Rhizosphere activity in an old-growth forest reacts rapidly to changes in soil moisture and shapes whole-tree carbon allocation.

Drought alters carbon (C) allocation within trees, thereby impairing tree growth. Recovery of root and leaf functioning and prioritized C supply to sink tissues after drought may compensate for drought-induced reduction of assimilation and growth. It remains unclear if C allocation to sink tissues during and following drought is controlled by altered sink metabolic activities or by the availability of new assimilates. Understanding such mechanisms is required to predict forests' resilience to a changing climate. We investigated the impact of drought and drought release on C allocation in a 100-y-old Scots pine forest. We applied 13CO2 pulse labeling to naturally dry control and long-term irrigated trees and tracked the fate of the label in above- and belowground C pools and fluxes. Allocation of new assimilates belowground was ca. 53% lower under nonirrigated conditions. A short rainfall event, which led to a temporary increase in the soil water content (SWC) in the topsoil, strongly increased the amounts of C transported belowground in the nonirrigated plots to values comparable to those in the irrigated plots. This switch in allocation patterns was congruent with a tipping point at around 15% SWC in the response of the respiratory activity of soil microbes. These results indicate that the metabolic sink activity in the rhizosphere and its modulation by soil moisture can drive C allocation within adult trees and ecosystems. Even a subtle increase in soil moisture can lead to a rapid recovery of belowground functions that in turn affects the direction of C transport in trees.

Long-term GnRH-induced gonadotropin secretion in a novel hypothalamo-pituitary slice culture from tilapia brain.

Organotypic cultures, prepared from hypothalamo-pituitary slices of tilapia, were developed to enable long-term study of secretory cells in the pituitary of a teleost. Values of membrane potential at rest were similar to those recorded from acute slices, and cells presented similar spontaneous spikes and spikelets. Some cells also exhibited slow spontaneous oscillations in membrane potential, which may be network-driven. Long-term (6days) continuous exposure to GnRH induced increases in LH and FSH secretion. FSH levels reached the highest levels after 24h of exposure to GnRH, and the highest secretion of LH was observed in days 4 and 5 of the experiment. Since slices were viable for several weeks in culture, maintaining the original cytoarchitecture, electrical membrane properties and the ability to secrete hormones in response to exogenous GnRH, this technique is ideal for studying the mechanisms regulating cell-to-cell communication under conditions resembling the in vivo tissue organization.

Electrotonic coupling in the anterior pituitary of a teleost fish.

The anterior pituitary of teleost fish contains a variety of endocrine cells, which, under control from the hypothalamus, release trophic hormones and thereby play a major role in reproduction, social behavior, and growth. In fish, hypothalamic fibers directly innervate the pituitary. The hypothalamic hormones released from these fibers bind to membrane receptors on pituitary cells, triggering action potentials, a rise in cytosolic calcium, and exocytosis. It is unclear whether these activities are confined to the stimulated cell or propagate to adjacent cells. We addressed this issue using whole cell and perforated patch-clamp techniques in a novel, hypothalamo-pituitary slice preparation from the tilapia fish (Oreochromis niloticus). Pituitary cells at rest generated occasional spontaneous spikes and sharp depolarizations of lower amplitude. The latter probably represented spikes in neighboring, electrotonically coupled cells. The presence of electrotonic communication, probably mediated by gap junctions, was also supported by the finding that Lucifer Yellow diffuses between cells. To quantify this connectivity, we performed simultaneous recording from pairs of adjacent cells. Thirty-three percent of the cells exhibited strong reciprocal coupling. Coupling coefficients ranged between 0.18 and 0.31, and coupling resistances ranged between 16 and 39 GOhm. The electrical junctions were effective low pass filters, attenuating action potentials much more than low frequency waveforms. We conclude that electrical activities of anterior pituitary cells in teleost fish are synchronized by coupling through gap junctions. Regulation of this coupling may play a critical role in determining complex patterns of pituitary hormone secretion.