Aqp9 Gene Deletion Enhances Retinal Ganglion Cell (RGC) Death and Dysfunction Induced by Optic Nerve Crush: Evidence that Aquaporin 9 Acts as an Astrocyte-to-Neuron Lactate Shuttle in Concert with Monocarboxylate Transporters To Support RGC Function and Survival.

Aquaporin 9 (AQP9) is an aquaglyceroporin that can transport lactate. Accumulating evidence suggests that astrocyte-to-neuron lactate shuttle (ANLS) plays a critical role in energy metabolism in neurons, including retinal ganglion cells (RGCs). To test the hypothesis that AQP9, in concert with monocarboxylate transporters (MCTs), participates in ANLS to maintain function and survival of RGCs, Aqp9-null mice and wild-type (WT) littermates were subjected to optic nerve crush (ONC) with or without intravitreal injection of an MCT2 inhibitor. RGC density was similar between the Aqp9-null mice and WT mice without ONC, while ONC resulted in significantly more RGC density reduction in the Aqp9-null mice than in the WT mice at day 7. Positive scotopic threshold response (pSTR) amplitude values were similar between the two groups without ONC, but were significantly more reduced in the Aqp9-null mice than in the WT mice 7days after ONC. MCT2 inhibitor injection accelerated RGC death and pSTR amplitude reduction only in the WT mice with ONC. Immunolabeling revealed that both RGCs and astrocytes expressed AQP9, that ONC predominantly reduced astrocytic AQP9 expression, and that MCTs 1, 2, and 4 were co-localized with AQP9 at the ganglion cell layer. These retinal MCTs were also co-immunoprecipitated with AQP9 in the WT mice. ONC decreased the co-immunoprecipitation of MCTs 1 and 4, but did not impact co-immunoprecipitation of MCT2. Retinal glucose transporter 1 expression was increased in Aqp9-null mice. Aqp9 gene deletion reduced and increased the intraretinal L-lactate and D-glucose concentrations, respectively. Results suggest that AQP9 acts as the ANLS to maintain function and survival of RGCs.

[The disruption of aquaporin function in cell membranes as a cause of changes in germinability of pea seeds after exposure to low doses of gamma-radiation].

Pea seeds (Pisum sativum L.) from the seed lot with 80% germinability were separated in fractions according to room temperature phosphorescence: strong seeds were assigned to fraction I, and weak seeds formed to fraction II. During imbibition, the seeds of fraction II exhibited twofold higher rates of water uptake and experienced hypoxia. Some of these seeds suffocated from hypoxia, and other seeds produced seedlings with morphological defects (such seeds were considered incapable of germination). One week after irradiation with the dose of 3 Gy, germination percentage decreased to 45%, which was caused by the increase of number of weak seeds. The germinability of seeds subjected to gamma-irradiation at doses of 7 and 10 Gy was similar to that of control seeds. In these sub-lots, there appeared so-called "improved" seeds, which were similar to non-irradiated seeds in terms of phosphorescence level, the rate of water uptake and germination percentage. It was shown with the use of PCMB that the difference in the rates of water uptake by seeds of fraction I and II depended on the permeability of cell membranes. The permeability was determined by the state of aquaporins ("open"-"closed"). The experiments with phosphatase inhibitor (NaF) shown that in seeds irradiated with dose of 3 Gy (fraction II), the mechanism of aquaporins closing was broken (phosphatase was inactivated). In "improved" seeds (after irradiation with dose of 10 Gy), aquaporins were closed irreversibly in air-dry state, when aquaporin dephosphorylation was unlikely. It was concluded that the abnormal increase (following the initially decrease) in germination of pea seeds after irradiation can be explained without invoking the hypothesis on hyper function of reparatory mechanism of at low doses of irradiation.

The plasma membrane aquaporin NtAQP1 is a key component of the leaf unfolding mechanism in tobacco.

Epinastic leaf movement of tobacco is based on differential growth of the upper and lower leaf surface and is distinct from the motor organ-driven mechanism of nyctinastic leaf movement of, for example, mimosa species. The epinastic leaf movement of tobacco is observed not only under diurnal light regimes but also in continuous light, indicating a control by light and the circadian clock. As the transport of water across membranes by aquaporins is an important component of rapid plant cell elongation, the role of the tobacco aquaporin Nt aquaporin (AQP)1 in the epinastic response was studied in detail. In planta NtAQP1-luciferase (LUC) activity studies, Northern and Western blot analyses demonstrated a diurnal and circadian oscillation in the expression of this plasma membrane intrinsic protein (PIP)1-type aquaporin in leaf petioles, exhibiting peaks of expression coinciding with leaf unfolding. Cellular water permeability of protoplasts isolated from leaf petioles was found to be high in the morning, i.e. during the unfolding reaction, and low in the evening. Moreover, diurnal epinastic leaf movement was shown to be reduced in transgenic tobacco lines with an impaired expression of NtAQP1. It is concluded that the cyclic expression of PIP1-aquaporin represents an important component of the leaf movement mechanism.

Diurnal regulation of water transport and aquaporin gene expression in maize roots: contribution of PIP2 proteins.

In maize (Zea mays) roots, xylem water transfer supported by root pressure occurs during the day and is less important at night. Diurnal modifications of osmotic pressure gradient between medium and xylem could not explain the oscillation of water flux in young maize roots during the day-night cycle. We observed a high turgor pressure of root cortical cells associated with a high flux. In maize roots, ZmPIP transcripts oscillate during the day-night cycle exhibiting some characteristics of genes regulated by a circadian mechanism. The PIP protein level profile is different from the mRNA pattern. Moreover, ZmPIP1 and ZmPIP2 protein levels are differentially regulated during the light and dark period and in response to continuous darkness suggesting different roles for both classes of PIP. Finally, our results suggest that aquaporins from ZmPIP2 subgroup may contribute to root water transfer by cellular pathway that occurs during the light and the dark period of the day-night cycle.