The Role of Tissue-type Plasminogen Activator in Rat Corpus Luteum

It is here reported for the first time that luteal cells are capable of secreting plasminogen activators(PA),(both tissue-type,tPA,and urokinase-type,uPA),and plasminogen activator inhibitor type-1(PAl-1).Using organ culture model,we have demonstrated that tPA,but not uPA,showed markedchange during luteolytic period in rat corpus luteum.A great amount oftPA was secreted in corpusluteum on D 14 and D 17 while very low level of tPA activity was detected before D 12.Correspondingly,the progesterone production in the corpus luteum increased gradually in a time-dependent manner from D 1 to D 12 but dropped abruptly to a very low level on D 14.Additionof exogenous tPA to the CL culture caused considerable decrease in progesterone secretion whileinclusion of purified monoclone tPA antibodies in the culture augmented progesterone productionof CL.It is therefore suggested that tPA may play an important role in luteolytic process.     

人工合成寡肽对大鼠黄体细胞孕酮生成的调节及其可能作用机制

本实验比较了合成寡肽抗孕酮生成作用,并进行了相应机制探讨。发现当ＰＨ７．３－７．５时,能较强抑制孕酮分泌的寡肽其结构有共同特点;活性寡肽可对ＰＬＣ信使传递系统产生抑制作用。也可能通过调节黄体细胞内钙离子浓度降低了ｈＣＧ致孕酮的生成作用,甘－丝－赖还升高黄体细胞中ＰＫＣ活性,而降低了ＰＫＡ。可见人工合成寡肽的抗孕酮作用分子机制十分复杂,有待于深入探讨。     

Reflections on the Evolution of Piscine Viviparity

Viviparity first makes its evolutionary appearance within thecraniate-vertebrate line among fishes. We estimate that it hasindependently evolved at least 42 times in five of the ninemajor groups of fishes. Viviparity is the dominant mode of reproductionamong the cartilaginous sharks and rays, i.e., 55% of approximately900 living species. It is less prevalent among the five majorgroups of bony fishes, i.e., 2–3% of an estimated 20,000or more species. The evolution of viviparity from oviparityinvolves: 1) a shift from external to internal fertilization;2) retention of embryos in the female reproductive system; 3)utilization of the ovaryor oviduct as sites of gestation; 4)structural and functional modification of the embryo and thefemale reproductive system and; 5) modification of extant endocrinemechanisms controlling reproduction. Viviparity offers selectiveadvantages to parents and offspring, such as: 1) enhanced survivalof offspring; 2) compensation for low fecundity; 3) amplificationof reproductive niches to reduce competition; 4) exploitationof pelagic niches; 5) colonization of new habitats; and 6) increasedenergetic efficiency in viviparous matrotrophes. Its principaldisadvantages include: 1) reduced fecundity; 2) cost to thefemale; and 3) risk of brood loss through maternal death. Acquisitionof viviparityestablishes new maternal-embryonic relationships,namely: 1) trophic; 2) osmoregulatory and excretory; 3) respiratory;4) endocrinological; and 5) immunological. In sharks, rays,and the coelacanth, gestation takes place in the oviduct, butin teleosts gestation occurs either in the ovarian follicleor ovarian lumen. The cystovarian teleostean ovary is hypothesizedto function both as ovary and oviduct. Oviductal, ovarian lumenal,andfollicular epithelial cells are the maternal sites of metabolicexchange. Metabolic exchange inembryos takes place across theepithelia of the general body surface and its derivatives oracross the gut epithelium and its derivatives. Four patternsof piscine placentation have evolved,namely: 1) yolk sac; 2)follicular; 3) branchial; and 4) trophotaenial placentae. Thepericardial amniochorion, the embryonic portion of the follicularplacenta, occurs in poeciliids and several other teleosteangroups. Developmental, it is nearly identical to the anterioraminochorionic fold of tetrapod vertebrates. Trophotaeniae areexternal rosette or ribbon-like structuresthat have evolvedin four orders of teleosts by heterochrony, i.e., acceleratedoutgrowth and differentiation of the embryonic hind gut. Withthe possible exception of the coelacanth, theyolk sac placentaoccurs only in sharks. We estimate that it has independentlyevolved between 11 and 20 times. It displays considerable diversity.Evolution of the yolk sac placenta entails retention of theyolk sac and secondary differentiation of its distal portionfor implantation and maternal tissue-embryonic tissue metabolicexchange and its proximal portion for oviductal fluid-embryonictissue exchange. The yolk stalk lengthens, is modified intoan umbilical stalk, and establishes a site of autotomy at theembryo-umbilical stalk junction. The lumenal wall of the oviductbecomes competent to function as a site of implantation.     

Evolutionary Scenarios, Homology and Convergence of Structural Specializations for Vertebrate Viviparity

Viviparity has evolved many times in many lineages of vertebrates.Many evolutionary scenarios for the evolution of viviparity,or structures associated with it, have been proposed. Many ofthese are testable using methodologies developed recently. Somestructures that facilitate viviparity (e.g., corpora lutea andoviducts) are homologous, for they are inherited from an ancestorin a lineage. More such structures (e.g., placentas and placentalanalogues) are convergences, for they appear in diverse lineagesbut are not traceable to a common ancestor.Conservatism andconstraint characterize many, but not all, such convergences.A phylogenetic hypothesis facilitates identification of homologiesand convergences, and the testing of evolutionary scenarios.     

人类黄体组织抑制素与激活素构型的研究

本文观察了抑制素／激活素α、βＢ与βＢ亚基基因在人早（３天）、中（３天）及晚（１５天）期黄体组织的表达。α－ｍＲＮＡｓ在早期黄体最高;βＡ－ｍＲＮＡｓ仅在早期出现,而βＢ－ｍＲＮＡｓ从黄体早期至晚期逐步增高。提示人黄体抑制素可能为抑制素Ｂ。晚期黄体可能有激活素Ｂ的生成。     

Behavioral Dominance and Corpus Luteum Function in Red DeerCervus elaphus

Times of the ovulatory LH surge and characteristics of the rise in circulating progesterone concentrations after ovulation in red deer hinds were investigated in relation to each animal's dominance status. Observations were made during the 1992 (experiment 1) and 1993 (experiment 2) breeding seasons, while the same 12 hinds were held in a pen in the absence of a stag. Ovulation was synchronized by administration of progesterone followed by luteolytic prostaglandin F_2αanalogue. Social status was determined for each hind by noting dyadic agonistic interactions during the period of progesterone treatment. Hinds were weighed before and after the experiments. Time of onset of estrus (lordosis) was recorded while handling at 3-hr intervals (for 81 hr in experiment 1; 96 hr in experiment 2) after progesterone withdrawal, and blood samples were collected at these times to characterize the preovulatory LH surge. Subsequently, daily blood samples were collected for up to 11 days for measurement of progesterone concentrations. There was a tendency for weight change to be related to dominance status in experiment 1 and this was significant in experiment 2 (p< 0.01). The rate of increase in circulating progesterone concentration after ovulation was related to status (data of experiments 1 and 2 combined;p< 0.02), but was not correlated with the time of estrus or with the time or height of the LH surge. A third experiment (carried out in 1993), when the same hinds were kept with a stag after induced ovulation, showed that time of estrus (mating) was not related to dominance status. The data suggest that corpus luteum function is affected by social status. The results are discussed in the context of mechanisms by which dominance status influences the sex of a hind's calves.     

Noggin Expression in the Adult Retina Suggests a Conserved Role during Vertebrate Evolution

Vertebrates share common mechanisms in the control of development and in the maintenance of neural and retinal function. The secreted factor Noggin, a BMP inhibitor, plays a crucial role in neural induction during embryonic development. Moreover, we have shown its involvement in retinal differentiation of pluripotent cells. Here we show Noggin expression in the adult retina in three vertebrate species. Four Noggin genes are present in zebrafish (Danio rerio; ZbNog1, 2, 3, 5), three in frog (Xenopus laevis; XenNog1, 2 and 4), and one in mouse (Mus musculus; mNog). Quantitative RT-PCR experiments show the presence of ZbNog3 and ZbNog5 mRNAs, but not ZbNog1 and ZbNog2, in the adult zebrafish retina. All three genes are expressed in the frog retina, and mNog in the mouse. Immunohistochemistry data show that Noggin proteins are predominantly localized in the Golgi apparatus of photoreceptors and in the fibers of the outer plexiform layer. Lower expression levels are also found in inner plexiform layer fibers, in ganglion cells, in the ciliary marginal zone, and in retinal pigmented epithelium. Our results show that Noggin has a specific cellular and sub-cellular expression in the adult vertebrate retina, which is conserved during evolution. In addition to its established role during embryonic development, we postulate that Noggin also exerts a functional role in the adult retina.     

Intraspecific Phylogeography of Lacerta vivipara and the Evolution of Viviparity

The lacertid lizard Lacerta vivipara is one of the few squamate species with two reproductive modes. We present the intraspecific phylogeny obtained from neighbor-joining and maximum-parsimony analyses of the mtDNA cytochrome b sequences for 15 individuals from Slovenian oviparous populations, 34 individuals from western oviparous populations of southern France and northern Spain, 92 specimens from European and Russian viviparous populations, and 3 specimens of the viviparous subspecies L. v. pannonica. The phylogeny indicates that the evolutionary transition from oviparity to viviparity probably occurred once in L. vivipara. The western oviparous group from Spain and southern France is phylogenetically most closely related to the viviparous clade. However, the biarmed W chromosome characterizing the western viviparous populations is an apomorphic character, whereas the uniarmed W chromosome, existing both in the western oviparous populations and in the geographically distant eastern viviparous populations, is a plesiomorphic character. This suggests an eastern origin of viviparity. Various estimates suggest that the oviparous and viviparous clades of L. vivipara split during the Pleistocene. Our results are discussed in the framework of general evolutionary models: the concept of an oviparity–viviparity continuum in squamates, the cold climate model of selection for viviparity in squamates, and the contraction–expansion of ranges in the Pleistocene resulting in allopatric differentiation.     

Activity Physiology and Evolution of the Vertebrate Skeleton

SYNOPSIS. Vertebrates frequently rely on intramuscular glycolysisas the major source of ATP utilized during bouts of intenseexercise. This is often followed by extended periods of markedsystemic pH fluctuation. Such a pattern of activity physiologyis unique among the Metazoa and probably dates back to the veryearliest vertebrates. The origin of bone may have been necessitated by requirementfor a skeletal matrix with chemical stability over the broadrange of tissue pH associated with vertebrate exercise physiology.     

The Molecular Evolution of the Vertebrate Trypsinogens

We expand the already large number of known trypsinogen nucleotide and amino acid sequences by presenting additional trypsinogen sequences from the tunicate (Boltenia villosa), the lamprey (Petromyzon marinus), the pufferfish (Fugu rubripes), and the frog (Xenopus laevis). The current array of known trypsinogen sequences now spans the entire vertebrate phylogeny. Phylogenetic analysis is made difficult by the presence of multiple isozymes within species and rates of evolution that vary highly between both species and isozymes. We nevertheless present a Fitch-Margoliash phylogeny constructed from pairwise distances. We employ this phylogeny as a vehicle for speculation on the evolution of the trypsinogen gene family as well as the general modes of evolution of multigene families. Unique attributes of the lamprey and tunicate trypsinogens are noted. Received: 12 July 1997     

Matrix Metalloproteinase 2 Is Required for Ovulation and Corpus Luteum Formation in Drosophila

Ovulation is critical for successful reproduction and correlates with ovarian cancer risk, yet genetic studies of ovulation have been limited. It has long been thought that the mechanism controlling ovulation is highly divergent due to speciation and fast evolution. Using genetic tools available in Drosophila, we now report that ovulation in Drosophila strongly resembles mammalian ovulation at both the cellular and molecular levels. Just one of up to 32 mature follicles per ovary pair loses posterior follicle cells (“trimming”) and protrudes into the oviduct, showing that a selection process prefigures ovulation. Follicle cells that remain after egg release form a “corpus luteum (CL)” at the end of the ovariole, develop yellowish pigmentation, and express genes encoding steroid hormone biosynthetic enzymes that are required for full fertility. Finally, matrix metalloproteinase 2 (Mmp2), a type of protease thought to facilitate mammalian ovulation, is expressed in mature follicle and CL cells. Mmp2 activity is genetically required for trimming, ovulation and CL formation. Our studies provide new insights into the regulation of Drosophila ovulation and establish Drosophila as a model for genetically investigating ovulation in diverse organisms, including mammals.     

Evolution of Vertebrate Indoleamine 2,3-Dioxygenases

Indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) are tryptophan-degrading enzymes that catalyze the same reaction, the first step in tryptophan catabolism via the kynurenine pathway. TDO is widely distributed among life-forms, being found not only in eukaryotes but also in bacteria. In contrast, IDO has been found only in mammals and yeast to date. However, recent genome and EST projects have identified IDO homologues in non-mammals and found an IDO paralogue that is expressed in mice. In this study, we cloned the frog and fish IDO homologues and the mouse IDO paralogue, and characterized their enzymatic properties using recombinants. The IDOs of lower vertebrates and the mouse IDO paralogue had IDO activity but had 500–1000 times higher K _m values and very low enzyme efficiency compared with mammalian IDOs. It appears that L-Trp is not a true substrate for these enzymes in vivo, although their actual function is unknown. On the phylogenetic tree, these low-activity IDOs, which we have named “proto-IDOs,” formed a cluster that was distinct from the mammalian IDO cluster. The IDO and proto-IDO genes are present tandemly on the chromosomes of mammals, including the marsupial opossum, whereas only the proto-IDO gene is observed in chicken and fish genomes. These results suggest that (mammalian) IDOs arose from proto-IDOs by gene duplication that occurred before the divergence of marsupial and eutherian (placental) mammals in mammalian evolutionary history.     

Melatonin's Role in Vertebrate Orcadian Rhythms

The circadian secretion of melatonin by the pineal gland and retinae is a direct output of circadian oscillators and of the circadian system in many species of vertebrates. This signal affects a broad array of physiological and behavioral processes, making a generalized hypothesis for melatonin function an elusive objective. Still, there are some common features of melatonin function. First, melatonin biosynthesis is always associated with photoreceptors and/or cells that are embryonically derived from photoreceptors. Second, melatonin frequently affects the perception of the photic environment and has as its site of action structures involved in vision. Finally, melatonin affects overt circadian function at least partially via regulation of the hypothalamic suprachiasmatic nucleus (SCN) or its hofnologues. The mechanisms by which melatonin affects circadian rhythms and other downstream processes are unknown, but they include interaction with a class of membrane-bound receptors that affect intracellular processes through guanosine triphosphate (GTP)-binding protein second messenger systems. Investigation of mechanisms by which melatonin affects its target tissues may unveil basic concepts of neuromodulation, visual system function, and the circadian clock.     

The Origin and Evolution of Vertebrate Glycine Transporters

In the vertebrate central nervous system, glycinergic neurotransmission is regulated by the action of the glycine transporters 1 and 2 (GlyT1 and GlyT2)—members of the solute carrier family 6 (SLC6). Several invertebrate deuterostomes have two paralogous glycine carrier genes, with one gene in the pair having greater sequence identity and higher alignment scores with respect to GlyT1 and the other paralog showing greater similarity to GlyT2. In phylogenetic trees, GlyT2-like sequences from invertebrate deuterostomes form a monophyletic subclade with vertebrate GlyT2, while invertebrate GlyT1-like proteins constitute an outgroup to both the GlyT2-like proteins and to vertebrate GlyT1 sequences. These results are consistent with the hypothesis that vertebrate GlyT1 and GlyT2 are, respectively, derived from GlyT1- and GlyT2-like genes in invertebrate deuterostomes. This implies that the gene duplication which gave rise to these paralogs occurred prior to the origin of vertebrates. GlyT2 subsequently diverged significantly from its invertebrate orthologs (i.e., through the acquisition of a unique N-terminus) as a consequence of functional specialization, being expressed principally in the lower CNS; while GlyT1 has activity in both the lower CNS and several regions of the forebrain.