The properties of nerve cell precursors in hydra

Two signals, the head activator and an injury stimulus, control differentiation of nerve cells from uncommitted stem cells in hydra [Th. Holstein, H. C. Schaller, and C. N. David, (1986) Dev. Biol. 115, 9–17]. The time of action of these signals in the precursor cell cycle was determined. Methanol extracts of hydra containing 10−13 M head activator cause nerve cell commitment in S phase of the precursor cell cycle. Committed precursors complete the cell cycle, divide, and arrest in G1. Injury relieves the G1 block and precursors differentiate nerve cells. Under these conditions the time from commitment to nerve differentiation is 12 hr, the time from the end of S phase to nerve differentiation is 9 hr, and the time from the G1 block to nerve differentiation is 4 hr. Committed precursors blocked in G1 are unstable, decaying with a half-life of 12 hr if not stimulated to differentiate by an injury stimulus

Putative intermediates in the nerve cell differentiation pathway in hydra have properties of multipotent stem cells

We have investigated the properties of nerve cell precursors in hydra by analyzing the differentiation and proliferation capacity of interstitial cells in the peduncle of Hydra oligactis, which is a region of active nerve cell differentiation. Our results indicate that about 50% of the interstitial cells in the peduncle can grow rapidly and also give rise to nematocyte precursors when transplanted into a gastric environment. If these cells were committed nerve cell precursors, one would not expect them to differentiate into nematocytes nor to proliferate apparently without limit. Therefore we conclude that cycling interstitial cells in peduncles are not intermediates in the nerve cell differentiation pathway but are stem cells. The remaining interstitial cells in the peduncle are in G1 and have the properties of committed nerve cell precursors (Holstein and David, 1986). Thus, the interstitial cell population in the peduncle contains both stem cells and noncycling nerve precursors. The presence of stem cells in this region makes it likely that these cells are the immediate targets of signals which give rise to nerve cells

Tentacle morphogenesis in hydra

Stimulation of tentacle-specific cell differentiation by the neuropeptide head activator was investigated in Hydra magnipapillata. Tentacle-specific sensory nerve cells were identified by a monoclonal antibody, NV1. Treatment of hydra with 1pM head activator for 18h stimulated differentiation of NV1+ nerve cells and tentacle epithelial cells in tissue from the distal gastric region. Head tissue and tissue from the proximal gastric region did not respond to head activator treatment with increased NV1+ differentiation. Hence the distal gastric region appears to be the site of tentacle formation in hydra. Tentacle precursors in head tissue seem to be committed since they fail to respond to head activator or to changes in tissue size with altered amounts of tentacle formation. We suggest that NV1 precursors form a complex with tentacle epithelial cell precursors, which then moves distally through the head region into the tentacles. The signal for NV1+ differentiation appears to be formation of this complex

Mini-Collagens in Hydra Nematocytes

We have isolated and characterized four collagen-related c-DNA clones (N-COL 1, N-COL 2, N-COL 3, N-COL 4) that are highly expressed in developing nematocytes in hydra. All four c-DNAs as well as their corresponding transcripts are small in size (600-1,000 bp). The deduced amino acid sequences show that they contain a central region consisting of 14 to 16 Gly-X-Y triplets. This region is flanked amino-terminal by a stretch of 14-23 proline residues and carboxy-terminal by a stretch of 6-9 prolines. At the NH2- and COOH-termini are repeated patterns of cysteine residues that are highly conserved between the molecules. A model is proposed which consists of a central stable collagen triple helix of 12-14 nm length from which three 9-22 nm long polyproline II type helices emerge at both ends. Disulfide linkage between cysteine- rich segments in these helices could lead to the formation of oligomeric network structures. Electrophoretic characterization of nematocyst extracts allows resolution of small proline-rich polypeptides that correspond in size to the cloned sequences

Cell cycle length, cell size, and proliferation rate in hydra stem cells

We have analyzed the cell cycle parameters of interstitial cells in Hydra oligactis. Three subpopulations of cells with short, medium, and long cell cycles were identified. Short-cycle cells are stem cells; medium-cycle cells are precursors to nematocyte differentiation; long-cycle cells are precursors to gamete differentiation. We have also determined the effect of different cell densities on the population doubling time, cell cycle length, and cell size of interstitial cells. Our results indicate that decreasing the interstitial cell density from 0.35 to 0.1 interstitial cells/epithelial cell (1) shortens the population doubling time from 4 to 1.8 days, (2) increases the [3H]thymidine labeling index from 0.5 to 0.75 and shifts the nuclear DNA distribution from G2 to S phase cells, and (3) decreases the length of G2 in stem cells from 6 to 3 hr. The shortened cell cycle is correlated with a significant decrease in the size of interstitial stem cells. Coincident with the shortened cell cycle and increased growth rate there is an increase in stem cell self-renewal and a decrease in stem cell differentiation

A quantitative method for separation of living hydra cells

We describe a rapid method for the isolation of large numbers of livingHydra cells of defined cell type in an isotonic cell medium (Gierer et al. 1972). Intact animals are enzymatically dissociated into a single cell suspension and the various cell types separated in less than one hour by counterflow centrifugation elutriation. Cell loss is minimal. RNA isolated from various fractions can be probed with cell type specific cDNA-clones

Two plus one is almost three: a fast approximation for multi-view deconvolution

Multi-view deconvolution is a powerful image-processing tool for light sheet fluorescence microscopy, providing isotropic resolution and enhancing the image content. However, performing these calculations on large datasets is computationally demanding and time-consuming even on high-end workstations. Especially in long-time measurements on developing animals, huge amounts of image data are acquired. To keep them manageable, redundancies should be removed right after image acquisition. To this end, we report a fast approximation to three-dimensional multi-view deconvolution, denoted 2D+1D multi-view deconvolution, which is able to keep up with the data flow. It first operates on the two dimensions perpendicular and subsequently on the one parallel to the rotation axis, exploiting the rotational symmetry of the point spread function along the rotation axis. We validated our algorithm and evaluated it quantitatively against two-dimensional and three-dimensional multi-view deconvolution using simulated and real image data. 2D+1D multi-view deconvolution takes similar computation time but performs markedly better than the two-dimensional approximation only. Therefore, it will be most useful for image processing in time-critical applications, where the full 3D multi-view deconvolution cannot keep up with the data flow

A view to kill

Genome and proteome data from Hydra magnipapillata have opened the way for the molecular analysis of an ancient nervous system, which includes stinging cells, an unusual neurosensory and neurosecretory cell type. They hold some surprises for the mechanisms and evolution of sensory transduction that could not have been anticipated from what has been learned from flies and vertebrates. Research in BMC Biology now implicates the ancient opsin-mediated transduction pathway in the neuronal control of stinging cell discharge

Chiral corrections to the vector and axial couplings of quarks and baryons

We calculate chiral corrections to the semileptonic vector and axial quark coupling constants using a manifestly Lorentz covariant chiral quark approach up to order O(p4) in the two- and tree-flavor picture. These couplings are then used in the evaluation of the corresponding couplings which govern the semileptonic transitions between octet baryon states. In the calculation of baryon matrix elements we use a general ansatz for the spatial form of the quark wave function, without referring to a specific realization of hadronization and confinement of quarks in baryons. Matching the physical amplitudes calculated within our approach to the model-independent predictions of baryon chiral perturbation theory (ChPT) allows one to deduce the connection between our parameters and those of baryon ChPT.Comment: 28 pages, 4 figure