Microtubule-organelle associations during generative cell polarization in plumbago zeylanica

1986 ◽  
Vol 44 ◽  
pp. 280-281
Author(s):  
S. D. Russell ◽  
T. W. Mislan
Keyword(s):  
Sexual Reproduction ◽  
Sperm Cell ◽  
Generative Cell ◽  
Embryo Sac ◽  
Cell Polarization ◽  
Precursor Cell ◽  
Flowering Plant ◽  
Sperm Cells ◽  
Plumbago Zeylanica ◽  
Preferential Fertilization

Sexual reproduction in the flowering plant Plumbago zeylanica is characterized by pollen in which the two sperm cells differ significantly with respect to size, shape and organellar content. The sperm cell with the majority of mitochondria (ave. 256) typically fuses with the central cell to form the nutritive endosperm in which the embryo will develop. The sperm cell containing essentially all of the numerous plastids (ave. 24) and few mitochondria (ave. 40) fuses with the egg to form the embryo. This system of preferential fertilization results in each sperm cell having a predetermined fate upon arrival at the embryo sac. Features determining the distribution of organelles in the sperm cells originate in the polarity of their precursor cell, the generative cell.

Immunofluorescent Localization of Myosin on the Sperm Cells of Plumbago Zeylanica

1997 ◽  
Vol 3 (S2) ◽  
pp. 183-184
Author(s):  
Zhaojie Zhang ◽  
Scott D. Russell
Keyword(s):  
Immunofluorescence Microscopy ◽  
Embryo Sac ◽  
Indirect Evidence ◽  
Sperm Cells ◽  
Immunofluorescent Localization ◽  
Plumbago Zeylanica ◽  
Actomyosin Interaction ◽  
Rabbit Igg ◽  
Sigma Chemical ◽  
Phosphate Buffered Saline

Sperm cells in flowering plants are non-motile and are passive participants in their movement to the female reproductive cells. It is believed that actomyosin interaction may play a key role for sperm cell transmission in the pollen tube as well as in the embryo sac. However, indirect evidence has shown that the surface of sperm cells lacks amounts of myosin sufficient to support movement. Immunofluorescence microscopy was used in this study to further assess the presence of myosin on the surface of sperm cells of Plumbago Zeylanica.Sperm cells of Plumbago Zeylanica were isolated according to published methods. Isolated sperm cells were blocked 20 min with 1% bovine serum albumin and 2% normal goat serum in phosphate buffered saline (PBS) (pH 7.3, in 15% sucrose), incubated 2 hrs in anti-myosin antibody (M-7648, Sigma Chemical Co., St. Louis, MO) diluted 1:10 with blocking solution, washed three times (5 min) with blocking solution and incubated 1 hr in 1:30 FITC-conjugated anti-rabbit IgG as secondary antibody (EY Labs, Inc., San Mateo, CA) in blocking solution. Samples were rinsed in PBS and mounted in an anti-fading solution with 1:1 PBS:glycerol with 3% n-propyl gallate.

Cytoskeletal organisation and modification during pollen tube arrival, gamete delivery and fertilisation in Plumbago zeylanica

Zygote ◽  
1993 ◽  
Vol 1 (2) ◽  
pp. 143-154 ◽  
Author(s):  
Bing-Quan Huang ◽  
Elisabeth S. Pierson ◽  
Scott D. Russell ◽  
Antonio Tiezzi ◽  
Mauro Cresti
Keyword(s):  
Pollen Tube ◽  
Embryo Sac ◽  
Central Cell ◽  
Egg Cell ◽  
Target Cells ◽  
Sperm Cells ◽  
Plumbago Zeylanica ◽  
Rhodamine Phalloidin ◽  
Male Gametes ◽  
Pollen Tube Penetration

The cytoskeletal organisation of the isolated embryo sac and egg cells of Plumbago zeylanica was examined before, during and after pollen tube penetration into the embryo sac to determine the potential involvement of microtubules and actin filaments in fertilisation. Material was singly and triply stained using Hoechst 33258 to localise DNA, fluorescein isothiocyanate (FITC)-labelled anti- α-tubulin to detect microtubules and rhodamine-phalloidin to visualise F-actin. Microtubules in the unfertilised egg cell are longitudinally aligned in the micropylar and mid-lateral areas, aggregating into bundles near the filiform apparatus. In the perinuclear cytoplasm of the egg cell, microtubules become more or less randomly aligned. F-actin bundles form a longitudinally aligned mesh in the chalazal cytoplasm of the egg cell. In the central cell, microtubules and F-actin are distributed along transvacuolar strands and are also evident in the perinuclear region and at the periphery of the cell. During pollen tube penetration, sparse microtubule bundles near the pathway of the pollen tube may form an apparent microtubular ‘conduit’ surrounding the male gametes at the delivery site. Actin aggregates become organised near the pathway of the pollen tube and at the delivery site of the sperm cells. Subsequently, actin aggregates form a ‘corona’ structure in the intercellular region between the egg and central cell where gametic fusion occurs. The corona may have a role in maintaining the close proximity of the egg and central cell and helping the two sperm cells move and bind to their target cells. The cytoskeleton may also be involved in causing the two nuclei of the egg and central cell to approach one another at the site of gametic fusion and transporting the two sperm nuclei into alignment with their respective female nucleus. The cytoskeleton is reorganised during early embryogenesis.

Sporogenesis, gametogenesis, and progamic phase in Alstroemeria

1996 ◽  
Vol 74 (8) ◽  
pp. 1354-1361
Author(s):  
Marjo J. De Jeu ◽  
Francesc Garriga Calderé ◽  
Jacques L. Van Went
Keyword(s):  
Sexual Reproduction ◽  
Interspecific Hybridization ◽  
Developmental Stages ◽  
Generative Cell ◽  
Embryo Sac ◽  
Mitotic Division ◽  
Polygonum Type ◽  
Male And Female ◽  
Inca Lily ◽  
Reproduction Biology

The sexual reproduction biology of Alstroemeria was studied histologically. The processes of sporogenesis and gametogenesis were described in relation to the development of the male and female organs. Comparative developmental stages in Alstroemeria take much longer than they do in tobacco. Alstroemeria has the monosporic Polygonum type of embryo sac development. Bicellular pollen is formed, which after germination on the receptive stigma immediately undergoes the mitotic division of the generative cell, thus finalizing gametogenesis. Part of the progamic phase has been studied as well. As early as 12 h after pollination, some pollen tubes enter the micropyle of the ovules. This knowledge is important for the application of post-fertilization ovule culture to rescue abortive embryos during interspecific hybridization. Keywords: Alstroemeria, Alstroemeria, Inca Lily, sporogenesis, gametogenesis, progamic phase.

scholarly journals Isolation of Sperm and Egg Cells from Pepper

2018 ◽  
Vol 143 (4) ◽  
pp. 310-315 ◽  
Author(s):  
Wei Deng ◽  
Yunling Xie ◽  
Yilan Qiu
Keyword(s):  
Generative Cell ◽  
Embryo Sac ◽  
Pollen Tubes ◽  
Enzymatic Digestion ◽  
Sperm Cells ◽  
Free Solution ◽  
In Vitro Method ◽  
Viable Embryo

Pepper (Capsicum annuum) pollen is bicellular and contains a vegetative cell and a generative cell, which divides in pollen tubes to form two sperm cells. Sperm cells of pepper were isolated using an in vivo–in vitro method. Hand-pollinated styles were first grown in vivo for several hours, then cut from their base and cultured in vitro until pollen tubes grew from the cut end. When the pollen tubes were transferred to a breaking solution, sperm cells were released from broken tubes. Viable embryo sac cells of pepper were isolated using enzymatic digestion and mechanical dissection. Isolated ovules were digested using cellulase and pectinase for 40 minutes and then transferred to an enzyme-free solution for mechanical dissection. Three cells of the egg apparatus and a central cell were released from a cut at the chalazal end of each ovule by pressing on the micropylar area of the ovule with a microneedle. Optimal isolation conditions included 11% mannitol, 0.04% CaCl2, 1% bovine serum albumin (BSA), 1% cellulase, 1% pectinase, and 0.3% pectolyase. Using this protocol, populations of pepper egg cells, synergids, and central cells were isolated.

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

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kazuki Motomura ◽  
Hidenori Takeuchi ◽  
Michitaka Notaguchi ◽  
Haruna Tsuchi ◽  
Atsushi Takeda ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Pollen Tube ◽  
Sperm Cell ◽  
Pollen Tubes ◽  
Vegetative Nucleus ◽  
Sperm Cells ◽  
Apical Region ◽  
Basal Region ◽  
Specific Expression ◽  
Directional Growth

AbstractDuring 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.

scholarly journals Calcium: A Critical Factor in Pollen Germination and Tube Elongation

2019 ◽  
Vol 20 (2) ◽  
pp. 420 ◽  
Author(s):  
Ren Zheng ◽  
Shun Su ◽  
Hui Xiao ◽  
Hui Tian
Keyword(s):  
Calcium Ion ◽  
Pollen Germination ◽  
Higher Plants ◽  
Male Gametophyte ◽  
Embryo Sac ◽  
Critical Factor ◽  
Sperm Cells ◽  
Critical Element ◽  
Major Function ◽  
Spatial And Temporal Characteristics

Pollen is the male gametophyte of higher plants. Its major function is to deliver sperm cells to the ovule to ensure successful fertilization. During this process, many interactions occur among pollen tubes and pistil cells and tissues, and calcium ion (Ca2+) dynamics mediate these interactions among cells to ensure that pollen reaches the embryo sac. Although the precise functions of Ca2+ dynamics in the cells are unknown, we can speculate about its roles on the basis of its spatial and temporal characteristics during these interactions. The results of many studies indicate that calcium is a critical element that is strongly related to pollen germination and pollen tube growth.

scholarly journals Influence of Campylobacter fetus subsp. fetus on ram sperm cell quality

2008 ◽  
Vol 57 (11) ◽  
pp. 1405-1410 ◽  
Author(s):  
Tidhar Zan Bar ◽  
Ronen Yehuda ◽  
Tomer Hacham ◽  
Sigal Krupnik ◽  
Benjamin Bartoov
Keyword(s):  
Sperm Cell ◽  
Sperm Cells ◽  
Male Factor ◽  
Motility Index ◽  
Fas Receptor ◽  
Campylobacter Fetus ◽  
Fetus Subsp ◽  
Ram Sperm ◽  
Female Sheep

Campylobacter fetus subsp. fetus infection can occur in female sheep, causing infertility or abortion. Despite extensive research on the effect of these bacteria on female fertility, little research has been done on the influence of C. fetus subsp. fetus on the male factor. Our objective was to examine the influence of C. fetus subsp. fetus on ram sperm. Motility index, percentage of live spermatozoa, mean αt value (an indication of the chromatin stability of the sperm cell) and percentage of sperm cells expressing the FAS receptor were measured in sperm incubated in the presence or absence of C. fetus subsp. fetus. The motility index and viability of sperm incubated with the bacteria were lower than those of untreated sperm samples after 5 h. In bacteria-incubated sperm cells, the percentage expressing FAS receptor was already significantly elevated at 15 min. Bacteria-incubated sperm showed a greater prevalence of morphological damage. The bacteria were attached to tail and acrosome regions, and the sperm damage was concentrated in both the motility and chromatin regions. Bacteria-infected sperm cells showed a decrease in motility, increase in early acrosome reaction and chromatin damage. Similar effects were induced by incubation of the sperm with supernatants from C. fetus subsp. fetus cultures. Thus this study demonstrates that C. fetus subsp. fetus has a detrimental effect on the quality of ram sperm.

scholarly journals A Study of Aflatoxin on Sperm Cell Quality for Elderly People

2018 ◽  
Vol 6 (2) ◽  
pp. 45-49
Author(s):  
Akila CR ◽  
Dinesh Babu J ◽  
Sravan Kumar P ◽  
Vinaya B
Keyword(s):  
Elderly People ◽  
Sperm Cell ◽  
Sperm Cells

Mycotoxins represent poisonous materials produced via certain types of growths. Among different mycotoxins, aflatoxins are viewed as plainly hazardous, taking into account that they are portrayed as cancer-causing for creatures and individuals. The admission of aflatoxins by means of feeds or nourishments should cause pernicious results on creatures' or people's wellbeing. Exploration on creatures has indicated that the overall casing circumstance just as a portion of the blood boundaries, particularly those of the liver may be adversely influenced with aflatoxin the executives. As to conceptive device, in spite of the fact that not remarkably examined, a few specialists bolster the helpless impacts of aflatoxins both on women or on grown-up guys. More precisely, in male, the scale and weight of the genital organs, the spermatogenesis, the number, the motility and the morphology of sperm cells just as hormones' fixations can be influenced after presentation of the creatures to aflatoxins, making barrenness inconveniences additional normal. Most examination allude to lab and significantly less to viable creatures, while least complex two investigations look for counsel from the doable issues of barrenness on men in view of aflatoxins. Since imitation is made one out of the greatest fundamental areas of creature cultivation, exceptional intrigue should be paid to supplements with the goal that the chance of the aflatoxin admission by creatures could be evacuated, the creature wellbeing extraordinarily in regards to the conceptive gadget may be covered and financial misfortunes could be enhanced.

scholarly journals Expression and function of ras proto-oncogene proteins in human sperm cells

1992 ◽  
Vol 102 (3) ◽  
pp. 487-494 ◽  
Author(s):  
R.K. Naz ◽  
K. Ahmad ◽  
P. Kaplan
Keyword(s):  
Sperm Cell ◽  
Acrosome Reaction ◽  
Cell Function ◽  
Beat Frequency ◽  
Human Sperm ◽  
Sperm Cells ◽  
Ras Protein ◽  
Head Displacement ◽  
Sperm Penetration ◽  
And Function

The presence and role of c-ras proteins were investigated in mature human sperm cells. The v-H-ras monoclonal antibody (mAb) against the c-ras protein, p21, reacted specifically with the acrosomal region of methanol-fixed as well as unfixed-live capacitated and non-capacitated human sperm cell in the indirect immunofluorescence technique. The v-H-ras mAb predominantly recognized c-ras protein of 21 kDa on the Western blot of lithium diiodosalicylate (LIS)-solubilized human sperm preparation. The incubation of sperm cells with v-H-ras mAb affected the sperm cell function in the human sperm penetration assay. The antibody significantly reduced the acrosome reaction and release of acrosin activity from the sperm cells. There was no effect of the mAb on percentage motility, although the mAb significantly affected various motility characteristics such as linearity, amplitude of lateral head displacement (ALH) and beat frequency, the motility parameters involved in the hyperactivation phenomenon of sperm cells leading to capacitation and acrosome reaction. These results suggest that the c-ras or c-ras-like proteins are present in mature sperm cell and may have a role in capacitation and/or acrosome reaction of human sperm cell.

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