[Reproductive function of the rat with individual behavioral traits during maintenance in isolation and crowding].
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Animals↗
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Reproduction and ratio-production of brief duration under conditions of sensory isolation.
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Adult↗
T-cell receptor heterogeneity of gamma delta T-cell clones from human female reproductive tissues.
gamma delta T cells were isolated from human decidua parietalis, decidua basalis and cervix and cloned in the presence of interleukin-2 (IL-2). T-cell receptor (TcR) expression was then analysed and compared with that of a panel of gamma delta T-cell clones from peripheral blood. Only 17/40 (42.5%) clones from decidua parietalis were V gamma 9+/V delta 2+ as compared to 68/94 (72%) of peripheral blood clones (P < 0.005). Conversely, 50% of clones from decidua parietalis but only 15% of clones from peripheral blood were V delta 1+ (P < 0.001). At least seven distinct TcR types were identified among the panel of clones from decidua parietalis and at least six different types were expressed by the panel of 17 clones from cervix. This receptor heterogeneity was not a result of interdonor variation as in all instances where more than one clone was obtained from a single sample, individual clones having between two and five receptor types were identified. However, 23/24 (95.8%) of clones from decidua basalis were V gamma 9+/V delta 2+. Most clones from decidua parietalis and cervix, whether V gamma 9+/V delta 2+ or V delta 1+, were positive for the mucosal lymphocyte marker, HML-1, but expression was often heterogeneous within a single clone. In contrast, almost all gamma delta T-cell clones from peripheral blood were HML-1-. Thus, unlike the mouse, gamma delta T cells within these human female reproductive tissues have a diverse TcR repertoire which, in decidua parietalis, is distinct from that of peripheral blood.
Antigens, Neoplasm↗
MHC class II expression and antigen presentation by human endometrial cells.
It has been demonstrated previously that mixed cell suspensions from the female reproductive tract consisting of human epithelial and stromal cells were capable of presenting foreign antigen to autologous T cells. There have been, however, no reported studies examining antigen presentation by isolated epithelial cells from the human female reproductive tract. It is now shown that freshly isolated epithelial cells from the uterine endometrium constitutively express MHC class II antigen and that class II was upregulated on cultured epithelium by interferon gamma (IFNgamma). Using a highly purified preparation, it was demonstrated that these epithelial cells were able to process and present tetanus toxoid recall antigen driving autologous T cell proliferation. Cells isolated from the basolateral sub-epithelium stroma were also potent antigen presenting cells in this model system. Thus, isolated endometrial epithelial cells were able to directly process and present antigen to T cells and may be responsible for the transcytosis and delivery of antigen to professional antigen presenting cells found in the sub-epithelial stroma.
Adult↗
Serum immune responses to the proteins of porcine reproductive and respiratory syndrome (PRRS) virus.
The antibody responses of pigs to porcine reproductive and respiratory syndrome virus (isolate VR-2332) were evaluated by indirect immunofluorescence, virus neutralization, and immunoblotting. All pigs in each group were positive by indirect immunofluorescence 14-21 days postexposure (DPE), and antibodies to specific viral proteins (15, 19 or 26 kD) were initially demonstrated by immunoblotting at 7-21 days DPE. Neutralizing antibodies were detected in only 2 pigs that were inoculated intranasally and given additional parenteral injections with adjuvant. These antibodies appeared much later, 51-70 DPE, than did antibodies detected by indirect immunofluorescence. The titer of the neutralizing antibodies increased until 127 DPE, after which the titers decreased, and 1 animal became seronegative for neutralizing antibody by 262 DPE.
Animals↗
Expression of CCAAT-binding factor antisense transcripts in reproductive tissues affects plant fertility.
We have previously isolated a CCAAT-binding factor B subunit gene ( BnCBF-B) from Brassica napus that is widely expressed in different plant tissues and whose role is still unknown. To investigate the importance of this transcription factor subunit in plant reproductive tissues, we targeted antisense BnCBF-B transcripts to the tapetum of transgenic B. napus plants. Of the 24 independent transformants, 13 yielded reduced quantities of viable pollen, of which five were unable to produce the elongated siliques indicative of normal seed set. The decrease in pollen viability probably resulted from the precocious degeneration of the tapetal cell layer observed in these plants. Surprisingly, the male-sterile phenotype was also accompanied by a decrease in female fertility, which could be due to the expression of the antisense BnCBF-B transcripts in the female reproductive structures of the transgenic plants. These results suggest that the BnCBF-B gene plays a critical non-redundant role in plant reproductive tissues.
Antisense Elements (Genetics)↗
The role of growth hormone in fetal mouse reproductive tract differentiation.
Although GH plays a key role in postnatal growth and differentiation, its role in fetal differentiation is not clear at the present. The aim of the present study was to investigate whether GH plays a role in fetal sexual differentiation, and we used in vitro organ culture assay of sexual differentiation to determine this. The results showed that anti-rGH antibody blocked Wolffian duct differentiation specifically in the presence of fetal testes. Exogenous GH supplemented in the above experiment reversed the blocking effect of anti-GH. Among the other related products, insulin-like growth factor I was highly effective in reversing the anti-GH effect, insulin-like growth factor II was partially effective, but PRL was unable to reverse the anti-GH effect. GH itself was found to produce some masculinizing effect, as demonstrated by its ability to stabilize the Wolffian duct in female fetuses. The role of GH was further demonstrated by the observation that GH-immunoreactive material of the size of authentic GH was detected in the 18-day fetal reproductive tract, and the concentration of this material increased in response to progression of sexual differentiation. Determination of androgen-binding activity using Scatchard analysis on the cells isolated from the 18-day fetal reproductive tract indicated that androgen-binding activity increased after GH treatment of the cells. Thus, it may be concluded that GH influences male sexual differentiation and alters the androgen-binding activity of the fetal reproductive tract.
Androgens↗
Dysregulation of cytochrome P450 17alpha-hydroxylase messenger ribonucleic acid stability in theca cells isolated from women with polycystic ovary syndrome.
Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder characterized by ovarian hyperandrogenism. Theca interna cells isolated from the ovaries of women with PCOS are characterized by increased expression of cytochrome P450 17alpha-hydroxylase (CYP17) [steroid 17alpha-hydroxylase/17,20 lyase (P450c17)], a steroidogenic enzyme obligatory for the biosynthesis of androgens. Augmented expression of the gene encoding P450c17 (CYP17) in PCOS theca has been attributed, in part, to differential transcriptional regulation of the CYP17 promoter in normal and PCOS cells. The present studies examine whether CYP17 gene expression is also posttranscriptionally regulated at the level of mRNA stability in normal and PCOS theca cells maintained in long-term culture. Determination of endogenous CYP17 mRNA half-life by pharmacological inhibition of transcription demonstrated that the half-life of CYP17 mRNA increased 2-fold in PCOS theca cells, compared with normal theca cells. Forskolin treatment also prolonged CYP17 mRNA half-life in both normal and PCOS theca cells. In vitro mRNA degradation studies demonstrated that the 5'-untranslated region confers increased stability to CYP17 mRNA in PCOS theca cells and showed that the 5'-untranslated region of CYP17 also confers forskolin-stimulated stabilization of CYP17 mRNA. These studies indicate that a slower rate of CYP17 mRNA decay contributes to increased steady-state mRNA accumulation and augmented CYP17 gene expression in PCOS theca cells.
3' Untranslated Regions↗
Poliomyelitis surveillance in England and Wales, 1969-1975.
Poliomyelitis continued to be a rare disease in England and Wales in the period 1969-75. Only 31 paralytic and 44 cases of possible non-paralytic poliomyelitis were recorded during the 7 years. Of the 31 paralytic cases approximately one third were vaccine-associated; 3 were patients who had recently received oral poliovaccine and 7 had been in contact with a vaccinated person. Five of these 7 patients were parents of recently vaccinated children. The rate of vaccine-associated poliomyelitis was estimated in recipients to be 0.2 and in contacts 0.4 per million doses of vaccine given. Marker test results were reported on 555 strains of poliomyelitis virus isolated during 1969-75, using the reproductive capacity temperature test. Forty-eight (8.6%) resembled wild virus in this property, 15 strains being type 1, 8 type 2 and 25 type 3. Most of these isolations of apparently wild virus were from excreters with no symptoms of poliomyelitis, although 3 of the 15 type 1 strains were from patients with paralytic poliomyelitis and 3 from possible cases of non-paralytic poliomyelitis. None of the 8 apparently wild type 2 viruses was from a case of paralytic illness and only 1 of the 39 type 3 strains. Eleven of the 31 paralytic cases were in patients in whom the infection was likely to have been acquired abroad.
Adolescent↗
Mitochondrial heterogeneity in the parasitic nematode, Ascaris suum.
The anaerobic metabolism of Ascaris suum body wall muscle mitochondria has been well characterized, but little is known about the metabolism of other adult tissues. The present study was designed to further characterize the metabolism of mitochondria isolated from A. suum male reproductive tissues, which contain predominately sperm, and to compare it with that of muscle. Cytochrome oxidase activity could not be detected in muscle, testis, or sperm mitochondria either by diaminobenzidine staining or enzymatic assays. However, the activities of several tricarboxylic acid cycle enzymes, including citrate synthase and isocitrate dehydrogenase, were about 100-fold higher in testis/seminal vesicle mitochondria than muscle mitochondria. In contrast, malic enzyme activity in testis/seminal vesicle mitochondria was about 12-fold lower than that in muscle mitochondria. The incorporation of 32Pi into organic phosphate by either muscle or testis/seminal vesicle mitochondria appeared to be dependent on malate and pyruvate, and incorporation was inhibited by rotenone but not cyanide. Thus, the metabolism of testis/seminal vesicle mitochondrial preparations appears to be similar to that of ascarid muscle, despite the elevated levels of tricarboxylic acid cycle enzyme activities present in testis/seminal vesicle mitochondria. The function of these elevated enzymes is unclear, but the possibility that they are used later in the aerobic metabolism of the fertilized egg has not been excluded.
Animals↗
Growth of a Japanese isolate of Taenia crassiceps in intermediate and definitive hosts.
Higher infection rates were observed in gerbils and voles than in ICR mice after oral inoculation with eggs of a Japanese isolate of Taenia crassiceps. Asexual reproduction of T. crassiceps cysticerci was observed in all gerbils and voles infected i.p. with the cysticerci. However, ICR mice and Wistar rats were not suitable for the asexual proliferation of T. crassiceps. The hooks of cysticerci from mice were smaller than those from gerbils. In experimentally infected puppies, parasite development was noted as follows: strobilation and initial differentiation of the genital primordia on day 7 postinoculation (p.i.), appearance of the testes on day 9, observation of the ovaries on day 10, and development of the lateral branches of the uterus on day 15. The prepatent period was 27-31 days. After day 15 p.i., most of the worms were recovered from the middle third of the small intestine. The number of proglottids shed per day by each strobila was about 1. The number of eggs contained in a gravid segment was about 13,000.
Animals↗
Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line.
Two different cell populations, high- (MARC-145) and low-permissive cell clones (L-1) to porcine reproductive and respiratory syndrome (PRRS) virus, were derived from MA-104 cell line (parent cell: P) by cell cloning. Maximum virus yields in MARC-145, P, and L-1 cell clones were 10(8.5), 10(3.5), and 10(2.5) tissue culture infective dose 50 (TCID50)/0.1 ml, respectively. The MARC-145 cell clone supported replication of all 11 different porcine reproductive and respiratory syndrome virus isolates that were tested. These results indicated that the MARC-145 cells will be useful for PRRS virus replication.
Animals↗
The Ste16 WD-repeat protein regulates cell-cycle progression under starvation through the Rum1 protein in Schizosaccharomyces pombe.
The haploid cells of the fission yeast, Schizosaccharomyces pombe, are arrested in the G1-phase by nitrogen starvation and are committed to sexual reproduction (mating and sporulation). We isolated the sterile mutants which were defective in G1 arrest following nitrogen starvation. Genetic analysis of these mutants defined a single locus designated as ste16. The nucleotide sequence revealed that ste16+ encodes an 82-kDa protein containing eight WD40-repeats in its carboxy terminal half. The ste16 disruptant was viable, but arrested the cell cycle in the G2-phase after the nutritional down-shift. When transferred to fresh growth medium, the G2-arrested ste16Delta haploids resumed the mitotic cycle from the S-phase, resulting in diploidization. This diploidization phenomenon was completely suppressed by the null mutation of rum1 encoding the inhibitor of Cdc2 kinase. As the Rum1 protein level was remarkably elevated in the ste16Delta, the Ste16 protein negatively controls the Rum1 level. The loss of function of ste16 disturbs the cell-cycle progression and impairs the mechanism for the maintenance of ploidy.
Cell Cycle↗
Identification of an allatostatin from adult Diploptera punctata.
A peptide (allatostatin) causing strong and rapid inhibition of juvenile hormone synthesis in vitro by corpora allata from reproductively active females has been isolated from brain/retrocerebral complexes of the cockroach Diploptera punctata. The primary structure of this 13-residue peptide has been determined: Ala-Pro-Ser-Gly-Ala-Gln-Arg-Leu-Tyr-Gly-Phe-Gly-Leu-NH2. Removal of the terminal amide group caused at least a ten thousandfold loss of activity. This neurohormone has no sequence similarity with any other known neuropeptide. Its target in the biosynthetic pathway is located prior to the conversion of farnesol to juvenile hormone.
Animals↗
Anti-Müllerian hormone (AMH/AMH) in the European sea bass: its gene structure, regulatory elements, and the expression of alternatively-spliced isoforms.
In mammals, a multitude of studies have shown that anti-Müllerian hormone (AMH/AMH), apart from inducing Müllerian duct regression during male sexual differentiation, exerts inhibitory effects on male and female gonadal steroidogenesis and differentiation. However, in lower vertebrates like teleost fish, the function of AMH/AMH has been far less explored. As a first step to unravel its potential role in reproduction in teleost fish, we isolated and characterised the AMH gene in the European sea bass (sb), Dicentrachus labrax, determined putative regulatory elements of its 5'-flanking region, and analysed its gene expression and those of alternatively-spliced transcripts. The characterisation of sb-AMH revealed distinct features that distinguishes it from mammalian and bird AMH, suggesting a high rate of diversification of AMH during vertebrate evolution. It contained 7 exons that were divided by 6 introns, of which the last intron (intron vi) was localised only a few nucleotides upstream of the putative peptide cleavage site. The guanine and cytosine content of the open reading frame (ORF) was 52.7% and thus notably lower than that of bird and mammalian AMH. Sb-AMH cDNA was 2045 base pairs (bp) long, containing an ORF of 1599 bp encoding 533 amino acids. Deduced amino acid similarities of the conserved, carboxyterminal domain were highest with AMH in Japanese flounder (84.2%) and lowest with chicken AMH (45.5%). In the proximal promoter sequence of sb-AMH, a steroidogenic factor-1 (SF-1) binding site was present; however other regulatory sequences essential for transcriptional activation of AMH in mammals were absent. Likewise, there was no sequence homology to an SF3A2 sequence within the first 3200 bp upstream of the sb-AMH translation start site. Gene expression of sb-AMH and of alternatively-spliced sb-AMH transcripts were analysed in male and female juvenile and adult gonads as well as in somatic tissues of juvenile males. sb-AMH expression was highest in juvenile testis, but still remarkably high in juvenile ovaries and adult testis, as well as in brain, pituitary, and heart of juvenile male sea bass. Apart from adult ovary, levels of alternatively-spliced sb-AMHexonII/-99 were marginal in comparison with sb-AMH. In contrast, the transcript variant sb-AMHexonVII/+5 was expressed to a similar extent as sb-AMH in all tissues examined. The results of this work have provided the basis for future studies concerning the regulation and function of AMH/AMH in this species.
5' Flanking Region↗
Insulin-like growth factors I and II in the sole Solea senegalensis: cDNA cloning and quantitation of gene expression in tissues and during larval development.
Insulin-like growth factors I and II (IGF-I and IGF-II) play an important role as modulators of development, growth, and reproduction. This study aimed to isolate the IGF-I and IGF-II cDNAs and determine their temporal expression pattern in different organs and throughout larval development in Senegal sole. The rapid amplification of cDNA ends (RACE) was used to obtain both full-length IGFs sequences. A high sequence similarity with other teleosts sequences was observed. Domains B and A revealed as the most evolutionary conserved. Steady-state copy numbers of IGF-I and IGF-II were also quantified in different Senegal sole tissues by real-time PCR. IGF-I and IGF-II expressed ubiquitously with the highest mRNA levels in liver (88 x 10(6) molecules/microg total RNA) and gills (14.0 x 10(6) molecules/microg total RNA) respectively. IGF-II mRNA levels were higher than IGF-I in prehatching embryos and premetamorphic larvae with a significant drop before the commencement of eye migration in metamorphosis. The abundance of IGF-II transcripts correlated positively with the growth rate during larval development. The putative role of IGF-II on metamorphosis and larval growth is discussed.
Amino Acid Sequence↗
Structural and functional effects of endometrial photodynamic therapy in a rat model.
OBJECTIVE: Our purpose was to determine the optical dose required for irreversible endometrial destruction and prevention of implantation by photodynamic therapy with topical 5-aminolevulinic acid. STUDY DESIGN: Three hours after drug application 74 female Sprague-Dawley rats received varying doses of 630 nm of light delivered by an intrauterine cylindric diffusing fiber. RESULTS: A 64 J/cm2 in situ optical dose resulted in long-term irreversible endometrial destruction; 43 J/cm2 damaged endometrial stroma and myometrium but not glandular epithelium 1 day after photodynamic therapy. At this lower light dose endometrium regenerated to full thickness within 3 weeks; however, implantation sacs were significantly reduced. CONCLUSIONS: Photodynamic destruction of glandular epithelium accompanies irreversible endometrial ablation, whereas isolated stromal damage leads to reproductive impairment only. The optical dose required for endometrial ablation is approximately 1.5-fold higher than for reproductive impairment (functional damage) because of differential cell photosensitivity.
Aminolevulinic Acid↗
Reinforcement of genetic coherence: a single-locus model.
Genetic coherence and genetic separation are the outcome of evolutionary mechanisms which maintain genetic variation within populations through recombination on the one hand, and which divide this variation via speciation between reproductively (recombinatorically) more or less isolated populations on the other. While mechanisms of speciation have received considerable attention in biology, their counterpart, mechanisms of genetic coherence, are addressed only implicitly, if at all. Usually, genetic coherence is intuitively associated with the forces maintaining genetic polymorphisms and thus potential for flexible adaptational reaction of populations. However, so far no models seem to exist which explain the evolution of genetic coherence as the natural counterpart of genetic separation or speciation. In this paper a single-locus model is analyzed, in which a mutant allele is introduced into a resident stable diallelic polymorphism, and where this allele is equivalent to one of the resident alleles in all respects with the exception of mating relations. The conditions for replacement of the resident allele by its selectively equivalent mutant are obtained with reference to the associated mating relations. It turned out that for heterozygote advantage the mutant replaces the selectively equivalent resident allele if it increases the mating preferences for carriers of other alleles. The evolution of lower such preferences requires heterozygote inferiority, which confirms the Wallace effect of speciation (by reinforcement). It is argued that this observation suggests that non-selective constituents of the mating system form the section of the genetic system that is responsible for moderating the genetic load implied by adapting selection while simultaneously securing the adaptational potential embodied in the resident allelic variation. Mating systems thus serve the preservation of adaptability.
Adaptation, Physiological↗