C van den Berg

Short-range control of cell differentiation in the Arabidopsis root meristem

Meristems are distinctive regions of plants that have capacity for continuous growth. Their developmental activity generates the majority of plant organs. It is currently unknown how cell division and cell differentiation are orchestrated in meristems, although genetic studies have demonstrated the relevance of a proper balance between the two processes. Root meristems contain a distinct central region of mitotically inactive cells, the quiescent centre, the function of which has remained elusive until now. Here we present laser ablation and genetic data that show that in Arabidopsis thaliana the quiescent centre inhibits differentiation of surrounding cells. Differentiation regulation occurs within the range of a single cell, in a manner strikingly similar to examples in animal development, such as during delamination of Drosophila neuroblasts. Our data indicate that pattern formation in the root meristem is controlled by a balance between short-range signals inhibiting differentiation and signals that reinforce cell fate decisions.

Play is indispensable for an adequate development of coping with social challenges in the rat

In this study, young rats were deprived of early social interactions during weeks 4 and 5 of life. Different behavioral tests were conducted in adulthood to study the behavioral responses of rats lacking early social experiences. Juvenile deprivation resulted in decreased social activity and an altered sexual pattern, but did not affect locomotor activity or the performance in the elevated plus maze. Furthermore, behavioral and neuroendocrine responses of juvenile isolated rats were dramatically altered when they were confronted with territorial aggression. Juvenile deprived rats did not readily display a submissive posture in response to the resident and showed no immobility behavior after being returned to the residents territory, while their plasma corticosterone and adrenaline concentrations were significantly increased compared to nonisolated controls. In contrast, behavioral responses in the shock prod test were not affected by previous isolation. The results suggest that early social experiences are vital for interactions with conspecifics later in life, i.e., aggression, sexual, and social interactions.

Isolation during the play period in infancy decreases adult social interactions in rats

The effects of 1 or 2 weeks of social isolation immediately after weaning on social activity in adulthood were investigated in rats. In addition, it was studied whether these effects were influenced by social experiences of the cagemate when rehoused after the isolation period. Isolation during weeks 4 and 5 of age caused a reduction of social activity as compared to non-isolated controls. Previous social experiences of the cagemate (isolated or non-isolated) did not affect this decreased social activity. Isolation during week 4 of age resulted in similar effects, but the reduced social activity was not present when the rats were rehoused with non-isolated rats. Isolation during week 5 of age did not influence social activity patterns in adulthood. These findings support the idea of a sensitive period in infancy for subsequent social behavior in rats. It is suggested that especially deprivation of acquiring play behavior underlies the social disturbances in adulthood.

Roots Redefined: Anatomical and Genetic Analysis of Root Development

The postembryonic development of plants is fueled by apical meristems, which are the local production sites of new cells that form a pattern of different cell types within an organ. The regularity of this pattern in the root yielded ideas on its formation from the meristem well before critical studies on the shoot apex were performed (e.g. Hanstein, 1870). The model plant Arabidopsis thaliana, which allows genetic dissection of root development, is a paragon of this regularity. In this Llpdate, we review recent studies on root anatomy and genetics that are allowing us to refine, and perhaps redefine, our understanding of organ development. We will focus on two pivotal aspects of root development: pattern formation and cell proliferation. Important work on other aspects of Arabidopsis roots, such as cell elongation/ morphogenesis, tropism, and cell size and shape, has been covered in recent reviews (Benfey and Schiefelbein, 1994; Dolan and Roberts, 1995) and will not be discussed here.

Cell fate and cell differentiation status in the Arabidopsis root

Abstract. Post-embryonic development in plants is mainly achieved by its meristems. Within the Arabidopsis root meristem, both the fate and origin of its cells can be predicted with high accuracy. Mutants defective in the determination of root cell fates show that the corresponding genes are first required during embryogenesis. The sub-specification of cell fates, such as during epidermal root hair formation, involves transcription factors and phytohormones. In the Arabidopsis root, initial cell fate specification events must be followed by position-dependent reinforcement of cellular identity. A major question remains as to whether the signals that are involved in initiating the cellular pattern are the same or different from the signals used to reinforce it. The integrity of the root meristem is kept by balancing cell proliferation and cell differentiation, and differentiation-inhibiting signals originating from the quiescent centre are involved.

Light-regulated expression of the Arabidopsis thaliana ferredoxin gene requires sequences upstream and downstream of the transcription initiation site

The effect of light on the expression of the Arabidopsis thaliana ferredoxin gene (fedA) was studied in mature tobacco plants. In light-treated leaves of tobacco plants transformed with a full-length ferredoxin gene, fedA-specific mRNA levels were more than twenty fold higher than in dark-treated controls. This indicates that all components for regulation of the Arabidopsis ferredoxin gene are present in tobacco. To identify light-regulatory elements in the fedA gene, we have tested a set of chimeric genes containing various parts of the fedA gene for light-dependent expression in mature tobacco plants. A fedA promoter-GUS fusion gene was not light-responsive, indicating that the 5′-upstream promoter region is not sufficient for light regulation. Fusion genes in which different transcribed regions of the fedA gene were expressed from the CaMV 35S promoter showed only limited light regulation, if any at all. This indicates that, like the fedA upstream region, the region downstream of the transcription start site is also not sufficient for full light regulation. The combined results suggest that for full light-regulated expression of the fedA gene, both the promoter region and sequences downstream of the transcription start site are required.

Effect of sarcomere length and filament lattice spacing on force development in skinned cardiac and skeletal muscle preparations from the rabbit

SummarySkinned cardiac and skeletal muscle freeze-dried preparations were activated in solutions strongly buffered for Ca2+. The response of single skeletal muscle fibres or thin strips of papillary muscle was investigated in relation to changes in Ca content of the perfusate. Sarcomere length was set and controlled during the experiments. The relation between the negative logarithm of the Ca concentration, the pCa, and the normalized developed force proved to be sigmoidal. The exact position of these curves proved to be dependent upon both sarcomere length and the distance between the filaments. The latter was shown by means of osmotic compression of the fibres using dextran. As a consequence of these observations. it was concluded that the length-tension relation is dependent upon the actual Ca concentration. The results are discussed in terms of cross-bridge interaction.

Root anatomy and development, the basis for nematode parasitism

Plant parasitic nematodes appear to rely on very specific interactions with root cells to establish a feeding site. To understand these interactions in detail, it is of advantage to achieve a basic understanding of root development. Arabidopsis thaliana is a suitable plant to investigate root development genetically and molecularly, and it can act as a host plant for plant parasitic nematodes. The anatomy and the ontogeny of the Arabidopsis root can be described in considerable detail. Despite the rigorous lineage relationships in the root, laser ablation experiments demonstrate the presence of continuous information in the root meristem. This information guides cells to differentiate appropriately, according to position. A large spectrum of promoter traps that are specifically expressed in roots are examined in detail, and put into four categories. These expression patterns can be complex, and a relation between the tagged gene and cell type is not always obvious. As a complementary approach, genetic analysis, using specific mutants, is now beginning to unravel key genes that are involved in setting up the pattern of cell differentiation in the root. Combining promoter trap analyses with mutant analysis may create novel strategies for nematode control.

SU-E-J-07: A Functional MR Protocol for the Pancreatic Tumor Delineation

PURPOSE Pancreatic cancer is one of the cancers with the poorest survival prognosis. At the time of diagnosis most of pancreatic cancers are unresectable and those patients can be treated by radiotherapy. Radiotherapy for pancreatic cancer is limited due to uncertainties in CT-based delineations. MRI provides an excellent soft tissue contrast. Here, an MR protocol is developed to improve delineations for radiotherapy treatment of pancreatic cancer. In a later stage this protocol can also be used for on-line visualization of the pancreas during MRI guided treatments. METHODS Nine pancreatic cancer patients were included. The MR protocol included T2 weighted(T2w), T1 weighted(T1w), diffusion weighted(DWI) and dynamic contrast enhanced(DCE) techniques. The tumor was delineated on T2w and T1w MRI by an experienced radiation oncologist. Healthy pancreas or pancreatitis (assigned by the oncologist based on T2w) areas were also delineated. Apparent diffusion coefficient(ADC), and area under the curve(AUC)/time to peak(TTP) maps were obtained from DWI and DCE scans, respectively. RESULTS A clear demarcation of tumor area was visible on b800 DWI images in 5 patients. ADC maps of those patients characterized tumor as an area with restricted water diffusion. Tumor delineations based on solely DCE were possible in 7 patients. In 6 of those patients AUC maps demonstrated tumor heterogeneity: a hypointense area with a hyperintense ring. TTP values clearly discriminated the tumor and the healthy pancreas but could not distinguish tumor and the pancreatitis accurately. CONCLUSION MR imaging results in a more pronounced tumor contrast than contrast enhanced CT. The addition of quantitative, functional MRI provides valuable, additional information to the radiation oncologist on the spatial tumor extent by discriminating tumor from the healthy pancreas(TTP, DWI) and characterizing the tumor(ADC). Our findings indicate that tumor delineation in pancreatic cancer can greatly benefit from the addition of MRI and especially functional MR techniques.

SU‐E‐J‐57: MRI‐Linac (MRL) Guided Treatment for Esophageal Cancer

For radiotherapy, oesophageal cancer is located in a difficult area. Spatial control of the dose distribution is difficult to achieve with current CT-based radiation techniques, as on CT, soft-tissue contrast is too low. Furthermore, the oesophagus moves and organs at risk (e.g. lung, heart, liver, spinal cord) are in close proximity. An 1.5 T MRI-accelerator (MRL) has sufficient soft-tissue tumour visualization possibilities to allow for precise real-time, online, position verification and for dose escalation without organ at riskoverdose. Our research consists of the preparatory work for the first clinical study on the MRL for patients with oesophageal cancer. To improve image quality and reduce the motion artefacts, the benefit of cardiac triggering and breath holds is evaluated on fifteen oesophageal patients. Results show the superb image quality of these MRI sequences. The use of this high quality MRI gives the possibility for non-invasive real-time visualization andtracking of the tumour. We quantify oesophageal tumour motion on cineMRI. The tumour is tracked on sequential mixed T1/T2w images (acquisition time: 60s, temporal resolution: 0.5s, slice thickness: 7mm) of a single coronal and sagittal slice using a Minimum Output Sum of Squared Error (MOSSE) adaptive correlation filter. Tumour registration within the individual images can typically be done at a millisecond time scale. Motion of oesophageal tumours can well be tracked and is highly variable between patients. The greatest mobility is seen in cranio-caudal direction, with amaximum peak-to-peak amplitude of tumour movement of 24.5mm followed by the dorso-ventral and the medio-lateral direction. Movement seems greatest in tumours located in the lower part of the oesophagus. This study shows both the superb image quality for GTV localisation and the possibility for on-line and real time tumour tracking. The study opens thepossibility for tracked radiation delivery with a 1.5T MRI accelerator. Partial funding has been obtained by Elekta and Philips.