated. Expression of PTGS2/COX2, PTGES and PTGER4 was independent of the luteal stage in the investigated species. High levels of luteotrophic PGE2 in perCL might be associated with persistence of luteal function in lynxes. Signals for PGFS/AKR1C3 expression were weak in mid and late luteal stages of cats but were absent in lynxes, concomitant with low PGF2α levels in these species. Thus, regulation of CL regression by luteal PGF2α seems negligible. In contrast, expression of PTGFR was evident in nearly all investigated CL of cat and lynxes, implying that luteal regression, e.g. at the end of pregnancy, is triggered by extra-luteal PGF2α
Morphological appearance of the Iberian lynx ovary.
<p>Ovaries of the Iberian lynx 1 and 2 (Ib.lynx 1 and Ib.lynx 2, respectively), containing frCL and perCL. Black arrows indicate frCL with ovulation scars, gray arrows indicate perCL from previous cycle/s. Scale–mm.</p
Intraluteal relative mRNA levels during pregnant and non-pregnant luteal phases in the domestic cat.
<p>Box plots depict the distribution of data; whiskers indicate maximum and minimum values; black dots indicate outliers; horizontal lines indicate the median; P-values are calculated from the Kruskal-Wallis rank sum test; letters (a, b) identify significant differences between stages and are calculated from <i>post-hoc</i> pairwise comparisons (P-value adjustment: Benjamini-Hochberg). PR: pregnancy; NP: non-pregnant luteal phase; pri: pre-implantation period; poi: post-implantation period; r: regression; f: formation; d/m: development/maintenance; er: early regression; lr: late regression; CA: <i>corpus albicans</i>.</p
Immunohistochemical localization of CASP3 and TNF proteins in CL of the domestic cat.
<p>Active CASP3 was localized in luteal cells at the stages of: CL formation (A), development/maintenance (B), early regression (C) and late regression (D). The protein expression was consistently present throughout the CL life span, except for CA (E). TNF protein expression was identified in non-steroidogenic cells, presumably macrophages, but not in luteal cells (F). Arrows indicate positive staining. NC: negative control. Scale bar– 50 μm.</p
List of analyzed factors, sequences of PCR primers used for sequence analysis and expression studies, annealing temperatures, and product sizes.
<p>* GenBank sequences were obtained in this study</p><p>bp, base pair; fw, forward; rv, reverse; T<sub>A</sub>, annealing temperature; a, used for sequence analysis; b, used for gene expression studies. Intrinsic/extrinsic pathway refers to the apoptosis pathways</p
Intraluteal concentrations of estrogens and progesterone in Iberian and Eurasian lynx.
<p>Intraluteal concentrations of estrogens and progesterone in the domestic cat can be found in Amelkina <i>et al</i> [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0143414#pone.0143414.ref024" target="_blank">24</a>]</p