Endothelial and steroidogenic cell migration are regulated by WNT4 in the developing mammalian gonad

The signalling molecule WNT4 has been associated with sex reversal phenotypes in mammals. Here we show that the role of WNT4 in gonad development is to pattern the sex-specific vasculature and to regulate steroidogenic cell recruitment. Vascular formation and steroid production in the mammalian gonad occur in a sex-specific manner. During testis development, endothelial cells migrate from the mesonephros into the gonad to form a coelomic blood vessel. Leydig cells differentiate and produce steroid hormones a day later. Neither of these events occurs in the XX gonad. We show that WNT4 represses mesonephric endothelial and steroidogenic cell migration in the XX gonad, preventing the formation of a male-specific coelomic blood vessel and the production of steroids. In the XY gonad, Wnt4 expression is downregulated after sex determination. Transgenic misexpression of Wnt4 in the embryonic testis did not inhibit coelomic vessel formation but vascular pattern was affected. Leydig cell differentiation was not affected in these transgenic animals and our data implies that Wnt4 does not regulate steroidogenic cell differentiation but represses the migration of steroidogenic adrenal precursors into the gonad. These studies provide a model for understanding how the same signalling molecule can act on two different cell types to coordinate sex development.     

Parameters of photosynthesis: definitions, theory and interpretation of results

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Myocardium at the base of the aorta and pulmonary trunk is prefigured in the outflow tract of the heart and in subdomains of the second heart field

Outflow tract myocardium in the mouse heart is derived from the anterior heart field, a subdomain of the second heart field. We have recently characterized a transgene (y96-Myf5-nlacZ-16), which is expressed in the inferior wall of the outflow tract and then predominantly in myocardium at the base of the pulmonary trunk. Transgene A17-Myf5-nlacZ-T55 is expressed in the developing heart in a complementary pattern to y96-Myf5-nlacZ-16, in the superior wall of the outflow tract at E10.5 and in myocardium at the base of the aorta at E14.5. At E9.5, the two transgenes are transcribed in different subdomains of the anterior heart field. A clonal analysis of cardiomyocytes in the outflow tract, at E10.5 and E14.5, provides insight into the behaviour of myocardial cells and their progenitors. At E14.5, most clones are located at the base of either the pulmonary trunk or the aorta, indicating that these derive from distinct myocardial domains. At E10.5, clones are observed in subdomains of the outflow tract. The distribution of small clones indicates proliferative differences, whereas regionalisation of large clones, that derive from an early myocardial progenitor cell, reflect coherent cell growth in the heart field as well as in the myocardium. Our results suggest that myocardial differences at the base of the great arteries are prefigured in distinct progenitor cell populations in the anterior heart field, with important implications for understanding the etiology of congenital heart defects affecting the arterial pole of the heart.     

Shear stress-induced Ca2+ mobilization in MDCK cells is ATP dependent,no matter the primary cilium

Primary cilium has emerged as mechanosensor to subtle flow variations in epithelial cells, but its role in shear stress detection remains controversial. To probe the function of this non-motile organelle in shear stress detection by cells, we compared calcium signalling responses induced by shear stress in ciliated and unciliated MDCK cells. Cytosolic free Ca²⁺ ([Ca²⁺]_i) was measured using Fura-PE3 video imaging fluorescence microscopy in response to shear stress due to laminar flow (385 μl s^?1). Our results show that both unciliated and ciliated MDCK cells are shear stress sensitive via ATP release and autocrine feedback through purinergic receptors. However, purinergic calcium signals differed in response intensity and receptor subtypes. In unciliated cells, shear stress-induced elevation in [Ca²⁺]_i was predominantly mediated through P2X receptors (P2XR). In contrast, calcium mobilization in ciliated MDCK cells resulted from P2YRs and store-operated Ca²⁺-permeable channels besides P2XRs. These findings lend support to the hypothesis that ATP release in response to shear stress is independent of the primary cilium and that transduction of mechanical strain into a specific biochemical responses stems on the mobilization of different sets of purinergic receptors.     

An attempt to simulate the subsurface chlorophyll maximum using populations of unphased oscillating cells

A model is proposed to represent the vertical distribution ofchlorophyll in tropical waters. It assumes that the phytoplanktonpopulations are composed of cells which oscillate around a meanlevel at a density _t = B (close to the nutricline), with anamplitude A; A and B are normally distributed variables; theoscillations are unphased so that the number of cells in a waterlayer does not vary with the time. This model is tested on datafrom the eastern tropical Atlantic and from the southwesterntropical Pacific. The results agree with the observations whenthe surface mixed layer is nutrient-exhausted. However, themodel seems to account only for a deep population, and is thenconsistent with the hypothesis of a distinct mixed layer floralassociation. The agreement between the results and the observationsis difficult when the surface mixed layer is not nutrient-limited,in areas of upwelling or intense vertical mixing. The modelaccounts for a splitting of the subsurface chlorophyll maximumafter the passage of a front in the Cape Lopez area (GuineaGulf). It also accounts for the relations between the subsurfacechlorophyll maximum, the pycnocline, and the nutricline at 48stations from the cruise PREFIL 2 in the southwestern tropicalPacific. The permanent subsurface chlorophyll maximum of oligotrophicareas seems to be more satisfactorily explained by the unphasedoscillations invoked by the model than by the sinking of thecells. These unphased oscillations have not yet been observed;their possible consequences concerning the primary productionand grazing are postulated.     

Water-column chlorophyll in an oligotrophic environment: correction for the sampling depths and variations of the vertical structure of density, and observation of a growth period

The chlorophyll content of a water column (WCC), which is commonlyused as an index of the phytoplankton abundance, is affectedby the choice of the sampling depths and by the variations ofthe vertical structure of density. For instance, the thicknessof the water layer, between two sigma-t values, which containsthe deep chlorophyll maximum, can vary with internal waves.The resulting noise often dominates the mesoscale variationsof the observed water-column chlorophyll (OWCC). Sigma-t dependentstatistics (mean, standard deviation) of the chlorophyll concentrationare computed using the observations at 29 casts from a 22-day-longfixed station in an oligotrophic environment at 15•S, 173°E.For each cast, these statistics, the sampling depths, and thewater density at these sampling depths, allow the estimationof a station-dependent ‘expected water-column chlorophyll’(EWCC). The ratio of EWCC to the overall likelihood of WCC duringthe fixed station (i.e. the mean of all OWCC) is a measure ofthe effect of sampling and variable density structure at eachcast. When this effect is removed, the noise in WCC estimatesdecreases significantly. The time variations of WCC during thefixed station then show a trend with relatively high valuesduring the first days, followed by a 12-day-long period withlow values. A regular increase occurred from 1 October, whichwas accompanied by high carbon fixation rates and was mainlydue to an increase of the chlorophyll concentration betweenthe surface and the deep chlorophyll maximum. New productionduring this active phase was estimated to be 535 mgC m^–2day^–1, corresponding to 62% of the total production. Breakingof internal waves which were recorded at the beginning of thegrowth phase and vertical mixing of nutrients can explain theobservation.     

Wnt4 is required for proper male as well as female sexual development

Genes previously implicated in mammalian sexual development have either a male- or female-specific role. The signaling molecule WNT4 has been shown to be important in female sexual development. Lack of Wnt4 gives rise to masculinization of the XX gonad and we showed previously that the role of WNT4 was to inhibit endothelial and steroidogenic cell migration into the developing ovary. Here we show that Wnt4 also has a function in the male gonad. We find that Sertoli cell differentiation is compromised in Wnt4 mutant testes and that this defect occurs downstream of the testis-determining gene Sry but upstream of Sox9 and Dhh, two early Sertoli cell markers. Genetic analysis shows that this phenotype is primarily due to the action of WNT4 within the early genital ridge. Analysis of different markers identifies the most striking difference in the genital ridge at early stages of its development between wild-type and Wnt4 mutant embryos to be a significant increase of steroidogenic cells in the Wnt4 -/- gonad. These results identify WNT4 as a new factor involved in the mammalian testis determination pathway and show that genes can have a specific but distinct role in both male and female gonad development.     

Parameters of photosynthesis: definitions, theory and interpretation of results

A global assessment of carbon flux in the world ocean is oneof the major undertakings of the Joint Global Ocean Flux Study(JGOFS). This has to be undertaken using historical in situdata of primary productivity. As required by the temporal andspatial scales involved in a global study, it can be convenientlydone by combining, through appropriate models, remotely sensedinformation (chlorophyll a, temperature) with basic informationabout the parameters related to the carbon uptake by phytoplanktonicalgae. This requires a better understanding as well as a moreextended knowledge of these parameters which govern the radiativeenergy absorption and utilization by algae in photosynthesis.The measurement of the photosynthetic response of algae [ photosynthesis(P) versus it-radiance (E) curves], besides being less shiptimeconsuming than in situ primary production experiments, allowsthe needed parameters to be derived and systematically studiedas a function of the physical, chemical and ecological conditions.The aim of the present paper is to review the sig nificanceof these parameters, especially in view of their introductioninto models, to analyze the causes of their variations in thelight of physiological considerations, and finally to providemethodo logical recommendations for meaningful determinations,and interpretation, of the data resulting from P versus E determinations.Of main concern are the available and usable irradiance, thechloro phyll a-specific absorption capabilities of the algae,the maximum light utilization coefficient (cs), the maximumquantum yield (