In flowering plants, sperm cells develop in the pollen cytoplasm and are transported through floral tissues to an ovule by a pollen tube, a highly polarized cellular extension. After targeting an ovule, the pollen tube bursts, releasing two sperm that fertilize an egg and a central cell. Here, we identified the gene encoding Arabidopsis HAP2, demonstrating that it is allelic to GCS1. HAP2 is expressed only in the haploid sperm and is required for efficient pollen tube guidance to ovules. We identified an insertion (hap2-1) that disrupts the C-terminal portion of the protein and tags mutant pollen grains with the ␤-glucuronidase reporter. By monitoring reporter expression, we showed that hap2-1 does not diminish pollen tube length in vitro or in the pistil, but it reduces ovule targeting by twofold. In addition, we show that the hap2 sperm that are delivered to ovules fail to initiate fertilization. HAP2 is predicted to encode a protein with an N-terminal secretion signal, a single transmembrane domain and a C-terminal histidine-rich domain. These results point to a dual role for HAP2, functioning in both pollen tube guidance and in fertilization. Moreover, our findings suggest that sperm, long considered to be passive cargo, are involved in directing the pollen tube to its target.

In flowering plants, the egg develops within a haploid embryo sac (female gametophyte) that is encased within the pistil. The haploid pollen grain (male gametophyte) extends a pollen tube that carries two sperm cells within its cytoplasm to the embryo sac. This feat requires rapid, precisely guided, and highly polarized growth through, between, and on the surface of the cells of the stigma, style, and ovary. Pollen tube migration depends on a series of long-range signals from diploid female cells as well as a short-range attractant emitted by the embryo sac that guides the final stage of tube growth. We developed a genetic screen in Arabidopsis thaliana that tags mutant pollen with a cell-autonomous marker carried on an insertion element. We found 32 haploid-disrupting (hapless) mutations that define genes required for pollen grain development, pollen tube growth in the stigma and style, or pollen tube growth and guidance in the ovary. We also identified genomic DNA flanking the insertion element for eleven hap mutants and showed that hap1 disrupts AtMago, a gene whose ortholog is important for Drosophila cell polarity.

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Despite considerable progress in the knowledge about pathophysiology of cardiac allograft vascular disease (CAVD), only few systematic studies are available, characterizing the natural course during long-term follow-up after heart transplantation (HTX). Therefore, we analyzed in 354 heart transplant recipients (305 male, 45.9 +/- 11.2 years, mean observation period 5.8 +/- 3.0 years, range 0.9-12.4 years) the results of 1129 coronary angiograms under the aspects of development, severity, localization, and progression of disease related to the prognosis of patients. As expected an increasing prevalence was found over time with a luminal obstruction > or = 30% in 83% of all patients more than 10 years after HTX. Coronary artery stenosis (> or = 50%) at initial diagnosis was predominantly localized in the LAD (46%) followed by RCX (31%) and RCA (23%). Angiographic risk profiles with an impaired prognosis could be identified in the form of an early development (< 4 years post HTX) of disease (p = 0.03), luminal obstruction > 50% (p = 0.001), and multivessel appearance at first diagnosis (p = 0.02) as well as in progressive forms of CAVD (p = 0.001). Summarizing, CAVD is a frequent finding in HTX recipients. Especially in patients with early onset, progressive, and advanced stages of disease it represents a prognostically limiting complication following HTX. Identification of the natural course is of major importance defining the need and efficacy of future palliative therapeutical approaches.

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