Persistent directional growth capability in Arabidopsis thaliana pollen tubes after nuclear elimination from the apex

During the double fertilization process, pollen tubes deliver two sperm cells to an ovule containing the female gametes. In the pollen tube, the vegetative nucleus and sperm cells move together to the apical region where the vegetative nucleus is thought to play a crucial role in controlling the direction and growth of the pollen tube. Here, we report the generation of pollen tubes in Arabidopsis thaliana whose vegetative nucleus and sperm cells are isolated and sealed by callose plugs in the basal region due to apical transport defects induced by mutations in the WPP domain-interacting tail-anchored proteins (WITs) and sperm cell-specific expression of a dominant mutant of the CALLOSE SYNTHASE 3 protein. Through pollen-tube guidance assays, we show that the physiologically anuclear mutant pollen tubes maintain the ability to grow and enter ovules. Our findings provide insight into the sperm cell delivery mechanism and illustrate the independence of the tip-localized vegetative nucleus from directional growth control of the pollen tube.

A Tug-of-War Model Explains the Saltatory Sperm Cell Movement in Arabidopsis thaliana Pollen Tubes by Kinesins With Calponin Homology Domain

Upon pollination, two sperm cells are transported inside the growing pollen tube toward the apex. One sperm cell fertilizes the egg cell to form the zygote, while the other fuses with the two polar nuclei to form the triploid endosperm. In Arabidopsis thaliana, the transport of the two sperm cells is characterized by sequential forward and backward movements with intermediate pauses. Until now, it is under debate which components of the plant cytoskeleton govern this mechanism. The sperm cells are interconnected and linked to the vegetative nucleus via a cytoplasmic projection, thus forming the male germ unit. This led to the common hypothesis that the vegetative nucleus is actively transported via myosin motors along actin cables while pulling along the sperm cells as passive cargo. In this study, however, we show that upon occasional germ unit disassembly, the sperm cells are transported independently and still follow the same bidirectional movement pattern. Moreover, we found that the net movement of sperm cells results from a combination of both longer and faster runs toward the pollen tube apex. We propose that the observed saltatory movement can be explained by the function of kinesins with calponin homology domain (KCH). This subgroup of the kinesin-14 family actively links actin filaments and microtubules. Based on KCH's specific properties derived from in vitro experiments, we built a tug-of-war model that could reproduce the characteristic sperm cell movement in pollen tubes.

Characterization of ALBA Family Expression and Localization in Arabidopsis thaliana Generative Organs

ALBA DNA/RNA-binding proteins form an ancient family, which in eukaryotes diversified into two Rpp25-like and Rpp20-like subfamilies. In most studied model organisms, their function remains unclear, but they are usually associated with RNA metabolism, mRNA translatability and stress response. In plants, the enriched number of ALBA family members remains poorly understood. Here, we studied ALBA dynamics during reproductive development in Arabidopsis at the levels of gene expression and protein localization, both under standard conditions and following heat stress. In generative tissues, ALBA proteins showed the strongest signal in mature pollen where they localized predominantly in cytoplasmic foci, particularly in regions surrounding the vegetative nucleus and sperm cells. Finally, we demonstrated the involvement of two Rpp25-like subfamily members ALBA4 and ALBA6 in RNA metabolism in mature pollen supported by their co-localization with poly(A)-binding protein 3 (PABP3). Collectively, we demonstrated the engagement of ALBA proteins in male reproductive development and the heat stress response, highlighting the involvement of ALBA4 and ALBA6 in RNA metabolism, storage and/or translational control in pollen upon heat stress. Such dynamic re-localization of ALBA proteins in a controlled, developmentally and environmentally regulated manner, likely reflects not only their redundancy but also their possible functional diversification in plants.

POSSIBLE NEUROGENIC MECHANISMS SUDDEN INFANT DEATH SYNDROME

Sudden infant death syndrome (SIDS) is an unexpected non-violent death of apparently healthy child aged 7 days to 1 year, in which the anamnesis and autopsy data do not allow to find out the cause of the fatal outcome. Researchers have not come to a consensus on the thanatological role of morphological changes in the brain. Aim. To determine pathomorphological changes in the brain in the case of sudden death syndrome in children. Material and methods. Forensic medical studies of 118 deaths of healthy children who died suddenly for the period 2008-2017 were carried out and analyzed on the territory of the Stavropol region. Autopsy material was divided into groups: the main group (1) consisted of 74 observations (62.8%), where SIDS was the main final sectional diagnosis. The comparison group (2) consisted of 44 (37.2%) observations; the children died suddenly at home as a result of viral-bacterial pneumonia. For control, the third group was formed (3), which included 45 observations, where the death of children occurred as a result of drowning and carbon monoxide poisoning. Results and its discussion. A histological examination of pieces of the brain in the case of sudden death syndrome revealed the foci of angiomatosis of the vessels of the pia mater and brain matter with the normal structure of the elastic membrane of the arteries, the effects of glial proliferation in the subependymal region of the lateral ventricles and subcortical formations. In the medulla oblongata, in a detailed study of the posterior (vegetative) nucleus of the vagus nerve, degenerative changes in neurons were detected in the form of chromatolysis and karyolysis, a glial reaction of the type of neuronophagy. Conclusion. Pathomorphological changes in the brain stem may be due to combined intrauterine hypoxic and intrapartum traumatic factors, which, potentiating each other, lead to dysregulation of the respiratory rhythm with the development of a fatal outcome during sleep.

The nuclear envelope protein KAKU4 determines the migration order of the vegetative nucleus and sperm cells in pollen tubes

A putative component protein of the nuclear lamina, KAKU4, modulates nuclear morphology in Arabidopsis thaliana seedlings, but its physiological significance is unknown. KAKU4 was highly expressed in mature pollen grains, each of which has a vegetative cell and two sperm cells. KAKU4 protein was highly abundant on the envelopes of vegetative nuclei and less abundant on the envelopes of sperm cell nuclei in pollen grains and elongating pollen tubes. Vegetative nuclei are irregularly shaped in wild-type pollen. However, KAKU4 deficiency caused them to become more spherical. After a pollen grain germinates, the vegetative nuclei and sperm cells enter and move along the pollen tube. In the wild type, the vegetative nucleus preceded the sperm cell nuclei in >90% of the pollen tubes, whereas, in kaku4 mutants, the vegetative nucleus preceded the sperm cell nuclei in only about half of the pollen tubes. kaku4 pollen was less competitive for fertilization than wild-type pollen after pollination. These results led us to hypothesize that the nuclear shape in vegetative cells of pollen grains affects the orderly migration of the vegetative nucleus and sperm cells in pollen tubes.

Vegetative nuclear positioning is required for calcium and ROS signaling in Arabidopsis pollen tubes

Efficient transport and delivery of sperm cells (SCs) is vital for angiosperm plant fertility. In Arabidopsis thaliana, SCs are transported through the growing pollen tube by a connection with the vegetative nucleus (VN). During pollen tube growth, the VN leads the way and maintains a fixed distance from the pollen tube tip, while the SCs lag behind the VN. Upon reception at the ovule, the pollen tube bursts and the SCs are released for fertilization. In pollen tubes of Arabidopsis mutants wit12 and wifi, deficient in the outer nuclear membrane component of a plant LINC complex, the SCs precede the VN and the VN falls behind. Subsequently, pollen tubes frequently fail to burst upon reception. In this study, we sought to determine if the pollen tube reception defect observed in wit12 and wifi is due to decreased sensitivity to reactive oxygen species (ROS). Here we show that wit12 and wifi are hyposensitive to exogenous H2O2, and that this hyposensitivity is correlated with decreased proximity of the VN to the pollen tube tip. Additionally, we report the first instance of nuclear Ca2+ spikes in growing pollen tubes, which are disrupted in the wit12 mutant. In the wit12 mutant, nuclear Ca2+ spikes are reduced in response to exogenous ROS, but these spikes are not correlated with pollen tube burst. This study finds that VN proximity to the pollen tube tip is required for both response to exogenous ROS, as well as internal nuclear Ca2+ fluctuations.SummaryMutants deficient in outer nuclear membrane proteins display defects in reactive oxygen species-induced pollen tube burst and nuclear Ca2+ signatures that correlate with the position of the vegetative nucleus.

KAKU4-mediated deformation of the vegetative nucleus controls its precedent migration over sperm cells in pollen tubes

A putative nuclear lamina protein, KAKU4, modulates nuclear morphology in Arabidopsis thaliana seedlings but its physiological significance is unknown. KAKU4 was strongly expressed in mature pollen grains, each of which has a vegetative cell and two sperm cells. KAKU4 protein was highly abundant on the envelopes of vegetative nuclei (VNs) and less abundant on the envelopes of sperm cell nuclei (SCNs) in pollen grains and elongating pollen tubes. VN is irregularly shaped in wild-type pollen. However, KAKU4 deficiency caused it to become more spherical. These results suggest that the dense accumulation of KAKU4 is responsible for the irregular shape of the VNs. After a pollen grain germinates, the VN and SCNs migrate to the tip of the pollen tube. In the wild type, the VN preceded the SCNs in 91–93% of the pollen tubes, whereas in kaku4 mutants, the VN trailed the SCNs in 39–58% of the pollen tubes. kaku4 pollen was less competitive than wild-type pollen after pollination, although it had an ability to fertilize. Taken together, our results suggest that controlling the nuclear shape in vegetative cells of pollen grains by KAKU4 ensures the orderly migration of the VN and sperm cells in pollen tubes.HighlightThe nuclear envelope protein KAKU4 is involved in controlling the migration order of vegetative nuclei and sperm cells in pollen tubes, affecting the competitive ability of pollen for fertilization.

Facial nerve: Essential aspects from the biomedical sciences

This article is a review of the anatomical literature about of the facial nerve. It has historical aspects, functional components, periphery route inside and outside of the petrous portion of the temporal bone. It describes the nerve branches and territories of innervation. It also describes its nervous connections to the brain`s stem, specifically to the pons, which is the cerebral center, where facial nerves’ motor and vegetative nucleus is located. The information presented here could be useful to students of medicine, dentistry and others health programs at level of undergraduate and postgraduate. In this article have been used known books of anatomy and neuroanatomy. It has been also used articles of the last five years written by professional in neurosurgery, otorhinolaryngology and morphology. This article presents a correlation among the facial nerve anatomy and its implications in pathology, trauma and surgery of head and neck region.