Soluble laminin and arginine-glycine-aspartic acid containing peptides differentially regulate type IV collagenase messenger RNA, activation, and localization in testicular cell culture

Rat testicular cells in culture produce several metalloproteinases including type IV collagenases (Sang et al. Biol Reprod 1990; 43:946-955, 956-964). We have now investigated the regulation of testicular cell type IV collagenase and other metalloprotemases in vitro. Soluble laminin stimulated Sertoli cell type IV collagenase mRNA levels. However, three peptides corresponding to different domains of the laminin molecule (CSRAKQAASIKVASADR, FALRGDNP, CLQDGDVRV) did not influence type IV collagenase mENA levels. Zyniographic analysis of medium collected from these cultures revealed that neither soluble laminin nor any of the peptides influenced 72-Wa type IV collagenase protein levels. However, peptide FALRGDNP resulted in both, a selective increase in two higher molecular-weight metalloprotemnases (83 kDa and 110 Wa and in an activation of the 72-Wa rat type IV collagenase. Interleukin-1, phorbol ester, testosterone, and FSH did not affect collagenase activation, lmmunocytochemical studies demonstrated that the addition of soluble laminin resulted in a redistribution of type IV collagenase from intracellular vesicles to the cell-substrate region beneath the cells. Peptide FALRGDNP induced a change from a vesicular to peripheral plasma membrane type of staining pattern. Zymography of plasma membrane preparations demonstrated triton-soluble gelatinases of 76 Wa, 83 Wa, and 110 Wa and a triton-insoluble gelatinase of 225 Wa, These results indicate that testicular cell type IV collagenase mRNA levels, enzyme activation, and distribution are influenced by laminin and RGD-containing peptides.

Transmembrane/cytoplasmic domain-mediated membrane type 1-matrix metalloprotease docking to invadopodia is required for cellinvasion

The invasion of human malignant melanoma cells into the extracellular matrix (ECM) involves the accumulation of proteases at sites of ECM degradation where activation of matrix metalloproteases (MMP) occurs. Here, we show that when membrane type 1 MMP (MT-MMP) was overexpressed in RPMI7951 human melanoma cells, the cells made contact with the ECM, activated soluble and ECM-bound MMP-2, and degraded and invaded the ECM. Further experiments demonstrated the importance of localization of the MT-MMP to invadopodia. Overexpression of MT-MMP without invadopodial localization caused activation of soluble MMP-2, but did not facilitate ECM degradation or cell invasiveness. Up-regulation of endogenous MT-MMP with concanavalin A caused activation of MMP-2. However, concanavalin A treatment prevented invadopodial localization of MT-MMP and ECM degradation. Neither a truncated MT-MMP mutant lacking transmembrane (TM) and cytoplasmic domains (ΔTM(MT-MMP)), nor a chimeric MT-MMP containing the interleukin 2 receptor α chain (IL-2R) TM and cytoplasmic domains (ΔTM(MT-MMP)/TM(IL-2R)) were localized to invadopodia or exhibited ECM degradation. Furthermore, a chimera of the TM/cytoplasmic domain of MT-MMP (TM(MT-MMP)) with tissue inhibitor of MMP 1 (TIMP-1/TM(MT- MMP)) directed the TIMP-1 molecule to invadopodia. Thus, the MT-MMP TM/cytoplasmic domain mediates the spatial organization of MT-MMP into invadopodia and subsequent degradation of the ECM.

Localization of fibronectin in the rat testis : effects of serum and dibutyryl cyclic AMP on fibronectin deposition by peritubular myoid cells in culture

The peritubular zone of the rat testis has an extensive extracellular matrix (ECM). Fibronectin (FN) is distributed primarily in the basal lamina of the seminiferous tubule boundary tissue and is synthesized by peritubular myoid cells. Several extracellular changes are mediated by growth factors and these changes occur at the time of hormone mediated testicular development, particularly in the peritubular zone. The effects of serum or dibutyryl cyclic AMP (cAMP) on FN production by the mesenchymal peritubular myoid cells were evaluated. Rats of various ages (10, 15, 20, 40 and 80 days) were employed for immunofluorescent localization of rat testicular FN in frozen sections. In all age groups tested, FN was primarily present in a broad layer around each seminiferous tubule, and blood vessel, and in variable distribution throughout the interstitial stroma. By day 20 there was no clear distinction in FN staining between the peritubular zone and the interstitial tissue. This indicates an involvement of FN in the ECM developments which occur in the peritubular zone of the testis at this time. The peritubular myoid cells were isolated from 20-22 day old rat testis and cultured on glass coverslips. These cells were grown to confluence with 10% fetal calf serum (FCS) in medium until day 4 and then subcultured to have secondary monocultures maintained with or without serum. By means of immunofluorescence and cytochemistry using avidin-biotin peroxidase complex it was observed that peritubular myoid cells were positive for FN and most of the FN was localized in the perinuclear region. Subcultured peritubular myoid cells maintained for 4 days in medium containing FCS developed an extensive interconnecting FN matrix. In the presence of 0.5 mM cAMP in culture, FN became localized along the filamentous process of peritubular myoid cells and more prominently in the areas of triangulated multi-cell aggregates as well as on the surface of the contracted small spherical cells. The addition of cAMP in the presence of FCS, also caused a noticeable change in the staining pattern; FN was detected along the filamentous process developing into a complex network of cells encased in an extensive matrix. It would appear that the translocation of FN in the cytoplasmic extensions of peritubular myoid cells may be a direct consequence of morphological changes associated with metabolic regulation of cAMP. This may also be related to the puberty associated development of in vivo changes in the ECM produced by peritubular myoid cells.

Estimating local stream fish assemblage attributes: Sampling effort and efficiency at two spatial scales

As part of a wider study to develop an ecosystem-health monitoring program for wadeable streams of south-eastern Queensland, Australia, comparisons were made regarding the accuracy, precision and relative efficiency of single-pass backpack electrofishing and multiple-pass electrofishing plus supplementary seine netting to quantify fish assemblage attributes at two spatial scales (within discrete mesohabitat units and within stream reaches consisting of multiple mesohabitat units). The results demonstrate that multiple-pass electrofishing plus seine netting provide more accurate and precise estimates of fish species richness, assemblage composition and species relative abundances in comparison to single-pass electrofishing alone, and that intensive sampling of three mesohabitat units (equivalent to a riffle-run-pool sequence) is a more efficient sampling strategy to estimate reach-scale assemblage attributes than less intensive sampling over larger spatial scales. This intensive sampling protocol was sufficiently sensitive that relatively small differences in assemblage attributes (<20%) could be detected with a high statistical power (1-β > 0.95) and that relatively few stream reaches (<4) need be sampled to accurately estimate assemblage attributes close to the true population means. The merits and potential drawbacks of the intensive sampling strategy are discussed, and it is deemed to be suitable for a range of monitoring and bioassessment objectives.

Spatial prediction on a river network

This article develops methods for spatially predicting daily change of dissolved oxygen (Dochange) at both sampled locations (134 freshwater sites in 2002 and 2003) and other locations of interest throughout a river network in South East Queensland, Australia. In order to deal with the relative sparseness of the monitoring locations in comparison to the number of locations where one might want to make predictions, we make a classification of the river and stream locations. We then implement optimal spatial prediction (ordinary and constrained kriging) from geostatistics. Because of their directed-tree structure, rivers and streams offer special challenges. A complete approach to spatial prediction on a river network is given, with special attention paid to environmental exceedances. The methodology is used to produce a map of Dochange predictions for 2003. Dochange is one of the variables measured as part of the Ecosystem Health Monitoring Program conducted within the Moreton Bay Waterways and Catchments Partnership.

Identifying the spatial scale of land use that most strongly influences overall river ecosystem health score

Catchment and riparian degradation has resulted in declining ecosystem health of streams worldwide. With restoration a priority in many regions, there is an increasing interest in the scale at which land use influences stream ecosystem health. Our goal was to use a substantial data set collected as part of a monitoring program (the Southeast Queensland, Australia, Ecological Health Monitoring Program data set, collected at 116 sites over six years) to identify the spatial scale of land use, or the combination of spatial scales, that most strongly influences overall ecosystem health. In addition, we aimed to determine whether the most influential scale differed for different aspects of ecosystem health. We used linear-mixed models and a Bayesian model-averaging approach to generate models for the overall aggregated ecosystem health score and for each of the five component indicators (fish, macroinvertebrates, water quality, nutrients, and ecosystem processes) that make up the score. Dense forest close to the survey site, mid-dense forest in the hydrologically active nearstream areas of the catchment, urbanization in the riparian buffer, and tree cover at the reach scale were all significant in explaining ecosystem health, suggesting an overriding influence of forest cover, particularly close to the stream. Season and antecedent rainfall were also important explanatory variables, with some land-use variables showing significant seasonal interactions. There were also differential influences of land use for each of the component indicators. Our approach is useful given that restoring general ecosystem health is the focus of many stream restoration projects; it allowed us to predict the scale and catchment position of restoration that would result in the greatest improvement of ecosystem health in the regions streams and rivers. The models we generated suggested that good ecosystem health can be maintained in catchments where 80% of hydrologically active areas in close proximity to the stream have mid-dense forest cover and moderate health can be obtained with 60% cover.

Intrinsic frames of reference in haptic spatial learning

It has been proposed that spatial reference frames with which object locations are specified in memory are intrinsic to a to-be-remembered spatial layout (intrinsic reference theory). Although this theory has been supported by accumulating evidence, it has only been collected from paradigms in which the entire spatial layout was simultaneously visible to observers. The present study was designed to examine the generality of the theory by investigating whether the geometric structure of a spatial layout (bilateral symmetry) influences selection of spatial reference frames when object locations are sequentially learned through haptic exploration. In two experiments, participants learned the spatial layout solely by touch and performed judgments of relative direction among objects using their spatial memories. Results indicated that the geometric structure can provide a spatial cue for establishing reference frames as long as it is accentuated by explicit instructions (Experiment 1) or alignment with an egocentric orientation (Experiment 2). These results are entirely consistent with those from previous studies in which spatial information was encoded through simultaneous viewing of all object locations, suggesting that the intrinsic reference theory is not specific to a type of spatial memory acquired by the particular learning method but instead generalizes to spatial memories learned through a variety of encoding conditions. In particular, the present findings suggest that spatial memories that follow the intrinsic reference theory function equivalently regardless of the modality in which spatial information is encoded.

Peripheral vision benefits spatial learning by guiding eye movements

The loss of peripheral vision impairs spatial learning and navigation. However, the mechanisms underlying these impairments remain poorly understood. One advantage of having peripheral vision is that objects in an environment are easily detected and readily foveated via eye movements. The present study examined this potential benefit of peripheral vision by investigating whether competent performance in spatial learning requires effective eye movements. In Experiment 1, participants learned room-sized spatial layouts with or without restriction on direct eye movements to objects. Eye movements were restricted by having participants view the objects through small apertures in front of their eyes. Results showed that impeding effective eye movements made subsequent retrieval of spatial memory slower and less accurate. The small apertures also occluded much of the environmental surroundings, but the importance of this kind of occlusion was ruled out in Experiment 2 by showing that participants exhibited intact learning of the same spatial layouts when luminescent objects were viewed in an otherwise dark room. Together, these findings suggest that one of the roles of peripheral vision in spatial learning is to guide eye movements, highlighting the importance of spatial information derived from eye movements for learning environmental layouts.

Differential effects of aging on spatial learning through exploratory navigation and map reading

It has been shown that abilities in spatial learning and memory are adversely affected by aging. The present study was conducted to investigate whether increasing age has equal consequences for all types of spatial learning or impacts certain types of spatial learning selectively. Specifically, two major types of spatial learning, exploratory navigation and map reading, were contrasted. By combining a neuroimaging finding that the medial temporal lobe (MTL) is especially important for exploratory navigation and a neurological finding that the MTL is susceptible to age-related atrophy, it was hypothesized that spatial learning through exploratory navigation would exhibit a greater decline in later life than spatial learning through map reading. In an experiment, young and senior participants learned locations of landmarks in virtual environments either by navigating in them in the first-person perspective or by seeing aerial views of the environments. Results showed that senior participants acquired less accurate memories of the layouts of landmarks than young participants when they navigated in the environments, but the two groups did not differ in spatial learning performance when they viewed the environments from the aerial perspective. These results suggest that spatial learning through exploratory navigation is particularly vulnerable to adverse effects of aging, whereas elderly adults may be able to maintain their map reading skills relatively well.

Sequential versus simultaneous viewing of an environment : effects of focal attention to individual object locations on visual spatial learning

We investigated memories of room-sized spatial layouts learned by sequentially or simultaneously viewing objects from a stationary position. In three experiments, sequential viewing (one or two objects at a time) yielded subsequent memory performance that was equivalent or superior to simultaneous viewing of all objects, even though sequential viewing lacked direct access to the entire layout. This finding was replicated by replacing sequential viewing with directed viewing in which all objects were presented simultaneously and participants’ attention was externally focused on each object sequentially, indicating that the advantage of sequential viewing over simultaneous viewing may have originated from focal attention to individual object locations. These results suggest that memory representation of object-to-object relations can be constructed efficiently by encoding each object location separately, when those locations are defined within a single spatial reference system. These findings highlight the importance of considering object presentation procedures when studying spatial learning mechanisms.

Path information effects in visual and proprioceptive spatial learning

Objects in an environment are often encountered sequentially during spatial learning, forming a path along which object locations are experienced. The present study investigated the effect of spatial information conveyed through the path in visual and proprioceptive learning of a room-sized spatial layout, exploring whether different modalities differentially depend on the integrity of the path. Learning object locations along a coherent path was compared with learning them in a spatially random manner. Path integrity had little effect on visual learning, whereas learning with the coherent path produced better memory performance than random order learning for proprioceptive learning. These results suggest that path information has differential effects in visual and proprioceptive spatial learning, perhaps due to a difference in the way one establishes a reference frame for representing relative locations of objects.

Visual and proprioceptive representations in spatial memory

It has been shown that spatial information can be acquired from both visual and nonvisual modalities. The present study explored how spatial information from vision and proprioception was represented in memory, investigating orientation dependence of spatial memories acquired through visual and proprioceptive spatial learning. Experiment 1 examined whether visual learning alone and proprioceptive learning alone yielded orientation-dependent spatial memory. Results showed that spatial memories from both types of learning were orientation dependent. Experiment 2 explored how different orientations of the same environment were represented when they were learned visually and proprioceptively. Results showed that both visually and proprioceptively learned orientations were represented in spatial memory, suggesting that participants established two different reference systems based on each type of learning experience and interpreted the environment in terms of these two reference systems. The results provide some initial clues to how different modalities make unique contributions to spatial representations.

Qualitative spatial reasoning about directions in a corridor containing a single turn

An experiment was conducted to investigate the process of reasoning about directions in an egocentric space. Each participant walked through a corridor containing an angular turn ranging in size from 0° to 90°, in 15° increments. A direction was given to participants at the entrance of the corridor and they were asked to answer this direction at the end of this corridor. Considering the fact that participants had to reason the direction in the featureless corridor, two hypotheses were proposed: (i) reasoning about directions falls into qualitative reasoning by using a small number of coarse angular categories (four 90° categories or eight 45° categories: 90° categories consist of front, back, left, right; 45° categories consist of 90° categories and the four intermediates) that reference axes generate; (ii) reasoning about directions would be done by recalling the rotation angle from the traveling direction to the direction that participants tried to answer. In addition, the configuration of reference axes that participants employed was examined. Both hypotheses were supported, and the data designated that reference axes consisted of eight directions: a pair of orthogonal axes and diagonals.